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It was aimed to evaluate and compare the primary lymphoid organs bursa Fabricius and thymus tissues of chukar partridge (Alectoris chukar) and pheasant (Phasianus colchicus) histologically. In the study, three chukar partridges and three pheasants were provided from Süleyman Demirel University Department of Animal Science Poultry Breeding Unit. Bursa Fabricius and thymus tissue samples were taken by dissection. It was performed hematoxylin-eosin and Masson's trichrome for general histological examination, and periodic acid schiff (PAS) for the neutral mucosubstance and carbohydrate content, methyl green-pyronin staining for the plasma cells. Van gieson with Weigert's hematoxylin staining was performed for the major cells of thymus. Toluidine blue and alcian blue/safranin O combination procedure was performed to show the mast cells. The sizes of plasmocyte, mastocyte and lymphocyte cells in the cortex and medulla regions of the thymus and bursa Fabricius of chukar partridge and pheasant were measured and their average values were calculated. In addition, photographs of tissues were taken with a Leica DM 500 microscope. When the bursa Fabricius and thymus tissues of chukar partridge and pheasant were compared histologically, it was concluded that there was no significant difference., Kınalı keklik (Alectoris chukar) ve sülün (Phasianus colchicus)’ün primer lenfoid organlardan bursa Fabricius ve timus dokularının histolojik yönden değerlendirilmesi ve karşılaştırılması amaçlandı. Çalışmada, Süleyman Demirel Üniversitesi Zootekni Bölümü Kanatlı Hayvan Yetiştirme Biriminden 3 adet, sağlıklı kınalı keklik ve sülün temin edildi. Bursa Fabricius ve timus doku örnekleri diseksiyon ile alındı. Genel histolojik incelemeler için hematoksilen-eosin ve masson trikrom boyamaları, doku ve hücrelerdeki nötral mukosubstansı ve karbonhidrat içeriğini belirleyebilmek için periodic acid schiff (PAS) reaksiyonu, plazma hücrelerini belirlemek için methyl green-pyronin boyaması uygulandı. Timusun major hücreleri için Weigert’s hematoksilenli van gieson boyaması gerçekleştirildi. Mast hücrelerini göstermek için ise toludin blue ve alcian blue/safranin O kombinasyon boyama teknikleri yapıldı. Kınalı keklik ve sülün timus ve bursa Fabricius’unun korteks ve medulla bölgelerindeki plazmosit, mastosit ve lenfosit hücrelerinin büyüklüklerinin ölçümleri yapılıp ortalama değerleri hesaplandı. Ayrıca dokuların fotoğrafları Leica DM 500 mikroskobunda çekildi. Kınalı keklik ve sülün bursa Fabricius ve timus dokuları histolojik açıdan karşılaştırıldığında önemli bir farklılığın olmadığı sonucuna varıldı.

Miastenija gravis je bolest koja zahvaća neuromišićnu spojnicu uzrokujući abnormalnu slabost, umor i propadanje muskulature. Dolazi do gubitka ili disfunkcije acetilkolinskih receptora u najvećem slučaju zbog prisutnosti protutijela na nikotinske acetilkolinske receptore (NAChR). Timus i timom imaju posebnu ulogu u patofiziologiji miastenije gravis zbog uloge timusa u sazrijevanju i distribuciji limfocita gdje može doći do križne reaktivnosti i posljedične autoimunosti na NAChR. Najčešće korištena dijagnostička imunološka pretraga za dijagnozu miastenije gravis je mjerenje razine protutijela na NAChR što je visokospecifični test za dijagnostiku miastenije gravis. CT je metoda izbora u dijagnostici timoma. Zbog važnosti timusa u patofiziologiji miastenije gravis bolest se može liječiti kirurški, uklanjanjem timusa. Ukoliko je prisutan timom za planiranje operativog zahvata služe Masaoka i TNM klasifikacija. Metode kojima se može odstraniti timus/timom su transsternalni pristup, transcervikalna timektomija, video asistirana torakoskopska kirurgija (VATS – video-assisted thoracoscopic surgery) te robotska kirurgija. Neki od faktora koji utjeću na prognozu nakon timektomije su dob i spol, prisutnost timoma, kirurška tehnika, obujam resekcije i težina i duljina trajanja simptoma miastenije gravis. Nove metode timektomije su minimalno invazivne i s brzim oporavkom, a većina pacijenata pokazuje remisiju bolesti nakon uklanjanja timusa., Myasthenia gravis is a disease that affects the neuromuscular junction causing abnormal weakness, fatigue, and muscle decay. Loss or dysfunction of acetylcholine receptors occurs mostly due to the presence of antibodies to nicotinic acetylcholine receptors (NAChR). The thymus and thymoma have a special role in the pathophysiology of myasthenia gravis due to the role of the thymus in the maturation and distribution of lymphocytes where cross-reactivity and consequent autoimmunity to NAChR can occur. The most commonly used diagnostic method for myasthenia gravis is the measurement of antibody levels to NAChR which is a highly specific test for the diagnosis of myasthenia gravis. CT is the method of choice in the diagnosis of thymoma. Due to the importance of the thymus in the pathophysiology of myasthenia gravis the disease can be treated surgically, by removing the thymus. If there is thymoma present, to plan the operation, Masaoka and TNM classification are used. Methods that can be used to remove the thymus / thymoma are transsternal thymectomy, transcervical thymectomy, video-assisted thoracoscopic surgery (VATS) and robotic surgery. Some of the factors influencing the prognosis after thymectomy are age and sex, presence of thymoma, surgical technique, extent of resection and severity and duration of symptoms of myasthenia gravis. New methods of thymectomy are minimally invasive and with rapid recovery, and most patients show remission of the disease after removal of thymus.

Mucoepidermoid carcinoma of the thymus is a rare malignant neoplasm of the anterior mediastinum. There are less than 30 cases described in the English literature. We report a case of a 77-year- old man with right chest pain. Thorax-CT scan showed a lobulated cystic mass having a focal solid portion with direct invasion of the adjacent pericardium in the anterior mediastinum. He underwent surgical resection for his anterior mediastinal mass via right thoracoscopic surgery without any complication. He had an uneventful recovery and was discharged on the sixth postoperative day.

