Churg-Straussov sindrom: prikaz slučaja

Churg-Strauss syndrome (CSS) is a necrotizing small vessel vasculitis characterized by the presence of asthma, hypereosinophilia and sinusitis. Other common manifestations are pulmonary infiltrates, skin, gastrointestinal and cardiovascular involvement. Although not a criterion for the diagnosis of CSS, the presence of antineutrophil cytoplasmic autoantibodies (ANCA) is now established as being associated with CSS. In this report, a 31-year-old male with a history of difficult-to-control asthma is presented. It was associated with peripheral and bronchoalveolar eosinophilia, sinusitis, and high level of ANCA (perinuclear labeling pattern). Clinical manifestations observed at the time of relapses were palpable purpura, erythema nodosum, arthralgia, musculoskeletal complications, and episcleritis. In spite of vasculitis remission, low doses of steroids to control asthma were necessary for 10 years. The aim of this case report is to point to the possibility of CSS in patients with severe persistent asthma and atypical allergic diathesis.Churg-Straussov sindrom (CSS) je granulomatozni vaskulitis posredovan antineutrofilnim citoplazmatskim autoantitijelima perinuklearne fluorescencije (P-ANCA). Očituje se kliničkim simptomima ustrajne astme i sinusitisa uz izraženu eozinofiliju, što kod progredirajućih oblika bolesti prethodi migrirajućim plućnim infiltratima uz izvanplućne pojavnosti na perifernim živcima, koži, središnjem živčanom, probavnom i krvožilnom sustavu. Prikazujemo bolesnika s CSS koji se očitovao teškom ustrajnom astmom, perifernom i tkivnom eozinofilijom, sinusitisom i izrazito visokim titrom P-ANCA. Kontrola astme postignuta je tek nakon 10-godišnje steroidne terapije uz mnogobrojne nuspojave. Cilj prikaza je upozoriti na mogućnost CSS kod težih oblika astme i ukazati na mogućnost autoimunih pojavnosti u bolesnika s atopijskom konstitucijom

Regulacija rasta i diferencijacije embrija sisavaca faktorima rasta FGF i NGF u kulturi organa in vitro

