Vargas Zamora, Fernando Rodolfo de Jesus (22 September 1928 – 20 February 1989)

Raised a Catholic, Vargas renounced Catholicism when as a teenager he left Costa Rica. The co-founder of Variety Recording Studio in New York City, Vargas was a significant figure in show business from the late 1950s until his death in 1989.

Vargas was the son of Manuel Elias Vargas Cordero, a department store owner, and Elena Zamora Paniagua. Their children were Aura Vargas de Moreno, Eugenia Vargas Zamora, Nery Vargas Zamora, Elena Vargas Zamora, and Otto Vargas Zamora. Fernando was the last-born.

When 16 and a student at Colegio Los Angeles in San José, Vargas earned his best grades in biology and history, his weakest in religion and English. His father owned a large department store, in which he worked and was allowed to handle money. His youth was spent excelling in sports, riding a bicycle, visiting relatives in Cartago, Heredia, and Alajuela, and spending time near Teatro Melico Salazar and the Parque Central, the park on Calle 14 that he knew the best.

His hard-working mother supervised several servants and looked after her large family. When her husband died, a big tomb was erected at Cementerio Obreros, for he was an important person who once had dreams of becoming President. MANUEL ELIAS VARGAS CORDERO was written on the tomb, which had an angel atop. Later, members of the family were to be buried there next to Father and Mother.

On 3 May 1946, at the passport office Fernando received his passport and on 22 May 1946 he, his sister Elena, and his mother arrived in Miami. His Alien Registration Receipt Card was #6312922. Quickly, Selective Service on 3 Dec 1946 gave him a registration card that showed he had green eyes, was white, had brown hair, was 5' 7", and weighed 132.

When he was 8 months older than 17, Vargas arrived at 175 Claremont Avenue, New York City. His mother, sister, and he lived in a one-bedroom apartment between 123rd and 125th Street near the International House and the Manhattan School of Music. It was just west of Harlem, and he and his sister were not allowed to walk in the neighborhood, which their mother thought was dangerous. Little by little, they were allowed to go together around the immediate neighborhood. Much of his time was spent watching TV or going shopping with his sister for groceries or supplies. In the evenings they eventually got tired of supervising him and allowed him to go out alone. He spent lots of time around the corner at Grant’s Tomb, 122nd Street.

Almost every time that he went to Grant’s Tomb and walked around Riverside Park, he found people who were Columbia University professors, students, and professional types. Also, as was the case at Parque Central near his home in San José, he noticed some younger and older males who were looking for sex. Although they were relatively rare at Parque Central, they were quite numerous in that section of Riverside Park. If you sat on a park bench alone here, someone might sit down next to you and if you just sat in a slouched position, your hands clasped behind your head, someone would unzip your pants and play with you.

His first introduction to sex had been when he was an altar boy and a Catholic priest, who pretended it was accidental, touched his crotch. Asking a childhood friend whose family was German and imported toys if the priest had ever pulled his pants down to see his penis, he found that had happened to him, also. The German boy was a best friend, was heavier and easily beaten in races. But on one occasion his friend claimed he had something bigger than Vargas did and, bragging, pulled out his pene and "testículos." Then, his friend had put one of Vargas's hands on his own crotch, which felt good, and he reciprocated. Not knowing whether to tell an older brother or anyone else, he decided he might get in trouble both with the church and his family and said nothing.

Between Riverside Drive and the Hudson River, the park was extensive. There was a tennis court, a skating area, and there was a big sports area before you got as far downtown as 96th Street. Whatever transpired while cruising the area, he had to make sure he got home before 10 p.m.

Harold Bonilla, who was Costa Rica’s consul in the city, accidentally met him in the park and asked him to move in with him nearby on 103rd Street. It was not difficult to obtain his mother's approval, for here was a person who wrote history books, was a Catholic, and mentioned that he might be able to get Fernando a job and schooling.

Now exactly 20, finding no jobs but not having to pay any rent, Vargas moved in. But when "Major" Bonilla was at work, Vargas hung out in the nearby Riverside Park.

Just up from 103rd Street one day in September 1948, he saw a college student sitting alone. The guy was friendly, said hello, and Vargas's response sounded to the stranger as if he were French. What transpired that day, the first week in which the stranger had arrived on the GI Bill to study English at Columbia University, has been described in Warren Allen Smith's biography/autobiography of their 40 years together. Bonilla helped Vargas get a job in Manhattan's Garment District, one in which he used a sewing machine to stitch garments and was paid by the piece.

But in 1949 Vargas moved from Bonilla's place with Smith to a furnished room on 109th Street, where their Austrian landlady, Sophie Likar, liked them so much she allowed them alone of her various renters to share her refrigerator. Smith now had earned his M.A. at Columbia - Vargas took a photo after Eisenhower handed him the sheepskin - and began teaching English at Bentley School on 86th Street. He answered an ad for Vargas that led to his being interviewed by R. E. L. Lab, where he made a favorable impression on Edwin Armstrong after being coached about how to weld a certain item. Hired, he found Armstrong avuncular and confiding his problems. Telling Armstrong he lived on 109th near Columbia, he learned that his boss taught there and in 1913 had discovered regeneration. With talk about amplifiers, feedback, and oscillators, the work turned out to be educational and enjoyable, particularly when Edwin - they soon were on a first-name basis - told of being paid only $1 by Columbia but was earning money from his patents. He allowed the military to use royalty-free his findings about FM - frequency modulation - but when the war ended RCA claimed that they, not he, had invented it and they were suing each other. Edwin complained that he became nearly bankrupt from defending his patents, that his wife had left him in the middle of all this, and in 1953 his licenses and patents would all expire. He did not confide everything all at once, but on some days when agitated he would tell bits and pieces about having made the first portable radio and advanced AM radio and on other days he was like a friendly uncle.

In 1954, Edwin dressed neatly, walked to a 13th story window, and jumped out. His body hit a third story overhang. Vargas was devastated. Later, Armstrong's wife settled with RCA for over a million dollars. Vargas then studied electrical engineering at RCA Labs in Greenwich Village, West 4th and Greenwich Street, receiving a certificate in a year.

When Major Bonilla told them that a 1 1/2 room apartment adjacent to his own on 103rd Street was vacant (and had once been lived in by a mistress of Costa Rican President Rafael Calderón Guardia), the two moved into their first apartment together.

Upon hearing from one of his many gay friends that Audiosonic Recording Studio was looking for someone who knew about electrical engineering, Vargas applied for the job at the Brill Building on Broadway and was hired as a temporary the same day by Bob Guy, the studio's owner.

When one of the engineers mentioned that there was a vacant 2 1/2 room apartment at 425 West 45th in Hell's Kitchen, Vargas and Smith moved into Apartment 3FW where the rent of $194.40/month was only slightly more than the one back on 103rd Street.

By 1961, Audiosonic was floundering. Guy, the bi-sexual owner, was kiting checks and had put a prima donna trick on the payroll, Vargas complained. Fellow engineer Joe Cyr warned that they might all need to look for a job somewhere else. Vargas's and others' paychecks began to bounce and, at a quickly called meeting the owner said, yes, the place was bankrupt, he was sorry the paychecks were no good, and Eaton Factors that was owed money was going to shut the place down.

Cyr and Vargas, explaining this to Smith, suggested the three together might try to salvage the company by working with Eaton Factors. It was agreed that the three would be equal owners of a Sub-Chapter S corporation that would be set up. Space at Variety Arts, a major rehearsal building on 46th Street, became available. With many difficulties, the three not only founded Variety Sound Corporation but also included Guy as a partner because he had contacts with all the clients and would include Ad-Lib, his company that made radio jingles for stations.

In April of 1961, Variety Sound Corporation was formed. Smith and Vargas also started Variety Recording Service, a d/b/a that separated Variety Sound Corporation's recording income from Vargas's wholly owned dub-cutting business. The agreement with Guy included the stipulation that only Smith could sign checks, and eventually it was arranged that Guy would exchange his one-fourth interest in Variety Sound for his entire Ad-Lib company. With Guy no longer associated with the business, Variety continued profitably until Vargas's death in 1989.

Philosopher Corliss Lamont came early one morning to Vargas's 103rd Street Manhattan apartment to speak with Vargas’s roommate, Warren Allen Smith. Finding Vargas in red pajamas, he said, “Oh, your roommate rooms with the Devil?” It was Vargas’s first introduction both to a Columbia University professor and to a bona fide naturalistic humanist.

On another occasion Vargas attended Charles Francis Potter’s humanist “church,” laughing at the lecture on the joys of sex. But astronomy, not religion or academic philosophy, was his major diversion. A nominal member of the Bertrand Russell Society, he was only mildly interested in the various philosophers. For its founding meeting in 1989, he allowed the New York Chapter of Secular Humanists to meet in his recording studio and became its first member.

To have lived a great life, no matter how long, is life’s purpose, he believed. To that end he mastered acetate disks, using his own inventive modification of a Scully lathe and being one of the few in the Greater New York Area who could operate such a machine; recorded Broadway plays, working with Arthur Miller ("After the Fall," 1964), Paddy Chayevsky ("Joseph D," 1964], Robert Whitehead ("Medea," 1982), Hal Prince, "Fiddler on the Roof," 1964; David Amram ("Joseph D," 1964); worked with internationally known songwriters and performers (he recorded Liza Minnelli’s first demo record, at which Marvin Hamlisch was accompanist in 1963); worked with songwriter Jerry Bock on “Fiorello” (1959) and “Fiddler on the Roof” (1964); completed master acetates from 1961 to 1990 for Sun Ra, arguing over wine with him about mysticism; and was well-known among a who’s who of Broadway and Latino musicians and artists.

Vargas produced a collector’s LP of “Costa Rica’s Caruso, "Manuel Salazar."

He sang “I-gotta-be-me” and fearlessly ventured on life’s less-traveled roads.

Vargas, who died six months after being diagnosed with having Kaposi’s sarcoma in the lungs, was resigned to his condition and spent much of his final and painful weeks studying the latest developments in astronomy. Up to 2007, over 25,000,000 had died of AIDS and in 2007 an estimated 33,000,000 adults and children were living with HIV/AIDS.

A portion of his cremains were scattered in the Hell’s Kitchen and Times Square neighborhoods of New York City, where he had spent most of his life. Some ashes were deposited behind the walls of the Variety Recording Studio during a time in which a door was being replaced and they were easily inserted. Another portion was saved to be mixed with the cremains of his companion of forty years, Warren Allen Smith, to be buried at the Spencer Smith plot in Waukee, Iowa. The bulk was returned by Smith to Costa Rica, where his cremains are buried in the family tomb next to his father (Elias) and mother (Elena) at San José’s Central Cemetery. In his honor, an Agua Buena support group was formed in Costa Rica.

