Connectivity of neuronal populations within and between areas of primate somatosensory cortex.

Functions of the cerebral cortex emerge via interactions of horizontally distributed neuronal populations within and across areas. However, the connectional underpinning of these interactions is not well understood. The present study explores the circuitry of column-size cortical domains within the hierarchically organized somatosensory cortical areas 3b and 1 using tract tracing and optical intrinsic signal imaging (OIS). The anatomical findings reveal that feedforward connections exhibit high topographic specificity, while intrinsic and feedback connections have a more widespread distribution. Both intrinsic and inter-areal connections are topographically oriented across the finger representations. Compared to area 3b, the low clustering of connections and small cortical magnification factor supports that the circuitry of area 1 scaffolds a sparse functional representation that integrates peripheral information from a large area that is fed back to area 3b. Fast information exchange between areas is ensured by thick axons forming a topographically organized, reciprocal pathway. Moreover, the highest density of projecting neurons and groups of axon arborization patches corresponds well with the size and locations of the functional population response reported by OIS. The findings establish connectional motifs at the mesoscopic level that underpin the functional organization of the cerebral cortex.

Layer-specific activity of tissue non-specific alkaline phosphatase in the human neocortex.

The ectoenzyme tissue non-specific alkaline phosphatase (TNAP) is mostly known for its role in bone mineralization. However, in the severe form of hypophosphatasia, TNAP deficiency also results in epileptic seizures, suggesting a role of this enzyme in brain functions. Accordingly, TNAP activity was shown in the neuropil of the cerebral cortex in diverse mammalian species. However in spite of its clinical significance, the neuronal localization of TNAP has not been investigated in the human brain. By using enzyme histochemistry, we found an unprecedented pattern of TNAP activity appearing as an uninterrupted layer across diverse occipital-, frontal- and temporal lobe areas of the human cerebral cortex. This marked TNAP-active band was localized infragranulary in layer 5 as defined by quantitative comparisons on parallel sections stained by various techniques to reveal the laminar pattern. On the contrary, TNAP activity was localized in layer 4 of the primary visual and somatosensory cortices, which is consistent with earlier observations on other species. This result suggests that the expression of TNAP in the thalamo-recipient granular layer is an evolutionary conserved feature of the sensory cortex. The observations of the present study also suggest that diverse neurocognitive functions share a common cerebral cortical mechanism depending on TNAP activity in layer 5. In summary, the present data point on the distinctive role of layer 5 in cortical computation and neurological disorders caused by TNAP dysfunctions in the human brain.

Caveolae and caveolin isoforms in rat peritoneal macrophages.

Caveolea are special (highly hydrophobic) plasma membrane invaginations with a diameter of 50-100 nm. Their characteristic features are the flask- or omega-shape and the lack of basket-like coat composed of clathrin. Caveolin-an integral membrane protein-is the principal component of caveolae membranes in vivo. Multiple forms of caveolin have been identified: caveolin-1alpha, caveolin-1beta, caveolin-2 and caveolin-3. They differ in their specific properties and tissue distribution. In this paper we summarize the morphological and biochemical data providing strong evidence about the existence and function of caveolae in rat peritoneal macrophages. When studied electron microscopically, the surface of both resident and elicited macrophages exhibited omega- or flask-shaped plasma membrane invaginations. There was a significant difference, however, in the number of these profiles: whereas in resident cells only a small amount of them was found on the cell surface, in elicited cells they were abundantly present on the plasma membrane. Using an antibody against the VIP21/caveolin-1 isoform we showed that these plasma membrane pits were indeed caveolae. The number and the appearance of caveolae were found to be in close correlation with the functional activity of these phagocytotic cells, indicating that the formation of caveolae is a highly regulated process. Using Western blot analysis two different proteins ( approximately 29 and approximately 20 kDa)-both labelled with anti-caveolin antibodies-were identified in resident and elicited macrophages that have been isolated from rat peritoneal cavity. The approximately 20 kDa protein was labelled specifically only by anti-VIP21/caveolin-1, while the approximately 29 kDa protein was labelled by both anti-VIP21/caveolin-1 and anti-caveolin-2 antibodies. The presence of the approximately 29 kDa protein was highly characteristic of resident cells, and only a small amount of approximately 20 kDa protein was detected in these cells. Elicitation has resulted in a significant increase in the amount of approximately 20 kDa protein labeled only with anit-VIP21/caveolin-1. Our morphological (confocal and electron microscopical) studies have shown that in resident cells caveolin was present in the cytoplasm, in smaller vesicles and multivesicular bodies around the Golgi area. Only a very small amount of caveolae was found on the cell surface of these cells. In elicited macrophages, caveolae (labelled with anti-VIP21/caveolin-1 antibody) appeared in large numbers on the cell surface, but caveolin detected by anti-caveolin-2 was also found in small vesicles and multivesicular bodies. These data support the idea that the expression of the approximately 29 kDa (caveolin-related) protein is insufficient for caveolae formation in resident cells, it can function as a modified, macrophage-specific caveolin-2 isoform. Our results strongly suggest that caveolin-1 plays a crucial role in the formation of caveolae: it is the amount of caveolin-1 that regulates the appearance of caveolae on the plasma membrane. Studying the endocytotic processes of resident and elicited macrophages we have found that elicited macrophages bound and internalized significantly larger amounts of fluid phase marker (HRP) and immune complex (peroxidase-antiperoxidase-PAP) than resident cells. Serial section analysis, double labelled immunocytochemistry, and filipin treatment were used to demonstrate that caveolae can pinch off from the plasma membrane and can take part in endocytotic processes as alternative carriers in elicited macrophages.

