Surgical microscope may enhance root coverage with subepithelial connective tissue graft: a randomized-controlled clinical trial.
Minimally invasive techniques have broadened the horizons of periodontal plastic surgery to improve treatment outcomes. Thus, the purpose of this clinical trial was to compare root coverage, postoperative morbidity, and esthetic outcomes of subepithelial connective tissue graft (SCTG) technique with or without the use of a surgical microscope in the treatment of gingival recessions. In this split-mouth study, twenty-four patients with bilateral Miller's Class I or II buccal gingival recessions ≥2.0 mm in canines or premolars were selected. Gingival recessions were randomly designated to receive treatment with SCTG with or without the assistance of the surgical microscope (test and control groups, respectively). Clinical parameters evaluated included the following: depth (RH) and width (RW) of the gingival defect, width (WKT) and thickness (TKT) of keratinized tissue, probing depth (PD), and clinical attachment level (CAL). Postoperative morbidity was evaluated by means of an analog visual scale and questionnaire. Patient satisfaction was also evaluated with a questionnaire. Descriptive statistics were expressed as mean ± SD. Repeated-measures analysis of variance was used for examination of differences regarding PD, CAL, and TKT. The Wilcoxon test was used to detect differences between groups and the Friedman test to detect differences within group regarding WKT, RH, and RW. The average percentages of root coverage for test and control treatments, after 12 months, were 98.0% and 88.3%, respectively (P <0.05). Complete root coverage was achieved in 87.5% and 58.3% of teeth treated in test and control groups, respectively. For all parameters except recession height, there was an improvement in the final examination but without difference between treatments. For the RH, a lower value was found in the test group compared to the control group (P <0.05). In the test group, all patients were satisfied with the esthetics obtained, and 19 patients (79.1%) were satisfied in the control group. For postoperative morbidity, 14 patients in each of the two treatment groups did not use analgesics for pain control. Both approaches were capable of producing root coverage; however, use of the surgical microscope was associated with additional clinical benefits in the treatment of teeth with gingival recessions.


Facile synthesis of nanostructured perovskites by precursor accumulation on nanocarbons.
Perovskite-type oxides have impacted various research fields, including materials and energy science. Despite their vast potential in various applications, general and simple synthesis methods for nano-perovskites remain limited. Herein, various nano-perovskites were synthesized by a facile approach involving the use of nanocarbons. The calcination of the nanocarbon deposited with metal salts yielded nano-perovskites, emulating the morphology of nanocarbons. The accumulation of precursors (i.e., metal salts) on the surface of the nanocarbon during the evaporation of the solvent is the key step in which the precursors are homogeneously mixed prior to calcination. The homogeneity of the precursors facilitated low-temperature calcination that resulted in the formation of nano-perovskites. Various nano-perovskites, including LaMnO3, LaCoO3, LaFeO3, LaNiO3, LaAlO3, LaGaO3, CaMnO3, BaMnO3, SrMnO3, La0.7Sr0.3FeO3, La2CuO4, and Ca2Fe2O5, were successfully synthesized, demonstrating the simplicity and novelty of the method for the general synthesis of nano-perovskites.


Relationship between conformational changes in the dopamine transporter and cocaine-like subjective effects of uptake inhibitors.
Cocaine exerts its stimulatory effect by inhibiting the dopamine transporter (DAT). However, novel benztropine- and rimcazole-based inhibitors show reduced stimulant effects compared with cocaine, despite higher affinity and selectivity for DAT. To investigate possible mechanisms, we compared the subjective effects of different inhibitors with their molecular mode of interaction at the DAT. We determined how different inhibitors affected accessibility of the sulfhydryl-reactive reagent [2-(trimethylammonium)ethyl]-methanethiosulfonate to an inserted cysteine (I159C), which is accessible when the extracellular transporter gate is open but inaccessible when it is closed. The data indicated that cocaine analogs bind an open conformation, whereas benztropine and rimcazole analogs bind a closed conformation. Next, we investigated the changes in inhibition potency of [(3)H]dopamine uptake of the compounds at a mutant DAT (Y335A) characterized by a global change in the conformational equilibrium. We observed a close relationship between the decrease in potencies of inhibitors at this mutant and cocaine-like responding in rats trained to discriminate cocaine from saline injections. Our data suggest that chemically different DAT inhibitors stabilize distinct transporter conformations and that this in turn affects the cocaine-like subjective effects of these compounds in vivo.