Among many functions of the body affected by the complex process of aging, the immune system, primarily due to declining of thymic function, the ancient and conserved evolutionary process, undergoes complex remodelling in the second part of life, with recapitulation of inversely evolutionary pattern of the immune system. In approaching the complex analysis of age-associated derangement of the immune system, homeostasis, and its clinical consequences, classical monoclonal lymphoproliferative syndrome (CLL) accompanied by a myriad of cellular and humoral defects, has been selected as appropriate and useful model for studies of impact T-cell and B-cell defects on appearance, evolution of clinical manifestations and outcome of CLL syndrome. Therefore imbalance in cascade secretion of a number of Th-1 (pro-inflammatory cytokines) and/or Th-2 (anti-inflammatory cytokines) in CLL patients with their pleiotropy, redundancy, synergistic and antagonistic activity and parallelism can cause variety of clinical manifestations as recurrent infections, systemic inflammation/sepsis, immunodeficiency, autoimmune disorder, indolent antiself malignancy, and/or other diverse secondary tumours., Fiziološkom involucijom timusa i slabljenjem njegove kompleksne funkcije u drugom dijelu životnog vijeka dolazi do evolucijski uvjetovanog funkcionalnog remodeliranja imunosnog sustava. U analizi starosno uvjetovane poremećene funkcije imunosnog sustava odabrali smo kao klinički model kroničnu limfocitnu leukemiju (KLL). KLL je vrlo pogodan model za longitudinalno prećenje uloge pojedinih imunoloških defekata u evoluciji patogeneze sindroma. Naime, neuravnotežena sekrecija proupalnih Th-1 citokina, protuupalnih Th-2 citokina, u kombinaciji s njihovom pleiotropijom, redundancijom, sinergističkom i antagonističkom aktivnošću i paralelizmom, može se klinički manifestirati kao rekurentne infekcije, sistemska upala/sepsa, imunodeficinjencije autoimuni procesi i sekundarni tumori i tako bitno utjecati na kliničke manifestacije, tok i ishod bolesti.

Svrha ovog preglednog rada je prikaz timektomije kao oblika liječenja u osoba oboljelih od miastenije gravis. Miastenija gravis je kronična, autoimuna, neuromuskularna bolest karakterizirana slabošću i zamaranjem skeletnih , poprečnoprugastih, mišića. U 80% oboljelih od miastenije gravis nalazi se abnormalnost timusa od čega 10-20% ima timom, a preostali dio hiperplaziju timusa. Timus je parenhimatozni organ sivocrvenkaste boje smješten u prednjem gornjem dijelu medijastinuma, iza prsne kosti. To je središnji organ limfatičnog sustava čija je glavna uloga prethodna obrada limfocita T. Poremećaji u obradi limfocita T imaju ulogu u patogenezi miastenije gravis. Liječenje miastenije gravis je simptomatsko, a cilj liječenja je potpuni nestanak simptoma. Timektomija, uz farmakološko liječenje, predstavlja oblik terapije za oboljele od miastenije gravis. Timektomijom se želi postići značajno poboljšanje simptoma kod bolesnika, prvenstveno smanjiti slabost mišića, zatim u najvećoj mogućoj mjeri reducirati farmakološku terapiju te u idealnom slučaju postići potpunu remisiju bolesti. Više različitih čimbenika, kao što su dob i spol bolesnika, težina simptoma, prisutnost timoma, prisutnost protutijela na AChR ili MuSK receptore, oblik bolesti, pa čak i trudnoća utječu na odluku o tome da li će osoba biti podvrgnuta timektomiji ili ne.Razlikujemo nekoliko glavnih kirurških pristupa: transcervikalnu, transsternalnu i VATS timektomiju. Nije potvrđeno da VATS timektomija ima veću učinkovitost u remisiji bolesti od transsternalne timektomije, ali minimalno invazivni postupci imaju niže stope morbiditeta i mortaliteta, manji broj komplikacija te je kraće vrijeme oporavka bolesnika., The purpose of this thesis is to present timectomy as a form of treatment in people suffering from myasthenia gravis. Myasthenia gravis is a chronic, autoimmune, neuromuscular disease characterized by weakness and fatigue of skeletal muscles. In 80% of patients with myasthenia gravis there is an abnormality of thymus, of which 10-20% has a thymoma, and the rest of them the thymus hyperplasia. Timus is a parenchymal organ of reddish-brown color placed in the front upper part of the mediastinum behind the chest bone. It is the central organ of the lymphatic system whose main role is the prior processing of lymphocytes T. Thymus plays a role in the pathogenesis of myasthenia gravis. The treatment of myasthenia gravis is symptomatic, and the goal of treatment is complete disappearance of symptoms. Timectomy, along with pharmacological treatment, is a form of therapy for people suffering from myasthenia gravis. Timectomy seeks to significantly improve clinical symptoms of the patient, primarily to reduce the muscle weakness, then to reduce the pharmacological therapy and ideally achieve complete remission of the disease. Multiple factors, such as age and sex, symptom weight, thymoma presence, presence of antibodies to AChR or MuSK receptors, the form of the disease, and even the pregnancy, influence the decision whether or not the subject will be operated. Major surgical approaches are: transcervical, transcernal, and VATS timectomy. It has not been shown that VATS timectomy has greater efficacy in remission of myasthenia gravis, but minimal invasive procedures have lower rates of morbidity and mortality, fewer complications, and shorter recovery time of the patient.

Thymolipoma is a rare and benign lesion originating from the anterior mediastinum. Thymolipoma is mostly diagnosed incidentally. However, cough, dyspnea and chest pain can be seen. A 35-year-old male patient presented to our hospital with the complaint of shortness of breath lasting for one month. The physical examination revealed a massive pleural effusion on the right. In computed tomography (CT) imaging of the thorax, a soft tissue density of 7.5x5x5 cm in size, containing calcifications were observed in the anterior mediastinum. A right thoracotomy was performed for the treatment and to make a definite diagnosis. The histopathological examination resulted in the diagnosis of a thymolipoma. We presented our case with its differential diagnosis because of its rarity.

