Distribution of N-cadherin and NCAM in neurons and endocrine cells of the human embryonic and fetal gastroenteropancreatic system

When the thoracic spinal cord of the North American opossum is transected early in development, supraspinal axons grow through the lesion. In the experiments reported here, we asked whether regeneration of cut axons contributes to such growth. Fast Blue (FB) was injected into the lumbar cord on postnatal day (PD)5, 8, 15, or 20. Five days later, FB was removed by gentle suction, and the spinal cord was transected at thoracic levels. Fourteen days later, rhodamine B dextran was injected between the site of the FB injection and the lesion. The pups were maintained for an additional 7-10 days before killing and perfusion. We assumed that supraspinal neurons that contained FB survived axotomy and those that contained both FB and rhodamine B dextran supported regenerating axons. In the PD5 group (lesioned at PD10), regenerative growth was documented for axons originating in all of the supraspinal nuclei that innervate the lumbar cord by PD10. When the injections were made at the later ages, however, neurons that supported regenerative growth were fewer in number and regionally restricted. In some cases, they were limited primarily to the red nucleus, the medullary raphe, and the adjacent reticular formation. Our results show that regeneration of cut axons contributes to growth of supraspinal axons through the lesion after transection of the thoracic cord in developing opossums and that the critical period for regenerative growth is not the same for all axons.     

Quality of life in patients with endocrine tumors of the gastrointestinal tract: patient and staff perceptions

Myo-inositol (mI) as a precursor in the phosphatidylinositol second messenger system has been reported to be reduced in depression. By means of proton-magnetic resonance spectroscopy (1H-MRS) the mI levels in the frontal brain were investigated in vivo in the present study. Twenty-two patients (mean age: 42.8 +/- 10.7 years) with depressive episodes according to ICD 10 (HAMD score > 17) were compared to 22 healthy subjects (28.0 +/- 5.3 years). Two voxels (30 x 20 x 20 mm3) in the frontal lobes were examined in a Siemens Magnetom SP 4000 at 1.5 T (STEAM sequence: TR = 3500 ms, TE = 55 ms). With the total creatine (Cr) as an internal standard, mI/Cr ratios were calculated to follow the mI levels. In the left frontal lobe, mI/Cr was 0.43 +/- 0.06 in depressive patients and 0.46 +/- 0.07 in healthy subjects; concerning the right frontal lobe, mI/Cr was 0.46 +/- 0.08 and 0.48 +/- 0.06, respectively. There were neither significant differences between the two groups nor between the hemispheres. Since there was a significant positive correlation (R = 0.6) between the age and the mI/Cr in the right frontal lobe of depressed patients, age matched pairs analysis was performed (n = 2 x 10, in each group: nine females, one male, < 40 years). In the right frontal lobe, the patients' mI/Cr of 0.40 +/- 0.05 was now significantly lower than the controls' mI/Cr of 0.45 +/- 0.06. However, most of the patients were on antidepressive medication. Interestingly, it was exactly this group of patients which showed significantly lower mI levels. We regard our investigation as a pilot study which suggests an influence of age and antidepressants on mI levels and should be taken into consideration in further investigations in depressive patients.     

Immunocytochemical characterisation of rabbit and mouse embryonic fibroblasts

By using indirect immunofluorescence we demonstrated the localisation of extracellular matrix (ECM) proteins (laminin--LAM, collagen IV--COL IV, fibronectin--FN) and the basic fibroblast growth factor (bFGF) in rabbit and mouse primary embryonic fibroblasts (PEF). Proliferating mitotically arrested PEF (by mitomycin C) were compared in both species. The stability of protein expression was ascertained during the first five successive passages. In addition, STO cells (i.e. permanent line of irradiated mouse fibroblasts) were similarly analysed. Rabbit PEF showed very high extracellular staining for FN and a negligible cytoplasmic positivity for LAM and COL IV. A totally reversed staining pattern for ECM proteins was found in mouse PEF. A dense cytoplasmic granulation (concentrated around the nucleus) was revealed for LAM and COL IV and almost no reaction for FN. The staining patterns were very stable at the culture conditions we applied. They were maintained during the first five successive passages in proliferating as well as non-proliferating mouse and rabbit PEF and were independent of cell concentration (individually dispersed cells versus cells in a confluent layer). STO cells showed the same staining for ECM proteins as the mouse PEF, thus confirming their origin from the same animal species. Light granular staining for bFGF was found in the cytoplasm of proliferating and mitotically arrested rabbit and mouse PEF and STO cells. The differences in expression of ECM proteins between the rabbit and mouse PEF, as well as the synthesis of bFGF, should be taken into consideration when these cells are used in vitro as a feeder layer for various cells (e.g. embryonic stem cells).     