Low Serum Vitamin C Status Among Pregnant Women Attending Antenatal Care at General Hospital Dawakin Kudu, Northwest Nigeria.
Vitamin C levels are low in pregnancy. The purpose of this study was to determine serum Vitamins C levels among pregnant women attending antenatal care at a General Hospital in Dawakin Kudu, Kano, and this can help further research to determine the place of Vitamin C supplementation in pregnancy. This was a prospective study of 400 pregnant women who presented for antenatal care in General Hospital Dawakin Kudu, Kano, Nigeria. Research structured questionnaire was administered to 400 respondents. Determination of serum Vitamin C was done using appropriate biochemical methods. Vitamin C deficiency was found in 79.5% of the participants. The values for Vitamin C were 0.20 ± 0.18 mg/dl during the first trimester, 0.50 ± 0.99 mg/dl in the second trimester, and 0.35 ± 0.36 mg/dl in the third trimester and P = 0.001. There is a significant reduction in the serum Vitamins C concentration throughout the period of pregnancy with the highest levels in the second trimester. Therefore, Vitamin C supplementation is suggested during pregnancy, especially for those whose fruit and vegetable consumption is inadequate.


Mitogenomic sequences effectively recover relationships within brush-footed butterflies (Lepidoptera: Nymphalidae).
Mitogenomic phylogenies have revealed well-supported relationships for many eukaryote groups. In the order Lepidoptera, 113 species mitogenomes had been sequenced (May 14, 2014). However, these data are restricted to ten of the forty-three recognised superfamilies, while it has been challenging to recover large numbers of mitogenomes due to the time and cost required for primer design and sequencing. Nuclear rather than mitochondrial genes have been preferred to reconstruct deep-level lepidopteran phylogenies, without seriously evaluating the potential of entire mitogenomes. Next-generation sequencing methods remove these limitations by providing efficiently massive amounts of sequence data. In the present study, we simultaneously obtained a large number of nymphalid butterfly mitogenomes to evaluate the utility of mitogenomic phylogenies by comparing reconstructions to the now quite well established phylogeny of Nymphalidae. We newly obtained 30 nymphalid mitogenomes via pyrosequencing on the Roche 454 GS Junior system, and combined these sequences with publicly accessible data to provide a 70-taxa dataset covering 37 genes for a 15,495 bp alignment. Polymorphic sites were not homogeneously distributed across the gene. Two gene regions, nad6 and 3' end of nad5, were most variable, whereas the cox1 and 5' ends of rrnL were most conserved. Phylogenetic relationships inferred by two likelihood methods were congruent and strongly supported (>0.95 posterior probability; ML bootstrap >85%), across the majority of nodes for multiple partitioning strategies and substitution models. Bayes factor results showed that the most highly partitioned dataset is the preferred strategy among different partitioning schemes. The most striking phylogenetic findings were that the subfamily Danainae not Libytheinae was sister of the remaining brush-footed butterflies and that, within Limenitidini, the genus Athyma was clearly polyphyletic. None of the single-gene phylogenies recovered the highly supported topologies generated on the basis of the whole mitogenomic data. Thirty mitogenomes were assembled with 89% completeness from the contigs of pyrosequencing-derived reads. Entire mitogenomes or higher-quality sequences could be obtained by increasing pyrosequencing read coverage or by additional Sanger sequencing. Our mitogenomic phylogenies provide robust nodal support at a range of levels, demonstrating that mitogenomes are both accurate and efficient molecular markers for inferring butterfly phylogeny.


