Risk factors and mortality associated with multi-drug-resistant Gram-negative bacterial infection in adult patients following abdominal surgery.

BACKGROUND: Multi-drug-resistant (MDR) Gram-negative bacterial (GNB) infection remains a significant cause of morbidity and mortality among surgical patients. The objective of this study was to recognize the risk factors for MDR GNB infection in patients following abdominal surgery, and determine the predictors independently associated with death. METHODS: From 2010 to 2017, a retrospective cohort study was conducted among patients with abdominal surgery admitted to the surgical intensive care unit (ICU). Patients with GNB infection were included for analyses. RESULTS: In total, 364 patients experienced GNB infection following abdominal surgery. Of these, 117 (32.1%) were MDR GNB infection. Of 133 MDR GNB isolates, the most common isolate was Escherichia coli (45.1%). Patients with MDR GNB infection had significantly longer ventilator-days and hospital stay, as well as higher 30-day and in-hospital mortality compared with non-MDR GNB patients. Multi-variable analysis showed that longer length of pre-ICU stay, surgical re-exploration, receipt of group 2 carbapenems (e.g. imipenem, meropenem and doripenem) and fluoroquinolones, and higher total bilirubin were independent risk factors for the acquisition of MDR GNB infection. Predictors for 30-day mortality among patients with MDR GNB infection were chronic kidney disease, receipt of group 2 carbapenems and inappropriate empirical antimicrobial therapy. CONCLUSIONS: This study provides important information about the risk factors for MDR GNB infection and 30-day mortality among patients following abdominal surgery.

Amitriptyline modulated Ca2+ signaling and induced Ca2+-independent cell viability in human osteosarcoma cells.

Amitriptyline is a widely used tricyclic antidepressant, which acts primarily as a serotonin-norepinephrine reuptake inhibitor. This study examined the effect of amitriptyline on Ca2+ homeostasis and its related mechanism in MG63 human osteosarcoma cells. Amitriptyline evoked cytosolic-free Ca2+ concentrations ([Ca2+]i) rises concentration dependently. Amitriptyline-evoked Ca2+ entry was confirmed by Mn2+-induced quench of fura-2 fluorescence. This entry was inhibited by Ca2+ entry modulators nifedipine, econazole, SKF96365, the protein kinase C (PKC) activator phorbol 12-myristate 13 acetate but was not affected by the PKC inhibitor GF109203X. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin (TG) inhibited amitriptyline-evoked [Ca2+]i rises by 95%. Conversely, treatment with amitriptyline abolished TG-evoked [Ca2+]i rises. Inhibition of phospholipase C (PLC) with U73122 inhibited amitriptyline-evoked [Ca2+]i rises by 70%. Amitriptyline killed cells at 200-500 µM in a concentration-dependent fashion. Chelating cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane- N, N, N', N'-tetraacetic acid/AM did not reverse amitriptyline-induced cytotoxicity. Collectively, our data suggest that in MG63 cells, amitriptyline induced [Ca2+]i rises by evoking PLC-dependent Ca2+ release from the endoplasmic reticulum and Ca2+ entry via PKC-regulated store-operated Ca2+ entry. Amitriptyline also induced Ca2+-disassociated cell death.

Evaluation of wound healing effect on skin-defect nude mice by using human dermis-derived mesenchymal stem cells.

We report that human dermis-derived mesenchymal stem cells (hDMSCs) possess differentiation potential of epidermis facilitating wound healing in skin-defect nude mice in combination with the treatment using gelatin/thermosensitive poly N-isopropylacrylamide (pNIPAAm)/polypropylene (PP). The results showed that the rate of cell growth and wound recovery in the hDMSC and gelatin/pNIPAAm/PP-treated group was significantly greater than those in the gelatin/pNIPAAm/PP-treated only group (P < .01). The reepithelialization marker of human pan-cytokeratin was also significantly increased on days 14 and day 21 in the wound site of hDMSCs and gelatin/pNIPAAm/PP-treated group. Furthermore, the stem cell marker of human CD13 gradually decreased during the period of wound healing. In sum, this novel method provided a transferring system for stem cell therapy, maintaining its temperature-sensitive property of easy peeling by lower temperature treatment.

Systemic administration of a water-soluble hexasulfonated C(60) (FC(4)S) reduces cerebral ischemia-induced infarct volume in gerbils.