The aim was to analyze the regulation of growth and tissue differentiation in a unique in vitro4culture model of gastrulating mammalian embryo by fibroblast growth factor (FGF) and nerve growth factor (NGF) during two weeks. They both play a crucial role during embryogenesis and the purpose of this study was to test their possible synergistic influence in the period when mammalian embryos are extremely sensitive to external factors. We cultivated 9.5-day-old rat embryos on metal grid supported lens paper, at the air-liquid interface in culture medium (Eagle\u27s minimal essential medium (MEM) with 50% of homologous serum) with the addition of FGF, NGF and a combination of FGF/NGF in a time frame of 9 days. Other three groups of embryos were for 24 hours pretreated with 5-azacytidine (5azaC), an agent that can activate repressed genes. A parallel group of nontreated control embryos were cultivated with each experimental group. During 14 days of culture embryos grown in teratoma-like explants and growth rate were evaluated by measuring average size of explants using an eyepiece micrometer on days 5, 7, 11 and 14 after the addition of growth factors. Differences between respective groups were estimated by Student\u27s t-test. Differentiated tissues were estimated on serial histologic sections. c2-test or Fisher exact test were used to compare the proportion of tissues between respective groups. In embryo-derived teratomas NGF or FGF/ NGF combination used within the 9 day time frame did not stimulate differentiation of any kind of tissues; moreover, FGF/NGF inhibited maturation of epidermis, while FGF stimulated differentiation of neural tissue, hemopoiesis and myotubes. We did not observe any kind of stimulative cooperative action of FGF and NGF in differentiation processes. So it seems that NGF hinders the stimulating effect of FGF. NGF alone impaired growth of explants, but in combination with FGF acted synergistically, thus improving the growth rate of cultivated embryos. Additional activation of genes with 5azaC had no the effect on possible NGF influence on neural tissue differentiation, but resulted in improved myotube differentiation. The activation of genes with 5azaC/FGF signal and 5azaC/FGF/NGF combination improved the proportion of neural tissue and myotubes as well as hemopoiesis. Obviously, these results supported the role of FGF as neural inducer and mesoderm inducer. Anyway, FGF or NGF induced differentiation at least partially depends on the status of gene methylation.Namjera je bila istražiti regulaciju rasta i diferencijacije pomoću fibroblastnog faktora rasta (FGF) i faktora rasta živaca (NGF) u jedinstvenom in vitro modelu za kultiviranje embrija sisavaca u kritičnoj fazi razvoja, gastrulaciji, tijekom dva tjedna. Oba faktora imaju ključnu ulogu u embriogenezi sisavaca, a svrha je bila istražiti njihov mogući sinergistični učinak kada su embriji sisavaca najosjetljiviji na djelovanje pojedinih vanjskih čimbenika. Embrije štakora stare 9,5 dana kultivirali smo na metalnoj mrežici s lećnim papirićem, na granici plinske i tekuće faze, u temeljnom mediju za kultiviranje (Eagleovom minimalnom esencijalnom mediju (MEM) s 50% homolognog seruma) uz dodatak faktora rasta FGF, NGF i kombinacije FGF/NGF tijekom 9 dana kulture. Druge tri skupine embrija su prethodno bile tretirane 24 sata 5-azacitidinom (5azaC), agensom koji može aktivirati gene. Paralelne skupine embrija kultivirane su u temeljnom mediju kao kontrolne. Tijekom 14 dana kultiviranja embriji izrastu u teratoidne strukture, a pritom se rast procjenjuje određivanjem prosječene veličine eksplantata pomoću okularnog mikrometra u određene dane (5, 7, 11, 14) nakon dodavanja faktora rasta. Razlike u rastu između pojedinih skupina ispitane su Studentovim t-testom. Prisutnost diferenciranih tkiva u teratomima utvrđena je analizom na serijskim histološkim rezovima. Razlike u proporcijama diferenciranih tkiva između pojedinih skupina utvrđene su c2-testom ili Fisherovim egzaktnim testom. U embrijskim teratomima razvijenim in vitro niti NGF niti FGF/NGF kombinirani tretman nisu stimulirali diferencijaciju bilo koje vrste tkiva, dapače, FGF/NGF kombinacija je inhibirala sazrijevanje epidermisa. FGF je stimulirao diferencijaciju živčanog tkiva (p<0,05), miotuba i hemopoezu. Nismo mogli utvrditi sinergistični učinak FGF i NGF u procesu diferencijacije bilo koje vrste tkiva. Dapače, čini se da NGF smanjuje stimulacijski učinak FGF u procesu diferencijacije. NGF suzbija rast eksplantata, ali u kombinaciji s FGF djeluje sinergistično, značajno povećavajući rast eksplantata. NGF nije imao učinka na očekivano povećanje diferencijacije živčanog tkiva, nakon dodatne aktivacije gena pomoću 5azaC, međutim, diferencijacija miotuba je značajno povećana. Kombinirano djelovanje 5azaC/FGF i 5azaC/FGF/NGF na aktivnost gena povećalo je učestalost diferencijacije živčanog tkiva, miotuba i hemopoezu u eksplantatima. Ovaj rezultat potvrđuje djelovanje FGF kao induktora živčanog tkiva i mezoderma. U svakom slučaju diferencijacija potaknuta FGF i NGF signalima djelomice ovisi o stupnju metilacije gena

Neurološke manifestacije celijakije u odraslih: prikaz slučaja

Celiac disease is more common than previously thought, and a high index of suspicion is important in its diagnosis. Typically, cases of celiac disease present at the age of 5-24 months with the symptoms of intestinal malabsorption, growth retardation, abnormal stools, abdominal distension, muscle wasting, hypotonia, poor appetite and low spirit. In adults, the symptoms of celiac disease may be highly varied. The incidence of malignancies is also increased. Most of these are small bowel lymphomas and carcinomas of the esophagus and colon. In some patients, the disease is associated with clinical dysfunction of the nervous system, manifesting variably as encephalopathy, cerebellar abnormalities, seizures, cerebral atrophy and dementia, brain stem encephalitis, cerebral vasculitis, myopathy, quadriparesis (metabolic), myelopathy, peripheral neuropathy, multifocal leukoencephalopathy, and psychiatric disorders. Presentation is made of a 47-year-old woman with misdiagnosed celiac disease, who first developed pain in the back which was treated with spinal support. She also complained of very severe bone pains. Finally, she was admitted to the hospital with a history of increasing difficulty on walking, weakness in her legs, urine incontinence, and 5-kg weight loss in four months. Celiac disease should therefore be considered on differential diagnosis in patients presenting with unexplained neurologic symptoms.Celijakija je učestalija nego što se misli i to treba uzeti u obzir pri diferencijalnoj dijagnostici nejasnih neuroloških sindroma. Obično se javlja u dobi od 5-24 mjeseca sa znacima crijevne malapsorpcije, zaostajanjem u rastu, nadutošću uz učestale stolice, slabost mišića, hipotoniju, slab apetit i neraspoloženje. U odraslih znaci celijakije mogu biti različiti. Učestalost malignoma također je povećana. U nekih je bolesnika celijakija povezana sa znacima oštećenja živčanoga sustava poput encefalopatije, cerebelarne simptomatologije, cerebralne atrofije, demencije, cerebralnog vaskulitisa, miopatije, oduzetosti ekstremiteta, periferne neuropatije, višežarišne encefalopatije i psihijatrijskih poremećaja. Opisana je 47-godišnja bolesnica s nedijagnosticiranom celijakijom, u koje je prvi simptom bila intenzivna bol u leđima, zbog koje je provodila fizikalnu rehabilitaciju. Nakon toga javili su se difuzni bolovi u kostima uz teškoće pri hodu, a potom napredujuća slabost donjih ekstremiteta. Zbog flacidne parapareze, smetnji mokrenja i gubitka tjelesne težine bolesnica je primljena na bolničko liječenje. Dakle, u bolesnika s nejasnom neurološkom simptomatologijom treba u obzir uzeti i mogućnost da se primarno radi o celijakiji