Captopril and stress-induced hypertension in the borderline hypertensive rat.
Daily exposure to air-jet stress (AJS) causes sustained elevations of blood pressure in borderline hypertensive rats (BHR). It is known that the renin-angiotensin system is activated during episodes of behavioral stress, and the purpose of this study was to assess the involvement of renin-angiotensin system in the development of stress-induced hypertension in the BHR. Four groups of 8- and 9-week-old rats were studied: they received, respectively, oral captopril and AJS; oral captopril without AJS; AJS without captopril; and neither AJS nor captopril. After 10 days of AJS and captopril conditions, femoral and jugular catheters were implanted for the measurement of mean arterial pressure and pressor responses to norepinephrine and tyramine. Blood samples were collected for the measurement of norepinephrine and plasma renin activity. Ten days of AJS caused a significant elevation of blood pressure in BHR exposed to AJS without receiving captopril but not in those animals given captopril concurrently with AJS. Circulating norepinephrine and blood pressure responses to exogenous norepinephrine and tyramine were similar across the four groups. Plasma renin activity was highest in BHR given captopril, and was significantly elevated during an acute episode of AJS. The renin-angiotensin system may be important in the development of stress-induced hypertension in the BHR.


The specificity of disrupted processes in families of adult children of alcoholics.
Children of alcoholics (COAs) have been characterized as an at-risk population in part, because of the dysfunctional family environments that disrupt psychosocial development among offspring exposed to parental alcoholism. This study examined the specificity of problematic family environments to children of alcoholics vs children exposed to other significant family stressors that included parental death, unemployment, separation, divorce, or major illness. University students completed self-report measures of family stressors, family relationship problems, family communication quality, family conflict, and relationship with parents. Based on a family stressor checklist, 20 students exposed only to parental alcoholism and no other family stressors were compared to several other groups exposed to specific family stressors, and to 50 control subjects who had reported no family stressors. Results showed that students from families where a parent is or was an alcoholic, and where there were no other family stressors, recall disturbed family relations no more commonly than students specifically exposed to other family stressors such as parental divorce, death, or major illness, and no more commonly than those who reported no family stressors. Further analyses suggest that the disruption of COAs' family environments may be explained by their increased likelihood of experiencing additional family stressors that can have a disruptive effect, such as parental separation, divorce and unemployment.


Polymerase gene products of hepatitis B viruses are required for genomic RNA packaging as wel as for reverse transcription.
All reactions involving reverse transcription of RNA are segregated from the cytosol within a subviral particle or capsid composed of the major capsid protein, the polymerase and the RNA template. A key step in the formation of these particles is the selective encapsidation of the RNA template. Although an important general feature of the reverse transcription pathway, encapsidation has been carefully studied only for retroviruses. We have now examined the encapsidation reaction in a family of enveloped DNA viruses that replicate by reverse transcription--the hepatitis B viruses (hepadnaviruses). Our results indicate that the hepadnaviral polymerase (P) gene product is required for RNA packaging, and that the encapsidation function of the enzyme can be separated from its DNA polymerase activity. To our knowledge, this is the first description of a role for polymerase gene products in this step of the reverse transcription pathway.


Molecular Survey of Bacterial Zoonotic Agents in Bats from the Country of Georgia (Caucasus).
Bats are important reservoirs for many zoonotic pathogens. However, no surveys of bacterial pathogens in bats have been performed in the Caucasus region. To understand the occurrence and distribution of bacterial infections in these mammals, 218 bats belonging to eight species collected from four regions of Georgia were examined for Bartonella, Brucella, Leptospira, and Yersinia using molecular approaches. Bartonella DNA was detected in 77 (35%) bats from all eight species and was distributed in all four regions. The prevalence ranged 6-50% per bat species. The Bartonella DNA represented 25 unique genetic variants that clustered into 21 lineages. Brucella DNA was detected in two Miniopterus schreibersii bats and in two Myotis blythii bats, all of which were from Imereti (west-central region). Leptospira DNA was detected in 25 (13%) bats that included four M. schreibersii bats and 21 M. blythii bats collected from two regions. The Leptospira sequences represented five genetic variants with one of them being closely related to the zoonotic pathogen L. interrogans (98.6% genetic identity). No Yersinia DNA was detected in the bats. Mixed infections were observed in several cases. One M. blythii bat and one M. schreibersii bat were co-infected with Bartonella, Brucella, and Leptospira; one M. blythii bat and one M. schreibersii bat were co-infected with Bartonella and Brucella; 15 M. blythii bats and three M. schreibersii bats were co-infected with Bartonella and Leptospira. Our results suggest that bats in Georgia are exposed to multiple bacterial infections. Further studies are needed to evaluate pathogenicity of these agents to bats and their zoonotic potential.