Comparative study on the appearance of various bioactive peptides in foregut derivates during the ontogenesis.

Bioactive peptides have an important multifunctional role in the gastrointestinal tract. In the present study we have investigated the dynamism of the appearance of PACAP (pituitary adenylate cyclase activating polypeptide), VIP (vasoactive intestinal polypeptide), gastrin, and secretin immunoreactivities in human foregut derivates during the ontogenesis using an immunohistochemical approach. None of these peptides were observed in the foregut derivates of an 8-week-old embryo. VIP immunoreactive nerve fibers appeared by the 11th week in the smooth muscle layers of the stomach. No other peptide immunoreactivities were observed of this stage. In 18- and 20-week old fetuses PACAP, secretin, and gastrin immunoreactive cells appeared in the developing glands of the stomach. In the duodenum gastrin immunoreactivity was present in the Lieberkühn's glands and secretin immunoreactive cells were seen between the surface epithelial cells. In the pancreas secretin immunoreactivity was found in the Langerhans islets; however, PACAP immunreactivity was observed in the exocrine portion. The distribution of VIP fibers did not change during the fetal life and it was similar to the adult pattern. According to our results the appearance of PACAP, secretin, and gastrin in the developing glands suggests their role in the proliferation and differentiation of the epithelial derivates.

Pituitary adenylate cyclase activating polypeptide (PACAP) is present in human and cat gastric glands.

In the present work we have studied the occurrence of pituitary adenylate cyclase activating polypeptide (PACAP) in human and cat stomach mucosa using immunohistochemistry. As seen under a light microscope, there were many large rounded and ovoid cells that were PACAP immunopositive, mainly in the neck of the gastric glands of both species. The immunopositive material was predominant in the perinuclear area. The PACAP immunolabeling was specific because the preincubation of the antiserum with PACAP abolished the immunostaining. In human samples under electron microscope, the PACAP immunoreactive cells have shown the characteristics of parietal cells. In faintly stained cells, the localization of DAB reaction product was associated with the surface of the intracellular canaliculi. Cell labeling could not be observed besides parietal cells.

PACAP colocalizes with luteinizing and follicle-stimulating hormone immunoreactivities in the anterior lobe of the pituitary gland.

Pituitary adenylate cyclase activating polypeptide (PACAP) and its close relative vasoactive intestinal polypeptide (VIP) were demonstrated in the anterior pituitary gland. The cells which exhibited PACAP immunoreactivity were oval or round shaped. Their distribution was similar to that of gonadotropes but the number of PACAP immunoreactive cells was less. Double labeling revealed that PACAP immunoreactivity partially colocalized with luteinizing and follicle-stimulating hormone; however, colocalization with other pituitary hormone immunoreactivities was not demonstrated. Our results suggest an autocrine or paracrine role of PACAP in the regulation of pituitary functions.

[Pulmonary function tests in children and adolescents]. / Testes de função pulmonar em crianças e adolescentes.