Timus je mjesto formacije adaptivnog imunosnog sustava, gdje epitelne stanice upravljaju sazrijevanjem limfocita T. Matične stanice i progenitori nalaze se u nišama unutar tkiva odraslog organizma i sudjeluju u regeneraciji i održavanju homeostaze. Postojanje matičnih stanica koje sudjeluju u održavanju timusne strome prethodno je pokazano u odraslome mišu, no ne i u čovjeku. Cilj ovoga rada bio je u timusu ljudi pronaći matične epitelne stanice. Sekvencijskom razgradnjom tkiva timusa i nasađivanjem stanica u uvjetima prilagođenima uzgoju matičnih stanica u tri dimenzije dosljedno smo uspostavili metodu stvaranja organoida kao pokazatelja prisutnosti matičnih stanica. Dokazali smo da organoidi nisu agregati stanica, već su nastali iz pojedinačnih stanica koje imaju svojstva matičnosti - samoobnavljanje, umnožavanje i diferencijacija. Otklanjanje limfocita iz suspenzije stanica pomoću magnetske deplecije ili inkubacije s 2'-deoksiguanozinom povećava broj organoida. Stanice organoida eksprimiraju gene nužne za funkciju epitela, no i gene aktivne tijekom organogeneze timusa. Proteinska ekspresija markera zrelosti epitelnih stanica pronađena je na manjem dijelu stanica organoida. Dobiveni organoidi su sposobni usmjeravati diferencijaciju hematopoetskih matičnih stanica u zrele limfocite T. The thymus is the central hub of T lymphocyte differentiation, where epithelial cells guide the process of their maturation. Progenitors and stem cells are found in niches deep within the tissues of an adult organism. Their function is tissue regeneration and maintenance of homeostasis. The existence of stromal stem cells within the postnatal mouse thymus has been shown, but these have not been proven to exist in the human thymus. The aim of this research was to find epithelial stem cells within the human thymus. Sequential tissue digestion and plating the obtained cells in a manner suited for 3D cultures, we established a method that consistently leads to a formation of organoids indicating the presence of stem cells. The organoids are not mere cell aggregates, they are formed from individual cells with the capability of self-renewal, as well as other properties of stem cells, such as maintaining their immature state while giving rise to differentiated cells. The removal of lymphocytes from the cell suspension by either magnetic depletion or incubation with 2'-deoxyguanosine leads to an increase of organoid number. The cells within the organoid express genes necessary for thymic epithelial function, but also genes active during thymic organogenesis. The expression of protein markers of mature epithelial cells has been found on a smaller proportion of organoid cells. The organoids are capable to lead the differentiation of a hematopoietic stem cell into mature single positive T lymphocytes.

Bu çalışmanın amacı yumurtaya verilen Bisfenol A'nın, tavuklarda timusun gelişimi ve perifer kan alfa naftil asetat esteraz pozitif lenfosit oranı üzerindeki etkilerinin histolojik ve enzimhistokimyasal metodlar kullanarak belirlemektir. Bu amaçla Isa Brown ırkı yumurtacı tavuklara ait 310 adet her biri 50-55 gr ağırlağında olan döllü yumurta kontrol, taşıyıcı kontrol, 50, 100, 250 μg/yumurta BPA olmak üzere 5 gruba ayrıldı. Test solüsyonları inkübasyondan önce yumurta sarısına enjekte edildi. Kuluçkanın 13, 18 ve 21. günlerinde her gruptan 10'ar adet yumurta açıldı ve elde edilen embriyolardan kan ve timus doku örnekleri alındı. Kuluçkanın 13. 18. ve 21. günlerinde BPA uygulanan gruplarda kontrol grubuna göre timus dokusunun embriyonik gelişiminin geri olduğu tespit edildi. Aynı zamanda BPA verilen gruplarda lenfoid dokunun daha az hücre yoğunluğuna sahip olduğu ve ANAE pozitif lenfositlerin sayıca azaldığı dikkati çekti. Perifer kan ANAE pozitif lenfosit oranlarının da kontrol gruplarına göre BPA verilen gruplarda önemli oranda düştüğü tespit edildi (p, The aim of this study is to the determination of effects of in ovo administrated Bisphenol A on the development of thymus and proportion of alpha-naphthyl acetate esterase enzyme lymphocyte by means of histological and enzymehistochemical methods in chicken.For this purpose, 310 fertile eggs of Isa Brown laying parent stock were divided into 5 groups as control, vehicle-vehicle-control, 50,100, and 250 μg/egg BPA. Test solutions were injected into yolk before incubation. At the 13th, 18th and 21th days of incubation, 10 eggs were opened from each group and blood and thymus tissue samples were taken from the obtained embryos. On the 13th, 18th and 21th days of incubation, BPA-treated groups were found to be retarded embryonic development of thymus tissue compared to the control group. At the same time, in BPA-treated groups, lymphoid tissue had less cell density and the number of ANAE positive lymphocytes decreased. The percentage of peripheral blood ANAE positive lymphocytes was significantly lower in the BPA-treated groups than in the control groups (p

Primary thymic carcinoma is a rare heterogeneous group of tumors of the anterior mediastinum with a poor overall prognosis. Lymphoepithelioma like carcinoma is an extremely rare subtype of thymic carcinoma. A 14-year-old boy who was presented with chest pain and dyspneae had a mass lesion in the anterior mediastinum. The patient underwent right thoracotomy with mediastinal mass resection and wedge resection of the middle lobe. Macroscopically the resected mass measured 8x7x4 cm and surrounded partially by a capsule. Microscopic examination showed a syncytial growth of carcinoma cells accompained by a lymphoplasmacyctic infiltration in the thymic tissue. Lung parenchyma was infiltrated by the tumor cells but the cut margins were safe. Following operation the patient had chemotherapy and radioterapy. Three years after operation, he is well without sign of recurrence. Because of its rarity we presented this case with literature. Key words: Thymus; Lymphoepithelioma Like Carcinoma; Differential Diagnosis., Primer timik karsinomlar, ön mediastenin nadir görülen, kötü prognozlu heterojen tümörleridir. Lenfoepitelyoma benzeri karsinom ise timik karsinomun oldukça nadir görülen alt tipidir. Göğüs ağrısı ve nefes darlığı şikayetiyle kliniğe başvuran 14 yaşında erkek hastada çekilen bilgisayarlı tomografide ön mediastende kitle tesbit edildi. Hastaya sağ torakotomi ile mediastinal kitle eksizyonu ve sağ orta lobdan wedge rezeksiyon uygulandı. Makroskopik incelemede tümöral kitle 8x7x4 cm ölçülerinde olup, bazı alanlarda kapsül yapısı ile çevriliydi. Mikroskopik incelemede timus dokusu içinde sinsityal gelişim göstermiş karsinom hücrelerine eşlik eden lenfoplazmositik infiltrasyon izlendi. Akciğer parankimi tümör hücreleri ile infiltreydi ancak cerrahi sınırlarda tümör mevcut değildi. Hasta operasyon sonrası kemoterapi ve radyoterapi aldı. Üç yıllık takibinde hastada rekürrens bulgusu saptanmadı. Oldukça nadir olması nedeniyle bu olgu literatür bilgileri eşliğinde sunuldu. Anahtar kelimeler: Timus; Lenfoepitelyoma Benzeri Karsinom; Ayırıcı Tanı.