Crystallinity and porosity are of central importance for many properties of covalent organic frameworks (COFs), including adsorption, diffusion, and electronic transport. We have developed a new method for strongly enhancing both aspects through the introduction of a modulating agent in the synthesis. This modulator competes with one of the building blocks during the solvothermal COF growth, resulting in highly crystalline frameworks with greatly increased domain sizes reaching several hundreds of nanometers. The obtained materials feature fully accessible pores with an internal surface area of over 2000 m2 g–1. Compositional analysis via NMR spectroscopy revealed that the COF-5 structure can form over a wide range of boronic acid-to-catechol ratios, thus producing frameworks with compositions ranging from highly boronic acid-deficient to networks with catechol voids. Visualization of an −SH-functionalized modulating agent via iridium staining revealed that the COF domains are terminated by the modulator. Using functionalized modulators, this synthetic approach thus also provides a new and facile method for the external surface functionalization of COF domains, providing accessible sites for post-synthetic modification reactions. We demonstrate the feasibility of this concept by covalently attaching fluorescent dyes and hydrophilic polymers to the COF surface. We anticipate that the realization of highly crystalline COFs with the option of additional surface functionality will render the modulation concept beneficial for a range of applications, including gas separations, catalysis, and optoelectronics.

pubs.acs.org/doi/abs/10.1021/jacs.5b10708

We present a method of rendering aerial and volumetric graphics using femtosecond lasers. A high-intensity laser excites physical matter to emit light at an arbitrary 3D position. Popular applications can then be explored especially since plasma induced by a femtosecond laser is safer than that generated by a nanosecond laser. There are two methods of rendering graphics with a femtosecond laser in air: producing holograms using spatial light modulation technology, and scanning a laser beam using a galvano mirror. The holograms and workspace of the system proposed here occupy a volume of up to one cubic centimeter; however, this size is scalable depending on the optical devices and their setup.

Credit: Yoichi Ochiai

Complete back view of set. Two aerial sockets might seem strange to some, but is quite normal for 405 line sets post 1953. The initial BBC allocations were all Band 1, but when ITV joined the scene, Band 3 was opened up. Multiband aerials were not popular in the UK, possibly because they were seen as a compromise, except for good signal areas, and also that transmitters were not always co-sited.

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

amorphica.com/networked.html

www.facebook.com/amorphica

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

TNS 400 serie Servo Motor is designed to meet almost all basic light duty requirements of various commercial sewing machines, including lockstitch sewing machines, overlock sewing machines and interlock sewing machines. It utilizes Ferrite permanent magnets.The motor produces almost no noise, saves energy (60-80%) and is brushless, speed djustable and durable. It provides a high starting torque even at low speed or from a complete stop. By using a modern technologically advanced microprocessor,Hall sensor and Pulse-Width Modulation technology,the TNS400 series motor can be set to rotate at different maximum speeds, in either normal or reverse directions, and can start with different accelerating speeds. It will stop automatically with any interruption such as in-line voltage, electrical surge, radio frequency interference or overloading. It is fully protected by the software and will give error messages indicating which problem is encountered. It even works well in environments with an unstable electrical power supply.

Features:

Features & Settings:

1.Motor rotating direction setting

2.Slow starting speed

3.Maximum speed setting

Supplied with:

1.Instruction manual / parts list

2.Mounting hardware

3.Pulley cover (belt guard)

4.65mm pulley

5.Push-button on/off wire harness

Specification:

1.110V Singe Phase

2.Cycles: 50/60

3.Variable Speed to 5000 RPM

4.Power: Up to 1000W

Application:

All kinds of lockstitch sewing machines, overlock sewing machines and interlock sewing machines.

www.tnservomotor.com

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

amorphica.com/networked.html

www.facebook.com/amorphica

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

Microbiology Test Piece Medi-Ca of DNP Medi·Ca Culture Dish Media

Make food microbiological testing much easier and more effective

Compared with the agar medium, our bacterial culture media plate makes the detection of food microorganisms easier, more standardized, and more efficient, but without pollution.

1. Culture media ready to be used after opening;

2. Colonies easy to see;

3. Reduce the culture area and waste.

Notes

1) The product is used for the microbiological examination of foods and beverages, and cannot be used clinically.

2) Do not open the tectorial membrane before transplanting the sample.

3) Do not use expired products.

4) Do not use products that are damaged, deformed, discolored, dirty, or foreign matters.

5) Do not expose the product to ultraviolet light or sunlight.

6) Do not press the film after the sample liquid is dropped. This will cause the sample solution to overflow to the outside of the culture area.

7) When the sample solution overflows from the culture area, replace it with a new one.

8) If the product enters the eye or mouth, immediately wash it with water and go to the hospital.

9) When using the product, it is susceptible to microbial contamination. Perform specific operations under the guidance of relevant qualified personnel.

10) Please note that the sample or the product that has been in contact with the sample liquid is a contaminated item.

Preservation method

Keep in cold storage (2~8°C), fold it twice at the opening of the bag after breaking, and fix it with tape.

Validity period

The validity period of the product is written on the aluminum bag and the upper part of the product (the date described after "EXP" is the validity period). Please use the aluminum bag within 3 months after opening.

Abandonment method

The product has a risk of secondary pollution after use. Therefore, after proper sterilization, it should be disposed of according to the waste standards of the respective samples and related facilities.

Guarantee responsibility

When the product defect is obvious, replace the corresponding quantity with the new product. The judgment and application of the inspection results are the user’s responsibility, and the manufacturing company and the agent of the product are exempt from liability.

Quantity per carton

Medi-Ca AC and Medi-Ca CC: 1000 sheets

Medi-Ca EC and Medi-Ca SA: 500 sheets

Performance Characteristics of DNP Medi·Ca Culture Dish Media

Medium ready to be used after opening

The product does not require pre-modulation and autoclaving, just check it. No need for mixed release. Double layer coverage is easy to operate which will improve working efficiency.

Colonies easy to see

Due to the use of the color former, the colonies are clearly visible and easy to discriminate. It is beneficial to improve the working efficiency of counting bacteria and working standardization. In addition, since Medi-Ca AC is a bacillus, it is one of the characteristics that the coloring penetration is relatively small.

Reduce the culture area and waste

Compared with the culture dish, it can be reduced to about 1/15~1/20, which can reduce the storage space and the waste after use.

Variety introduction

The Culture Media Plate Medi-Ca is not considered for personal visual differences but produced according to the coloring universal design that most people can easily see, and is also certified by the NPO corporate coloring general design agency.

Usage Method of DNP Medi·Ca Culture Dish Media

1. Sample liquid preparation

Add appropriate sterile dilutions to the sample and homogenize with a homogenizer.

2.Pipetting into the culture area

3.Culture

Put into the incubator, the product can overlap up to 25 pieces.

4．Judgment

SA uses chromogenic enzymes to easily determine and count bacteria.

When the bacteria number is large, please perform statistics in the grid on the film (1cm × 1cm).

Multiply the bacteria number in a grid by the total grids number

5. Fishing for colonies which can be re-cultured

Remove the bacteria by opening the membrane.

Colony Count System

By using the colony counting system (dedicated scanner and application), high-resolution images can be analyzed, and automatic counting and inspection result image storage management can be realized in a short time, which can improve the counting efficiency.

Tear off the foil pouch and remove the required amount.

Medium TypeCulture temperatureCulture time

Media-Ca AC

(Measurement of general bacteria amount)35 ± 1 ℃48±2 hours

Media-Ca CC

(Measurement of Coliforms)35 ± 1 ℃24±1 hours

Media-Ca EC

(Measurement of Escherichia coli and Coliforms)35 ± 1 ℃24±1 hours

Media-Ca SA

(Measurement of Staphylococcus)35 ± 1 ℃24±1 hours

www.elexbio.com/dnp-medica-culture-dish-media.html

28th February 2015 at Half Moon, Putney, London SW5.

Effects Pedals modify the sound of a musical instrument such as an Electric Guitar by means of changes like distortion, modulation, and feedback. They are often found on the floor on a pedalboard, and are operated with the feet.

The photo includes (top to bottom): Boss Chromatic Tuner TU-2, Boss Delay DM-3 (a 1980s analog delay pedal), Durham Electronics Zia Drive (an overdrive pedal, which distorts the wave form of the audio signal) and Danelectro Tuna Melt (a tremelo pedal).

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

amorphica.com/networked.html

www.facebook.com/amorphica

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

Ogundipe Fayomi's monument for Dr. Ronald Erwin McNair (1950–1986) combines a traditional bust with a uniquely shaped pedestal. McNair was the African-American astronaut, physicist, teacher, and musician who died aboard the Space Shuttle Challenger when it exploded on January 28, 1986.

This park, formerly known as Guider Park, was named for Dr. McNair in the same year as the Challenger disaster. The City’s Department of Cultural Affairs sponsored a competition through its Percent-for-Art program to choose an artist to create a central sculpture. They ultimately selected the Nigerian-born sculptor Fayomi, who fashioned a sensitive bronze portrait, set within a nine-foot tall polished red-granite pedestal resembling a modified rocket ship. The pyramidal base features bronze relief with images relating to Dr. McNair’s achievements and interests.

Dr. McNair was born on October 21, 1950, in Lake City, South Carolina. He graduated from Carver High School in Lake City in 1967, and received a B.S. degree in physics from North Carolina A & T State University in 1971. In 1976, Dr. McNair completed his Ph.D. in physics at the Massachusetts Institute of Technology (MIT). After graduating from MIT, Dr. McNair was employed as a staff physicist at Hughes Research Laboratories in Malibu, California. His work there involved developing lasers for isotope separation and photochemistry, using non-linear interactions in low-temperature liquids. He also conducted research on electro-optic laser modulation for satellite-to-satellite space communications and explored the scientific foundations of the martial arts. A member of numerous scientific organizations and a visiting lecturer in physics at Texas Southern University, Dr. McNair also taught karate as a fifth-degree black belt and was a performing jazz saxophonist.

In 1978, the National Aeronautics and Space Administration (NASA) selected Dr. McNair as an astronaut candidate. He completed his training the following year, and became eligible as a mission specialist astronaut on Space Shuttle flight crews. He first flew as a mission specialist on Mission STS-41-B on February 3, 1984, which featured the first untethered spacewalk. Serving as a mission specialist on Mission STS-51-L, his life was tragically cut short when the space shuttle exploded one minute and 13 seconds into the launch. After his death, the Dr. Ronald E. McNair Foundation for Science, Technology & Space Education was established in Atlanta, Georgia.

When this monument was dedicated on June 14, 1994, family, friends, former colleagues, community representatives, city officials and hundreds of school children gathered in memory of Dr. McNair’s legacy. The monument and the park, which was renovated at the time of the sculpture’s installation, evoke a mood in keeping with Dr. McNair’s wish inscribed on the pedestal. It reads, “that we should allow this planet to be the beautiful oasis that she is, and allow ourselves to live more in the peace she generates.”