OBJECTIVE: Evaluation of pulmonary function in pediatric patients. METHODS: Bibliographic review of the main techniques of pulmonary function testing. CONCLUSIONS: Pulmonary function tests are an important complement to the diagnosis of respiratory illnesses in children. They can also be of help in the follow-up and treatment of these illnesses, and since they can reproduce physiopathological phenomena under laboratory conditions they can also be of help in the study of several diseases. The basic spirometric tests most often used are the volume-time curve (V-T curve) and the flow-volume curve (F-V curve). Bronchial hyperreactivity can be assessed through provocation and effort tests.

Effect of the lack of light impulses on the hypothalamic PACAP and C-fos immunoreactivities in rats.

Pituitary adenylate cyclase activating polypeptide (PACAP) is a member of the secretin family. It is widely distributed in the central and peripheral nervous systems. The highest concentration of PACAP was found in the hypothalamus. In the present work it has been studied whether PACAP is involved in the mediation of photic stimuli to the anterior pituitary gland. We have examined the effect of the lack of light impulses on the hypothalamic PACAP and C-fos immunoreactivities. In adult rats 10 days after the removal of the eyes (surgical enucleation) and in those received monosodium glutamate treatment neonatally (chemical enucleation). The PACAP immunostaining enhanced in the hypothalamic magnocellular nuclei and in the extemal zone of the median eminence. C-fos immunoreactivity also enhanced in a few hypothalamic nuclei 2 hours after the surgical enucleation indicating that the lack of light impulses activated hypothalamic neurons which, in turn, might stimulate the release of PACAP into the portal circulation. It has been concluded that PACAP may be involved in photoendocrine regulations.

[Hypodense eosinophils in blood of pediatric patients with asthma]. / Eosinófilos hipodensos no sangue de pacientes pediátricos com asma.

OBJECTIVE: Determine the number of circulating hypodense eosinophils in the blood of atopic asthmatics. METHODS: The hypothesis was that hypodense eosinophils, the activated cells, were higher in asthmatics than in non-atopic children. Granulocytes were separated from mononuclear cells by Ficoll-hypaque. Gradients of density were generated through dilutions of Percoll for fractionations of cells. RESULTS: Asthmatics had blood eosinophilia (768 +/- 75/mcl) when compared to normal control (200 +/- 28/mcl), and higher mean hypodense eosinophils (39%) as compared to normal children (11.7%). CONCLUSION: Eosinophils accumulate in asthmatic, especially the low density activated variant, suggesting its active role in the allergic inflammatory process.

[Peripheral blood eosinophils and serum level of eosinophil cationic protein in asthmatics]. / Contagem de eosinófilos circulantes e níveis séricos de proteína catiônica eosinofílica em asmáticos.

OBJECTIVE: Determine the number of circulating eosinophils and serum eosinophil cationic protein levels in asthmatics, and the relationship between them. METHODS: We studied 26 atopic asthmatics aged 7-16 years divided in two groups according to severity of asthma. Eosinophil counts were obtained by multiplying leukocyte number by eosinophil percent. Serum eosinophil cationic protein levels were determined by fluoroenzymatic method, CAP System, Pharmacia Diagnostics. RESULTS: Asthmatics had eosinophil counts (810 +/- 103/mcl) significantly higher than non atopic children (158 +/- 30/mcl). Mean eosinophil cationic protein levels were higher in severe asthmatics (p < 0,001) and correlated with eosinophil counts. CONCLUSIONS: Serum eosinophil cationic protein levels could be useful in assessing the allergic inflammation and probably reflect the activation of eosinophils.

New aspects of the neuroendocrine role of PACAP.

The presence of PACAP was revealed in the anterior pituitary with RIA, HPLC, and with the demonstration of its mRNA. The level of PACAP mRNA in the anterior pituitary is the highest during the proestrous LH surge. In our immunohistochemical studies we were able to demonstrate PACAP immunoreactive cells in the anterior pituitary. The shape and the distribution of PACAP immunoreactive cells were very similar to that of the gonadotropes; however, the number of PACAP cells was less than that of LH cells. Additionally, another PACAP-positive cell population with small diameter appeared in the proestrous stage, during pregnancy and lactation. Double labeling revealed that the major part of large PACAP cells exhibited LH immunoreactivity and those with a small diameter contained PRL. It is not clear whether the pituitary- or the hypothalamic-born PACAP, or both, influence pituitary LH and PRL secretion. I.c.v. administration of PACAP just prior to the critical period in the proestrous stage inhibited the expected ovulation and blocked the proestrus LH and PRL surge, although i.v. administration of PACAP had no effect. PACAP antiserum did not interfere with ovulation when i.c.v. or i.v. injection was used. Our results support the view that PACAP has a role in the control of LH and PRL secretion during the estrous cycle, pregnancy, and lactation. The inhibitory effect of PACAP on ovulation is mediated through the hypothalamus.

