Colocalization of calretinin and glucagon in rat pancreatic A cells.

Calretinin is a calcium-binding protein whose major functions are assumed to be buffering, transport of Ca2+, regulation of various enzyme systems and cellular protection. Moreover, calretinin does not seem to be a specific marker for any particular cell since it has been discovered in various mammalian and avian organs. In order to give clue to its function(s), we investigate its distribution in the rat pancreas using immunocytochemistry and transmission electron microscopy. We have found calretinin immunoreactive cells in rat pancreas only in the islets of Langerhans. These cells correspond to A cells producing glucagon. The colocalization of calretinin and glucagon was confirmed with double-labelling immunofluorescence. Nevertheless, the role of calretinin and its relationship with the production and/or the exocytosis of glucagon granules remains to be elucidated.

The value of ultrasound in staging the severity of acute pancreatitis.

The authors asses the accuracy and role of early ultrasound examination in staging the severity of acute pancreatitis. A total of 110 consecutive patients with acute pancreatitis were included into the study. The ultrasound findings were categorized into six categories and compared with a modification of multiple prognostic criteria, computerized tomography, operative findings (when available) and clinical outcome. The probability of a positive ultrasound finding in a patient with clinically severe acute pancreatitis was 89.6% (sensitivity). In comparison to computerized tomography the sensitivity of ultrasound in discovering CT diagnosed moderate and severe forms of acute pancreatitis was 86.6%. The sensitivity of ultrasound in discovering moderate and severe forms of acute pancreatitis as defined at laparotomy was 77.8%. The specificity of ultrasound was low (44.0%) in comparison with modified prognostic criteria, but high in comparison with computerized tomography (87.5%) and staging at laparotomy (85.7%). There was a good correlation between US defined moderate and severe forms of acute pancreatitis and clinical outcome (average number of hospital days and case fatality). The authors conclude that early ultrasound examination in acute pancreatitis is indicated and can help stage the severity of the disease and affect decision making.

Calretinin immunoreactivity in pineal gland of different mammals including man.

Calretinin (CR) is a calcium-binding protein, found in a variety of organs and systems such as the central nervous system and the pineal gland. It was first thought to be a specific neuronal marker but this selectivity is now in question since CR has been demonstrated in avian thymus, rat ovary, rat and guinea pig inner ear, rat testis, and chicken and rat pineal gland. To contribute to the knowledge of the presence of CR-positive cells in the pineal parenchyma of rat and other mammalian including man, we performed immunocytochemistry on pineal glands of gerbils, rats, goats, cows, and humans, using a CR anti-serum. To confirm it was actually CR that was demonstrated, we performed Western Blot analyses. Finally, to precisely identify the nature of CR-positive cells we accomplished double-labelling immunofluorescence, using antisera against some nerve cell specific cytosquelettal proteins such as MAP-5, MAP-2, NF-L, NF-M, and NF-H. CR-positive cells were found in all pineal glands studied. These cells all possess a round, oval, or polygonal-shaped perikaryon sending one or more processes of different lengths into the glandular parenchyma. There is a lack of CR immunoreactivity in the nucleus and cell organelles while the cytosol contains a high concentration of this protein. Nevertheless, there are some slight differences between species, especially concerning the number of reactive cells and their relationships with different parenchymal structures such as blood vessels or acervuli. Among the CR-positive cells, only a few were actually nerve cells, contributing probably to an intrinsic innervation of the gland. The remaining CR-reactive cells seem to correspond mostly to pinealocytes in a specific histophysiological state and possibly to neuron-like cells. The significance of the CR-positive cells in the pineal glands remains to be elucidated.

Ductal morphometry of ventral pancreas in pancreas divisum. Comparison between clinical and anatomical results.

Aim of the investigation is to precisely study the morphological features of ventral pancreas ductography in pancreas divisum, in order to improve the radiological interpretation and differential diagnosis of this frequent pancreatic anomaly. The clinical part of the study was based on 610 endoscopic retrograde pancreatograms, with pancreas divisum diagnosed in 14 (2.3%) cases; while the anatomical part consisted of 203 postmortem pancreatograms of human pancreas obtained at autopsy, where pancreas divisum was found in 12 (5.9%) cases. The following ductal features of the ventral pancreas were studied: length and calibre of the main duct, number of side branches, calibre of the common bile duct and the biliopancreatic junction angle. No significant differences were detected between the results from the 2 groups, with the exception of side branches, which were more numerous in the anatomical series, probably because of higher injection pressure and consequent better opacification. These results underline the potential of the anatomical pancreatography serving as a model for studying the ductal system of pancreas divisum.