Bu araştırma Selçuk Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü tarafından 13202034 proje numarası ile desteklenmiştir., Bu çalışmada, kuluçka başlangıcında kuluçkalık yumurtaların hava kamarasına enjekte edilen farklı dozlardaki Sunset Yellow FCF'nin (E110) tavuk embriyolarında timus ve bursa Fabricii'nin embriyonik gelişimi üzerindeki etkileri histolojik, histomorfometrik ve enzim histokimyasal yöntemlerle belirlendi. Çalışmada, Bursa Hastavuk İşletmesinden temin edilen Ross 508 hattı broyler anaçlardan elde edilen; 250 adet kuluçkalık yumurta kullanıldı. Yumurtalar, her birinde 50 yumurta bulunan 5 gruba ayrıldı. Gıda boyası olarak yaygın biçimde kullanılmakla birlikte, son yıllarda alerjik reaksiyonlara yol açtığı konusunda şikâyetler artan Sunset Yellow FCF (E110), merkezi lenfoid organlar olan tavuk timus ve bursa Fabriciisinin embriyonik gelişiminde geriliğe neden olmaktadır. Her ne kadar bu çalışmada elde edilen morfolojik bulguların fonksiyonel bulgularla desteklenmesine gerek olmakla birlikte, son yıllarda alerjik rahatsızlıkların çocuklarda artış göstermesi dikkate alındığında, gıda katkı maddelerinin tek başlarına ya da kombine etkilerinin bu artıştan sorumlu olabileceği düşünülmektedir. Konunun aydınlatılmasında memeli hayvan modeli üzerinde daha detaylı araştırmalara gerek olmakla birlikte, en azından puberta öncesi çocuklarda ve gebeliğin erken döneminde sentetik gıda katkı maddesi ve özellikle gıda boyaları içeren hazır gıdaların kullanılmaması ya da kullanımının sıkı denetim altına alınması gerektiği önerilebilir., In this study, the effects of different doses of Sunset Yellow FCF (E110) on embryonic development of thymus and bursa of Fabricius were determined by means of histological, histomorphometrical and histochemical methods. The substance was injected into air sac of fertilized chicken eggs prior to incubation process. The Ross 508 eggs obtained form Hastavuk, Bursa were divided into 5 groups, each having 500 eggs. Although Sunset Yellow FCF (E110) has largely been used as food coloring agent, which complaints accumulated about the possibility of its allergenicity, caused to developmental delay in central lymphoid organs, thymus and bursa of Fabricius of the chicken. Although morphological results of this study require functional support, in regarding the increase in childhood allergic reactions, it has been suggested that food additives might be questioned for the increase. Detailed studies including the mammalian animal model is required to elucidate the topic, it might be recommended that consuption of fast food containing food additives and especially food colors containing foods, should be limited or used under strictly- controlled conditions.

It is widely accepted that the progressive decline in the efficiency of thymopoiesis, starting at puberty, is causally related to increasing concentrations of gonadal hormones. In addition, thymic level of catecholamines, which act, via β- adrenoceptors, as tonic inhibitors of thymopoiesis, increases around puberty. There is a pile of evidence that mature immune cells (T-lymphocytes, macrophages and dendritic cells) as well as different types of epithelial cells, synthesize catecholamines. Furthermore, it has been shown that in various tissues male gonadal hormones affect catecholamine synthesis and β-adrenoceptor expression. These findings imposed the question whether thymocytes and/or thymic stromal cells synthesize catecholamines, and, if so, whether testicular hormones influence the synthesis of catecholamines and/or expression of β2-adrenoceptors, and possibly, consequently, the modulatory action of catecholamines on thymopoiesis. Considering all the aforementioned, the dissertation was aimed to: 1) explore whether thymocytes and/or thymic stromal cells in adult male rats synthesize catecholamines, and to define the types of thymic cells expressing β2-adrenoceptor; 2) examine whether orchidectomy at the age of 30 days, when rat thymus is the most sensitive to testicular hormone deprivation, affects catecholamine levels and β2- adrenoceptor expression in the thymus of adult rats, and, if so, 3) determine whether the modulatory effects of catecholamines on thymopoiesis differ in orchidectomized and non-orchidectomized animals. It has been found that thymocytes, thymic epithelial cells and macrophages in adult rats contain and synthesize catecholamines. In addition, it was shown that subsets of these cells also express the β2-adrenoceptors. Orchidectomy at the age of 30 days decreased the concentration of noradrenaline in the thymus of adult animals by diminishing the density of noradrenergic nerve fibers and catecholamine-containing thymic stromal cells and their noradrenaline content... Danas je široko prihvaćeno mišljenje da je progresivno smanjenje efikasnosti timopoeze, koje počinje u pubertetu, kauzalno povezano sa porastom koncentracije polnih hormona. Pored toga, i koncentracija kateholamina, toničnih inhibitora timopoeze posredstvom β-adrenergičih receptora, se u timusu povećava, počevši od pubertetnog perioda. Poznato je da zrele ćelije imunskog sistema (T-limfociti, makrofage i dendritske ćelije), i različiti tipovi epitelnih ćelija, imaju sposobnost sinteze kateholamina. Konačno, u različitim tkivima, muški polni hormoni utiču na sintezu kateholamina i ispoljavanje β-adrenergičkih receptora. Ova saznanja su nužno nametnula pitanje da li timociti i/ili ćelije strome timusa poseduju sposobnost sinteze kateholamina, i, ukoliko je odgovor potvrdan, da li i u kojim tipovima ćelija timusa hormoni testisa utiču na sintezu kateholamina i/ili ispoljavanje β2-adrenergičkog receptora i time, moguće, na modulatorno delovanje kateholamina na proces timopoeze. Imajući u vidu sve prethodno navedene nalaze, postavljeni su ciljevi ove disertacije: 1) da se utvrdi da li timociti i/ili ćelije strome timusa odraslih mužjaka pacova sintetišu kateholamine, i da se precizno definišu tipovi ćelija u ovom organu koji ispoljavaju β2-adrenergički receptor, 2) da se ispita da li orhidektomija u uzrastu od 30 dana, kada je timus pacova najosetljiviji na efekte kastracije, utiče na koncentraciju kateholamina i ispoljavanje β2-adrenergičkog receptora u timusu odraslih pacova, i, ako je odgovor potvrdan, 3) da se utvrdi da li se modulatorno delovanje kateholamina na timopoezu razlikuje kod orhidektomisanih i neorhidektomisanih životinja. Nađeno je da timociti, timusne epitelne ćelije i makrofage sadrže i sintetišu kateholamine. Osim toga, subpopulacije ovih tipova ćelija, takođe, ispoljavaju β2- adrenergički receptor. Orhidektomija u uzrastu od 30 dana je smanjila koncentraciju noradrenalina u timusu odraslih životinja, usled smanjenja gustine noradrenergičkih nervnih vlakana i ćelija strome timusa koje sadrže kateholamine i sadržaja noradrenalina u njima. Pored toga, u timusu ovih životinja je detektovano i smanjenje ekspresije iRNK za β2-adrenergički receptor, kao i površinske gustine ovog receptora na membrani timocita...