Clear cell tumors of the alimentary tract and abdominal cavity

In a population of male workers in two ship maintenance companies (n = 32), a workplace survey was conducted in order to quantify their physical load. Postural load was measured by using the Ovako Working posture Analyzing System. During 7480 observations, working postures, exertion of force and working activities were recorded. Awkward postures of the back occurred in 38% of the worktime, stress on the neck/shoulder region due to one or both arms above shoulder level was present in 25% of the worktime. Forceful exertions during lifting, pushing and pulling activities sometimes exceeded published guidelines for manual material handling. Determinants of physical load could be identified and a hazard evaluation procedure was designed by applying rating schemes to weight various patterns of physical load. Ship maintenance work compared well with other strenuous occupations. Considering the high prevalence of back pain (80%) and neck/shoulder pain (60%), as well as the results of the observation method ergonomic improvements are warranted. Physical load can be reduced by several technical adaptations and applications, and by enlarging task rotation.     

A simple method for purification of adult pig pancreatic endocrine cells

Adult pig pancreatic endocrine cells were harvested by autodigestion without added enzymes. The isolated, crude cells were purified by mono-poly resolving medium (MPRM). The purity of the harvested cells was determined by dithizone staining and the number of pancreatic endocrine cells was counted. A large number of the cells were stained red with dithizone and showed a high viability and a good insulin secretory response to glucose stimulation. The average number of cells purified by MPRM was 3.40 +/- 1.32 x 10(5) cells/g pancreas and the number of dithizone-stained cells was 2.81 +/- 1.09 x 10(5) cells/g pancreas. The insulin secretion from the pancreatic endocrine cells was maintained throughout a 40-day observation period and high glucose stimulation induced an increase in insulin secretion from the cultured cells. In the cells purified by MPRM, light and electron microscopic studies showed the cells to be typical pancreatic endocrine cells. The present purification method using MPRM allowed us quickly to obtain a large amount of adult pig pancreatic endocrine cells from the unpurified preparations. It is thought to be useful for transplantation and biochemical or biological studies of adult pig pancreatic endocrine cells.     

Expression of nerve growth factor and its high-affinity receptor Trk-A in the rat pancreas during embryonic and fetal life

We recently reported that nitric oxide (NO), which is produced by chondrocytes treated with interleukin-1beta (IL-1), releases basic fibroblast growth factor (bFGF) stored in the matrix of articular chondrocytes. To clarify the mechanism of the IL-1-induced bFGF release, we investigated the production and gene expression of bFGF, matrix metalloproteinases (MMPs), syndecan 3, and inducible NO synthase (iNOS) by IL-1-treated rabbit articular chondrocytes. IL-1 stimulated not only the release of bFGF but also the production of it. Gelatin and casein zymography revealed that IL-1 stimulated the production of not only MMP-9 but also MMP-3. The increase in the production of these MMPs preceded the IL-1-stimulated bFGF release. An MMP inhibitor partially suppressed the release of bFGF, indicating that matrix degradation is at least partially involved in the IL-1-stimulated bFGF release even if increased production of bFGF is related to the release. IL-1 sequentially stimulated mRNA expression of iNOS, membrane type 1-MMP, MMP-9 and -3, and bFGF, in that order. NG-Monomethyl-L-arginine, an inhibitor of NO production, inhibited gene expression of MMP-9 and bFGF. These findings suggest that elevation of the NO level via iNOS mRNA expression stimulated by IL-1 mediates gene expression and production of MMPs and bFGF, resulting in the release of bFGF, and also reveal molecular mechanisms implicating the degradation of articular cartilage followed by angiogenesis in the synovium in arthritic joints.     