The peony is named after Paeon (also spelled Paean), a student of Asclepius, the Greek god of medicine and healing. Asclepius became jealous of his pupil; Zeus saved Paeon from the wrath of Asclepius by turning him into the peony flower.

Over 262 compounds have been obtained so far from the plants of Paeoniaceae. These include monoterpenoid glucosides, flavonoids, tannins, stilbenes, triterpenoids and steroids, paeonols, and phenols. Biological Activities include Antioxidant, Antitumor, Antipathogenic, Immune-System-Modulation Activities,Cardiovascular-System-Protective Activities and Central-Nervous-System Activities.

Turbocoleoptozoide a modulation cosmique urbexologique n°10

Blacktron Gold - Listening and Assault Unit

Spacecraft equipped with:

- stereo cockpit

- optoechoic head

- white noise generator

- modulation metronome

- dual megabass cannon

- large aperture antenna with phrase scanning

- dual IR (iridium) jam-session-er

- powerful pro-tone torpedo

- dual frequency Hi-Fi-per sonic missiles

I've not found the correct 9 volt battery for this yet, but it does play when I apply 9 volts to the terminals.

amorphica.com/networked.html

Group 4_

Aaron Onchi, Betty Sanchez, Roberto Gutierrez, Frank Durán , Belén Olaya García

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

The Dirty Carter Electronic Sound Generating Instrument was designed by John Richards (Dirty Electronics) and Chris Carter from legendary Industrial pioneers Throbbing Gristle. It was produced for a special performance by Carter and the 25 strong Dirty Electronics Ensemble in 2010. It was originally designed as a touch controlled instrument with the player's skin resistance completing the circuit. This hard wired modification by A.S.M.O. gives more control and predictability by wiring all to the touch contacts to pots and switches. An additional low pass resonant filter has been added, LFO and an external CV socket for filter modulation.

The case is made of stained ply and the front panel is covered with black leatherette.

amorphica.com/networked.html

Group 4_

Aaron Onchi, Betty Sanchez, Roberto Gutierrez, Frank Durán , Belén Olaya García

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

amorphica.com/networked.html

Group 4_

Aaron Onchi, Betty Sanchez, Roberto Gutierrez, Frank Durán , Belén Olaya García

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

amorphica.com/networked.html

Group 4_

Aaron Onchi, Betty Sanchez, Roberto Gutierrez, Frank Durán , Belén Olaya García

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

2 RED LFOs and a Proto of the GREEN SEQ (here in black as the green Plexi didnt make it in time...)

all running Temposynchron, the left LFO modulating PWM, the 2nd modulating Filter Cutoff, the Sequencer OSC Frequency (via a Quantizer).

You cant imagine how much fun it is just to step thru the LFO Waveforms and adjust the PhaseShift for the Filter Modulation......

Sample image taken with a Fujinon XF 56mm f1.2 R mounted on a Fujifilm XT1 body; each of these images is an out-of-camera JPEG with Lens Modulation Optimisation enabled. These samples and comparisons are part of my Fujinon XF 56mm f1.2 R review at:

cameralabs.com/reviews/Fujifilm_Fujinon_XF_56mm_f1-2_R/

Feel free to download the original image for evaluation on your own computer or printer, but please don't use it on another website or publication without permission from www.cameralabs.com/

Echo Base Delay / Modulation Pedal PT2399

An infographic detailing the history of guitar effects, starting all the way back to the 1930s with Rickenbacker’s Vibrola Spanish Guitar and progressing to the present day. Along the way, we’ll see iconic guitar-effect breakthroughs like gain and reverb in the 1940s to 1950s’ effects like fuzz (discovered by accidentally dropping a Fender Bass amp on a rainy street!) and distortion (the fortuitous discovery when Link Wray stabbed a hole in his amp’s speaker). The guitar effects hit their stride in the 1960s with the first transistor-powered guitar pedals, including the first wah-wah pedals and the first octave effect pedal among glorious others. The explosion of effects pedals in the 1970s reverberates today: signal alterations, distortion, modulation, time-based effects, and filter effects. A timeline of all these effects along with the iconic musicians who used what—Bo Diddley’s Trem Trol 800 Tremolo, Jimi Hendrix’s Leslie rotating speaker, the Rolling Stones’ Maestro Fuzz Tone, etc.

Feel free to use this infographic but please give credit with a link to www.songsimian.com. The original infographic, with embed codes, can be found here.

The red side button was transplanted from the original two-way radio. It turns on the "robot" pseudo ring modulation.

Most recently dug out of the basement from the radio boxes, a citizen's band transceiver; it provides AM, USB and LSB operating modes over the usual 40 channels. It can be modified to do more, but that's not of interest to me. It has very good audio quality on both receive and transmit, and enough capabilities that along with a good antenna, any reasonable communications scenario can be addressed.

Notable are three different types of noise reduction; instant access to channels 9 and 19 without disturbing the main dial; courtesy beep; RF gain; RF power; tone; SWR, power and modulation metering; transmit monitoring; full panel dimming; and PA capability, which will be useful to us as a quick way to address the back yard, as we have no windows back there, only cameras.

Canon EOS 50D [modified IR response in Hα range], Canon EF-S 18-55mm ƒ/3.5-5.6 IS zoom [ø58mm] @ 23mm, tripod, IS off, available light (indirect sunlight through closed opaque blinds), ƒ/3.5, ISO 100, 2.5s exp. Taken in SRAW; this is a 1:1 clip.

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

amorphica.com/networked.html

www.facebook.com/amorphica

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

WORMOD Art 330 : By passing from a basic design with various effects and modulations, a Kaleidoscopic Chrominance is obtained.

The title “Unitxt” could be read as “unit extended” referring to a unit of a rhythmic grid or universal text referring to a universal language, e.g. mathematics: units, constants, measurements, prefix-, SI-system of units and is represented in spoken word and by codes in sound itself.

In collaboration with derivative’s touch designer software the original visuals, which are based on real-time manipulation/modulation of software-generated test patterns, has been expanded to a multi-screen set-up with a highly elaborated visual outfit to form an installation of an almost immersive quality.

This special set-up has been developed for more large scale venues and festivals of high demand. So far it has had presentations at Transmediale Berlin, followed by shows at Mutek Montréal and Sonár Barcelona.

credit: rubra

The first commercially available synth to implement MIDI!! It's a fun synth. Its big brother is the legendary Prophet 5. The P600 is very affordable today and is a great buy. Models with the newest software will enjoy polyphonic MIDI implementation and up to 100 memory patches to store their own sounds! The sound of the Prophet 600 is brighter and harsher than that of a Juno 106 but still just as funky.

The P600 has two oscillators per voice with sawtooth, triangle and variable pulse waveforms. The oscillators can be individually tuned or synced together. Similar quality VCF and VCA sections from the Prophet 5 can be found here too! The P5's Poly-Mod section has also been passed onto the P600.

The P600 is extremely versatile and easy to use! Its best functions include the onboard arpeggiator, 2-track sequencer and poly-modulation. The P600 is great for creating analog effects, swells and drones. It has a cool glide effect and has very flexible modulation possibilities!

Please visit www.winecountrysequential.com for spare parts and software upgrades.

Space Baby modulated beat-synced digital delay. This was the project for Handmade Music Austin #5 (along with the Bleep Labs PicoPao), which was on Feb 28, 2010. More information is available on woosteraudio.com/space-baby.html

amorphica.com/networked.html

Group 4_

Aaron Onchi, Betty Sanchez, Roberto Gutierrez, Frank Durán , Belén Olaya García

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

§ Eventually, after a long sequence of successive impressions that incessantly compassed into our experiences, the journey is terminated to a dissolution of serenity when we stand confronting the frontage of central sikhara • The incremental aspect in the anthropometrical scale of sikhara is inverse to the decremental determination in the sensational space of our Self • When our expectation was aroused in each step of experiences, we finally can substantiate the salient characteristics of this destination • The architect articulated the modulation of scale and space as cogent mean to stimulate our delectation while enforcing us simultaneously to be aware of our deterioration before affirming them into emancipation at end •

§ The galleries that conjoin to each cardinal axis of the central sikhara project our eyes to elevate from the horizontal line to the finial • The massive form of God proposes incisively an effect to our senses and elicits an emotion to its plastic form that wakes a profound reverberation in us • The ever-diminishing tiers of the curvilinear sikhara assist our eyes to comprehend the finial, and thereby the infinite atmosphere above the sky and the Universe • The formal diminution of its superstructure, by the dimensional depth of its perspective in compliant with the inventive space of the vaporous cloud that oscillates about the finial, reveals the ingenious articulation of the architect in maturating our contemplation with deepest relaxation •

§ Angkor Vat is not an autotelic architecture • The manifestation of the Universe, the objective world of reality, the analogy of God to the subjective realm of Truth in Self, all were simply existed here • A brief tenancy of ego (ahamkara) in the world of human egocentrism in the past, the present, and the future is a mere delusion; only when we can deliberate ourselves from ignorance • Time is a reflexive force of space which contains in all animate beings with ignorance • Only quintessence of our Self-Realisation will terminate time and space of here and there everlastingly •

Crown XLS1000 DriveCore Series Power Amp Description:

Crown's XLS1000 power amplifier is a premiere portable PA system with unmatched performance, technology, and affordability. It includes multiple inputs so you can plug in anything and play anywhere, along with several system setup configurations. This high-performance Crown power amp provides enormous power and flexibility thanks to the integrated DriveCore Technology, PureBand Crossover System and Peakx limiters. Weighing less than 12 pounds, The Crown XLS1000 power amp is much easier to set up and move from show to show.

A Power Amp with Integrated DriveCore Technology Class D amplifiers are notable for extraordinarily high efficiency and being well suited for driving difficult reactive loads such as subwoofers. However, their performance can suffer impaired performance on marginal and unstable AC line supplies. To overcome this obstacle, Crown engineers developed DriveCore Technology—a proprietary hybrid analog-digital integrated circuit (IC) developed with Texas Instruments that drives the "front end" of the Class D output stage. Over 60 years of Crown's design knowledge and experience went into the development of this technology resulting in truly remarkable benefits. The DriveCore Technology incorporated in the Crown XLS1000 power amp provides an extremely wide tolerance with regards to sagging or "dirty" AC line conditions, providing consistent performance without affecting audio quality. This means the Crown power amp will not compromise your performance by fluctuating generator power, or overload from lighting rigs, backline gear, etc.

In addition, DriveCore Technology's patented feedback and PWM modulation circuits enable fast recovery on peak transients, accurate reproduction of low-level detail, and precise tracking of low-frequencies at high power levels for maximum subwoofer output.

Advanced Switched-Mode Power Supply

The advanced power supply in the Crown power amp is highly efficient and optimized for maximum power transfer from the AC line through the Class D output stage to the loudspeakers. A benefit to this is substantial weight reduction when compared to older 60Hz transformer-based power supplies.