Introduction. Submandibular salivary gland is a part of the neuro-immune-endocrine system. It contains biological factors which regulate a number of functions in the body including the modulation of thymus function. Objective. The aim of the study was to investigate immunophenotypic characteristics of submandibular salivary glands of rats during ontogenesis, using the panels of monoclonal antibodies and to compare with the phenotypic characteristics of epithelial components of the thymus. Methods. Submandibular salivary glands and thymus were obtained from 1, 30 and 60 days old male AO (Albino, Oxford) rats. Streptavidin-biotin peroxidase method was used for staining. Results. Immunohistochemical analysis of rat submandibular salivary glands showed phenotypic heterogeneity of particular components of this gland during the postnatal development. We demonstrated that rat submandibular salivary glands share common antigens with rat thymic epithelial cells, but the observed phenotypic similarity between the individual regions was considered much more significant. Our data showed that the phenotypic similarity between duct epithelial cells and subcapsular epithelial cells and most medullary cells, whereas cortical epithelial cells are phenotypically similar to acinar cells. Conclusion. This immunohistological study showed phenotypic complexity of the submandibular salivary gland and similarity to the thymus that opens new perspectives in studying phenotypic similarities between this gland and lymphatic organs., Uvod. Submandibularna pljuvačna žlezda je deo neuroimunoendokrinog sistema koja posredstvom bioloških faktora reguliše mnoge funkcije u organizmu, uključujući i modulaciju funkcije timusa. Cilj rada. Cilj rada bio je da se ispitaju imunofenotipske odlike submandibularne pljuvačne žlezde pacova tokom ontogeneze pomoću panela monoklonskih antitela i uporede s fenotipskim svojstvima epitelnih komponenti timusa. Metode rada. Istraživanja su rađena na pacovima soja AO (Albino Oxford), muškog pola, starosti od jednog, 30 i 60 dana. Za fenotipsku analizu korišćena je streptavidin-biotin- peroksidazna metoda bojenja. Rezultati. Ova imunohistološka studija ukazuje na heterogenost fenotipskih odlika pojedinih komponenti submandibularne pljuvačne žlezde tokom postnatalnog razvoja. Takođe je ustanovljeno da submandibularna pljuvačna žlezda i timus imaju zajedničke antigene, ali je mnogo značajnija fenotipska sličnost između pojedinih regiona u oba ispitivana organa. Epitelne ćelije izvodnih kanala submandibularne pljuvačne žlezde su fenotipski slične epitelnim ćelijama subkapsularne i većini ćelija u meduli timusa. Epitelne ćelije acinusa su fenotipski slične epitelnim ćelijama korteksa timusa. Zaključak. Fenotipska kompleksnost submandibularne pljuvačne žlezde i sličnost s timusom otvara nove mogućnosti za analizu fenotipske sličnosti između ove žlezde i limfatičnih organa.

Imazalil, cipermetrin i karbendazim su rezidualno detektibilni u namirnicama u Republici Hrvatskoj. Cilj rada je utvrditi na koji se način mijenja broj leukocita u perifernoj krvi ovisno o toksičnom djelovanju pojedinog pesticida u Swiss miševa izloženih dozama NOAEL pojedinog pesticida i njihovim kombinacijama tijekom 28 dana. Broj leukocita povećan je izlaganjem imazalilu, krabendazimu a smanjen je pri kombinaciji imazalila + karbendazima. U svih skupina izloženih pesticidima i njihovim kombinacijama promijenjeni su postotni udjeli granulocita i agranulocita. Broj limfocita smanjen je u grupi izloženoj imazalilu + krabendazimu a u svih ostalih skupina je povećan, najviše u tretmanu s imazalilom. Broj neutrofila je povećan u svim skupinama od kojih najviše u grupi izloženoj imazalilu i cipermetrinu. Slično je i sa brojem monocita, iako cipermetrin nije jednako jako povećao broj neutrofila. Vjerojatno je u tih skupina povećana potreba za tkivnom makrofagnom aktivnošću. Broj bazofila povećan je u svih skupina ali najviše nakon izloženosti karbendazimu što upućuje na njegov alergeni potencijal. Slično ukazuje i povećan broj eozinofila. Staničnost i težina imunosnih organa mijenjala se specifično s obzirom na pojedini pesticid i njihove kombinacije i ukazuje na dinamiku imunotoksičnosti tijekom pokusa. U skupina u kojima je staničnost i težina slezene, timusa i limfnih čvorova smanjena vjerojatno je već došlo do mobilizacije novo stvorenih stanica u krv gdje doprinose ukupnom povećanju staničnosti do 28 dana. U organima u kojima je broj stanica povećan one su najvjerojatnije još uvijek u fazi diobe potaknute pesticidima i nisu ispuštene u krv. Subkronične niske doze ovih pesticida i njihovih kombinacija imaju imunotoksični, imunostimulativni potencijal osim u skupine tretirane karbendazimom i cipermetrinom koji ima jači citotoksični potencijal pa je i generalno smanjen broj imunokompetentnih stanica u krvi. Imazalil, cypermethrine and carbendazime are rezidually detected in food-stuffs in Republic Croatia. The aim of reasearch is to establish in which way of the number of leukocytes in pheripheral blood depend on the toxicological activity individual pesticide on the immune organs of Swiss mice exposed to them for the 28 days. It is supposed that number of leukocytes in blood will follow pathophysiological conditions in thymus, spleen and lymph node during pesticide treatment. Analysis of leukocytes showed the increase in total number of leukocytes in treatment with imazalil, carbendazime and combination of carbendazime. In all other groups percentage of granulocytes and agranulocytes was changed. The number of lymphocytes was reduced after combination of imazalile and carbendazime, and after imazalile, cypermethrine and carbendazime (used individually) was increased compared to control, especially in group imazalil. A similar thing happened with monocyte number, although the cypermethrine did not strongly increase the number of neutrophils. Probably these pesticides increased macrophage activity. The number of basophils was increased after treatment with all pesticides, especially after carbendazime. Eosinophils showed the similar effect. Cellularity and wight of immune organs was changed specifically to certain pesticide and their combinations and points to the dynamics of immunotoxicity tests. In groups where the cellularity and weigth of spleen, thymus and limph node decreased, it is possible that mobilization of new cells had been ocured in the blood leading to overall cellularity increase up to 28 days. In the organs in which cell number increased, cells are probably still in the phase of separation and not yet released into the blood. Sub-chronic low doses of these pesticides and their combinations are immunotoxic and immunostimulating except the combination of carbendazime and cypermethrine, which showed a strong cytotoxic potential and reduced the number of immunocompetent cells in the blood.

