Clinical features, antimicrobial susceptibilities, and outcomes of Elizabethkingia meningoseptica (Chryseobacterium meningosepticum) bacteremia at a medical center in Taiwan, 1999-2006.

A total of 118 patients with Elizabethkingia meningoseptica bacteremia at a medical center in Taiwan from 1999 to 2006 were studied. Minimum inhibitory concentrations (MICs) of 99 preserved isolates were determined. The incidence (per 100,000 admissions) of E. meningoseptica bacteremia increased from 7.5 in 1996 to 35.6 in 2006 (p = 0.006). Among them, 84% presented with fever, 86% had nosocomial infections, and 60% had acquired the infection in intensive care units (ICUs). The most common underlying diseases were malignancy (36%) and diabetes mellitus (25%). Seventy-eight percent of patients had primary bacteremia, followed by pneumonia (9%), soft tissue infection, and catheter-related bacteremia (6%). Forty-five patients (38%) had polymicrobial bacteremia. Overall, the 14-day mortality was 23.4%. Multivariate analysis revealed E. meningoseptica bacteremia acquired in an ICU (p = 0.048, odds ratio [OR] 4.23) and presence of effective antibiotic treatment after the availability of culture results (p = 0.049, OR 0.31) were independent predictors of 14-day mortality. The 14-day mortality was higher among patients receiving carbapenems (p = 0.046) than fluoroquinolones or other antimicrobial agents. More than 80% of the isolates tested were susceptible to trimethoprim-sulfamethoxzole, moxifloxacin, and levofloxacin. The MIC(50) and MIC(90) of the isolates to tigecycline and doxycycline were both 4 µg/mL and 8 µg/ml, respectively.

Efficacy of chlorine dioxide disinfection to non-fermentative Gram-negative bacilli and non-tuberculous mycobacteria in a hospital water system.

BACKGROUND: Chlorinated tap water in hospitals often contains low levels of non-fermentative Gram-negative bacilli (NFGNB) and non-tuberculous mycobacteria (NTM). Measures are needed to ensure a safe water supply in hospitals to prevent nosocomial infections from these waterborne pathogens. AIM: To evaluate the efficacy of ClO2 treatment of a hospital water system on the levels of NFGNB and NTM in the water. METHODS: Our institution is a 1000-bed medical centre with two main buildings (B1 and B2). B1 has three intensive care units (ICUs) and transplant wards and polyethylene water pipes. B2 (control) has no ICUs and galvanized water pipes. A ClO2 generating unit was installed in the water system of B1 in April 2012 and water samples were collected in B1 and B2 before and eight times after installation. All samples were cultured for NFGNB and NTM. FINDINGS: The ClO2 concentration was significantly lower in the hot water than in the cold water (P<0.001). After 40 weeks of ClO2 use, the overall NFGNB colonies decreased significantly (hot water: 160±143 vs 2±4cfu/mL, P<0.001; cold water: 108±138 vs 3±7cfu/mL, P<0.001). Highly prevalent nosocomial NFGNB, such as Pseudomonas spp. and Stenotrophomonas spp., were undetected three months after ClO2 disinfection; Sphingomonas spp. persisted but had lower colony counts. NTM was present in 25% (three out of 12) of sampling locations initially, but was not detected at two weeks after ClO2 disinfection. The ICUs had no overall change in the number of NFGNB nosocomial infections after the intervention. CONCLUSION: Addition of a ClO2 disinfection unit to our hospital water system reduced the numbers of NTM and NFGNB in the hot and cold water systems.

Effect of diet-induced weight loss on muscle strength in adults with overweight or obesity - a systematic review and meta-analysis of clinical trials.