Immunocytochemical localization of desmin in human fetal skeletal muscle

INTRODUCTION: A comparative evaluation of four air samplers was performed using bioaerosol collection in the outdoor environment. METHODS: Test samplers used included a Rotorod, a Kramer-Collins suction trap, an all-glass impinger (AGI-30), and a high-volume cyclonic liquid impinger (SpinCon). All samples were analyzed microscopically for spores and pollen. The two collectors providing a liquid sample (AGI-30 and SpinCon) also were analyzed for specific allergen content by enzyme-linked immunoassay. RESULTS: The SpinCon collected a larger number of spores than the other devices. The number of spores collected by this unit per volume of air sampled was comparable to the AGI-30. The Rotorod and Kramer-Collins collected a lower number of spores per unit of air but collected a larger number of pollen grains per volume sampled. Alternaria allergens Alt a I and GP70 were collected by both liquid impingers; however, the SpinCon collected more Alt a I and the AGI-30 collected more GP70. CONCLUSIONS: The SpinCon is a device that is capable of efficiently sampling a high volume of air and concentrating it in a form that can be analyzed for the presence of spores and fungal allergens. It is less useful for collecting intact pollen grains. Pollen allergen quantitation has not yet been performed on the SpinCon effluent.     

A comparative study of spirochaetes from the porcine alimentary tract

Strains of Treponema hyodysenteriae capable of inducing swine dysentery in specific pathogen-free pigs were compared with other spirochaetes from the porcine alimentary tract by biochemical and serological tests and by electrophoresis of their proteins. Carbohydrate fermentation and esculin hydrolysis were similar in all the spirochaetes. Indole was produced by T. hyodysenteriae and by some of the other spirochaetes. Analysis of the fatty acids produced from glucose showed a difference between T. hyodysenteriae and other spirochaetes only in the amount of n-butyric acid produced. The indirect fluorescent antibody test showed extensive cross-reactions between all the spirochaetes unless antisera were first absorbed. A microtitre agglutination test and a growth-inhibition test were both more specific; strains of T. hyodysenteriae could be distinguished from the other spirochaetes using unabsorbed sera. Both tests revealed some antigenic heterogeneity among strains of T, hyodysenteriae. The cell proteins of a single strain of T. hyodysenteriae gave an electrophoretic pattern distinct from those of the other spirochaetes. Two of the six spirochaetes not associated with swine dysentery, PWS/B and PWS/C, were indistinguishable serologically and electrophoretically. The other four strains were serologically distinct from one another and from PWS/B and PWS/C. Only two of these spirochaetes were examined electrophoretically, but each gave a different pattern from PWS/B and PWS/C. The diversity observed among spirochaetes not associated with swine dysentery indicates that their suggested inclusion in a single species, T. innocens, may prove to be unjustified.     

The ontogeny of key endoplasmic reticulum proteins in human embryonic and fetal red blood cells

Recently, using immunohistochemical methods, we surprisingly found that endoplasmic reticulum glucose-6-phosphatase is present in human embryonic and fetal red blood cells (RBCs) but not in adult RBCs. The fact that an endoplasmic reticulum enzyme, whose major site of expression in adults is the liver, is present in human embryonic and fetal RBCs, particularly nucleated cells, indicated that it would be sensible to determine whether these cells also contain other endoplasmic reticulum enzyme systems normally found in adult liver. Therefore, we have studied the expression of other endoplasmic reticulum proteins and found that human embryonic and fetal RBC precursors contain other protein components of the glucose-6-phosphatase system, ie, the phosphate and glucose transport proteins as well as other enzymes (eg, uridine diphosphate-glucuronosyltransferases, cytochrome P450 isozymes, nicotinamide adenine dinucleotide phosphate cytochrome P450 oxidoreductase, and prostaglandin H synthase). In addition, we also found the predominantly cytosolic markers 15-hydroxyprostaglandin dehydrogenase, prostaglandins PGE2 and 13,14-dihydro-15-keto-PGE2. The expression of key enzymes that control glucose production, detoxification of endobiotics and xenobiotics, and the regulation of prostaglandin levels in embryonic and early fetal RBCs means that these cells may have an important role in protecting the developing conceptus before it establishes an efficient circulation and before all tissues fully express their normal complement of these enzymes.     