Antisense oligonucleotides containing modified bases inhibit in vitro translation of Leishmania amazonensis mRNAs by invading the mini-exon hairpin.
Complementary oligodeoxynucleotides (ODNs) that contain 2-aminoadenine and 2-thiothymine interact weakly with each other but form stable hybrids with unmodified complements. These selectively binding complementary (SBC) agents can invade duplex DNA and hybridize to each strand (Kutyavin, I. V., Rhinehart, R. L., Lukhtanov, E. A., Gorn, V. V., Meyer, R. B., and Gamper, H. B. (1996) Biochemistry 35, 11170-11176). Antisense ODNs with similar properties should be less encumbered by RNA secondary structure. Here we show that SBC ODNs strand invade a hairpin in the mini-exon RNA of Leishmania amazonensis and that the resulting heteroduplexes are substrates for Escherichia coli RNase H. SBC ODNs either with phosphodiester or phosphorothioate backbones form more stable hybrids with RNA than normal base (NB) ODNs. Optimal binding was observed when the entire hairpin sequence was targeted. Translation of L. amazonensis mRNA in a cell-free extract was more efficiently inhibited by SBC ODNs complementary to the mini-exon hairpin than by the corresponding NB ODNs. Nonspecific protein binding in the cell-free extract by phosphorothioate SBC ODNs rendered them ineffective as antisense agents in vitro. SBC phosphorothioate ODNs displayed a modest but significant improvement of leishmanicidal properties compared with NB phosphorothioate ODNs.


Neurodevelopmental disorder in an Egyptian family with a biallelic ALKBH8 variant.
Alkylated DNA repair protein AlkB homolog 8 (ALKBH8) is a member of the AlkB family of dioxygenases. ALKBH8 is a methyltransferase of the highly variable wobble nucleoside position in the anticodon loop of tRNA and thus plays a critical role in tRNA modification by preserving codon recognition and preventing errors in amino acid incorporation during translation. Moreover, its activity catalyzes uridine modifications that are proposed to be critical for accurate protein translation. Previously, two distinct homozygous truncating variants in the final exon of ALKBH8 were described in two unrelated large Saudi Arabian kindreds with intellectual developmental disorder and autosomal recessive 71 (MRT71) syndrome (MIM# 618504). Here, we report a third family-of Egyptian descent-harboring a novel homozygous frame-shift variant in the last exon of ALKBH8. Two affected siblings in this family exhibit global developmental delay and intellectual disability as shared characteristic features of MRT71 syndrome, and we further characterize their observed dysmorphic features and brain MRI findings. This description of a third family with a truncating ALKBH8 variant from a distinct population broadens the phenotypic and genotypic spectrum of MRT71 syndrome, affirms that perturbations in tRNA biogenesis can contribute to neurogenetic disease traits, and firmly establishes ALKBH8 as a novel neurodevelopmental disease gene.