Buckminsterfullerence and its derivatives have recently been shown to exhibit considerable in vivo biological activities. A water-soluble hexasulfonated C(60) (FC(4)S) has been shown to protect against oxidative stress. Neuroprotective effects of FC(4)S were investigated in the present study. Focal cerebral ischemia was produced by a permanent occlusion of the right middle cerebral artery in gerbils. Infarct volumes were determined by 2,3,5-triphenyltetrazolium chloride transcardiac perfusion 24 h after cerebral ischemia. Chronic pretreatment of FC(4)S (0.5 and 5.0 mg/kg/day, intraperitoneally for 2 weeks) significantly reduced the infarct volume (by 42% and 68%, respectively) when compared to that of the control group. Results revealed that chronic pretreatment of FC(4)S may protect the brain against focal cerebral ischemia.

A new method for true and spurious eigensolutions of arbitrary cavities using the combined Helmholtz exterior integral equation formulation method.

Integral equation methods have been widely used to solve interior eigenproblems and exterior acoustic problems (radiation and scattering). It was recently found that the real-part boundary element method (BEM) for the interior problem results in spurious eigensolutions if the singular (UT) or the hypersingular (LM) equation is used alone. The real-part BEM results in spurious solutions for interior problems in a similar way that the singular integral equation (UT method) results in fictitious solutions for the exterior problem. To solve this problem, a Combined Helmholtz Exterior integral Equation Formulation method (CHEEF) is proposed. Based on the CHEEF method, the spurious solutions can be filtered out if additional constraints from the exterior points are chosen carefully. Finally, two examples for the eigensolutions of circular and rectangular cavities are considered. The optimum numbers and proper positions for selecting the points in the exterior domain are analytically studied. Also, numerical experiments were designed to verify the analytical results. It is worth pointing out that the nodal line of radiation mode of a circle can be rotated due to symmetry, while the nodal line of the rectangular is on a fixed position.

Hepatitis C virus infection among teenagers in an endemic township in Taiwan: epidemiological and clinical follow-up studies.

The aim of the study was to elucidate the epidemiological features of Hepatitis C virus (HCV) infection among teenagers in an endemic area by conducting a mass screening study. We also investigated the clinical outcome of the anti-HCV-positive subjects by conducting subsequent short-term and long-term follow-up studies. All 2837 students of two junior middle schools in Tzukuan, aged 13-16 years, were invited to be screened for anti-HCV, HBsAg, AST and ALT in October 1995. A total of 2726 (96%) students responded. Anti-HCV, HCV RNA and aminotransferase levels were evaluated among anti-HCV-positive students 1 month and 30 months later, respectively. A total of 38 (1.4%; M/F = 22/16) participants were anti-HCV-positive. The anti-HCV-positive students had higher rates of exposures to transfusion, anti-HCV-positive families and surgery. The prevalence (2.8%) of the 7 maritime villages was markedly higher than that (0.7%) of the other 8 villages (P < 0.001). Subsequent follow-up studies demonstrated that there might be 5 cases of acute or recent HCV infection, and 6 cases who had recovered from chronic HCV infection.

Satellite videoconferencing for healthcare workers: audience characteristics and the importance of continuing education credits.

To assess the opinions of healthcare workers (HCWs) about a satellite videoconference as a means of earning continuing education credit, a telephone survey was conducted in September 1998, 1 month after a live interactive satellite videoconference on antimicrobial use and resistance. There were 180 registered sites in 45 states surveyed, representing 1,589 viewers: 764 nurses (48.1%), 201 physicians (12.6%), and 624 other HCWs (39.3%). Continuing education credit was requested by 51% of nurses, 31% of physicians, and 27% of all other HCWs. Although preferred learning formats varied, 70% of respondents said it was important to offer continuing education credit. Furthermore, 31% of the respondents stated that the videoconference influenced institutional strategies. We concluded that satellite videoconferences are a method to reach audiences around the world efficiently and effectively, provide the latest information, facilitate interaction, and meet some of the demand for continuing education credit for HCWs.

Endothelial cell regrowth and morphology after balloon catheter injury of alloxan-induced diabetic rabbits.