A modified technique for the reconstruction of the alimentary tract after pancreatoduodenectomy

The high death rate for those patients undergoing pancreatoduodenectomy, which is a result of leakage of biliary and pancreatic anastomoses in most instances, justifies the study of this technique for rebuilding the alimentary tract. The use of two separate intestinal loops for the biliary and pancreatic anastomoses is the basis of this method. This technique reduces the morbidity and the death rates which are the result of leakage in either anastomosis because it separates biliary and pancreatic fistulas.     

Autonomous cell death of mouse male germ cells during fetal and postnatal period

OBJECTIVE: To determine the utility of CT-determined main pulmonary artery diameter (MPAD) for predicting pulmonary hypertension (PH) in patients with parenchymal lung disease. DESIGN: Retrospective review of right-heart hemodynamic data and chest CT scans in 45 patients. SETTING: Tertiary-referral teaching hospital and VA hospital. PATIENTS: Between October 1990 and December 1995, 36 patients referred for evaluation of parenchymal lung disease or possible pulmonary vascular disease were found to have PH, as defined by mean pulmonary artery pressure (mPAP) > or =20 mm Hg. Nine control patients (mPAP <20 mm Hg) were also identified (4 from hospital records search, 5 after evaluation for possible PH). RESULTS: CT-determined MPAD was 35+/-6 mm in patients with PH and 27+/-2 mm in control subjects. In our group of patients, MPAD > or =29 mm had a sensitivity of 87%, specificity of 89%, positive predictive value (PPV) of 0.97, and positive likelihood ratio (LR) of 7.91 for predicting PH; in the subgroup of patients with parenchymal lung disease (n=28, PH and control subjects), MPAD > or =29 mm had a sensitivity of 84%, specificity of 75%, PPV of 0.95, and positive LR of 3.36 for predicting PH. The most specific findings for the presence of PH were both MPAD > or =29 mm and segmental artery-to-bronchus ratio > 1:1 in three or four lobes (specificity, 100%). There was no linear correlation between the degree of PH and MPAD (r=0.124). CONCLUSIONS: CT-determined MPAD has excellent diagnostic value for detection of PH in patients with advanced lung disease. Therefore, standard chest CT scans can be used to screen for PH as a cause of exertional limitation in patients with parenchymal lung disease. Because CT is commonly used to evaluate parenchymal lung disease, this information is readily available.     

Immunocytochemical localization of the NPY/PYY Y1 receptor in enteric neurons, endothelial cells, and endocrine-like cells of the rat intestinal tract

Neuropeptide Y (NPY) and peptide YY (PYY) are structurally related peptides that are considered to mediate inhibitory actions on gastrointestinal motility, secretion, and blood flow. Several receptor subtypes for these peptides have been identified and the Y1, Y2, Y4/PP1, Y5, and Y5/PP2/Y2b receptors have been cloned. In this article we report the immunocytochemical localization of the Y1 receptor to myenteric and submucosal nerve cell bodies, endothelial cells, and scattered endocrine-like cells of rat intestinal tract. Moreover, double immunofluorescence demonstrates that subpopulations of the Y1 receptor-positive nerve cell bodies are immunopositive for NPY, vasoactive intestinal polypeptide, and nitric oxide synthase. In part, such co-localizations were made possible by use of peroxidase-mediated deposition of tyramide, which permitted use of antisera derived from the same species. Our observations suggest the existence of multiple neuronal, endothelial, and endocrine target sites for NPY and PYY and that some of the actions of these regulatory peptides can be mediated by vasoactive intestinal peptide and nitric oxide synthase.     

Immunocytochemical localisation of parietal cells and G cells in the developing human stomach

Previous studies on the distribution of parietal cells and G cells in normal adult stomachs have shown that in about 20% of specimens parietal cells extended to the pylorus. This study aimed to examine the distribution of parietal cells and G cells in the body and antrum of the developing human stomach in relation to anatomical landmarks, using histological and immunocytochemical methods. In all 15 fetal stomachs examined, parietal cells extended to the pylorus and expressed intrinsic factor and hydrogen-potassium-ATPase activity from week 13 of gestation. By contrast, in only one of the five infant stomachs did parietal cells extend to the pylorus: this is identical to the distribution in the adult. G cells developed in the antrum from 18 weeks' gestation and their distribution did not differ between the fetal and infant stomachs. These findings indicate that parietal cells disappear from the antrum of the stomach in the third trimester of pregnancy, but this process fails to occur in approximately 20% of the population.     