The aim of this thesis is the characterization of a protein involved in apoptosis. Our group has identified an early step common to different forms of intrinsic apoptosis stimuli. This step requires de novo synthesis of a novel Cyclin, Cyclin O, that upon apoptosis induction in lymphoid cells forms active complexes, primarily with Cdk2. Cyclin O expression precedes glucocorticoid and gamma radiation-induced apoptosis in vivo in mouse thymus and its overexpression induces apoptosis in cultured cells. Knocking down the endogenous expression of Cyclin O by shRNA leads to the inhibition of glucocorticoid and DNA damage-induced apoptosis while leaving CD95 death receptor mediated apoptosis intact. This data demonstrates that apoptosis induction in lymphoid cells is one of the physiological roles of Cyclin O and it does not act by perturbing a normal cellular process such as the cell cycle. In addition we have identified c-Myb a substrate of Cdk2-Cyclin O complexes and we show that c-Myb is downregulated during apoptosis of lymphoid cells., L'objectiu d'aquesta tesi és la caracterització d'una proteïna involucrada en l'apoptosi. El nostre grup ha identificat un pas primerenc comú en diversos estímuls apoptòtics de la ruta intrínseca. Aquest pas requereix la síntesi de novo d'una nova Ciclina, Ciclina O, que quan s'indueix apoptosi en cèl·lules limfoides forma complexes actius majoritàriament amb Cdk2. L'expressió de la Ciclina O és prèvia a l'apoptosi induïda per glucocorticoids i radiació gamma i la seva sobreexpressió indueix apoptosi en cultius cel·lulars. La baixada dels nivells d'expressió de la Ciclina O endògena amb shRNA provoca una inhibició de l'apoptosi induïda per glucocorticoids o agents que danyen el DNA, mentre que l'apoptosi mediada pel receptor CD95 es manté intacta. Aquests resultats demostren que la inducció d'apoptosi en cèl·lules limfoides és una de les funcions fisiològiques de la Ciclina O i que no es deu a una pertorbació de processos cel·lulars normals com ara el cicle cel·lular. A més a més, hem identificat c-Myb com a substrat dels complexes Cdk2-Ciclina O i demostrem que els nivells de c-Myb baixen durant l'apoptosis de cèl·lules limfoides.

INTRODUCCIÓN: El tratamiento antirretroviral de gran actividad (TARGA) ha permitido reducir la morbilidad y la mortalidad de la infección por el VIH, pero no puede erradicar la infección. Por eso se buscan alternativas como las terapias inmunomediadas, que potencian la respuesta inmune del paciente, para poder retirar, al menos temporalmente, el TARGA. Estas respuestas inmunes, tanto CD4+ como CD8+, existen al inicio de la infección, pero desaparecen con el TARGA, probablemente debido a la pérdida de estimulación del sistema inmune. En este sentido, los pacientes que presentan episodios de viremia intermitente (EVI), es decir, elevaciones transitorias de la carga viral (CV), se ha demostrado que aumentan estas respuestas. Dichos EVI se han asociado a diferentes circunstancias, entre ellas las vacunaciones. Como los pacientes VIH son inmunodeprimidos, se aconseja que sean vacunados. Por tanto, si dichas vacunaciones inducen EVI, esto se podría asociar a dos efectos antagónicos: por un lado, un aumento en las respuestas anti-VIH con un potencial mejor control de la infección; pero por otro lado, un potencial riesgo de aparición de resistencias al TARGA y fracaso terapéutico. PACIENTES Y MÉTODOS: Para evaluar esta circunstancia se llevó a cabo un ensayo clínico unicéntrico, prospectivo, randomizado, doble ciego y controlado con placebo en el que 26 pacientes con infección por el VIH se randomizaron a recibir durante 12 meses su TARGA junto a, o bien un calendario vacunal de 10 vacunas comerciales diferentes (hepatitis B, hepatitis A, gripe, neumococo, varicela, triple vírica (sarampión-rubéola-parotiditis) y tétanos-difteria), o bien placebo, suspendiendo el TARGA a los 12 meses. Se estudió el efecto del calendario vacunal, tanto durante el TARGA como tras su interrupción, sobre la CV (incluyendo la aparición de resistencias), las subpoblaciones linfocitarias, las respuestas específicas anti-VIH, la función tímica y las respuestas vacunales humorales y celulares. Además, mediante tres subestudios se evaluó el efecto de los EVI (superiores a 20 y 200 copias) sobre los mismos aspectos, y el efecto de la interrupción del TARGA sobre la función tímica, el sistema de la IL-7 y su receptor y las respuestas adquiridas a los antígenos vacunales. RESULTADOS: Ni el calendario vacunal en general ni ninguna vacuna en particular se asoció a un aumento en la frecuencia de CV detectables, aunque sí provocaron un descenso en los linfocitos T CD4+, un aumento en la activación y maduración linfocitaria hacia células memoria, así como en las respuestas linfoproliferativas, sin cambios en las respuestas específicas CD8+ anti-VIH. Tras la interrupción, el grupo vacunado presentó un rebote igual al que recibió el placebo, sin más resistencias virales, aunque presentó mayores respuestas específicas CD8+ anti-VIH en el periodo sin tratamiento. Cuando se analizó la influencia de los EVI sobre el sistema inmune se observó que los pacientes detectables presentaban durante el tratamiento menor número de linfocitos CD4+ y mayor de CD8+, así como un aumento en las respuestas específicas anti-VIH CD8+. Tras la interrupción, los pacientes detectables presentaron también un mayor rebote viral y un aumento de las respuestas linfoproliferativas anti-VIH. Las diferencias entre pacientes detectables e indetectables disminuyeron cuando se definieron por un ensayo con límite de 20 c/mL. Cuando se analizó la función tímica tras la interrupción se observó un descenso inicial con una recuperación parcial posterior. Y las respuestas vacunales humorales descendieron tras la interrupción, aumentando las celulares. CONCLUSIONES: El calendario vacunal fue seguro y no se asoció a efectos secundarios valorables ni elevaciones de la CV, pero tampoco fue útil como inmunoterapia. Además, la estimulación continuada produjo un aumento de la activación linfocitaria y una disminución en el recuento de linfocitos T CD4+. PALABRAS CLAVE: Infección por VIH, vacunaciones, inmunoterapia, timo, interrupciones de tratamiento antirretroviral., "Study of the influence of immunological repeated stimuli with commercial vaccines over the Viral Load (VL), resistance development and specific immunological response against HIV in early stage HIV patients with undetectable VL after HAART"INTRODUCTION: As HIV-infected patients are considered immunocompromised, it is generally recommended that they have to receipt appropriate vaccines. However data are conflicting concerning potential harmful effects following the administration of commercial vaccines in HIV-infected patients. Transient increases ("blips") in the viral load have been described associated with a single dose of vaccine, with the potential risk of developing resistance to HAART. On the other hand, there has been described that patients with blips can have an increase in HIV-specific immune responses, which may help to improve the viral control. PATIENTS AND METHODS: We have performed a clinical trial to evaluate the effect of a vaccination program in successfully treated HIV-infected adults on HAART compared to placebo. Its influence over viral load (including resistances), lymphocyte subsets, specific anti-HIV responses, thymic function and responses to vaccinations were analysed. There were also studied the influence of blips over immunity and the influence of interrupting HAART over thymic function and vaccination responses. RESULTS: Vaccinations were not associated to more detectable viral loads, but induced an increase in lymphocyte activation and a loss of CD4+ T-cells. They were not useful as immunotherapy because viral load rebounded after HAART interruption in the same way in placebo and vaccinated group, although specific CD8+ responses against HIV were higher in the latest. Patients with blips had less CD4+ T-cells, more specific CD8+ responses against HIV, and a higher rebound of viral load after discontinuing HAART. After HAART interruption, thymic function decreased firstly, but later presented a partial recovery. Humoral responses to vaccines decreased after stopping HAART.CONCLUSIONS: Vaccinations in HIV patients were safe and did not associate with more detectable viral loads. However, they induce a loss in CD4 T cells. They were not useful as immunotherapy.