Neointimal thickening after catheter injury has been reported to be influenced by the integrity of the vascular endothelium. We have previously shown that neointimal thickening is significantly reduced in alloxan-induced diabetic New Zealand White rabbits after catheter injury compared with euglycemic rabbits. In the present study, it was hypothesized that endothelial cell regrowth, morphology, and endothelium-dependent vasoreactivity after catheter injury are improved in the diabetic rabbit (glucose >/=400 mg/dl) compared with the euglycemic rabbit. Two weeks after catheter injury, the percent endothelial regrowth was significantly increased in diabetic animals compared with euglycemic animals (32.1 +/- 2 and 15.6 +/- 1, respectively; P < 0.05). The endothelial cell morphology analyzed by scanning electron microscopy was also restored 2 wk after catheter injury in thoracic aortas from the diabetic animals compared with vessels from euglycemic animals. Endothelium-dependent relaxation to ACh in vessels from diabetic and euglycemic rabbits was attenuated 2 wk after injury, and, although improved by 4 and 8 wk, relaxation remained significantly depressed. These results suggest that endothelial cell regrowth and morphology in diabetic animals was improved compared with euglycemic animals; however, endothelium-dependent vasoreactivity remained impaired. Thus the attenuated neointimal thickening seen in the diabetic rabbit may be a function of the rate and degree of regrowth rather than the normalization of ACh-induced relaxation.

Ex vivo expression of nitric oxide synthase isoforms (eNOS/iNOS) and calmodulin in human penile cavernosal cells.

PURPOSE: Nitric oxide (NO) synthesized by nitric oxide synthase (NOS) is recognized as the central mediator of penile erection. This process appears to be mediated mainly by neuronal NOS (nNOS), which is localized to the nonadrenergic, noncholinergic innervation of the penis. However, the role of non-neuronal penile constituents (specifically the cavernosal smooth muscle), as well as other NOS isoforms in NO production in the human penis is not well understood. The present study evaluates the expression of non-neuronal (inducible and endothelial) isoforms of NOS in human penile cavernosal smooth muscle cells in culture. MATERIALS AND METHODS: Primary culture was initiated with explants of human corpora cavernosa. For gene expression studies, total RNA was extracted from cavernosal cells and subjected to reverse transcriptase polymerase chain reaction (RT-PCR). For NADPH-diaphorase histochemistry, the cells were incubated with 1 mM beta-NADPH and 0.5 mM nitrobluetetrazolium at 37C for 3 hours. For indirect immunofluorescence and electron microscopy, cells were incubated overnight at 4C with specific primary (eNOS; calmodulin) and secondary antibodies. A conventional avidin biotin complex technique was used for electron microscopy. RESULTS: The mRNA expression studies revealed that these cells express both endothelial (eNOS) and inducible (iNOS) forms. Localization studies showed positive signals for NADPH-diaphorase, eNOS, and calmodulin. The electron microscopic evaluation confirmed the localization of eNOS to the cytoplasm and small vesicles in the cells. CONCLUSIONS: These findings support the hypothesis that human cavernosal smooth muscle cells express both endothelial and inducible forms of NOS, which may significantly contribute to NO production in the penile architecture during the erectile process.

Mediation of gamma-hexachlorocyclohexane-induced DNA fragmentation in HL-60 cells through intracellular Ca2+ release pathway.

The cytotoxicity of gamma-hexachlorcyclohexane (gamma-HCC) was evaluated in HL-60 cells. Gamma-HCC dose-dependently induced cytotoxicity of HL-60 with an IC50 value of 60+/-5 microM. The gamma-HCC treated cells showed some characteristic changes of apoptosis, including blebbing of the membrane, condensation of the nuclear chromatin, vacuolation of cytoplasm and internucleosomal DNA fragmentation. Gamma-HCC induced DNA fragmentation of HL-60 cells in a dose-, time- and Ca2+-dependent manner. The DNA fragmentation induced was inhibited by intracellular Ca2+ chelator, calmodulin antagonist and Ca2+ sensitive endonuclease inhibitor. Gamma-HCC caused a steady increase in the cytosolic free Ca+ concentration due to release from intracellular stores. Neither the DNA fragmentation nor the increase of intracellular Ca2+ induced by gamma-HCC was inhibited by the removal of extracellular Ca2+. These data suggested that the cytotoxicity of gamma-HCC in HL-60 cells is mediated by the increase of intracellular Ca2+ concentration and the activation of Ca2+-dependent endonuclease, which triggers apoptosis in a Ca2+ and calmodulin-dependent manner.

Comparative studies on the induction of muscle contracture in mouse diaphragm and Ca2+ release from sarcoplasmic reticulum vesicles by organotin compounds.