One layer continuous anastomosis of the alimentary tract with absorbable polydioxanone suture

A prospective study using absorbable Polydioxanone (PDS) suture material in a one layer continuous technique for gastrointestinal anastomosis was conducted. There were 40 anastomoses constructed in 39 children and 61 anastomoses in 49 adults which were classified as "non-complicated anastomoses" without any clinical evidence of leakage or any other complications attributable to the anastomotic technique or to the suture materials. There were other 20 anastomoses created in 20 patients with malnutrition or those receiving chemotherapy and/or radiation or where there was tension at the anastomosis which were classified as "complicated anastomosis". Anastomotic leakage was observed in one patient (5%). The starvation period was 3.16 +/- 0.9 days compared to 3.46 +/- 1.0 days in the two layer technique "control" group. The rate of complications and the function of the GI tract in both non-complicated and complicated anastomoses after one layer continuous Polydioxanone anastomotic completion was not significantly different from those using conventional two layer anastomosis. The technique for one layer continuous suture is simple, easy and takes less time than the conventional method. This technique also theoretically provides better postoperative condition in which bowel anatomy and physiology can return to normal earlier, causing minimal tissue trauma, and less narrowing of the lumen although the evidence cannot be supported by this study. The Polydioxanone suture material is biodegraded by specific time, and hence allows normal growth of the anastomosed bowel; it is therefore suitable for both children and adults.     

Evaluation of bone mass density after extensive resections within the alimentary tract

The authors have examined a group of 30 patients at 24-36 months after extensive resections within the alimentary tract. The bone density assessment has been done. The results were compared to those obtained from another group of 30 patients awaiting sinister operations. Bone loss was found in all postoperative patients. However, it was statistically significant only in patients after pancreatoduodenectomy.     

Immunocytochemical localization of progesterone receptor in the reproductive tract of adult female rats

The distribution of the progesterone receptor (PR) was investigated immunocytochemically in female reproductive tracts of rats during the estrous cycle and early pregnancy through use of an anti-PR monoclonal antibody. PR was localized predominantly in the nuclei of epithelial, stromal, and muscle cells in the uterus and vagina during the estrous cycle. In the uterus, the nuclei of epithelial cells were stained intensively at diestrus, while the PR staining of the stromal cells was more intense at proestrus than at any other stage of the cycle. PR expression during the cycle in muscle cells of the myometrium was similar to that in the endometrial stromal cells. In the vagina, however, PR expression during the cycle was approximately the same among epithelial, stromal, and muscle cells, the nuclei of which were stained deeply at proestrus. Ovariectomy at various stages of the cycle altered the PR expression appearing in the uterus and vagina during the cycle. In ovariectomized rats, estrogen increased the PR immunoreaction of various types of cells examined in the uterus and vagina except for the uterine epithelial cells. The reaction of these uterine epithelial cells was decreased by estrogen but was increased by progesterone given after estrogen; however, progesterone given alone reduced the reaction. In the epithelial and stromal cells of the uterus, intensity of the staining was increased after mating, reaching maximum on Day 3 of pregnancy, and then decreased on Day 4 (day of implantation), while in epithelial and stromal cells of the vagina the staining remained weak during early pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)     

Histological and ultrastructural observations on the development of the lung of the fetal pig

Lungs of fetal pigs having gestational ages ranging from 80 to 115 days were examined histologically and by electron microscopy. At 80 days bronchial epithelium was ciliated but bronchiolar cells were not and bronchial mucosal glands were absent. Peripheral regions consisted predominantly of mesenchymal tissue with glandular alveoli. 92 days marked the transition from the immature to the more mature lung type. Bronchial glands appeared and began to grow from the epithelium into the lamina propria, bronchiolar epithelial cells acquired cilia, and alveoli were becoming irregular in shape and had thinner interalveolar septa. Close contact between capillaries and alveolar epithelium established the blood-air barrier at many points. Differentiation of alveolar epithelium into types I and II pneumonocytes occurred at this stage and lamellated osmiophilic inclusion bodies were present in type II cells for the first time. The number of lamellated bodies increased progressively to term at 115 days.