Purification of rabbit, rat and mouse protein C with the use of monoclonal antibody to human protein C, PC01

Ca(++)-dependent monoclonal antibody specific to gamma-carboxyglutamic acid (Gla) domain of protein C was produced. It did not cross-react to the other vitamin K-dependent plasma proteins but to protein C of the other species. Using this monoclonal antibody, PC01, rabbit (170 micrograms), rat (60 micrograms) and mouse (40 micrograms) protein Cs were isolated from 100 ml of their plasma by affinity chromatography. All of these protein Cs were two chain form linked by disulfide bond as well as human protein C and activated by thrombin-thrombomodulin complex. Rat and mouse protein Cs showed similar characters to human protein C. On the other hand rabbit protein C had different M(r) of heavy and light chains and showed lower anticoagulant activity compared with human protein C.     

Developmental patterns of intermediate filament gene expression in the normal hamster brain

We have examined the patterns of expression of the major intermediate filament (IF) protein mRNAs during development of the hamster brain. Quantitative northern blotting was used to examine changes in the levels of mRNAs for the low, middle and high molecular weight neurofilament proteins (NF-L, NF-M, NF-H) as well as peripherin, vimentin and glial fibrillary acidic protein (GFAP). Total RNA was isolated from hamster brains at embryonic (E) days 12 and 14 and postnatal (P) days 1, 3, 5, 7, 9, 11, 13, 15, 20, 28 and 60-90 (adult), and probed with specific IF cDNAs. Northern blotting revealed that NF-L and NF-M mRNAs were present at very low levels in embryonic brain and that significant expression of these genes only occurred postnatally when the levels increased dramatically until P28 and then declined again in the adult. Increases in NF-H mRNA levels were somewhat delayed relative to those of NF-L and NF-M. NF-H mRNA was not seen at embryonic stages and was expressed at very low levels prior to P9; after that time the levels increased rapidly until P28 and then declined in the adult. Two of the type III IF genes, peripherin and vimentin, followed a pattern of expression opposite that of the NF genes. Both peripherin and vimentin mRNAs were present in embryonic brain and were expressed at higher levels during early postnatal stages than at later times. The magnitude and rate of reduction in vimentin gene expression in the postnatal interval was much greater than that of peripherin. GFAP mRNA levels were extremely low prior to P9 after which a robust increase occurred, followed by a decline in the adult. We discuss the implication of the dramatic changes in IF isotype expression in brain to the pathways of both neuronal and glial development in vivo.     

Distribution of cytochrome oxidase and parvalbumin in the primary visual cortex of the adult and neonate monkey,Callithrix jacchus

The anatomical distributions of the mitochondrial enzyme cytochrome oxidase (CO) and of the calcium binding protein parvalbumin (PV) were studied in the striate cortex of adult and neonate New World monkeys (Callithrix jacchus). In the adult marmoset, both proteins were found in laminar arrangements similar to those described for the macaque monkey, with prominent bands of PV-like immunoreactive (PV-LI) puncta in layers IV and IIIb, and fairly evenly distributed PV-LI nonpyramidal neurons. Furthermore, the pattern of CO activity in area 17 of the neonate marmoset was almost identical to the CO pattern described in neonate macaque and squirrel monkeys. It came, therefore, as a surprise to find that the adult pattern of PV-like immunoreactivity (PV-LI) in the marmoset striate cortex arises from a neonatal pattern strikingly different from that seen in any developmental stage of the macaque, or in any other mammal studied so far. In the deep layers IV through VI of the neonate marmoset, a large number of PV-LI neurons was stained in bandlike patterns, their number in layers IV and V exceeding the number of PV-LI neurons present in these layers of the adult marmoset area 17. Staining of layers IV and VI was restricted to area 17 and involved nonpyramidal cells and their exceeding the number of PV-LI neurons present in these layers of the adult marmoset area 17. Staining of layers IV and VI was restricted to area 17 and involved nonpyramidal cells and their processes. The stained band of layer V, in contrast, continued throughout most of the neocortex. In area 17, an estimated 10 to 20% of the stained cells in layer V exhibited pyramidal shapes. The findings show that the expression of PV by visual cortical cells occurs before birth and suggest that the comparatively early onset of PV expression is not dependent on the onset of textured vision. The exuberant number of stained cells in some layers, and particularly the staining of pyramidal cells, in the neonate marmoset, suggest that a considerable number of cells possesses the stainability for PV-LI only transiently, i.e., in the marmoset, these cells have a specific demand for parvalbumin during this phase of their development. © 1994 Wiley-Liss, Inc.     