Inducirani sputum - suvremena metoda za citološku analizu bronhalnih uzoraka

Sputum induction by inhalation of hypertonic saline is a noninvasive method to obtain secretions from the lower respiratory tract for cell counting and bacteriologic analyses. The aim of this study was to introduce the method of induced sputum in our clinical practice and to get an insight into airway inflammation by examining cell count differences in 15 asthmatic patients, 30 chronic obstructive pulmonary disease (COPD) patients and 15 healthy controls. Gradually increasing concentrations (3%, 4%, 5%) of hypertonic saline were inhaled. Subjects were encouraged to cough in order to expectorate. The quality of the sample was scored on the volume of the plugs (considered to represent lower respiratory tract secretion) and salivary contamination (proportion of squamous cells in the slides). A sample score ≥4 was considered adequate, 3 intermediate, and ≤2 inadequate. Cytologic sputum analysis was done after microscopic selection of the plugs. The results showed that inhalation of hypertonic saline is a safe method to get adequate sample for investigating various inflammatory mechanisms in lower airways. There were differences in sputum from three groups of subjects, i.e. a higher proportion of neutrophils in COPD (32%) in comparison to asthmatics (15%) and healthy controls (10%). The percentage of alveolar macrophages also differed (COPD 56.5%, asthmatics 46%, healthy 40%). Asthmatic patients had a higher proportion of eosinophils (asthmatics 18.5%, COPD 1.9%, healthy 0.06%) and metachromatic cells (asthmatics 0.3%, COPD 0.039%, healthy 0.014%).Inducirani sputum je metoda potpomognutog iskašljavanja putem inhaliranja rastućih koncentracija (3%, 4%, 5%) hipertonične otopine NaCl. Tim neinvazivnim načinom mogu se izbjeći invazivnije dijagnostičke metode poput bronhoskopije, a dobiti prikladni uzorci sekreta iz donjih dišnih putova. Cilj ovoga istraživanja bio je: a) uvesti metodu induciranog sputuma u našu kliničku praksu, b) ispitati njenu sigurnost i uspješnost indukcije i c) usporediti ukupan i diferencijalni broj stanica u 15bolesnika s astmom, 30 s KOPB i 15 zdravih ispitanika. Kvaliteta sputuma procjenjivana je na temelju količine bronhalnih odljevaka (čepića koji dokazuju da se radi o sekretu iz donjeg dišnog sustava) te udjela stanica pločastog epitela (dokaz da uzorak čini slina). Bodovi ≥4 pokazuju da je uzorak primjeren, 3 da je srednje kvalitete, a bodovi ≤2 da nije prikladan za analizu. Rezultati su pokazali da je inhalacija hipertonične otopine sigurna metoda za dobivanje prikladnih uzoraka kojima se može dobiti uvid u različite upalne mehanizme kod bolesnika s opstruktivnim plućnim bolestima. U bolesnika s KOPB utvrđen je značajno veći udio neutrofila (KOPB 32%, astma 15%, zdravi 10%) i alveolarnih makrofaga (KOPB 56,5%, astma 46%,zdravi 40%) u odnosu na druge dvije ispitivane skupine. U bolesnika s astmom utvrđen je značajno veći udio eozinofila (astma 18,5%, KOPB 1,9%, zdravi 0,06%) i metakromatskih stanica (astma 0,3%, KOPB 0,039%, zdravi 0,014%)