PureBand Crossover System

The PureBand Crossover System in the XLS Series adds an enormous amount of flexibility and performance to any system. With this system, the crossover frequency is completely variable allowing the choice of any crossover point between 50Hz and 3kHz on 1/12 octave centers. The use of 4th order Linkwitz-Riley filters provides steep slopes for a seamless transition between high and low drivers. And with four crossover modes to choose from providing the ultimate in flexibility, all of your system needs are covered.

Peakx limiters

Peakx limiters provide the ultimate in performance and protection for your entire system. This advanced algorithm was specifically developed and tuned to work with this amplifier and power-supply to achieve higher SPL will less audible artifacts. This means less distortion, less shutdowns, and maximum safe power delivered to your speakers. The Peakx limiters can be easily turned on or off by channel right from the front panel eliminating the need to be digging around in the back of the dark rack.

Crown XLS1000 DriveCore Series Power Amp Features:

XLS High Performance, Lightweight Class D amp weighs less than 11 lb.

Integrated PureBand Crossover System for better performance and control

Peakx Limiters provide maximum output while protecting your speakers

XLR, 1/4", RCA inputs provide outstanding flexibility

1/4" Inputs can be used as loop-thrus to distribute signal to additional amplifiers Efficient forced-air fans prevent excessive thermal buildup

Electronically balanced XLR inputs; touchproof binding post and Speakon outputs

Precision detented level controls, power switch, power LED, and six LEDs indicate signal, clip and fault for each channel

Three-Year, No-Fault, Fully Transferable Warranty completely protects your investment and guarantees its specifications

SMS303 Tantek Tanrak (9 module Modular FX):

- Comp-Lim2

- Parametric Equaliser

- Enhancer

- Modulation Oscillator

Info:

Mid 1980's Tantek, Tanrak Studio Effects Rack which was available in kit form or ready built. These were bought as kits and put together by an electronics engineer. On the face of it, they're simple analogue effects - a bit old-fashioned, really - but that's the charm of them. They've perfectly useable and immediately accessible, so you'll have great fun fiddling with the settings - try sweeping the EQ frequency, or riding the delay time for on-the-fly munchkinisation, for instance.

Even better, you'll find new ways to patch the modules together. Everything - in, out and sidechain - is accessible from the rear panel (there's a default path from left to right across the rack if you don't want to use patch cords) so you can create LFO-modulated delay effects, frequency-sensitive compression ... you think of it, you can do it.

STEREO COMPRESSOR/LIMITER - A high quality stereo comp/limiter with variable input, slope, attack and release controls, and a switched 'key' input that can link both channels...handy for de-essing, ducking etc. It's pretty much 'invisible' when used as a limiter, only squeezing when the threshold is crossed (depending on the ratio setting). Great for laying vocal tracks, mix thickening, fattening up drums, percussions and bass. In fact, it can make anything sound 'phat' but still retains that important top-end clarity.

MODULATION OSCILLATOR - A CV modulation source whose features include sinewave output, variable duty cycle, key or CV controlled depth, triggerable sweeps and two independently variable outputs. Used with the muli-dealy to create chorus, flanging etc.

The Dirty Carter Electronic Sound Generating Instrument was designed by John Richards (Dirty Electronics) and Chris Carter from legendary Industrial pioneers Throbbing Gristle. It was produced for a special performance by Carter and the 25 strong Dirty Electronics Ensemble in 2010. It was originally designed as a touch controlled instrument with the player's skin resistance completing the circuit. This hard wired modification by A.S.M.O. gives more control and predictability by wiring all to the touch contacts to pots and switches. An additional low pass resonant filter has been added, LFO and an external CV socket for filter modulation.

The case is made of stained ply and the front panel is covered with black leatherette.

amorphica.com/networked.html

Group 4_

Aaron Onchi, Betty Sanchez, Roberto Gutierrez, Frank Durán , Belén Olaya García

Networked Fabrication for Urban Provocations.

Shifting Paradigms from Mass Production to Mass Customization

Computational architecture and design course

Conventional construction methods all depart from the basic premises of mass production: standardization, modulation and a production line. What these systems developed during the last two centuries fail to take into account are the evolutionary leaps and bounds the manufacturing industry has taken over the last decades. With the introduction of CNC technologies and rapid prototyping machines have altered the paradigms of fabrication forever. It is due to these new tools that it is now possible to create (n) amount of completely unique and different pieces with the same amount of energy and material that is required to create (n) identical pieces. The possibilities for implementation of new forms, textures, materials and languages are infinite due to the versatility that these new tools offer a growing network of architects, designers, fabricators that are integrating them into their professional practices to generate unique and precise objects that respond to countless data and real-life conditions.

Instructors:

Monika Wittig [ LaN, IaaC ]

Shane Salisbury [ LaN, IaaC ]

Filippo Moroni [ SOLIDO, Politecnico di Milano ]

MS Josh Updyke [ Advanced Manufacturing Institute, KSU, Protei ]

Aaron Gutiérrez Cortes [ Amorphica ]

Specification

Coach Model MAN 18.350 HOCL/R

Chassis Length 11,850 mm

Chassis Width 2,526 mm

GVW 18,200 kg

Engine Type

Vertical, Water Cooled 6-cylinder 4-stroke Diesel Engine

With Common Rail Injection,

Exhaust Turbocharger and Intercooler

ECR, Replaceable Cylinders Liners

Engine Model MAN D2066 LUH13 Euro 4

Displacement 10,518 c.c

Maximum Output 257 kW (350 hp) @ 1,700 rpm

Maximum Torque 1,750 Nm @ 1,000-1,400 rpm

Bore 120 mm

Stroke 155 mm

Fuel Capacity 300 dm³

Transmission ZF 6S 1900 BO 6-speed Synchromesh Manual Transmission

ZF 6 HP 504C 6-speed Automatic Transmission

Voith D864.5 4-speed Automatic Transmission

Drive Axle MAN HY-1336-B

Suspension Capacity 13,000 kg

Front Axle MAN V9-82 SL

Suspension Capacity 8,200 kg

Brake

Dual Circuit Air Brake System to ADR Directives by Wabco

Front and Rear Axle Disc Brakes

Electronic brake system EBS (ABS, TCS)

Auxiliary Brake Manual Transmission: Engine Brake Valve (EBV)

Automatic Transmission: Integrated Retarder and

Water Cooled with Electric Pressure Modulation

Suspension

Air suspension with 6 identical rolling seals

With Integrated Elastic Stroke Limiter

Electronically Controlled Constant Entrance Height

Suspension Characteristics Under All Load Conditions

Front Suspension 2 x Air Bellows

2 x Shock Absorbers

1 x Level Control Values

1 x Stabilizers

Rear Suspension 4 x Air Bellows

4 x Shock Absorbers

2 x Level Control Values

1 x Stabilizers

Blacktron Gold - Listening and Assault Unit

Spacecraft equipped with:

- stereo cockpit

- optoechoic head

- white noise generator

- modulation metronome

- dual megabass cannon

- large aperture antenna with phrase scanning

- dual IR (iridium) jam-session-er

- powerful pro-tone torpedo

- dual frequency Hi-Fi-per sonic missiles

Video here: www.youtube.com/watch?v=XrHkvvtrXhA

The Crazy Looper is a small handmade device that allows you the create real-time noise loops with a fast modulation metallic effect.

A regular location for taking photos at. I cross this point twice on bicycle commuting days.

The vehicle's rear lights appears as a dashed line. These are LED rear lights that derive their tail lamp illumination setting via Pulse Width Modulation control (they are turned on/off at a high frequency). The LEDs are operated 'full on' when used as a brake light.

From the early years of transistor radio history... this one has 6 whole transistors! They were very expensive in the late 1950's and started getting cheaper in the 1960's.

At the Cluny Museum, medieval culture showcases its ancestral knowledge. It took five centuries to discover that the thymus and the genitals are connected, as seen in this statue of the first man to experience desire, through a dream about a mythical serpent.....

Within the thymus, regulation of the cellular crosstalk directing T cell development depends on spatial interactions within specialized niches. To create a spatially defined map of tissue niches guiding human postnatal T cell development, we employed the multidimensional imaging platform co-detection by indexing (CODEX) as well as cellular indexing of transcriptomes and epitopes sequencing (CITE-seq) and assay for transposase accessible chromatin sequencing (ATAC-seq). We generated age-matched 4- to 5-month-old human postnatal thymus datasets for male and female donors, identifying significant sex differences in both T cell and thymus biology. We demonstrate a possible role for JAG ligands in directing thymic-like dendritic cell development, identify important functions of a population of extracellular matrix (ECM)− fibroblasts, and characterize the medullary niches surrounding Hassall’s corpuscles. Together, these data represent an age-matched spatial multiomic resource to investigate how sex-based differences in thymus regulation and T cell development arise, providing an essential resource to understand the mechanisms underlying immune function and dysfunction in males and females.

The thymus is the primary organ responsible for the generation and selection of mature, functional, and self-tolerant T cells.1 Effective T cell development is a critical component of our immune system’s ability to accurately and exclusively identify and kill foreign entities such as pathogens. During early postnatal T cell development—the period in life when T cell development is most active2—thymic seeding progenitors migrate to the thymus and mature into thymocytes. Thymic architecture is highly organized to provide spatially defined, stage-specific signaling cues to migrating thymocytes that guide development toward functional mature T cells.3,4,5,6

Recent single-cell sequencing resources demonstrating the diversity of human thymus tissue are incongruous with our current framework of thymus structure and organization,7,8,9,10,11,12,13,14,15,16,17,18,19 which describe a general migratory path thymocytes take through the cortex and medulla during conventional αβT cell development. Spatial transcriptomic sequencing of human thymus has demonstrated a deeper granularity of thymic niches and their evolution during fetal development to support different waves of non-conventional T cells.19,20 However, our understanding of how human postnatal thymus niches support conventional and non-conventional T cell development, T-lineage branching, and alternative lineage development remains limited.3,4,6 T cells generated at this stage of postnatal human development will become the foundation of our immune system, patrolling the body for decades.21 Thus, insights into early postnatal thymus niche biology are crucial to understand how our adaptive immune system is built and how perturbations in postnatal T cell development may emerge as immune dysfunction later in life.

To create a spatially defined map of tissue niches guiding human postnatal T and alternative lineage cell development, we employed multi-dimensional spatial proteomic imaging using co-detection by indexing (CODEX),22,23 single-cell transcriptomic-proteomic profiling using cellular indexing of transcriptomes and epitopes sequencing (CITE-seq),24 and single-cell assay for transposase accessible chromatin sequencing (ATAC-seq).25 Given the emerging recognition of sex differences in thymus gene expression and function,26,27,28,29,30,31 we collected and analyzed samples from male and female donors. Our analysis identifies significant sex differences during early postnatal development that affect T cell and thymus biology through common and cell type-specific mechanisms. Additionally, we highlight key cell types contributing to thymic involution that exhibit sex-based differences in thymic growth and early transition toward adipogenesis. These data suggest that kinetic differences in thymic involution are present between sexes and, importantly, that mechanisms driving thymic involution begin early in life. Altogether, these data represent a powerful age-matched spatial multiomic resource to investigate how sex-based differences in thymus biology and T cell development arise, and how they contribute to sex differences in diseases caused by immune dysfunction.