Practical application of wastewater reuse in tourist resorts.
A medium-scale membrane bioreactor was tested in a large tourist resort on the south-western coast of Turkey with the treated wastewater subsequently being used for irrigational purposes. The wastewater treatment system was designed to eliminate carbonaceous and nitrogenous substances. Treatment efficiency was monitored by means of regular chemical and microbiological analyses. Information was collected on water use at different locations of the hotel. Specific values based on the number of guests were determined. Wastewater streams from kitchen, laundry and rooms were analysed to investigate the various contribution from these points. The social acceptance of the guests concerning the on-site wastewater treatment and reuse in the hotel was analysed using a questionnaire. The investigations indicated that the treated wastewater provides the required chemical and hygienic conditions to satisfy requirement for its reuse in irrigation. The acceptance by guests was encouraging for such applications.


On the affinity and specificity of quercetin for DNA.
The potential of quercetin (QUE), being a member of the whole family of structurally different flavonoids, to serve as an anti-tumor agent has been recognized, but not fully understood. The interactions between DNA and a series of the flavonoids have so far been mainly investigated using a variety of experimental techniques. Herein, the specificity of QUE for DNA is explored using sophisticated density functional theory (DFT) methods employed to generate the optimized structure of QUE in complex with adenine (A), guanine (G), thymine (T) and cytosine (C), respectively. As far as a preference of QUE is concerned, structural and energetic as well as NMR chemical shift arguments clearly indicate a highest for G and a lowest for C. This observation is further substantiated by analyzing the binding modes of QUE docked in a quadruplex receptor structure of DNA and in a duplex receptor structure of DNA. Among all possible single point mutations of the DNA quadruplex and duplex residues, several critical ones causing a conspicuous stabilizing effect on the original complexes of QUE with the DNA receptors are identified. Consequently, several fundamental standpoints shedding new light on the molecular mechanisms underlying the interactions between QUE and DNA are discussed.


Ultrasound Shear Wave Elastography and Contrast Enhancement: Effective Biomarkers in Crohn's Disease Strictures.
Inflammation, fibrosis, and muscular hypertrophy lead to thickened bowel in Crohn's disease forming strictures. Ultrasound shear wave elastography (SWE) measures bowel wall stiffness. Contrast-enhanced ultrasound (CEUS) uniquely detects bowel wall inflammation. We aim to correlate SWE of ileal Crohn's disease in vivo to CEUS peak enhancement and pathology grades of inflammation, fibrosis, and muscular hypertrophy. In a prospective institutional review board-approved study, 105 consecutive ileal patients with Crohn's disease received ultrasound. At maximal bowel wall thickness (>4 mm), SWE and CEUS were performed. Fifteen patients had ileal resection within a mean time interval of 71.0 ± 66.9 days. Pathology scores for inflammation, fibrosis, and muscular hypertrophy were compared with SWE and CEUS measurements. Mean in vivo SWE velocity for patients with and without surgery was 2.8 ± 0.7 and 2.2 ± 0.8 m/s (P < 0.01), respectively. In all ileal specimens, chronic exceeded active inflammatory change (P < 0.001). There was an inverse relationship between CEUS peak enhancement and both fibrosis, r = -0.59, P = 0.02, and SWE velocity measurements, r = -0.61, P = 0.03. Strictured bowel specimens had more smooth muscle hypertrophy than fibrosis, P < 0.001. There was moderate correlation between SWE and muscular hypertrophy, r = 0.59, P = 0.02 and no significant relationship between SWE and fibrosis scores (P > 0.05). Stiffer bowel from smooth muscle hypertrophy increases SWE measurements. We report a novel relation of high SWE with muscle hypertrophy, and inverse relationship with CEUS peak enhancement; providing differentiation between active and chronic bowel wall inflammation to improve selection between medical therapy and surgery.