Analysis of Ikaros Family Splicing Variants in Human Hematopoietic Lineages

Transcription factors from the Ikaros family are involved in lymphocyte differentiation and have a critical role at specific check points of the haemopoietic pathway. However, how developmentally regulated changes are reflected in gene expression programs of lymphocyte differentiation is not well understood. It has been suggested that disregulation of transcription factors from the Ikaros family is associated with the development of different human leukemias. In this work we analyzed the state of Ikaros family members in different leukemic cells with the aim to explore the transcriptional control of human hematopoietic lineages and shed some new light on our understanding of transcription factor significance in human leukemias. By means of RT-PCR and specific primers we investigated the expression of Ikaros, Aiolos and Helios transcription factors and their splicing variants in seven leukemia cell lines derived from different types of leukemia (ALL, CML, AML) and lymphoma (histiocytic lymphoma, Burkitt lymphoma and anaplastic large cell lymphoma). In all of the cell lines examined Ikaros was present in dominant Ik1 to Ik4 isoforms and small Ik6 isoform was absent. Aiolos was expressed in the majority of the cell lines, of both, B and T origin, in the form of the full length Aio1. Helios was also present only in two long isoforms Hel1 and Hel2, and was absent in one third of the lines. Similar distribution of positive and negative expression of Aiolos and Helios found in various types of leukemias could implicate common pathways of their regulation

Neurogeneze u ljudi

Modified organ culture of mammalian postimplantation embryo developed in our laboratory provides favorable conditions for terminal differentiation of the main tissues of gastrulating embryo, including nerve tissue. Intracellular signals provided by growth and neurotrophic factors play a crucial role during neurogenesis. In the present study, the role of retinoic acid (RA), fibroblast growth factor (FGF) and nerve growth factor (NGF) in neural tissue differentiation was investigated in cultivated 9.5-day-old rat embryo in the critical period of mammalian development, gastrulation. The proper embryonic part of the whole embryo without extraembryonic membranes was cultivated in air-liquid interface for two weeks. To study the effect of neural tissue-inducing agents, various concentrations of RA, FGF, NGF and FGF/NGF combinations were added to the basic culture medium within the time frame of 9 days. The embryos developed into teratoma-like structures, and then they were fixed and histologically examined for the presence of neural tissue. RA and NGF did not improve neural tissue differentiation. However, its proportion was significantly higher in FGF-treated embryos. FGF is less effective in the presence of NGF at the very beginning of neural tissue differentiation. It seems that NGF hinders the stimulating effect of FGF. In our experimental model, neural tissue differentiation is probably initially regulated by FGF signal, although there is evidence that RA and NGF signals are also needed later in development. Our model system has the advantage of being simpler than the embryo in vivo but closer to normal development than cell cultures. Therefore, our results are in accordance with the idea that FGF is a neural inducer in the vertebrates.U našem laboratoriju je razvijena modificirana kultura organa za kultiviranje embrija sisavaca, koja osigurava najbolje uvjete za terminalnu diferencijaciju osnovnih tkiva zametka u stadiju gastrulacije, a između ostalog i živčanog tkiva. Unutarstanični signali, kao što su čimbenici rasta i neurotrofični čimbenici, imaju ključnu ulogu u procesu neurogeneze. Istraživali smo ulogu retinoične kiseline (RA), faktora rasta fibroblasta (FGF) i faktora rasta živaca (NGF) u diferencijaciji neuralnog tkiva u kultiviranim 9,5 dana starim embrijima sisavaca u kritičnom razdoblju razvitka sisavaca, gastrulaciji. In vitro su bili tijekom dva tjedna kultivirani samo embrionalni dijelovi zametaka, bez izvanembrionalnih membrana na granici zračne i tekuće faze. Kako bismo istražili učinak potencijalnih uzročnika koji induciraju neuralnu diferencijaciju, u temeljni medij za kultiviranje su tijekom 9 dana bile dodavane različite koncentracije RA, FGF, NGF i kombinacija FGF/NGF. Embriji su se razvili u teratome, tada su bili fiksirani i histološki ispitani na prisutnost neuralnog tkiva. RA i NGF nisu poboljšali diferencijaciju neuralnog tkiva. Međutim, zastupljenost neuralnog tkiva bila je značajno češća u embrijima tretiranim pomoću FGF. FGF je na samom početku neurogeneze manje učinkovit u prisutnosti NGF. Stoga se čini da NGF smanjuje stimulacijski učinak FGF. U našem eksperimentalnom modelu je diferencijacija neuralnog tkiva vjerojatno inicijalno regulirana signalom FGF, iako postoje podaci da su RA i NGF također, ali vjerojatno kasnije, potrebni za razvitak. Prednost našega eksperimentalnog modela je u tome što je to jednostavniji sustav nego embrij in vivo, ali ipak bliži normalnom embrionalnom razvitku od staničnih kultura. Prema tome, rezultati su u suglasju s idejom da je FGF induktor živčanog tkiva u kralješnjaka