We conducted a systematic review and meta-analysis to identify how diet-induced weight loss in adults with overweight or obesity impacts on muscle strength. Twenty-seven publications, including 33 interventions, most of which were 8-24 weeks in duration, were included. Meta-analysis of seven interventions measuring knee extensor strength by isokinetic dynamometry in 108 participants found a significant decrease following diet-induced weight loss (-9.0 [95% confidence interval: -13.8, -4.1] N/m, P < 0.001), representing a 7.5% decrease from baseline values. Meta-analysis of handgrip strength from 10 interventions in 231 participants showed a non-significant decrease (-1.7 [-3.6, 0.1] kg, P = 0.070), with significant heterogeneity (I(2) = 83.9%, P < 0.001). This heterogeneity may have been due to diet type, because there was a significant decrease in handgrip strength in seven interventions in 169 participants involving moderate energy restriction (-2.4 [-4.8, -0.0] kg, P = 0.046), representing a 4.6% decrease from baseline values, but not in three interventions in 62 participants involving very-low-energy diet (-0.4 [-2.0, 1.2] kg, P = 0.610). Because of variability in methodology and muscles tested, no other data could be meta-analyzed, and qualitative assessment of the remaining interventions revealed mixed results. Despite varying methodologies, diets and small sample sizes, these findings suggest a potential adverse effect of diet-induced weight loss on muscle strength. While these findings should not act as a deterrent against weight loss, due to the known health benefits of losing excess weight, they call for strategies to combat strength loss - such as weight training and other exercises - during diet-induced weight loss. © 2016 World Obesity.

Nociceptin/orphanin FQ knockout mice display up-regulation of the opioid receptor-like 1 receptor and alterations in opioid receptor expression in the brain.

The opioid receptor-like 1 receptor is a novel member of the opioid receptor family and its endogenous peptide ligand has been termed nociceptin and orphanin FQ. Activation of the opioid receptor-like 1 receptor by nociceptin/orphanin FQ in vivo produces hyperalgesia when this peptide is given supraspinally but analgesia at the spinal level. Nociceptin/orphanin FQ also reverses stress-induced analgesia, suggesting that the peptide has anti-opioid properties. Nociceptin/orphanin FQ knockout mice show alterations in pain sensitivity and stress responses and display increased morphine dependence, suggesting an interaction of the nociceptin/orphanin FQ system with classical opioid receptor function. To determine if the behavioural phenotype of nociceptin/orphanin FQ knockout mice reflects changes in either opioid receptor-like 1 or classical opioid receptor expression, we have carried out quantitative autoradiography of the opioid receptor-like 1, mu-, delta- and kappa-opioid receptors in the brains of these animals. Receptor density was measured on coronal sections from wild-type, heterozygous and homozygous mice using [(3)H]nociceptin, [(3)H][D-Ala(2)-N-methyl-Phe(4)-Gly(5) ol] enkephalin, [(3)H]deltorphin-I, or [(3)H](-)-N-methyl-N-[7-(1-pyrrodinyl)-1-oxospiro[4,5]dec-8-yl]-4-benzofuranacetamide to label opioid receptor-like 1, mu-, delta- and kappa-receptors, respectively. A region-specific up-regulation of the opioid receptor-like 1 receptor (up to 135%) was seen in brains from homozygous mice. Mu-Receptors also showed significant differences between genotypes whilst changes in delta- and kappa- receptors were minor. In conclusion the region-specific up-regulation of the opioid receptor-like 1 receptor indicates a tonic role for nociceptin/orphanin FQ in some brain structures and may suggest the peptide regulates the receptor expression in these regions. The changes in the opioid receptor-like 1 receptor may relate to the anxiogenic phenotype of these animals but the observed change in mu-receptors does not correlate with altered morphine responses.

Quantitative autoradiographic mapping of the ORL1, mu-, delta- and kappa-receptors in the brains of knockout mice lacking the ORL1 receptor gene.