Rechallenge for Patients With RAS and BRAF Wild-Type Metastatic Colorectal Cancer With Acquired Resistance to First-line Cetuximab and Irinotecan: A Phase 2 Single-Arm Clinical Trial.
Based on a small retrospective study, rechallenge with cetuximab-based therapy for patients with KRAS wild-type metastatic colorectal cancer (mCRC) who were previously treated with the same anti-epidermal growth factor receptor-based regimen might be efficacious. Recent data suggest the role of liquid biopsy as a tool to track molecular events in circulating tumor DNA (ctDNA). To prospectively assess the activity of cetuximab plus irinotecan as third-line treatment for patients with RAS and BRAF wild-type mCRC who were initially sensitive to and then resistant to first-line irinotecan- and cetuximab-based therapy. Multicenter phase 2 single-arm trial conducted from January 7, 2015, to June 19, 2017. Liquid biopsies for analysis of ctDNA were collected at baseline. Main eligibility criteria included RAS and BRAF wild-type status on tissue samples; prior first-line irinotecan- and cetuximab-based regimen with at least partial response, progression-free survival of at least 6 months with first-line therapy, and progression within 4 weeks after last dose of cetuximab; and prior second-line oxaliplatin- and bevacizumab-based treatment. Biweekly cetuximab, 500 mg/m2, plus irinotecan, 180 mg/m2. Overall response rate according to the Response Evaluation Criteria in Solid Tumors, version 1.1. Secondary end points included progression-free survival and overall survival and, as an exploratory analysis, RAS mutations in ctDNA. Twenty-eight patients (9 women and 19 men; median age, 69 years [range, 45-79 years]) were enrolled. Six partial responses (4 confirmed) and 9 disease stabilizations were reported (response rate, 21%; 95% CI, 10%-40%; disease control rate, 54%; 95% CI, 36%-70%). Primary end point was met because lower limit of 95% CI of response rate was higher than 5%. RAS mutations were found in ctDNA collected at rechallenge baseline in 12 of 25 evaluable patients (48%). No RAS mutations were detected in samples from patients who achieved confirmed partial response. Patients with RAS wild-type ctDNA had significantly longer progression-free survival than those with RAS mutated ctDNA (median progression-free survival, 4.0 vs 1.9 months; hazard ratio, 0.44; 95% CI, 0.18-0.98; P = .03). This is the first prospective demonstration that a rechallenge strategy with cetuximab and irinotecan may be active in patients with RAS and BRAF wild-type mCRC with acquired resistance to first-line irinotecan- and cetuximab-based therapy. The evaluation of RAS mutational status on ctDNA might be helpful in selecting candidate patients. ClinicalTrials.gov Identifier: NCT02296203.


Characteristics of Patients with Post-Colonoscopy Unplanned Hospital Visit: A Retrospective Single-Center Observational Study.
Colonoscopy, although a low-risk procedure, is not without associated adverse events. The rates of major adverse events such as perforation and bleeding after a colonoscopy are well reported. The rates of minor incidents following a colonoscopy, however, are less well examined. Recently the Centers for Medicare and Medicaid Services (CMS) started public reporting on the quality of outpatient endoscopy facilities by using a measure of risk-standardized rates of unplanned hospital visits within 7 days of colonoscopy. We intended to record and present the characteristics of our patient population who had an unplanned hospital visit within 7 days after undergoing colonoscopy in an outpatient setting. This is a retrospective single-center observational study. During the study period of July 2018 to December 2019, we reviewed charts of all patients who returned to the emergency room within a week of undergoing an outpatient colonoscopy. Patient demographics, clinical data and details of colonoscopy were collected and analyzed. Of the 5344 outpatient colonoscopies performed, our post-colonoscopy emergency room visit rate was 1.05% (n=56). The mean age of the participants was 58 years and 55% were male; 32% of our patients reported gastrointestinal symptoms such as abdominal pain or gastrointestinal bleeding. Patients with gastrointestinal symptoms had a higher rate of polypectomies performed (36.4% vs 11.8%, P = 0.04) and reported higher illicit drug use (31.9% vs 5.9%, P = 0.02) compared with those with non-gastrointestinal complaints. After colonoscopy, 41% of the patients reported reasons for emergency room visits that were entirely unrelated to the procedure. Our study highlights that unplanned visits within 7 days of colonoscopy are not necessarily related to the procedure, and those that are, tend to be due to unavoidable patient factors. Hence the CMS measure may not be an accurate determinant of the quality of procedure or facility care delivered.


