Hyperbaric oxygen induces endogenous neural stem cells to proliferate and differentiate in hypoxic-ischemic brain damage in neonatal rats.

BACKGROUND AND PURPOSE: Studies suggest that after brain injury, hyperbaric oxygen (HBO2) is neuroprotective by stimulating cell proliferation. We examine whether HBO2 promotes neural stem cells (NSC) to proliferate and differentiate in neonatal hypoxic-ischemic (HI) rats. METHODS: Seven-day-old rat pups were subjected to unilateral carotid artery ligation followed by 2 hours of hypoxia (8% O2). HBO2 was administered (2 ATA (atmospheres absolutes), once daily for 7 days) within 3 hours after HI. The proliferating neural stem cells in the subventricular zone (SVZ) and dentate gyrus (DG) were dynamically examined by 5-bromo-2-deoxyuridine (BrdU)/nestin immunofluorescence. Nestin protein was detected by western blot analysis at various time points (from 6 hours to 14 days) after HI. The migrating NSC were examined by BrdU/doublecortin (DCX) immunofluorescence 7 and 14 days after HI. The phenotype of the newborn cells was identified by BrdU/beta-tubulin, BrdU/ glial fibrillary acidic protein (GFAP) and BrdU/O4 (oligodendrocyte marker) immunofluorescence. Myelin basic protein (MBP) was examined by immunohistochemistry and pathological changes of the brain tissue were detected 28 days after HI. RESULTS: In neonatal HI rats treated with HBO2, the proliferation of endogenous NSC was observed in the SVZ and DG. Cell numbers peaked 7 days after HI and proliferating NSC migrated to the cerebral cortex at 14 d after HI. Twenty-eight days after HI, an increase in newly generated neurons, oligodendrocytes and MBP was observed in the HBO2 group compared to the untreated and HI-treated rats. CONCLUSIONS: This study suggests that HBO2 treatment may promote neurogenesis of the endogenous NSC in neonatal HI rats, contributing to repair of the injured brain.

Immune response, ciprofloxacin activity, and gender differences after human experimental challenge by two strains of enterotoxigenic Escherichia coli.

In order to test vaccines against enterotoxigenic Escherichia coli (ETEC)-induced diarrhea, challenge models are needed. In this study we compared clinical and immunological responses after North American volunteers were orally challenged by two ETEC strains. Groups of approximately eight volunteers received 10(9) or 10(10) CFU of E. coli B7A (LT+ ST+ CS6+) or 10(8) or 10(9) CFU of E. coli H10407 (LT+ ST+ CFA/I+). About 75% of the volunteers developed diarrhea after challenge with 10(10) CFU B7A or either dose of H10407. B7A had a shorter incubation period than H10407 (P = 0.001) and caused milder illness; the mean diarrheal output after H10407 challenge was nearly twice that after B7A challenge (P = 0.01). Females had more abdominal complaints, and males had a higher incidence of fever. Ciprofloxacin generally diminished or stopped symptoms and shedding by the second day of antibiotic treatment, but four subjects shed for one to four additional days. The immune responses to colonization factors CS6 and colonization factor antigen I (CFA/I) and to heat-labile toxin (LT) were measured. The responses to CFA/I were the most robust responses; all volunteers who received H10407 had serum immunoglobulin A (IgA) and IgG responses, and all but one volunteer had antibody-secreting cell (ASC) responses. One-half the volunteers who received B7A had an ASC response to CS6, and about one-third had serum IgA or IgG responses. Despite the differences in clinical illness and immune responses to colonization factors, the immune responses to LT were similar in all groups and were intermediate between the CFA/I and CS6 responses. These results provide standards for immune responses after ETEC vaccination.

Effect of phosphorus limitation on microbial floc structure and gene expression in activated sludge.

The effect of limiting phosphorus (P) in activated sludge was investigated in laboratory-scale sequencing batch reactors (SBRs). Correlative microscopy revealed that P-limitation (COD:N:P = 100:5:0.05) leads to morphological changes in floc structure and the composition of extracellular polymeric substances (EPS). This was found to be accompanied by expression of quorum-sensing in an acyl homoserine lactone bioassay. Differential gene expression in relation to P-limitation was examined in a global profile using the Affymetrix Escherichia coli antisense genomic microarray. Three separate experiments were conducted where the impact of P-limitation was examined under batch conditions and in SBRs at stable operating conditions and within 3-7 days following a down-shift in P. Significant changes in open reading frames (ORF) and intergenic regions based on the E. coli microarray were observed. Several genes associated with cell structure, including slt, wbbH, fimH, amB, rfaJ and slp were found to be expressed. Quorum regulated genes were also found to be expressed including psiF which is known to be induced by P-starvation (92% confidence level; 1.45 log ratio).

Glutamine utilization in activated lymphocytes from rats receiving endotoxin.