Until recently the opioid receptor family was thought to consist of only the mu-, delta- and kappa-receptors. The cloning of opioid receptor like receptor (ORL1) and its endogenous ligand nociceptin/orphanin FQ, which displayed anti-opioid properties, has raised the issue of functional co-operativity of this system with the classical opioid system. ORL1 receptor knockout mice have been successfully developed by homologous recombination to allow the issue of potential heterogeneity of this receptor and also of compensatory changes in mu-, delta- or kappa-receptors in the absence of ORL1 to be addressed. We have carried out quantitative autoradiographic mapping of these receptors in the brains of mice that are wild-type, heterozygous and homozygous for the deletion of the ORL1 receptor. ORL1, mu-, delta- and kappa-receptors were labelled with [(3)H] leucyl-nociceptin (0.4 nM), [(3)H] DAMGO (4 nM), [(3)H] deltorphin-I (7 nM), and [(3)H] CI-977 (2.5 nM) respectively. An approximately 50% decrease in [(3)H] leucyl-nociceptin binding was seen in heterozygous ORL1 mutant mice and there was a complete absence of binding in homozygous brains indicating the single gene encodes for the ORL1 receptor and any putative subtypes. No significant gross changes in the binding to other opioid receptors were seen across genotypes in the ORL1 mutant mice demonstrating a lack of major compensation of classical opioid receptors in the absence of ORL1. There were a small number of region specific changes in the expression of classical opioid receptors that may relate to interdependent function with ORL1.

The efficacy of intrathecal coadministration of morphine and bupivacaine for labor analgesia.

BACKGROUND: Intrathecal (i.t.) opioids can provide labor analgesia, but the onset of pain relief is slow. Bupivacaine has the beneficial property of less motor blockade than other local anesthetics. This study retrospectively examined the efficacy of concomitant use of i.t. morphine and bupivacaine for labor pain relief. METHODS: Fifty five nulliparas who requested analgesia in the active phase (IA group) prior to a cervical dilation 3.9 +/- 0.6 cm and received i.t. morphine 0.5 mg and 0.1% bupivacaine 2.5 mg served as the treatment group, and 88 similar nulliparas who did not request and receive analgesia served as the control group. RESULTS: The mean onset time of analgesia was 2.6 +/- 0.5 min, the duration was 4-12 h and 93% of parturients did not request additional analgesia after a single injection of i.t. morphine and bupivacaine. The active phase of the first and second stages of labor in the treatment group were significantly longer [318 +/- 214 min vs. 176 +/- 120 min; 74 +/- 29 min vs. 37 +/- 26 min]. Frequency of instrument-assisted vaginal delivery was higher compared with the control group (30.9% vs. 14.1%). However, there were no significant differences in the rate of cesarean section and the Apgar scores of newborns at 1 min and 5 min between the treatment and control group. The major side effects of the treatment group included pruritus (48%), nausea (40%), vomiting (37%), somnolence (27%), shivering (27%), urinary retention (21%), hypotension (15%), and bradycardia (13%). Most of the side effects were mild and could be alleviated by naloxone. Neither post-spinal headache nor respiratory depression was noted. CONCLUSIONS: Our results showed that a single injection of i.t. morphine and bupivacaine provided rapid onset and effective analgesia with manageable side effects and without major complications. Thus, i.t. morphine and bupivacaine provides an alternative to epidural analgesia for most women in labor.

Sequential time to positivity of blood cultures can be a predictor of prognosis of patients with persistent Staphylococcus aureus bacteraemia.

A short time to positivity (TTP) correlates with poor clinical outcome in patients with Staphylococcus aureus bacteraemia, but the association between sequential TTPs and the outcome of these patients is unclear. Sequential TTPs from patients with S. aureus bacteraemia persisting for >48 h were analysed with respect to clinical parameters and patient outcome at a tertiary hospital. During the 5-year study period, 87 patients (9.2%; mean age of 64 years) had persistent S. aureus bacteraemia, with an average Pittsburgh bacteraemia score of 2.7. Forty-eight patients (55%) had methicillin-resistant S. aureus infection, and 28 (32%) had nosocomial infection. The most common underlying disease was end-stage renal disease (43%). The most common type of infection was catheter-related infection (31%), followed by infective endocarditis (18%). The in-hospital mortality rate was 40%. Higher Pittsburgh scores (p 0.005; OR 1.37; 95% CI 1.1-1.7) and a second TTP/first TTP ratio of <1.5 (p 0.004; OR 0.2; 95% CI 0.07-0.6) were independent risk factors for mortality. Among patients receiving adequate empirical therapy, a second positive blood culture growing within 12 h was more frequent in patients who finally died. Factors associated with a second TTP/first TTP ratio of <1.5 included older age (p 0.02; OR 0.96; 95% CI 0.92-0.99) and inadequate empirical antimicrobial therapy (p 0.01; OR 3.53; 95% CI 1.42-8.78). Among patients with persistent S. aureus bacteraemia, a second TTP/first TTP ratio of <1.5 is a predictor of poor outcome. Physicians should search for interventions guaranteeing that all patients with S. aureus bacteraemia receive adequate empirical therapy.