Результати досліджень показали, що при дії хронічного радіоактивного опромінення малої інтенсивності відбуваються зміни морфологічного стану та морфометричних параметрів тимусу собак, що свідчить про розвиток вторинного імунодефіциту та прискорення процесу інволюції., Результаты исследований показали, что при действии хронического радиоактивного излучения малой интенсивности происходят изменения морфологического состояния и морфометрических параметров тимуса собак, что свидетельствует о развитии вторичного иммунодефицита и ускорении процесса инволюции., The results of the investigation conducted testify that under the low intensity permanent radioactive irradiation there occur changes in the morphological state and morphometric parameters in dogs. The above testifies to the development of secondary immunodeficiency as well as to the acceleration of involution process.

DiGeorge Sendromu (DGS), timus ve paratiroid bezinin konjenital yokluk sendromu olarak 1959’da bildirilmiştir. A.M. DiGeorge (1) 1965 yılında immünitede timusun önemli bir rolü olduğunu göstererek, hipoparatiroidizm, timik hipoplazi ve tekrarlayan enfeksiyon geçiren olgularını bildirmiş, böylece timik aplazi ve konjenital hipoparatiroidizm birlikteliği ‘DiGeorge Sendromu’ olarak adlandırılmıştır. La Chapelle (2) 1981’de bir ailenin dört etkilenen bireyinde 22. kromozom translokasyonunu göstererek bu sendromun genetik orjini için ilk ipuçlarını sağlamıştır.

Istraživanja izražaja neutralnih GSL i gangliozida u normalnim limfatičnim tkivima i stanicama miša dosad su uglavnom izvođena biokemijskom i imunokemijskom analizom izdvojenih i pročišćenih GSL razdvojenih tankoslojnom kromatografijom. Kako bi istražili raspodjelu globotriaozil-ceramida (Gb3Cer), globozida (Gb4Cer), gangliotriaozil-ceramida (Gg3Cer), gangliotetraozil-ceramida (Gg4Cer) i gangliozida GM3 i GalNAc-GM1b u mišjem timusu, slezeni i limfnom čvoru, rabili smo specifična poliklonalna protutijela za imunohistološku, protočno citometrijsku i Western blot analizu. Nezreli timociti izražavali su GSL biljege koje su prepoznala sva navedena protutijela osim Gb4Cer protutijela. Zreli timociti obilježeni su protutijelima na GalNAcGM1b, Gg4Cer i Gb4Cer. U sekundarnim limfatičnim organima, protutijela na globo-seriju neutralnih GSL obilježila su sinusoidne i vaskularne prostore, dok su protutijela na ganglio-seriju neutralnih GSL obilježila i područja koja sadržavaju limfocite. U Western blot analizi, samo se protutijelo protiv GalNAcGM1b gangliozida vezalo na specifičnu proteinsku vrpcu u sva tri ispitivana tkiva. Analiza limfocita iz sekundarnih limfatičnih organa protočnim citometrom pokazala je da limfociti B nose epitope koje prepoznaju sva navedena protutijela, dok je T-stanični izražaj GSL bio većinom usmjeren k ganglio-seriji neutralnih GSL i gangliozida tipa GM1b. Rezultati imunohistološke analize tkiva i analize stanica protočnim citometrom nisu se uvijek podudarali, vjerojatno zbog križne reaktivnosti GSL protutijela s glikoproteinski vezanim oligosaharidnim lancima i/ili zbog razlika u ugljikohidratnim profilima izoliranih stanica i stanica u tkivnom mikrookolišu. Stoga samo usporednim korištenjem biokemijskih metoda zajedno s imunohistološkom analizom i protočnom citometrijom može dati uvid u složenu tkivnu i staničnu raspodjelu GSL in vivo.