Results

Spatial multiomic profiling of human postnatal thymus identifies sex-based differences in T cells and thymus biology

We performed single-cell CITE-seq, ATAC-seq, and CODEX imaging on 4–33 months human postnatal thymuses, including 6 (3 female and 3 male) 4- to 5-month-old age-matched samples (Table S1). Each donor sample was processed simultaneously for CODEX imaging and sequencing (Figure 1A). We included a comprehensive 137 antibody panel (Data S1), allowing us to compare epigenomic, transcriptomic, and proteomic expression kinetics across developing thymocytes and enabling direct comparison of cells identified via phenotypic expression in CODEX with cells captured via CITE-seq. Prior to sequencing, we enriched CD45− non-hematopoietic cells and CD25+CD8− regulatory T (Treg) cells to ensure coverage of low-abundance cell types. After quality control and computational merging of individually sequenced patient datasets, we obtained a total of 74,334 cells with CITE-seq, including 19,434 non-T-lineage cells, and captured 25,717 nuclei with ATAC-seq. Importantly, cell proximity in CODEX tissue niches was used to screen predicted receptor-ligand interactions.

Figure 1 Spatial multiomic analysis identifies sex-biased characteristics of thymic niches

Show full captionFigure viewer

CITE-seq cells were clustered based on transcriptional expression and annotated based on marker gene and surface protein expression (Figure S1A; Table S2).7,8 ATAC-seq clusters were computationally labeled using CITE-seq reference cluster labels, which identified 34 ATAC-seq cluster transfer labels for dataset integration (Figures 1B and S1B). We captured 54,900 thymocytes spanning development from early thymic progenitors (ETPs) to mature single positive (SP) T cells, immature innate cells, innate-like cells, and Tregs. We identified three Treg populations expressing canonical lineage markers, namely Treg progenitors (Pro-Tregs), thymic Tregs (tTregs), and recirculating/resident Tregs (rrTregs).32 We also identified antigen-presenting cells, including B cells, mast cells, monocytes, and six populations of dendritic cells (DCs).33 In addition to the activated DCs (aDCs), plasmacytoid DCs (pDCs), DC1, and DC2/3 populations described by Park et al.,7 we found proliferating populations of pDCs and DC1. We also captured 7,093 epithelial cells, including cortical epithelial cells (cTECs), medullary epithelial cells (mTECs), activated mTECs, and mimetic TECs.

Importantly, we captured 7,721 mesenchymal cells, which contribute to negative selection and thymic involution.9,19,34,35,36 Subclustering identifies important mesenchymal cell types, including two populations of endothelial cells (ECs) defined by differential expression of Notch ligands (ECs, ECs (Notch)). Additionally, we identified lymphatic ECs (LECs), pericytes, vascular smooth muscle cells (VSMCs), and five distinct fibroblast cell types, including DPP4+ capsular fibroblasts (DPP4+ capFibs), capsule fibroblasts (capFibs), medullary fibroblasts (mFibs), KRT+ fibroblasts (KRT+ Fibs), and proliferating fibroblasts (Fibs (P)).

We imaged each tissue sample with a custom 48 antibody CODEX panel to study the architecture and function of niches guiding thymocyte development, aiming to define the niche characteristics guiding T-lineage branch points. Stage-specific thymocyte phenotyping markers (CD62L, CCR7, CD1A, CD5, CD7, CD4, CD8, CD3, CD45RO, CD45RA, FOXP3, and SATB1) identified CD3+ double positive cells (DPs) undergoing T-lineage commitment toward CD4 or CD8 T cells. Phenotyping markers for non-T-lineage hematopoietic cells (CD19, CD11c, CD11b, and CD68), epithelial cells (EPCAM and KRT5/8), mural cells (MCAM and SMA), ECs (CD31), and fibroblasts (PDGFRA) identified the remaining major cell types defining thymic niche architecture. Finally, we included functional markers to define patterns of antigen presentation (CD86), human leukocyte antigen (HLA) class I and II expression (HLA-ABC and HLA-DR,DP,DQ), adhesion ligands (ICAM and VCAM), Notch ligands (DLL1, DLL4, JAG1, and JAG2), T cell activation (PD-1), self-tolerance (PD-L1), proliferation (Ki67), and enzymatic regulation (15-PDGH). In sum, our CODEX panel enabled investigation of spatially regulated mechanisms directing human T cell development.

Using neural-network-driven cell segmentation and Leiden-based clustering,23 we identified individual cells within thymic tissue for each sample (Figure S1C). We annotated cell types based on tissue location and phenotypic expression compared with CITE-seq clusters (Figure 1C), performed proximity-based neighborhood clustering to identify niches,23 and annotated niches based on location and cell type composition (Figure 1D; Figure S1D). This analysis quantified proximity-based cell-cell interactions (Figure S1E) and served as a platform to interrogate spatially defined thymic niche biology via integrated sequencing-imaging analysis.

Because of known sex differences in thymus and T cell gene expression,31 we compared our age-matched male and female samples separately. In line with prior reports of sex-biased gene expression on autosomes,37,38,39,40 only 2% of male differentially expressed genes (DEGs) were found on the Y chromosome and 0.3% of female DEGs were found on the X chromosome (Tables S3 and S4). Gene set enrichment analysis (GSEA) on male vs. female cells for each cell type identified pathways commonly upregulated in either sex (Figure 1E; Data S1). Pathways differentially regulated across hematopoietic, epithelial, and stromal cells represent cell-intrinsic sex-based differences. Female cells have higher gene expression of transcription, energy regulation, and antigen presentation. Male cells, by contrast, have increased gene expression of proinflammatory signaling, amino acid metabolism, and G protein-coupled receptors (GPCR) signaling. The top differentially expressed energy regulation and metabolism pathways were similarly sex-biased in human kidney,41 suggesting multiple organs show consistent sex-biased enrichment of pathways linked to metabolism and energy production. Our data align with sex-biased trends identified in human induced pluripotent stem cell (iPSC) lines42 and other human organs,43 indicating these pathways often differ between male and female cells across various cell types.

By contrast, some pathways showed cell type-specific sex-biased enrichment. Female T and hematopoietic cells showed enrichment of interferon signaling, and female fibroblast and perivascular cells were enriched in extracellular matrix (ECM)-centric pathways (Figure 1E). Our dataset also identified differential sex-specific pathway enrichment between cell types. Gene expression indicated higher cytokine signaling in T cells and hematopoietic cells in females and in epithelial and mesenchymal cells in males (Figure 1E). These data show significant gene expression differences in male and female thymic cells. To demonstrate sex differences at the proteomic level, we identified genes with a log fold change greater than 1 that contributed to increased chemokine signaling in male T cells. CXCR4, an important chemokine receptor in thymocyte migration and development, had increased expression in male progenitor T (pro-T) cells, which we confirmed via flow cytometry (3 male, 3 female; p = 0.03; Figure S1F). As higher levels of cytokine and interferon signaling have been previously shown to influence thymus and T cell biology,44,45 our data suggest male and female T cells develop in different signaling environments and may respond differently to cytokine stimuli.

Next, we quantified cell type abundance within male and female tissues, demonstrating differences in cortical and medullary cell distributions between sexes. When normalized to the total number of cells per lobe, female thymus lobes contained significantly more DPs (p = 0.011) and cTECs (p = 0.0023). In males, we found significantly more SPs (p = 4.2 × 10−4), CD3+ DPs (p = 9.9 × 10−4), activated mTECs (p = 0.0014), and VSMCs (p = 2.4 × 10−6) (Figure 1F). Given that thymus lobules with more DPs and cTECs would have a greater proportion of cells undergoing positive selection and lobules with more medullary cells would have more cells undergoing negative selection, these data suggest that sex differences in cell type abundance may influence the resources directed toward specific stages of thymocyte selection. Alternatively, these results may suggest that male and female thymuses are developmentally asynchronous, with males exhibiting faster growth and involution kinetics, resulting in decreased cortical-to-medullary ratios even in early neonatal stages. We focused further analyses on sequential developmental niches, including analysis of sex differences in cell types and niches at each stage.

JAG1 skews ETP development toward thymic DCs

We first analyzed the cortico-medullary junction (CMJ) where cells home to the thymus (Figure 2A). This region recruits and supports ETPs10 and is composed of ECs, VSMCs, and pericytes expressing the Notch ligand JAG1 (Figures 2B and 2C). CITE-seq demonstrated that the cell adhesion molecule used by ETPs to enter the thymus, CD62L, is quickly downregulated upon CMJ entrance through the vasculature (Figure S2A). However, recently immigrated CD62L+ double negative cells are frequently located in the subcapsular zone (Figure S2B), suggesting that ETPs enter the thymus and rapidly migrate to a subcapsular niche where DLL4, a more potent Notch ligand, is highly expressed on fibroblasts and subcapsular epithelial cells (Figures 2D and S2C). However, the concentrated presence of JAG1 at the entry point indicates that ETPs are first exposed to this Notch ligand.

Figure 2 Thymic progenitors entering via the corticomedullary junction are exposed to a gradient of Notch ligands, which influence lineage specification

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CellChat46 pathway analysis showed that JAG1-NOTCH1 interactions between endothelial and perivascular cells are enriched with ETPs (Figure 2E), while JAG1-NOTCH2 and JAG1-NOTCH3 interactions are enriched with DC1, DC1 (P), DC2/3, and aDCs (Figures 2E–2G). These data suggest that JAG1 could induce commitment toward other hematopoietic lineages, such as pDCs, conventional DCs (cDCs), or macrophages, which are known to develop within the thymus.10 As JAG ligands induce weaker Notch induction,47,48,49,50 we hypothesized that early contact with ETPs could maintain T-lineage potential while cells migrate toward DLL4 in the subcapsular niche.

We first analyzed the ability of the four thymic Notch ligands to induce T-lineage commitment or alternative lineage development from cord-blood-derived CD34+ hematopoietic stem and progenitor cells (HSPCs) in a defined, feeder-free culture system44 (Figure 2H). We included titrated concentrations of granulocyte-macrophage colony-stimulating factor (GM-CSF), which is produced by mast cells at the CMJ, to support DC development.51 We found that only DLL1 and DLL4 ligands induce T-lineage commitment, whereas JAG ligands or no ligand controls supported myeloid cell development and did not induce T-lineage commitment (Figure S2D). Specifically, JAG ligands with GM-CSF skewed CD68+ DC development toward CD14− DC1 cells, while no ligand controls skewed CD68+ DC development toward CD14+ DC2/3 cells (Figures 2I and S2E).