Loss of the mu opioid receptor on different genetic backgrounds leads to increased bromodeoxyuridine labeling in the dentate gyrus only after repeated injection.

The endogenous opioid system is involved in various physiological processes, including neurogenesis in the dentate gyrus (DG) of the hippocampus. In the current study, we investigated the role of the mu opioid receptor (MOR-1) on DG neurogenesis and measured glucocorticoid levels following several injection paradigms to supplement the neurogenesis experiments. MOR-1 knockout (KO) mice on C57BL/6 and 129S6 backgrounds were injected with bromodeoxyuridine (BrdU) using either a single injection or two different repeated injection protocols and then sacrificed at different time points. The total number of BrdU and proliferating cell nuclear antigen (PCNA) positive cells in the DG is significantly increased in MOR-1 KO mice compared with wild type (WT) on both strains after repeated injection, but not after a single injection. Plasma corticosterone (CORT) levels increased similarly in MOR-1 KO and WT mice following both single and repeated injection, indicating that the stress response is activated following any injection protocol, but that the mechanism responsible for the increase in BrdU labeling in MOR-1 KO mice is CORT-level independent. Finally, WT 129S6 mice, independent of genotype, showed higher levels of plasma CORT compared with WT C57BL/6 mice in both noninjected controls and following injection at two separate time points; these levels were inversely correlated with low numbers of BrdU cells in the DG in 129S6 mice compared with C57BL/6 mice. In summary, these data demonstrate that loss of MOR-1 increases BrdU labeling in the DG independent of CORT levels, but only following a repeated injection, illustrating the capability of injection paradigms to influence cell-proliferative responses in a genotype-dependent manner.

Laminar-specific and developmental expression of aquaporin-4 in the mouse hippocampus.

Mice deficient in the water channel aquaporin-4 (AQP4) demonstrate increased seizure duration in response to hippocampal stimulation as well as impaired extracellular K+ clearance. However, the expression of AQP4 in the hippocampus is not well described. In this study, we investigated (i) the developmental, laminar and cell-type specificity of AQP4 expression in the hippocampus; (ii) the effect of Kir4.1 deletion on AQP4 expression; and (iii) performed Western blot and RT-PCR analyses. AQP4 immunohistochemistry on coronal sections from wild-type (WT) or Kir4.1-/- mice revealed a developmentally-regulated and laminar-specific pattern, with highest expression in the CA1 stratum lacunosum-moleculare (SLM) and the molecular layer (ML) of the dentate gyrus (DG). AQP4 was colocalized with the glial markers glial fibrillary acidic protein (GFAP) and S100ß in the hippocampus, and was also ubiquitously expressed on astrocytic endfeet around blood vessels. No difference in AQP4 immunoreactivity was observed in Kir4.1-/- mice. Electrophysiological and postrecording RT-PCR analyses of individual cells revealed that AQP4 and Kir4.1 were co-expressed in nearly all CA1 astrocytes. In NG2 cells, AQP4 was also expressed at the transcript level. This study is the first to examine subregional AQP4 expression during development of the hippocampus. The strikingly high expression of AQP4 in the CA1 SLM and DG ML identifies these regions as potential sites of astrocytic K+ and H2O regulation. These results begin to delineate the functional capabilities of hippocampal subregions and cell types for K+ and H2O homeostasis, which is critical to excitability and serves as a potential target for modulation in diverse diseases.

Risk factors for nosocomial infection during extracorporeal membrane oxygenation.