[Infiltration of the testes in acute lymphoblastic leukemia in childhood--personal experience]. / Infiltracija testisa u toku akutne limfoblastne leukemije decijeg uzrasta--nase iskustvo.

According to various literature data testicular leukemic infiltration occurs in 5-40%, while the mean time for testicular recurrence is 36 months. Hematologic recurrence usually occurs in the interval from 1-6 months. Although testicular leukemic infiltration clinically occurs as an isolated recurrence of leukemia, it usually presents only a clearly marked spot in the general recurrence of the disease. It is a painless enlargement of one or both testes. The diagnosis is made ultrasonographically and by histopathologic examination of the biopsied testicle tissue. This paper deals with 4 patients with acute lymphoblastic leukemia and testicular recurrence. In 2 patients the testicular recurrence occurred as a part of the general recurrence of the disease, while in the other 2 at the time of testicular recurrence diagnosis, the bone marrow was regular. All 4 patients were treated with combined radiotherapy and chemotherapy, in 3 cases successfully.

[Congenital leukemia--personal experience]. / Kongenitalna leukemija--nasa iskustva.

Congenital leukemia is a very rare form of acute leukemia and up to now more than 100 well documented cases have been published. Symptoms and signs of the disease may be faintly differentiated and be a differential diagnostic problem for a long period of time in comparison to other common diseases of the newborn. This is a case report on two newborns with congenital leukemia. It was an acute non-lymphoblastic leukemia in the first case, and acute lymphoblastic leukemia in the second. In both cases there were nonspecific manifestations of the disease (no weight gaining, hemorrhagic enterocolitis). In both cases polyhemotherapeutic protocols were performed. In the first case only partial remission occurred and death occurred during the hemathologic recidive of the disease. In the second case, 15 months after the diagnose of congenital leukemia has been established, complete remission is in course.

Nitric oxide synthase in the subfornical organ of Mongolian gerbil (Meriones unguiculatus), mouse and rat.

The present paper aims to contribute to the knowledge of the histophysiologic role of the subfornical organ. Using nicotinamide adenine dinucleotide phosphate diaphorase histochemistry (NADPH-diaporase) and nitric oxide synthase immunocytochemistry, we detected a high concentration of the neuronal isoform of nitric oxide synthase in neurons of the subfornical organ of the adult Mongolian gerbil (Meriones unguiculatus), mouse and rat. Our results suggest that neurons of the subfornical organ produce a considerable amount of nitric oxide which acts, not only as a neurotransmitter, but could also diffuse into cerebral blood vessels and cerebrospinal fluid.

[Congenital methemoglobinemia]. / Urodena methemoglobinemija.

This is a report on a case of inborn methemoglobinemia. The disease has been diagnosed in the first week of infant's life. At first it was assumed it was a case of inborn heart defect because of expressed generalized cyanosis, but very soon it was excluded. Values of methemoglobin were extremely high, so three days after birth hereditary methemoglobinemia was diagnosed. Our diagnosis was confirmed by disappearing of cyanosis after ascorbic acid was applied, by values of methemoglobin reductase of the infant and its father, anamnestic data revealing that mother had had temporary cyanosis during her infancy, and satisfactory clinical course of illness.

Presence of calmodulin and NADPH-diaphorase activity but not of neuronal nitric oxide synthase in the subcommissural organ of the Mongolian gerbil.

The subcommissural organ (SCO) and the ventricular ependyma of the third ventricle of the Mongolian gerbil (Meriones unguiculatus) exhibit a highly positive histochemical reaction to NADPH-diaphorase and a considerable immunocytochemical expression of calmodulin. The SCO, however, does not display an immunocytochemical response to the neuronal isoform of nitric oxide synthase (NOS) whereas it was demonstrated in the ventricular ependyma. Since the NADPH-diaphorase, which detects all isoforms of NOS, was clearly positive at the level of the SCO, the obtained results suggest that the SCO of the Mongolian gerbil produces nitric oxide but contains a NOS whose antigenic specificity differs from that of the neuronal isoform of this enzyme.

Mid-term results with cultured epidermal autografts, allogenic skin transplants and cyclosporin A medication.