Next, to test our hypothesis that Notch signals via JAG1 ligands could act as a bridge toward later DLL4 interactions, we analyzed cells grown on JAG1 for 3, 5, or 7 days prior to DLL4 transfer (Figure 2J). We found that cells cultured on JAG ligands or no ligands for 3 days maintained reduced T-lineage commitment compared with DLL1 or DLL4 cells (pJAG1 = 0.033; pJAG2 = 0.017), whereas cells cultured on JAG ligands for longer than 3 days lost T-lineage potential (Figure 2K), indicating that JAG ligands could not support T-lineage potential.

We next analyzed the contribution of different Notch ligands to the development of male and female ETPs (Figures S2F and S2G). Our data suggest that JAG ligand interactions are more abundant and diverse in females, with JAG1-NOTCH1 interactions enriched in female ETPs and DLL4 interactions enriched in male ETPs.

Together, these data suggest that timely migration from the CMJ to DLL4 ligands at the subcapsular zone is critical for T-lineage commitment, and exposure to JAG ligands at the CMJ can guide alternative lineage development toward thymic-derived DCs. Our data further demonstrate previously unrecognized sex-biased regulation by Notch ligands.

Analysis of the subcapsular zone identifies sex-based differences in fibroblast regulation of DP development and thymus growth

From the CMJ, ETPs migrate to the subcapsular zone via a CCL25-CCR9 chemokine gradient established by cTECs and directed to pro-T, DP (P), and DP2 (Q), but not DP1 (Q) cells (Figure 3A; Figure S3A). The subcapsular niche consists of JAG1+ VCAM1+ DCs, cTECs, capsular fibroblasts, DPP4+ capsular fibroblasts, and proliferating fibroblasts, which secrete and maintain spatially regulated ECM ligands to support sequential thymocyte development (Figures 3B and 3C; Figure S3B and S3C).

Figure 3 Fibroblasts in the subcapsular zone contribute to regulation of thymus biology and T cell progenitor development

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GSEA showed that DPP4+ capsule fibroblasts were enriched in HSP90 chaperone cycle for steroid hormone receptors (padjusted = 0.0065; 18/52 pathway genes significantly upregulated) (Data S1), suggesting an enhanced response to steroid hormones and supporting their role in sex hormone-based thymic involution.9 By contrast, capFibs were enriched for genes related to cytokine (interleukin [IL]-33, padjusted = 1.50 × 10−6; IL-34, padjusted = 3.56 × 10−7) and chemokine signaling (CCL2, padjusted = 5.10 × 10−40; CXCL3, padjusted = 0.020; CXCL12, padjusted = 1.78 × 10−8; CXCL14, padjusted = 3.63 × 10−15), functions previously attributed to TECs. Furthermore, CellChat identified cortical fibroblasts as major contributors to insulin growth factor (IGF) signaling through predicted signaling to cTECs, which are found in close proximity in the cortex (Figure S3D), via IGF2-IGF1R and IGF1-IGF1R axes, and to ETPs and β-selection cells, which were found under the capsule (Figure S2B), via an IGF2-IGF2R axis (Figures 3D–3F).

We next explored the role of proliferating fibroblasts. GSEA comparisons between capFibs and Fibs (P) showed marked differences in signal transduction pathways. CapFibs resembled traditional fibroblasts, which upregulate tyrosine kinase, angiogenesis, and ECM regulation and deposition pathways, whereas Fib (P) upregulates WNT signaling and cell sensing pathways, including genes involved in transient receptor potential (TRP) channels in the stimuli sensing channels pathway and taste receptors (TASRs) (Figure 3G; Data S1). Interestingly, CODEX images identified ECM− PDGFRa+ fibroblasts lacking extra domain A fibronectin (EDA-FN) expression, indicating that Fibs (P) are not involved in fibrotic matrix deposition unlike capFibs (Figure 3H; Figure S3B). Fibs (P) form a network of PDGFRa+ cells throughout the cortex that does not overlap with the cTEC network, yet maintain cell-cell contact in specific niches and often localize near cortical capillaries (Figure S3D).

We found sex-specific differences in vascular endothelial growth factor A (VEGFA) signaling within ECM− fibroblasts (Fib (P)) and other mesenchymal cells. Although all thymic fibroblasts produce the angiogenesis growth factor VEGFA, male fibroblasts express more than female cells (Fibs (P): padjusted = 0.0306; DPP4+ capFibs: padjusted = 0.0318; mFibs: padjusted = 1.85 × 10−6) (Figure 3I). Given that postnatal male thymuses are larger than female thymuses in humans and primates26 (Figure S3E), male fibroblasts may provide increased VEGFA to support angiogenesis and rapid thymic growth observed during postnatal development.52 Additionally, male mFibs have higher expression of FGF7 (padjusted = 0.0154), which regulates thymus size.53 CellChat predicts that male Fibs (P) are enriched in FGF10 compared with females, which supports cTEC proliferation and vascular growth,53,54 and only male VSMCs express FGF18 (Figures S3F–S3H). These sex biases in fibroblast growth factor (FGF) gene expression may contribute to the larger size of early postnatal male thymuses by stimulating epithelial and EC growth and proliferation.

Comparison of DEGs between male and female mesenchymal cells found increased expression of adipogenesis, cytokine, and GPCR signaling pathways in DPP4+ capFibs (Figure 3J). We also found increased expression of APOD, a gene associated with androgen, estrogen, progesterone, and glucocorticoid signaling,55,56 across male fibroblast populations (Fibs (P): padjusted = 2.18 × 10−26, mFibs: padjusted = 8.45 × 10−32) (Figure S3I). Given the association of hormone signaling with thymic involution,29,52,57 these findings suggest early initiation of thymic involution in postnatal males.

In sum, we identified three roles for fibroblasts within the subcapsular niche: maintaining tissue structure and organization via ECM and chemokine signaling, directly regulating cTEC maintenance and expansion, and potentially coordinating T cell development directly through growth factors and cell-cell interactions.

Human postnatal thymocytes may self-select in the cortex to support positive selection of conventional αβT cells

Upon exiting the subcapsular zone, DPs migrate into the inner cortex toward the medulla, where they receive positive selection signals that guide T-lineage branching toward CD4 or CD8 SP cells (Figure 4A). For DPs to transition toward the CD4 lineage, cells must receive T cell receptor (TCR) stimulation through HLA class II interactions, yet previous mouse studies have shown transcriptional downregulation of HLA class I and II in DPs.58,59 Low transcriptional expression is hypothesized to prevent thymocyte-thymocyte self-selection during positive selection, necessitating DP interactions with cTECs to receive positive selection signals.

Figure 4 HLA class I and II interactions may support thymocyte positive selection in the inner cortical zone

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Analogous to mouse literature, quiescent human DPs do not express HLA class II transcripts and have closed CIITA promoters (Figures 4B and 4C). Despite the lack of class II mRNA, thymocytes express low levels of HLA class II protein throughout development (Figure 4B). Additionally, in contrast to mouse data, we observe constitutive class I mRNA expression, which increased as cells transitioned toward SPs (Figure 4D). This is consistent with ATAC-seq data demonstrating that the B2M promoter is open throughout thymocyte development (Figure 4E). We confirmed HLA expression via flow cytometry and found that approximately 25% of DPs express both class I and II, and over 65% of DPs are class I+ (Figure S4A). Thus, thymocyte self-selection within the cortex could support positive selection. In support of this notion, CODEX enabled us to identify locations within the cortex devoid of epithelial, fibroblast, endothelial, or DCs but packed with DPs expressing class II+ molecules concentrated at cell junctions (Figure 4F). We confirmed the absence of spindle-like cTEC projections in this niche via confocal imaging (Figure 4G). Additionally, we quantified cell-cell interactions and identified a niche (positive selection niche 1) consisting of class II+ DPs and CD3+ DPs and a niche (self-selection niche) containing mainly class II+ DPs (Figure 1D). Finally, we sorted thymocytes to isolate immature DPs (CD4+CD8+CD3−TCR−) and mature DPs (CD4+CD8+CD3+TCR+) from three donors and cultured them for 7 days in a feeder-free assay. In the absence of epithelial cells, both immature and mature DPs upregulate HLA class II proteins (Figure 4H), and immature DPs continue to mature along their developmental pathway, as indicated by increased percentage of CD27+ DPs in culture after 7 days (Figure 4I).

Next, we identified a niche that directs T-lineage commitment toward CD4 or CD8SPs. We performed differential gene expression analysis on clusters representing this lineage branch point to identify markers for our CODEX panel (Figure S4B). We found SATB1 expression increased as DPs transitioned toward SPs (Figure S4C), and compared with CD8SP transition cells, CD4SP transition cells had higher expression of this master transcription factor60 (Figures S4D and S4E). Imaging analysis confirmed increased SATB1 expression coincides with CD3 upregulation, consistent with a role in late DP development and lineage branching (Figure 4J).7 Neighborhood analysis identified a niche enriched for mature CD3+ DPs in the inner cortex, suggesting that there either exists a niche specifically for late DP development and CD4 lineage transition or that cells are pre-disposed to CD4 lineage development through their TCR and migrate as clonal populations after proliferation at the outer cortex.

We compared cortical niche organization between sexes and found differences in niche organization supporting conventional T cell development, self-selection, and cross presentation. Females showed increased neighborhood interactions between the cortical DC niche containing JAG1+ VCAM+ DCs and the mature DP niche containing CD3+ DPs, the positive selection niche 1 containing class II+ DP cells and CD3+ DP cells, and the positive selection niche 3 containing DCs and DPs (Figure S4F) as well as increased cell-cell interactions between cTECs and class II+ DPs (Figures S4G and S4H). Conversely, males had increased cell-cell interactions between cTECs and CD3+ DPs (Figures S4G and S4H). These data suggest that the proportionally larger female cortex could increase cross presentation from DCs and cTECs to class II+ DPs, possibly facilitating greater use of self-selection as an alternative mechanism for positive selection.

Taken together, spatial multiomic analysis of the inner cortex identified cortical niches supporting specific stages of DP development, including three positive selection niches, a specialized niche for self-selection, and a mature DP niche thymocytes migrate through prior to entering the medulla.

Spatial multiomics identifies key mechanisms regulating negative selection niches in the medulla

Mature DPs enter the medulla, an environment specialized for negative selection, and transition toward CD4 or CD8 lineages (Figure 5A). Within the medulla, cells specialized for negative selection localize around keratinized structures called Hassall’s corpuscles (HCs).61 HCs appear during late prenatal development and are abundant in human postnatal thymuses but rare in mice.62 Here, we demonstrate that HCs can be divided into three major components: an external epithelial border of highly keratinized cells, an inner border of cells expressing prostaglandin-degrading enzyme 15-PGDH (HPGD), and a central PDGFRa+ mass (Figure 5B). HCs produce thymic stromal lymphopoietin (TSLP),61 an analog of IL-7, which activates DCs to increase expression of class II and co-stimulatory molecules CD80 and CD86. Importantly, subclustering stromal populations identified a population of KRT+ fibroblasts resembling cells undergoing epithelial-to-mesenchymal transition (EMT)63 (Figures S5A and S5B). CITE-seq identified TSLP and 15-PGDH mRNA expression in KRT+ Fibs, mFibs, mTECs, activated mTECs, and aDCs (Figure 5C), implicating these cell types as potential contributors to the function of HCs. Finally, given the inner layer of 15-PGDH+ cells, we explored the role of prostaglandin signaling regulation within the medulla. We found that DC1 cells express high levels of PGE2, whereas DC2/3 cells and monocytes express the PTGER2 and PTGER4 receptors, and aDCs express the PTGER3 receptor (Figure 5C), suggesting prostaglandin signaling is a major regulator of DC activity near HCs.