An increasing number of patients receive extracorporeal membrane oxygenation (ECMO) for life support. This study aimed to investigate the incidence and risk factors for nosocomial infection in adult patients receiving ECMO. We reviewed the medical records of adult patients who received ECMO support for more than 72h at Far Eastern Memorial Hospital from 2001 to 2007. ECMO-related nosocomial infections were defined as infections occurring from 24h after ECMO initiation until 48h after ECMO discontinuation. There were 12 episodes of nosocomial infection identified in 10 of the 114 (8.77%) patients on ECMO, including four cases of pneumonia, three cases of bacteraemia, three surgical site infections and two urinary tract infections. The incidence of ECMO-related nosocomial infection was 11.92 per 1000 ECMO-days. The length of ECMO use and intensive care unit (ICU) stay were significantly different between patients with, and without, nosocomial infection (P<0.001). More than 10 days of ECMO use was associated with a significantly higher nosocomial infection rate (P=0.003). Gram-negative bacilli were responsible for 78% of the nosocomial infections. In the univariate analysis, the duration of ICU stay and duration of ECMO use were associated with nosocomial infection. In the multivariate analysis, only the duration of ECMO was independently associated with nosocomial infection (P=0.007). Overall, the only independent risk factor for ECMO-related nosocomial infection identified in this study was prolonged ECMO use.

Correlation between time to positivity of blood cultures with clinical presentation and outcomes in patients with Klebsiella pneumoniae bacteraemia: prospective cohort study.

Time to positivity (TTP) of blood cultures in patients with bacteraemia is considered to be a predictor of outcome for some bacterial species. Two hundred and thirty-one patients with Klebsiella pneumoniae monomicrobial bacteraemia at a hospital from 1 January to 31 December 2007 were prospectively enrolled. TTP <7 h (46 patients, 19.9%) was associated with a higher Pittsburg bacteraemia score (6.2 +/- 5.5 vs. 3.7 +/- 4.3, p 0.002), fewer non-fatal diseases by the McCabe classification (39.1% vs. 64.9%, p 0.002), a higher percentage of patients with liver cirrhosis, active malignancy, and chemotherapy within 3 months (28.3% vs. 11.9%, p 0.007; 28.3% vs. 14.6%, p 0.031; 23.9% vs. 5.4%, p <0.001), more primary bacteraemia (45.7% vs. 22.2%, p 0.002), and a higher 30-day mortality rate (47.8% vs. 21.1%, p <0.001). Risk factors for 30-day mortality in the univariate analysis included higher Pittsburg bacteraemia score (5.8 +/- 5.3 vs. 3.7 +/- 4.3, p 0.002), primary bacteraemia (41.0% vs. 21.8%, p 0.004), TTP <7 h (36.1% vs. 14.1%, p <0.001), and the presence of active malignancy (29.5% vs. 12.9%, p 0.004). In the multivariate analysis, higher Pittsburg bacteraemia score (OR 1.07; 95% CI 1.01-1.14), TTP <7 h (OR 2.46; 95% CI 1.20-5.05) and active malignancy (OR 2.21; 95% CI 1.03-4.73) were the significant factors associated with 30-day mortality. In the Kaplan-Meier survival curve, short TTP was significantly associated with mortality at all time-points after admission. TTP of blood cultures, interpreted with a cut-off point of <7 h, in patients with K. pneumoniae bacteraemia can provide useful prognostic information.

Developmental expression of the mu, kappa, and delta opioid receptor mRNAs in mouse.