Transplantation of allogenic split thickness skin grafts (STSG) and immunosuppression with cyclosporin A enable early and definitive skin replacement of extensive, deep partial and full thickness burns. Covering burn defects with definitely engrafted, allogenic dermis and cultivated epithelial autografts (CEA) permits the subsequent withdrawal of cyclosporin A medication. Light-microscopy examination of biopsies, taken 12 and 24 months postgrafting, and electron microscopy of biopsies taken 12 months postgrafting, demonstrates a re-established, but somewhat reduced anchoring of the CEA as compared with a normal epidermal-dermal junction. Clinical inspection, 20 months postgrafting, confirms the histological observations that epifascial transplantation is qualitatively inferior to placing the allogenic STSG on the subcutaneous tissue. In the first situation, the dermis is inelastic and collagen deposition is excessive, whereas in the second case collagen deposition is comparatively reduced and the dermis shows clinically some elasticity.

[Whipple's disease. Case report]. / Morbus Whipple. Povodom jednog slucaja.

This is a case report on a 42 years old patient suffering from Whipple's disease. The patient had characteristic symptoms: high temperature, frequent bowel movement, melanoderma, arthralgia and enlarged lymph nodes. After detailed clinical researches, x-ray and endoscopic examination, bacteriology examination of faeces, absorptional tests, the diagnosis was made thanks to pathohistologic analysis of biopsy samples of the small intestine by light and electron microscopy. The differential diagnostic similarity with AIDS, especially at the early phase of the illness, required excluding HTLV III. We examined the aetiophathogenesis of the illness and discussed the clinical and laboratory presentation of the patient considering contemporary ideas regarding this disease.

Occurrence of large granular lymphocytes and natural killer cells in the epithelium of the gut distinguishes two different coeliac diseases.

In a longterm study, we have divided coeliac disease into two distinct entities (abortive and permanent) based on the occurrence of large granular lymphocytes and natural killer cells within the epithelium of the gut. The natural killer and large granular lymphocytes cells were characterised by either immunohistochemical or phase contrast microscopical procedures on the initial biopsies from 15 children with coeliac disease. They were compared with seven individuals with partial villus atrophy and eight with normal villous morphology. Although the histological findings were similar in the initial biopsies of all patients with coeliac disease, the patients with permanent coeliac disease had a significantly lower number (0.41(0.61)cells/mm2) of large granular lymphocytes and natural killer cells compared with those patients with abortive coeliac disease (11.93 (6.23) cells/mm2). Those in the permanent group developed a significantly more pronounced flat mucosa after gluten challenge or provocation compared with the abortive group and had to remain on a strict gluten free diet in contrast with those in the abortive group. Thus, the occurrence of intraepithelial large granular lymphocytes and natural killer cells characterises two distinctly different coeliac diseases. Based on our results neither the histological evaluation of the biopsy nor the utilisation of the revised European Society for Paediatric Gastroenterology and Nutrition (ESPGAN) Criteria are adequate in diagnosing the two types of coeliac disease.

Light and electron microscopic immunocytochemical localization of glutamine synthetase in the superficial pineal gland of the rat.

Glutamine synthetase (L-glutamate:ammonia ligase; EC 6.3.1.2), an enzyme catalysing the ATP-dependent formation of glutamine from glutamate and ammonia, was detected immunocytochemically only in glial (interstitial) cells of the superficial pineal gland of the rat. The results show the important role of pineal glial cells in the metabolism of the presumptive neurotransmitters, glutamate and gamma-aminobutyric acid (GABA) as well as in detoxification of ammonia.

Histoenzymological light and electron microscopic localization of carbonic anhydrase in the rat superficial pineal gland.

A histoenzymological method for demonstration of carbonic anhydrase (CA) was used for cryostat sections of superficial rat pineal gland. Light and electron microscopic analysis showed a high concentration of this enzyme in stellate cells, particularly heavily distributed under the organ capsule and scattered in the parenchyma, where they form a widespread three-dimensional network. Most of the pericapillary spaces were lined by CA-reactive cells. The reaction product, in the form of strongly electron-dark precipitates, was localized in the cytoplasmic matrix, whereas organelles, except for some mitochondrial intracristal spaces, were CA-negative. On the basis of localization and morphology of CA-positive cells, we conclude that these represent glial (interstitial) cells.

Use of human organs fixed long ago for the scanning electron microscopic analysis.

Normal human organs fixed 10-15 years ago in formol and Bouin as well as organs prepared from cadavers used for the anatomical dissections show a surprisingly well preserved structure in the scanning electron microscope. Such organs can be successfully used for the teaching of the 3-dimensional microanatomy.