PACAP participates in the regulation of the hormonal events preceeding the ovulation.

Pituitary adenylate cyclase activating polypeptide (PACAP) is a member of the secretin family. It was isolated and characterized in 1989. Its neuroendocrine role was demonstrated in vivo and in vitro systems. It seems that in vivo the effect of PACAP on the gonadotrop hormone secretion depends on the route of administration. It was reported that intravenous (i.v.) injection of PACAP elevated, while intra-cerebro-ventricular (i.c.v.) administration depressed plasma LH levels. In the present study it was demonstrated that PACAP, administered i.c.v. before the critical period of the proestrous stage, blocked the ovulation and prevented the proestrous LH surge in rats. The blocking effect of PACAP is not directly mediated by endogenous opioids because the antagonizing effect of Naloxone, an opioid receptor antagonist, was questionable. Under our experimental conditions we could not confirm the stimulating effect of i.v. administered PACAP.

Successful management of catastrophic gastrointestinal involvement in polyarteritis nodosa.

A patient with polyarteritis nodosa developed necrotizing enterocolitis, as indicated by pneumatosis intestinalis seen on computed tomographic scans of the abdomen. Despite immunosuppressive therapy and concomitant resolution of the intramural and portal venous gas and general clinical improvement, on 2 occasions (between 20 and 30 days later) the patient developed bowel infarctions and perforations that necessitated bowel resection. Leaks developed at anastomotic sites, but were not closed surgically. However, these sites and the lower quadrants of the abdomen were drained, and the patient was given total parenteral nutrition. Over a 2-month period the patient completely recovered from this nearly always fatal gastrointestinal complication of polyarteritis nodosa. The medical, surgical, and radiographic approach we used may be applicable to the management of similar cases in the future.

Fatal pneumonia in an adult due to respiratory syncytial virus.

Respiratory syncytial virus (RSV) infections in adults are generally mild, and no fatalities in uninstitutionalized adults have been reported. To our knowledge, we document herein the first case of fatal RSV pneumonia in a previously healthy elderly woman living at home, in whom complement fixation titers against RSV rose from less than 1:8 to reach 1:256 at death. Cytoplasmic inclusions characteristic of RSV were detected at autopsy.

Paraneoplastic retinopathy.

Paraneoplastic retinopathy appears to represent a nonmetastatic remote effect of carcinoma and is characterized by rapid visual deterioration accompanied by narrow arterioles seen on ophthalmoscopic examination and an extinguished electroretinogram. This syndrome has been described in postmenopausal women, and it is possible that it represents an autoimmune disorder. Corticosteroid therapy may arrest the visual deterioration.

Apolipoprotein B subspecies in chylomicrons isolated from a patient with chyluria.

The diagnosis and the clinical course of a 17-year-old white male with chyluria are reported. Cloudy, milky urine appeared spontaneously, in the absence of edema or any signs or symptoms of parasitic infection. Pedal lymphangiography demonstrated the presence of a lymphatic renal fistula, and digital subtraction angiography showed aneurysmal dilatation of the aorta at the level of the renal arteries. This case provided an opportunity to ascertain which of the forms of apolipoprotein B were present in lymph chylomicrons. Apolipoprotein B is needed for chylomicron secretion. It exists in several forms--B-100, B-74, B-48, and B-26. After a meal consisting of fat, chylomicrons in which apolipoprotein B-48 was virtually the only apolipoprotein B present appeared in the urine, while apolipoprotein B-100 was the only apolipoprotein B present in the plasma very low-density lipoproteins. Chyluria disappeared two weeks after institution of a low-fat diet. This case illustrates an interesting, rare cause of chyluria. Because of the presence of chyluria, it was also demonstrated that chylomicrons in which apolipoprotein B-48 is virtually the only apolipoprotein B present are a physiologically normal product of the intestine.