Figure 5 HCs represent scalable organizing centers for negative selection in the neonatal thymic medulla

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CODEX imaging suggests HCs act as sub-medullary organizational centers to segregate the inner medulla into specialized niches for negative selection. CD86+ APCs, a subset of which express the co-stimulatory ligand CD40, localize near HCs and in direct contact with CD45RA+ mature SPs (Figure 5D; Figure S5C). In addition, approximately 30% of medullary area is composed of CD19+ B cells,64 which cluster into niches surrounding HCs (Figure S5D). These B cells are found in close contact with—and are often enveloped within—mTECs, potentially facilitating cross presentation with epithelial cells (Figure 5E). These results suggest thymic B cells may comprise an important source of antigen presentation for negative selection.64,65 We quantified medullary neighborhoods and identified six niches, including an mTEC maturation niche, a cross-presentation niche, and four niches specialized for negative selection, which vary in relative location to HCs or the CMJ, as well as their composition of APCs, epithelial, and T cells (Figure 1D; Figure S1D).

Negative selection niches surrounding HCs play a key role in conventional T cell and tTreg development.61 We enriched CD25+ cells for sequencing and found a population of CD25hi pro-Tregs expressing canonical Treg markers CTLA-4, TNFRSF1B (TNFR2), and TNFRSF4 (OX40); positive/negative selection markers (ITM2A, RANBP1, NCL, NME1, MIF, and ATP5G1); Treg developmental long non-coding RNA (MIR155HG)66,67,68,69; and other markers described in mice (Figure S5E). Whereas pro-Tregs expressed high levels of pro-apoptotic gene BCL2L11, mature tTreg subsets expressed the anti-apoptotic gene BCL2. Gene network reconstruction via SCENIC70 identified transcription factor networks activated during pro-Treg to tTreg transition (Figure 5F).

The thymus also contains mature, highly activated Tregs, labeled as rrTregs, believed to have recirculated from the periphery.71,72 rrTregs lack expression of CCR7 or thymic egress markers (KLF2 and S1PR1) but express IL1R2 (Figure S5F), which sequesters the inflammatory cytokine IL-1β to reduce local concentrations.73 CODEX imaging identified tTregs and rrTregs dispersed throughout the medulla, with rrTregs primarily adjacent to CD68+ DCs (Figure 5G). CellChat supported the potential of rrTregs to sequester inflammatory cytokines through interactions with DC2/3 via an IL-1β-IL-1R2 axis (Figure S5G). rrTregs also exhibited a tissue resident Treg phenotype (BATFhigh CCR8+) associated with wound healing and tissue regeneration function,74 and expressed remodeling and tissue repair-related genes such as matrix metalloproteinase enzymes (MMP25 and ADAM19) (Figure S5H). Overall, these findings illustrate Treg diversity in the thymus with their developmental trajectories and functions yet to be elucidated.

Comparisons of male and female rrTregs showed that male rrTregs had higher expression of IL-4 and IL-13, heat shock factor protein 1 (HSF1), and IL-1 signaling pathways (Figure 5H), suggesting rrTreg-mediated regulation of IL-1R2-mediated anti-inflammatory feedback checkpoints is a more prominent mechanism in male tTreg development in early postnatal thymus. Notably, male-activated mTECs have higher expression of CD40 and tumor necrosis factor (TNF) inflammatory pathways than females, possibly resulting in higher rrTreg activity (Figure S5I).

Finally, as Tregs have been shown in mouse to contribute to thymic involution through JAG1,75 we explored sex-based differences in tTreg gene expression. GSEA showed male rrTregs and tTregs have higher expression of adipogenesis pathways (Figures 5H and 5I). Given the presence of cells undergoing EMT, our data underlie the aggressive timeline of thymic involution and suggest that sex-based differences in thymus functional decline begin early in life.

Our detailed examination of the medulla identifies several niches specialized for negative selection, cross presentation, and mTEC maturation around HCs and demonstrates sex biases in inflammatory pathways and thymic involution kinetics within these niches.

Discussion

We performed spatial multiomics to construct a tissue atlas of niches guiding T cell development in early human postnatal thymus. These datasets characterize how key developmental niches drive lineage branch decisions, identify a possible mechanism for conventional αβT cell development through self-selection, and suggest additional functions for mesenchymal cell types governing thymus biology. Furthermore, we discovered several sex-specific differences in thymus cell and niche biology. As T cell development is a dynamic migratory process, knowledge of cell position in combination with proteomic, transcriptomic, and epigenomic sequencing data provides an invaluable resource to predict niche-specific signaling cues directing T cell development, and mechanisms responsible for maintaining tissue structure and directing thymic involution.

We describe an approach to sequencing analysis using multidimensional imaging to establish benchmarks for the location, ligand expression, and composition of key niches in T cell development. This enables us to analyze cell-cell interactions guided by niche composition, identifying physiologically relevant ligand-receptor interactions based on cell proximity within the tissue. Ultimately, this approach maps epigenomic, transcriptomic, and proteomic data to distinct tissue niches at single-cell resolution. Furthermore, we included equal numbers of male and female age-matched thymus samples, enabling comparison between sexes across platform modalities. Our analysis of sex-matched human early postnatal thymus demonstrates the highly plastic nature of thymus lobule organization and resource dedication. Each niche responds to sex-biased developmental kinetics, supporting robust T cell development to ultimately produce functional immune systems in different manners (Figure 6). The findings herein describe only a subset of the data, and we encourage the community to capitalize on this resource to provide further insight into sex differences and targeted niche-specific inquiries.

Figure 6 The human early postnatal thymus lobule is spatially organized into sex-biased niches to support stage-specific T cell development

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In our analysis of Notch ligands, we complemented our in silico approach with in vitro analysis. Our analysis suggests that JAG1 at the CMJ cannot support T-lineage commitment as cells migrate toward the subcapsular zone but instead skew alternative lineage development toward a CD14− DC1 subset (Figure 6). CD14 expression on DCs is linked with increased inflammatory cytokine production,76 suggesting that JAG ligands promote non-inflammatory DC phenotypes. These results highlight the importance of precise Notch signaling strength and timing in the thymus and emphasize the need for strict spatial control of different Notch ligands within thymic niches. Our observation of high JAG1 expression in the medulla and decreased DLL4 expression on cTECs outside the subcapsular zone aligns with previous studies on human postnatal thymus.77

In the subcapsular zone, we characterize the important roles of specialized fibroblasts. DPP4+ capFibs, described in mouse as cells with progenitor and anti-fibrotic potential,78,79,80,81,82 are observed as a fibroblast subset responsive to changes in systemic hormone levels. Since thymic function and involution are regulated by sex hormone levels,57,83,84,85 DPP4+ capFibs likely control these processes and are potential targets for addressing age-related thymic involution.86 Previously, only medullary fibroblasts were linked to thymocyte development and selection in the medulla.82 We demonstrate that capFibs may directly support thymocyte development in the cortex by producing growth factors like IGF2 (Figure 6). Blocking IGF2 signaling arrests thymocytes at the double negative stage,87 and our data identify capFibs as the IGF2 source, suggesting capFibs as an additional cell source of cytokines and growth factors for in vitro developmental systems. Finally, we demonstrate that ECM profiles of thymic fibroblasts are tightly regulated based on spatial localization. Future work should characterize how tissue stiffness changes as thymocytes migrate through developmental thymic niches to improve biomaterial strategies for in vitro T cell development.88

Furthermore, we identify a population of ECM− cortical fibroblasts that are enriched in cell sensing pathways, such as TASRs and TRP channels. Interestingly, TASRs regulate cell responses to local soluble substances, such as glucose, modulating release of hormones and other signaling molecules.89 Similarly, TRP channels play roles in cell sensing, such as pheromone signaling, nociception, temperature sensation, and osmoregulation.90 Given the proximity of these cells to vasculature in the cortex, Fibs (P) may play a critical role as regulatory cells by sensing environmental changes and modulating thymus size (Figure 6). Their lack of ECM production and network-like structure resemble fibroblast reticular cells (FRCs) in the lymph node, which rapidly proliferate and remodel the cortex during infection.91 Our data are generated from early postnatal thymus samples, an age with active T cell development, suggesting these fibroblasts expand the thymic cortex similarly to FRCs during infection, signaling through FGF and IGF to stromal and epithelial cells to orchestrate remodeling.

While the dogma in thymocyte positive selection suggests that DPs downregulate class II RNA to prevent self-selection and force interactions with cTECs,58,59 several studies suggest that T-lineage cells can select off each other to support CD4 T cell development.20,92,93,94 Here, we describe an inner cortical niche where class II+ DPs reside that may support positive selection via DP-DP self-selection (Figure 6). We show that immature DPs cultured without epithelial cells upregulate HLA class II and continue to mature and receive positive selection signals. Additionally, upregulated SATB1 expression identifies mature DPs in an inner cortical niche and the CD4 branch of their progeny, suggesting it may determine early lineage specificity. Future work should investigate critical features of this niche and SATB1’s role in thymocyte development.

Within the medulla, we identified a niche adjacent to HCs specialized for negative selection and highlighted the role of rrTregs in modulating the medullary inflammatory environment (Figure 6). The abundance of HCs in human but not mouse, and their proximity to negative selection niches, suggests these structures evolved to provide niche-level organization within the larger human medulla or to regulate negative selection more stringently in longer-lived species.

Comparing male and female tissue showed sex differences in both T cell and thymus biology. Studies on post-pubertal males and females show that sex hormones differentially regulate thymic involution between sexes,26,27,28,29,30,52,57,84,86 and that androgen blockers increase FOXN1 expression, thymic involution, and increased rejuvenation.29,30,52,84,86 Additionally, older males produce fewer recent thymic emigrants and have smaller thymuses compared with females.26,28 Some studies describe decreased numbers of AIRE+ mTECs with age and in females,95 potentially predisposing females who maintain greater thymic function later in life to autoimmune disease.29 These studies also observe less interlobular fat in young female thymus,26 suggesting differences in thymic involution kinetics begin pre-puberty. However, current literature has not addressed transcript-level sex differences underlying functional differences in thymic and immune function. Our analysis uncovers that female thymic cells upregulate energy regulation, transcription, and antigen-presentation pathways, whereas male cells increase proinflammatory signaling, amino acid metabolism, and GPCR signaling. These cell metabolic differences align with transcript-level sex differences in other organs41,42,43 and highlight the need for sex-based cell culture optimization in in vitro T cell culture systems.