Effects of organotins, including triethyltin and tributyltin, on skeletal muscle were studied with diaphragm and isolated sarcoplasmic reticulum membrane vesicles. Triethyltin induced muscle contracture in mouse diaphragm while tributyltin had comparatively less potency and efficacy in inducing the muscle contracture. The contracture induced by tributyltin was inhibited when the diaphragm was pretreated with low Ca2+ medium or caffeine while the contracture induced by triethyltin persisted in the Ca2+-free medium but was inhibited by pretreatment of caffeine. Pretreatment of dithiothreitol blocked the contracture induced by tributyltin but not that by triethyltin. Triethyltin dose-dependently induced Ca2+ release from sarcoplasmic reticulum vesicles and inhibited the Ca2+-ATPase activity. These results suggested that triethyltin induced contracture in mouse diaphragm was mainly by induction of Ca2+ release and inhibition of Ca2+ uptake of the internal Ca2+ storage site the sarcoplasmic reticulum, while the tributyltin induced contracture might be due to enhancement of extracellular Ca2+ influx which further induce the release of internal Ca2+ through the Ca2+-induced Ca2+ release mechanism.

Effect of short-term treatment with a monoclonal antibody to P-selectin on balloon catheter-induced: intimal hyperplasia, re-endothelialization, and attenuation of endothelial-dependent relaxation.

The effects of an anti-P-selectin monoclonal antibody (MAb, PB1.3; Cytel Corporation) on neoendothelialization; neoendothelial function, as evidenced by acetylcholine-induced relaxation (nitric oxide formation); and intimal hyperplasia following embolectomy catheter-induced injury to the rabbit thoracic aorta were investigated. Catheter injury was induced in two groups of New Zealand White rabbits. One group received no treatment, while the second group received short-term treatment with the MAb (i.p., immediately before and 12 h after induction of catheter injury). A third group underwent a sham operation and served as uninjured controls. Following sacrifice at 2 weeks after injury, aortic rings were assessed for degree of intimal hyperplasia, neoendothelial morphology (scanning electron microscopy), and acetylcholine-induced relaxation. Aortic tissue from catheter-injured animals that received treatment exhibited improved neoendothelial morphology, as compared with tissue from untreated but catheterized animals; however, no statistically significant attenuation of the hyperplastic response or improvement in the attenuated neoendothelial-dependent acetylcholine-induced relaxant response that is characteristic of neoendothelium that forms after catheter denudation was observed. These data suggest that short-term attenuation of P-selectin-mediated polymorphonuclear leukocyte (PMN)/endothelium, PMN/platelet interactions, and/or thrombin formation beneficially affects neoendothelialization of the vascular wall following balloon catheter-induced injury.

Incidence of hepatitis C infection in a hepatitis C endemic township in southern Taiwan.

To realize the incidence of hepatitis C virus (HCV) infection in an HCV endemic township, Tzukuan, in Taiwan, we conducted a follow-up community-based survey. A total of 173 adults, 82 males and 91 females, with mean age of 55.5 +/- 9.9 years received initial and follow-up anti-HCV tests with one-year interval. One (1.2%, 95% CI: 0%-5.5%) of 84 anti-HCV-positive subjects was negative-seroconversion, and 4 (4.5%, 95% CI: 0.2%-8.8%) of 89 anti-HCV-negative subjects were positive-seroconversion. The results indicated that hepatitis C might be still spreading in the HCV-endemic area now.

Induction of calcium release from isolated sarcoplasmic reticulum by triphenyltin.

A direct peripheral myopathy has been found in organotin intoxication and suggested to be a significant factor in the development of muscle weakness following exposure. In this study, by using the isolated sarcoplasmic reticulum membrane vesicles, we have shown that triphenyltin dose-dependently induced Ca2+ release from the actively and passively loaded sarcoplasmic reticulum vesicles. Triphenyltin induced Ca2+ release in ruthenium red-sensitive and insensitive ways with EC50 values of 75 and 270 microM, respectively. The Ca2+-ATPase activity and Ca2+ uptake of sarcoplasmic reticulum were also inhibited by triphenyltin. Triphenyltin exerted dual effects on the apparent [3H]ryanodine binding. Triphenyltin (0.5-10 microM) dose-dependently potentiated the [3H]ryanodine binding; however, the [3H]ryanodine binding decreased as the concentration of triphenyltin increased. The dissociation of bound [3H]ryanodine was facilitated by triphenyltin. The present study suggested that the internal Ca2+ store of skeletal muscle could be depleted by triphenyltin through the inhibition of the Ca2+ uptake and the induction of Ca2+ release by acting on the Ca2+-ATPase and Ca2+ release channel, also known as the ryanodine receptor, of sarcoplasmic reticulum, respectively. These results could partly explain the development of muscle weakness in organotin intoxication; however, their relevance to the development of peripheral myopathy requires further examination.

Different viral aetiology of hepatocellular carcinoma between two hepatitis B and C endemic townships in Taiwan.