To characterize further the establishment of the opioid system during prenatal mouse development, we have examined the spatial and temporal expression patterns of mu, kappa, and delta opioid receptor mRNAs and find that the expression patterns of these mRNAs are distinct at all ages. Within the embryo, kappa is the first opioid receptor expressed, with transcripts detected in the gut epithelium as early as embryonic day 9.5 (E9.5). By E10.5, mu receptor expression is first detected in the facial-vestibulocochlear preganglion complex, whereas delta receptor mRNA is first detected at E12.5 in several peripheral tissues, including the olfactory epithelium, heart, limb bud, and tooth. In the brain, both mu and kappa mRNAs are first detected at E11.5 in the basal ganglia and midbrain, respectively. During mid-gestation and late gestation, the expression of both mu and kappa receptors extends to other brain regions that exhibit high expression in the adult, including the medial habenula, hypothalamus, pons, and medulla for mu and the basal ganglia, thalamus, hypothalamus, raphe, and ventral tegmental area for kappa. Thus by E17.5, many aspects of the adult expression patterns of mu and kappa receptors already have been established. Compared with mu and kappa, delta receptor mRNA expression in the brain begins relatively late, and the expression levels remain very low even at E19.5. In contrast to its late appearance in the brain, however, delta is the first opioid receptor expressed in the dorsal root ganglion, at E12.5, before its expression in the spinal cord begins at E15.5. Mu receptor is the first opioid receptor expressed in the spinal cord, at E11.5. These results extend previous ligand-binding data to significantly earlier ages and suggest that early developmental events in both neural and non-neural tissues may be modulated by opioid receptors. Several examples of possible autocrine and paracrine loops of opioid peptide and receptor expression have been identified, suggesting a role for these local circuits in developmental processes.

Insulin-like growth factor I and II and insulin-like growth factor binding protein-2 RNAs are expressed in adjacent tissues within rat embryonic and fetal limbs.

Since the rapid proliferation of cells in a directed manner is a necessary component of limb formation, the distribution of locally produced mitogenic molecules within the developing limb is of considerable interest. We have used in situ hybridization to localize transcripts for both insulin-like growth factor binding protein-2 (IGFBP-2) and its ligands, the insulin-like growth factors I and II (IGF-I and IGF-II), within limb buds of rat embryos 10-16 days after conception (equivalent to stages 1-12 of mouse limb morphogenesis, Wanek et al, 1989. J. Exp. Zool. 249, 41-49). The mRNA for IGFBP-2 is very abundant in an anterior-posterior strip of ectoderm along the distal edge of the limb bud (the progenitor of the apical ectodermal ridge or AER) from as early as limb stage 1 (Embryonic Day 10) and is much less abundant in the rest of the limb ectoderm. A high level of IGFBP-2 expression continues to characterize the AER following its definitive appearance (stage 3) and throughout its existence (until stage 7). This is a period of rapid outgrowth during which the rate of mesodermal cell division is highest in cells nearest to the AER. The AER is known to have mitogenic activity in vitro and to direct limb outgrowth in vivo, but, until recently, few putative molecular correlates of these activities have been detected. The transcripts for IGF-I and IGF-II are also present at high abundance in developing limbs, especially in mesodermally derived cells. IGF-I mRNA is abundant in presumptive limb mesoderm from the beginning of limb outgrowth (just before stage 1), but is very low or undetectable in much of the rest of the embryo, while IGF-II mRNA becomes very abundant in limb mesoderm at stage 2. The distribution in limbs of both IGF-I and IGF-II mRNA changes dramatically during outgrowth and differentiation, so that their expression characterizes complementary populations of cells by stage 11. Taken together, these data suggest that IGFs and the IGF binding proteins, which may modulate IGF action, contribute to limb outgrowth and patterning.

Subcellular localization of mannose 6-phosphate glycoproteins in rat brain.

The intracellular transport of soluble lysosomal enzymes relies on the post-translational modification of N-linked oligosaccharides to generate mannose 6-phosphate (Man 6-P) residues. In most cell types the Man 6-P signal is rapidly removed after targeting of the precursor proteins from the Golgi to lysosomes via interactions with Man 6-phosphate receptors. However, in brain, the steady state proportion of lysosomal enzymes containing Man 6-P is considerably higher than in other tissues. As a first step toward understanding the mechanism and biological significance of this observation, we analyzed the subcellular localization of the rat brain Man 6-P glycoproteins by combining biochemical and morphological approaches. The brain Man 6-P glycoproteins are predominantly localized in neuronal lysosomes with no evidence for a steady state localization in nonlysosomal or prelysosomal compartments. This contrasts with the clear endosome-like localization of the low steady state proportion of mannose-6-phosphorylated lysosomal enzymes in liver. It therefore seems likely that the observed high percentage of phosphorylated species in brain is a consequence of the accumulation of lysosomal enzymes in a neuronal lysosome that does not fully dephosphorylate the Man 6-P moieties.