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In this study, embryonic development of the chicken thymus were investigated light microscopically Nor- mal involutive changes occured at post hacthing period and effects of hydrocortisone acetate (HCA) treatment on this lymphoid organ were also determined. As materials, 80 fertilized eggs from Avian Bred and tissue samples taken from 200 chickens from the same strain were used. On the 6th day of incubation, a number of large and dark ba- sophilic cells were observed in the organ primordium, and on the 8th day these cells have formed acumulations. On the 10th day, the thymic tissue has become lobulated. On the day 12, interlobular the connective tissue has thickened and in the central region of lobuli the vaskularization has increased. On the day 13, a difference between cortical and medullar regions was appearent. In this period, both large and small lymphocytes were observed in both cortical and medullar region, and reticular cells have accumulated in the medullar region. There after 13th day of incubation, a number of ANAE-positive lymphocytes were observed in the medullar region. On the day 15, medullary cysts located intercellularly were observed in the reticular cell accumulations, and both their numbers and sizes increased gradually at the following periods. During the posthatching period, on the 2nd day following HCA treatment, a relative decrease in cortical thickness of thymus and a striking increase in the number of medullar cysts were observed. Four days after HCA treatment, the cortical region has almost disappeared, and in some of the sections most of the cortical lymphocytes had pyknotic nuclei. Interlobar and interlobular connective tissue septae have thickened, the cysts have enlarged and their numbers were increased. Degenerated reticular cells and their groups have also formed large ac cumulations. Involutive changes continued gradually also in the following periods of the experiment., Bu çalışmada, tavuk timusunun embryonal dönemdeki histogenezisi ile kuluçkadan çıkıştan sonra verilen hidrokortizon asetatın (HCA) bu doku üzerine etkileri ışık mikroskopik olarak incelendi. Materyal olarak, S.Ü. Veterinar Fakültesi Zootekni ve Hayvan Besleme Bölümü'nden temin edilen Avian Irkı'na ait 80 adet dollü yumurta ve aynı irka ait 200 civcivden alınan doku ömekleri kullanıldı. Embriyonal gelişimin incelendiği grupta, inkubasyonun 6. gününde organ taslağını oluşturan hücre topluluğu içinde gözlenen, az sayıda, ini ve koyu bazofilik hücrelerin, 8. günde yer yer gruplaştıklar, 10. günde de küçük lopçuklar oluşturduklan gözlendi. Onikinci günde interlobuler dokunun daralmış ve lopçuların sıkaca paketlenmiş olduğu ve lopçukların merkezi bölümlerinde vaskülarizasyonun arttığı tespit edildi. Or ganin korteksmedulla ayınmi 13, günde belirgindi. Bu dönemde korteks ve medullada küçük ve büyük lenfositlerin her kisi de gözlenirken, medullada retikulum hücrelerinin gruplar oluşturduğu dikkati çekti. Onüçüncü günden itibaren timusun medullasinda az sayıda ANAE-pozitif lenfosite rastlandı. Inkubasyonun 15. gününde medulladaki retikulum hücre gruplarında küçük ve az sayıda kistik yapılar gözlendi. Kistik yapıların büyüklük ve sayılarının sonraki do nemlerde gittikçe arttiği tespit edildi. HCA uygulanan grupta, uygulamanın 2. gününde timusun medullasındaki kistlerde kontrol grubuna göre belirgin bir artış ve korteksin kalınlığında bir azalma dikkati çekerken; uygulamanın 4 gü nünde, organin korteksinin hemen hemen ortadan kalkmış olduğu gözlendi. Kortekste bulunan az sayıdaki lenfositin piknotik çekirdeklerinin olduğu, loplar ve lopçuklar arası bağ dokusunun kalınlaştığı dikkati çekti. Medullada ise kistlerin hem say ve hem de büyüklüklerinde artış, retikulum hücre gruplarında çoğalma ile birlikte dejenarasyon belirlendi. Bu bulgular ileriki dönemlerde de tespit edildi.

Although it is well known that the thymic epithelium is the main cellular component in driving the maturation of thymocytes through cell-to-cell and extracellular matrix-mediated interactions, in the regulation of proliferation and differentiation of T cells seems to participate also the locally produced hormones and neuropeptides, which in an autocrine and paracrine manner influence the thymic functions. In an attempt to elucidate the role of somatostatin in these events, in this study we investigated in vivo and in vitro effects of long acting somatostatin analogue, SMS 201-995 (SMS), on dexamethasone (DEX) and activation (anti-CD3 monoclonal antibodies) induced apoptosis of thymocytes. The data were estimated by analysis of absolute cellularity, subdiploid peak of DNA and maturational stage of thymocytes, detecting the CD4 and/or CD8 and T cell receptor (TCR) expression on thymocytes. The results have shown that SMS, given in vivo may potentiate the early phase of DEX-induced nuclear fragmentation (at 24h), accelerating simultaneously the elimination of thymic cells with double positive (DP) CD4high CD8high phenotype (expressed both as percentage and absolute number). On the contrary, SMS, given both in vivo and in vitro, down-regulated the late process (at 72h) of nuclear fragmentation, induced by anti-CD3 mAb, minimizing, simultaneously, the elimination of DP cells (expressed both as percentage and absolute number), and retarding the TRC  / high expression on anti-CD3 triggered apoptotic cells. During the early phase of activation-induced apoptosis SMS, however, augmented the hypoplasia in thymus, enhancing simultaneously the decrease of absolute number of thymic DP cells in CD3-treated mice.

Budući da je opstruktivna žutica često praćena jakom imunodisfunkcijom u radu se testirala mogućnost da se primjenom peptidoglikan-monomera vezanog sa zinkom (PGM-Zn) korigiraju promjene u imunološkom sustavu i toksičko oštećenje nastalo u jetri uslijed kolestaze. U tu svrhu je u miševa, sa podvezanim duktus koledokusom praćena dinamika protoonkogenske ekspresije u jetri, fenotipske promjene u intrahepatičkim limfatičkim stanicama u jetri, timusu i slezene, te njihov stanični ciklus. Osim toga, određivan je i fenotipski status periferne krvi bolesnika sa opstrukcijskom žuticom, a in vitro je praćena i blastična transformacija humanih mononuklearnih stanica periferne krvi, inducirana aktivatorima različitih traansmembranskih putova u prisustvu ikteričnog seruma i/ili PGM-Zn. Rezultati su ukazali na značajno imunosupresivno djelovanje opstrukcijske žutice i na prisutnost inhibitora u plazmi bolesnika. Pretretman PGM-Zn rezultirao je značajnim porastom proporcije CD4-CD8- stanica u timusu, redukcijom CD4-CD8+ timičnih stanica 1.post operativnog dana, većom proporcijom timičnih i spleničnih stanica u S fazi staničnog ciklusa, te izrazitim porastom ekspresije MHC antigena klase II u jetri i stimulacijom fagocitoze peritonealnih makrofaga. In vitro je utvrđeno da PGM-Zn poništava inhibicijske učinke ikteričnog seruma na PMA i anti-CD3+PMA induciranu blastogenezu humanih limfatičkih stanica. Imunohistološkom analizom je usto utvrđeno da PGM-Zn dovodi i do značajnih promjena u ekspresiji protoonkogenskih produkata (c-fos, H-ras, K-ras, p53 i Ki67) u kolestazom oštećenoj jetri.

Timus novorođenčeta u stanju stresa podliježe akutnoj reverzibilnoj involuciji zbog limfocitolitičkog učinka izlučenih glukokortikoidnih hormona. Deplecija timocita zbiva se kroz nekoliko dana, posredovana je makrofagnom fagocitozom i apoptozom, a neovisna je o gestacijskoj dobi. Obducirana su mrtva tijela 100 novorođenčadi s posebnom histološkom analizom timusa. Iz nalaza prvog uzorka periferne krvi prikupljeni su podaci o broju leukocita i diferencijalnoj krvnoj slici. Produženi stres u neonatalnoj dobi ima utjecaj na timusnu histologiju ; očekivani posljedični utjecaj na funkciju bilo bi potrebno ispitati kvantitativnom analizom B i T stanične populacije u perifernoj krvi.