In Taiwan, we found two hepatitis B virus (HBV)- and hepatitis C virus (HCV)-endemic townships, Paisha and Tzukuan, with an anti-HCV prevalence of 19 and 37% in men, and 26 and 38% in women, respectively. The hepatitis B surface antigen (HBsAg)-positive rates were 25 and 18%, for men and women in Paisha, and 25 and 22% in Tzukuan, respectively. According to the national death certification database (1982 to 1991), the annual age-adjusted mortality rates per 100,000 population for liver cancer among men and women were 83.0 and 13.8, respectively, in Paisha, and 55.9 and 17.0 in Tzukuan compared with 30.9 and 9.1 in Taiwan as a whole. The male-to-female ratios were 6.0 in Paisha and 3.3 in Tzukuan. Aetiology of 11 cases of hepatocellular carcinoma (HCC) from Paisha and 14 cases from Tzukuan were analysed. All HCC cases from Paisha were HBsAg positive, while 13/14 HCC cases from Tzukuan were anti-HCV positive. The endemic duration of HCV in Tzukuan seemed long enough to induce HCC, but the HCV appeared to be a newly introduced infection in Paisha.

Morphometric study of specific heart granules in the grey seal (Halichoerus grypus).

Specific heart granules in the grey seal, Halichoerus grypus, a marine mammal, were studied and compared with those in the Wistar rat by ultrastructural morphometry. There is extreme paucity of these granules in atrial cells of the seal, even in the region of the Golgi apparatus, compared with those in the rat. By numerical concentration, granules in the seal are 23 times fewer in number than in the rat. They are also fewer in number than in bats and hamsters for which data is available. The mean (+/-SEM) diameter of heart granules in the seal is significantly less (188 +/- 5.3 nm) than that of the rat (226 +/- 4.6 nm). These observations, the first in a marine mammal, are of interest in relation to the need for conservation of water and electrolytes in pennipids.

The cellular localization of vasoactive intestinal peptide (VIP) in the mouse median eminence by immuno-electron microscopy.

The aim of this study was to examine, by use of pre-embedding immunocytochemistry, the ultrastructural localization of vasoactive intestinal peptide (VIP) immunoreactivity in the mouse median eminence. VIP immunoreactivity was observed in axonal profiles. The VIP-immunoreactive axonal profiles were in close proximity to non-immunoreactive axonal profiles that contained dense granular vesicles and clear vesicles and also to processes of tanycytes. VIP-immunoreactive terminals were observed in the proximity of the perivascular space and in the neuropil. Our results suggest that VIP-immunoreactive axon terminals may possibly interact with other non-immunoreactive axon terminals containing peptide and/or other transmitters at the level of the median eminence or may be released to the portal vasculature thereby to effect anterior pituitary cells.

Different ultrastructural localization of VIP and prolactin in anterior pituitary cells of rats chronically treated with estrogen.

In the present study we investigated the effect of a long-term estrogen treatment on the intracellular distribution of VIP immunoreactivity in pituitary prolactin cells using double-labeling immunocytochemistry. With the use of pre-embedding ABC method it was found that VIP immunoreactivity was associated with the outer surface of membrane-bound organelles, and was not found in secretory granules. However, prolactin immunoreactivity demonstrated by postembedding immunogold technique was mainly associated within the secretory granules of the same cells. The discrepancy between our and Hsu et al.'s results (1989), who observed VIP immunoreactivity in secretory granules of human anterior pituitary cells, may be owing to the overstimulation of VIP cells by estrogen. It is possible that estrogen treatment depleted the VIP content of the secretory granules and enhanced the cytosolic VIP. The appearance of an alternative form of VIP in estrogen-treated rats with preferential distribution in the cytosol cannot be excluded.

Prevalence of antibody to hepatitis E virus among healthy individuals in southern Taiwan.

The seroprevalence of hepatitis E virus (HEV) among 997 healthy individuals aged 6 to 84 years, collected between July 1993 and June 1994 at Kaohsiung-Pingtung area in Southern Taiwan was studied. Of the study populations of vegetable farmers, elementary school children, volunteer blood donors and college students, the prevalence of IgG anti-HEV ranged from 6.4% to 8.8%. In suburban elemantary school children of Mang-Chou Village at Pingtung-Hsien, the seroprevalence rate (9.6%) was significantly higher than the positive rate (1.5%) found in rural aboriginal elementary school of San-Min Village at Kaohsiung-Hsien. IgG anti-HEV antibodies were widely distributed among all age groups, with a significantly higher percentage (13.1%) in the age group of 46-55 years old.