In addition to changes common to other organs,40,41 we identify thymus-specific differences affecting key processes in thymocyte development and training. Females have a larger proportion of cortical cells per lobule, aligning with lower thymic involution rates and a larger cortex/medulla ratio.26,27,52 ETPs have enriched interactions with JAG1 as they migrate away from the CMJ, suggesting increased JAG1 interactions could skew ETP lineage commitment toward less inflammatory DC phenotypes (Figure 6). In the female cortex, we observe increased cTEC and class II+ DP interactions and increased interactions between cortical DC and positive selection niches, suggesting thymocyte self-selection may play a larger role during positive selection (Figure 6). Conversely, the female medulla shows decreased inflammatory pathway activation and fewer medullary cells. These data suggest females prioritize generating a larger repertoire of DPs over deleting autoreactive cells through negative selection, potentially contributing to sex differences in autoimmune disease prevalence in females.96

In males, we observe enriched DLL4 interactions with ETPs, which aligns with previous data demonstrating that androgen levels positively correlate with DLL4 on cTECs.29 The male cortex shows increased interactions with mature CD3+ DPs and cTECs, suggesting male thymocytes may have lower proliferation rates post β-selection, allowing sufficient space for positive selection. In the medulla, male-activated mTECs exhibit increased inflammatory pathway markers, and male Tregs exhibit higher inflammatory modulation and activate thymic involution pathways.75 Upregulation of inflammatory modulation by male rrTregs may regulate the higher proinflammatory signaling in male cells (Figure 6). Interestingly, post-pubertal males have more Tregs and fewer CD4 T cells than females, possibly due to a more inflammatory medullary environment skewing CD4 development toward the Treg lineage.31

We further explore sex differences in thymus size control mechanisms. Among fibroblast populations, we find significant differences in expression of growth and angiogenesis factors, such as VEGFA and FGFs, potentially contributing to the size difference in male and female thymuses at this age (Figure 6). These data align with and extend known sex differences in growth factor expression, including sex-biased expression of growth hormone and IGF-1 in regulating size of different tissues.97,98 Importantly, these results indicate sex-specific differences in early thymus structure maintenance and growth, which could skew T cell development. We also establish an early transition toward an adipogenic environment in males. These observations align with findings in model organisms, where young male rats exhibit higher rates of thymic involution52 and early postnatal male primates have a larger interlobular fat area.26 Together, these factors define two possible mechanisms contributing to a male-female difference in thymus size and involution kinetics.

Future studies should test how sex differences at the transcript, niche, and organ level impact differential T cell production and quality as well as explore how sex differences in other organs contribute to known differences in immune responses. Defined in vitro and organoid culture systems replicating the thymic microenvironment present powerful platforms to test if the cell type-specific and sex-specific differences identified here lead to increased autoimmune disease incidence among females and increased infection susceptibility in males. Furthermore, given the surprising sex-based differences at this early postnatal stage, future work should examine aged thymus to investigate how cellular level differences in thymic involution kinetics may translate to larger impacts on our immune system later in life.

Limitations of the study

Our analysis of intra-sex variation is limited by access to patient samples as well as the inability to conduct mechanistic experiments in the context of a whole organism. There is an opportunity for future work to further validate and expand on predicted ligand-receptor interactions.

The thymic epithelium is responsible for the secretion of thymic peptides, which intervene in some steps of intra- and extrathymic T cell differentiation. Recent data suggest that thymic hormone secretion is modulated by the neuroendocrine network, comprising thyroid, adrenals, and gonads. However, the role of the pituitary gland in this regulation is still poorly understood. In the present paper we studied the in vivo and in vitro influences of PRL on the secretion of thymulin, one of the chemically defined thymic hormones, by thymic epithelial cells (TEC). When injected daily (20-100 micrograms/20 g) in young or old C57BL/6 mice, PRL induced a specific increase in thymulin synthesis and secretion, respectively, measured by the number of thymulin-producing cells in the thymus and the peripheral levels of the hormone. This stimulation was dose dependent and reversible after the end of treatment. Similar findings have been made in animals with pituitary dwarfism, known to have low levels of circulating thymulin. This stimulatory effect was also observed in primary cultures of human and mouse TEC when PRL (10(-7) to 10(-8) M) was applied to culture supernatants, thus suggesting that PRL could act directly on TEC. In addition, we induced in vivo experimental hypoprolactinemia, treating mice with bromocriptine, a dopamine receptor agonist that inhibits pituitary PRL secretion. Bromocriptine treatment (100-200 micrograms/20 g) yielded a significant decrease in thymulin secretion that could be reversed by coincident treatment with PRL. In the light of previous observations that bovine GH can also increase thymulin production in aged dogs, we performed a series of experiments in vitro to evaluate whether GH has a direct effect on TEC. We observed that only human GH preparations that are known to have a PRL-like effect were efficient in stimulating thymulin biosynthesis and release into the culture supernatants. The effects of PRL on TEC were not restricted to thymic hormone production. We observed that TEC proliferation, as well as the numbers of a TEC subset defined by the expression of cytokeratins 3 and 10, could also be increased by PRL treatment. All these findings show that the pituitary gland directly affects TEC in terms of cytoskeletal and secretory protein expression as well as cell cycle.. This paper reviews the mechanism of sex hormone actions on the thymus, presenting mainly our data obtained at the cellular and molecular levels. First, data supporting the "genomic" action via the nuclear sex hormone receptor complexes are as follows: 1) sex hormone receptors and the thymic factor (thymulin) are co-localized in thymic epithelial cells, but not in T cells; 2) production/expression of thymic factors (thymulin, thymosin alpha 1) are remarkably inhibited by sex hormone treatment; 3) sex hormones cause changes in T cell subpopulations in the thymus; and 4) sex hormones strongly influence the development of thymus tumors in spontaneous thymoma BUF/Mna rats through their receptor within the tumor cells. Secondly, data indicating the "non-genomic" action of sex hormones via a membrane signal-generating mechanism are as follows: 1) the proliferation/maturation of thymic epithelial cells is mediated through protein kinase C activity introduced by sex hormones; 2) sex hormones directly influence DNA synthesis and cdc2 kinase (cell cycle-promoting factor) activity..

pubmed.ncbi.nlm.nih.gov/2737149/

www.cell.com/developmental-cell/fulltext/S1534-5807(24)00539-2

I'd wanted them for a couple of years, and finally I found a local source for PURPLE LED fairy (faerie? feri?? no.) lights, and hooboy was I happy! Yay! Purple LED lights, no shipping! As soon as I got home I strung them up.

But- wait! I know one or two things about semiconductor physics and engineering -- and there's NO SUCH THING as a purple LED! What's going on?!

Al Centro DINO CAMPANA non si scherza.. e si spazia anche sul SANITARIO……Donne in primo piano.. ed i Progressi della ONCOLOGIA

Stefano Tamberi

Professore a contratto a titolo gratuito

Scuola di Medicina e Chirurgia

Curriculum Vitae

Nato il 26 giugno 1967, ha conseguito la laurea in Medicina e Chirurgia nel 1992 con 110 e lode presso L'Università di Bologna. Nel 1997 si è specializzato in Oncologia con 70 e lode presso l'Università di Bologna e consegue nel 2001 il dottorato di ricerca in Oncologia Gastrointestinale presso l'Università di Modena e Reggio Emilia. Vincitore dei premi "E.Baroni" e "Evaristo Stefanelli" per la ricerca in oncologia presso l'Università di Bologna negli A.A 1997 e 1998. Dal 2010 è direttore dell'UO di Oncologia dell'Ospedale di Faenza.

Le principali aree di ricerca clinica oncologica sono state nel corso degli anni la patologia oncologica polmonare, gastrointestinale e ginecologica. Membro attivo dei principali gruppi di ricerca nazionali e internazionali di tali ambiti (Gruppo MITO, Gynecologic cancer tergroup, GISCAD etc) è stato principal investigator di numerosi studi internazionali sui tumori dell'ovaio, del polmone e dei tumori del colon.

Ha sviluppato negli ultimi anni un interesse particolare perl'approccio psico-sociale al paziente oncologico con un periodo di visita presso le principali istituzioni del Quebec (Canada) per lo sviluppo di un processo organizzativo di cure oncologiche centrate sul paziente. Ha sviluppato progetti di supporto clinico psiconcologico a pazienti affette da carcinoma mammario operato con l'istituzione di supporto psicologico di gruppo e la consulenza psicologica prevista nella quotidiana attività clinica. Tali progetti pilota sono stai presentati nell'ambito del progetto "Empatia" della Regione Emilia Romagna (RER). Ha partecipato al corso regionale sulla "Health Literacy" tenuto dalla Prof.ssa Rudd (Harvard - Boston) e ha sviluppato progetti innovativi di comunicazione nel personale medico e infermieristico da lui diretto.

Alcune sue Pubblicazioni

1. Carboplatin, Fluorouracil and L-Folinic Acid in the treatment of Advanced Squamous Cell Carcinoma of the Head and Neck (SCCHN) . Guaraldi M., Martoni A., Morelli F., Panetta A., Tamberi S., Pannuti F. Annals of Oncology 5 (suppl.8):123,1994

2. A phase I-II study on 5-day 5-Fluorouracil continuous infusion with levo-Folinic Acid modulation in advanced colo-rectal cancer. Martoni A., Angelelli B., Panetta A., Tamberi S., Pannuti F. Proceedings of the "XVI International Cancer Congress". New Delhi 30 ottobre-5 novembre 1994

3. Combined chemo-immuno-ormonotherapy of advanced renal cell carcinoma. Panetta A., Martoni A., Guaraldi M., Tamberi S., Casadio M., Lelli G., Pannuti F. Journal of Chemotherapy 6:5,349-353, 1994

4. Prevalenza e caratteristiche morfologiche di lesioni polipoidi del tratto digestivo superiore nella poliposi adenomatosa familiare. Santucci R., Volpe L., Calabrese C., Poggi B., Di Febo G., Tamberi S., Biasco G. "I Tumori gastrici. Bilanci e prospettive" Roma 9-11 novembre 1995

5. Terapia cronomodulata dei tumori solidi. Biasco G., Brandi G., Tamberi S. G.Biasco. "Trials clinici in oncologia". Bologna, 1995

6. Abnormal mucosal cell proliferation as a biomarker of gastrointestinal cancer: methodological issues and clinical applications Biasco G., G.M.Paganelli, U.Zannoni, G.Brandi, R.Santucci, M.Renga, P.Mordenti, S.Tamberi, L.Barbara. Journal of tumor marker oncology vol.10 ,2, pag.89, 1995