BACKGROUND: A beneficial effect of supplemental glutamine for lymphocyte function in patients under metabolic stress has been suggested. Nevertheless, it is not clear how glutamine is used by lymphocytes when under stress. This time course study investigated the effect of endotoxin-induced stress on in vitro glutamine utilization and glutamine-dependent proliferation of activated lymphocytes. METHODS: Metabolic stress was modeled by intraperitoneal (ip) administration of endotoxin (5 mg/kg body wt) to rats. Control animals were injected with sterile saline. Cervical lymph node lymphocytes collected from animals 6, 12, 24, and 48 h following injection were activated with concanavalin A. Proliferation of these activated lymphocytes in the presence of 0.1-2 mM glutamine was determined. The glutamine utilization rate and glutaminase activity in the activated lymphocytes were also determined. RESULTS: The proliferation rate of lymphocytes was not affected by ip administration of endotoxin 6 h following the insult, however, 12, 24, and 48 h following the insult, the maximal response was suppressed (P < 0.05). In addition, at 12, 24, and 48 h, the concentration of glutamine for the maximal response of lymphocytes was lower than that for the control group (P < 0.05). Throughout the investigation period, both the glutamine utilization rate and glutaminase activity in the activated lymphocytes were decreased time-dependently. CONCLUSION: The present study demonstrates that glutamine utilization by lymphocytes under a mitogenic challenge in vitro is significantly decreased in the late period after endotoxin injection. This is at least partly due to decreased glutaminase activity and is associated with decreased proliferation rate of mitogen-activated lymphocytes.

Molecular cloning, characterization and tissue expression of prophenoloxidase cDNA from the mosquito Armigeres subalbatus inoculated with Dirofilaria immitis microfilariae.

A cDNA encoding mosquito Armigeres subalbatus prophenol oxidase (As-pro-PO) was obtained by rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) after Dirofilaria immitis inoculation. The 2205 bp As-pro-PO cDNA contains a 32 bp 5'-noncoding region, a 2055 bp open reading frame (685 amino acids), and a 118 bp 3'-noncoding region. Hydrophobic signal peptide for the endoplasmic reticulum targeting is not found in the NH2-terminal region. Two potential copper-binding domains, amino acids 197-245 and 345-412, are highly homologous to those of the other insect pro-POs. A 2.2 kb As-pro-PO transcript was identified by Northern blot analysis using D. immitis microfilariae-inoculated A. subalbatus. Both in situ hybridization and Northern blot analysis demonstrated that As-pro-PO mRNA was synthesized in mosquito haemocytes but not in other tissues, i.e. fat bodies, midguts and ovaries, etc.

From san gu liu po to 'caring scholar': the Chinese nurse in perspective.

This study presents a re-evaluation of the origins of Chinese nursing from the earliest traditional carers to the mature, professionalized institution of the republican period. The influence of traditional cultural values on the nature of the modern profession is discussed and its emergence considered in historical context with particular attention to the role of missionaries, philanthropic institutions and other aspects of foreign intervention, both European and Japanese. The Chinese anti-imperialist response, as stimulated by the 'May 4th Movement' and later the Sino-Japanese war, is shown to be an important ingredient in the transition from foreign to national control of the profession and in its achieving respect. Some doubt is cast on the assumption that Chinese nursing has followed a process of rapid convergence to the Western model.

Immunologic studies on drug addiction: III. Antibodies reactive with cocaine metabolites and their use for drug detection.

Antisera which react with the major metabolites of cocaine have been prepared in rabbits and a hemagglutination-inhibition (HI) test which detects these metabolites in urine or serum in concentrations of 1 ng/ml is described. A comparison of this test with alternate detection procedures shows excellent agreement at comparable sensitivity levels. HI tests on simian and human biological specimens suggest that the use of cocaine remains detectable for at least 3 days after administration of a minimal pharmacologically active dose. Combination of physical separation of drug metabolites with immunoassay procedures indicates that benzoylecgonine and ecgonine are the immunoreactive cocaine metabolites found in human urine. While it was possible to apply the HI test at maximal sensitivity to human sera and to murine or simian urine specimens, interference was encountered with some human urine specimens. Preliminary data suggest that by reducing the sensitivity of the test to a threshold of 200 ng/ml this interference can be overcome.

Efficacy of different phosphorus supplements for sows during gestation and lactation.

PIP: This study was undertaken to compare dicalcium, Curacao, and soft ph sophates as supplemental feeding during gestation and lactation. In Exp eriment 1, 37 crossbred sows at 11 months of age were mated and each of 4 groups given an experimental diet. The basal diet was a 16% crude pro tein-fortified corn-soybean meal containing .34% of phosphorus and .72% of calcium. The test diets were fortified with either dicalcium, Curaca o, or soft phosphate to provide .52% of phosphorus. Limestone was given to maintain .72% of calcium. Animals were confined in a concrete feedin g area and fed 1.8 kg of diet per head per day until Day 94 of gestation . At that time the diet was increased to 2.3 kg/day. On Day 109 of gestation animals were moved to farrowing crates until 10 days postpartu m and then to wooden units until weening at 8 weeks. Sows were sacrific ed after 8 weeks lactation, or if they developed posterior paralysis. In this experiment, the different phosphate sources were without observed influence on the livability or performance of pigs up to 8 weeks of age (p less than .05). Gilts receiving no supplemental phosphorus had significantly lower bone ash and phosphorus than did those receiving either of the supplements. Posterior paralysis was observed in 5 of 9 sows fed the unsupplemented diet, in 3 of 9 fed the soft supplemented diet, and in 2 of 10 fed the Curacao phosphate supplement. None fed dicalcium phosphate developed paralysis. The paralysis usually occurred after 6 weeks of lactation and was often accompanied by a fractured femur. In Experiment 2, 70 sows were bred at 10 months of age and fed 1.8 kg of the experimental diets. During 36 days of lactation, phosphorus digestibility, retention, and balance were shown to be lower for those fed only the basal diet. Bone ash, bone phosphorus, and bone strength were lower in those fed the unsupplemented diet. Results of these studies indicate the need for supplemental phosphorus in sow feeding. When feeding dicalcium phosphate or Curacao phosphate the use of over .5% phosphorus in the sow's diet is unnecessary during gestation and lactation.^ieng