Atypical Allergic Colitis in Preterm Infants

ABSTRACT. Two atypical cases of colitis due to cow's milk protein intolerance (CMPI) are reported, affecting preterm infants. One developed a toxic dilatation of the colon and responded well to a casein hydrolysate based feed. The second presented insidiously and failed to tolerate a casein hydrolysate, but responded well to a chicken-based modular feed.     

Chronic ethanol intake may delay the onset of gossypol-induced infertility in the male rat

Summary. Parameters were obtained from the reproductive organs of ethanol-fed, gossypol-treated Sprague Dawley rats. The experimental animals were fed either on a normal (15.17%) or low protein (8.00%) diet. Measurements included reproductive organ weights, seminal characteristics, serum concentration of testosterone and histological, stereological and histomorphometric evaluation of the testis. The testis size, length and diameter of the seminiferous tubule had the least values in the protein-malnourished, gossy-pol-treated rats (3.01±0.26 g, 0.56 ± 0.03 m, 281.34±11.30 μn), in comparison to corresponding animals which had received ethanol simultaneously with gossypol (3.40 ± 0.25, 0.71±0.06m, 314.42 ± 11.61 μn). As gossypol and ethanol are both associated with reduced reproductive capacity, this unexpected but interesting finding lends support to the hypothesis that either a normal dietary protein or ethanol consumption may modify the action of gossypol on body tissues, including the testis. This effect, presumably mediated through changes caused to the bioavailability of gossypol, modifies its antifertility activity. The present observation further highlights the need to consider the concurrent administration of other drugs, such as alcohol, and the nutritional status in the evaluation of gossypol for various potential uses.     

Nuclear p53 Overexpression in Bladder, Prostate, and Renal Carcinomas

Background :
The aim of this study was to examine nuclear p53 overexpression in transitional cell carcinoma of the bladder, adenocarcinoma of the prostate, and renal cell carcinoma.
Methods :
Forty-four pathologic specimens from 39 bladder cancer patients, 41 prostatic adenocarcinoma, and 39 renal cell carcinoma specimens were analyzed immunohistochemically with D07 monoclonal antibody to detect the expression of the mutant p53 gene. Overexpression was said to occur when the number of positively-stained tumor nuclei were≥ 10% in each specimen. p53 overexpression was correlated with the clinical and histopathological features of these cancers.
Results :
Nuclear p53 overexpression occurred in 18.2% of transitional cell bladder cancer specimens, 12.2% of prostate cancer specimens, and 17.9% of renal cell cancer specimens. Statistical analyses showed that grade, vascular invasion, and necrosis in bladder cancer, a high Gleason score in prostate cancer, and the 1-year mortality rate in renal cancer were significantly related with p53 nuclear overexpression (P<0.05).
Conclusion :
Using the D07 monoclonal antibody, nuclear p53 overexpression is relatively uncommon in urologic malignancies, and moderately correlates with several histopathological and clinical features of urologic malignancies.     

Gastro-oesophageal reflux disease: Medical or surgical treatment? Report of an interactive workshop

Gastroesophageal reflux disease (GERD) is the most common disease of the upper gastrointestinal tract. With the introduction of proton pump inhibitors medical treatment of GERD has been significantly improved. However, the development of laparoscopic antireflux surgery resulted in an increasing interest of surgeons in this disease. An interactive meeting was organized in order to develop an agreement between gastoenterologists and surgeons regarding therapeutic decisions and this is the main topic of this paper.