Complex pediatric orbital fractures combined with traumatic brain injury: treatment and follow-up.
The treatment of orbital fractures aims at the restoration of orbital anatomy and prevention of posttraumatic sequels. The treatment of facial fractures in patients with traumatic brain injury may necessitate a postponement of fracture surgery to allow for brain recovery. However, such delay of reconstruction in complex orbital fractures may lead to inferior results. Fourteen pediatric patients with complex orbital fractures were retrospectively reviewed. Eleven patients (79%) had brain injury, with a mean Glasgow Coma Scale score of 8.6 at initial evaluation. Fracture patterns were highly complex with many bilateral and multiple orbital wall fractures. Six patients (43%) had injuries to the medial canthal tendon, as part of a naso-orbito-ethmoid fracture. Mean follow-up time was 4.7 years (range, 1.2-13.1 years). Eleven patients (79%) had reconstructive surgery early (within 3 weeks), and 3 (29%) patients had the surgery postponed. Four patients had secondary surgeries for enophthalmos and medial canthal dislocation. Eighty-two percent of patients with brain injury had a good overall neurologic outcome (Glasgow Outcome Scale score of 1 or 2). Sixty-seven percent of patients had a good aesthetic outcome without any need for revisional surgery. Medial canthal injury and late surgery both correlated with an inferior aesthetic result. In conclusion, good results with respect to mental outcome and aesthetics can be achieved in highly complex orbital fractures combined with brain injury. Delayed surgery and medial canthal tendon injury predispose for posttraumatic sequels and secondary surgery.


Evaluating the effects of eating disorder memoirs on readers' eating attitudes and behaviors.
More than 50 individuals have published eating disorder (ED) memoirs. The current study was the first to test whether memoirs affect readers' eating attitudes and behaviors, and whether they normalize and/or glamorize EDs. Fifty female undergraduates read an ED or control memoir. Before and afterward, participants completed the 26-item Eating Attitudes Test (EAT-26), the Eating Disorders Inventory (EDI) Drive for Thinness subscale, a measure of perceived ED symptom prevalence, and an Implicit Association Test (IAT) measuring associations between anorexia and glamour/danger. Participants in the ED condition did not demonstrate significant changes in the EAT-26, the EDI Drive for Thinness subscale, perceived symptom prevalence, or IAT associations compared with controls. Before reading, the EAT-26 and EDI Drive for Thinness subscale correlated positively with perceived symptom prevalence and strength of the IAT association between anorexia and glamour. ED memoirs appear to have little effect on undergraduates' eating attitudes and behaviors. Future research should investigate whether memoirs affect individuals with preexisting eating pathology, who may normalize and glamorize ED symptoms.


Transcriptional regulation of Wnt inhibitory factor-1 by Miz-1/c-Myc.
The Wnt signaling pathway is capable of self-regulation through positive and negative feedback mechanisms. For example, the oncoprotein c-Myc, which is upregulated by Wnt signaling activity, participates in a positive feedback loop of canonical Wnt signaling through repression of Wnt antagonists DKK1 and SFRP1. In this study, we investigated the mechanism of Wnt inhibitory factor-1 (WIF-1) silencing. Mapping of CpG island methylation of the WIF-1 promoter reveals regional methylation (-295 to -95 bp from the transcription start site) that correlates with transcriptional silencing. We identified Miz-1 as a direct activator of WIF-1 transcriptional activity, which is found at WIF-1 promoter. In addition, we show that c-Myc contributes to WIF-1 transcriptional repression in a Miz-1-dependent manner. Although the transient repression mediated by Miz-1/c-Myc is independent of de novo methylation, the stable repression by this complex is associated with CpG island methylation of the critical -295 to -95-bp region of the WIF-1 promoter. Importantly, Miz-1 and c-Myc are found at WIF-1 promoter in WIF-1 non-expressing cell lines DLD-1 and 209myc. Transient knockdown or somatic knockout of c-Myc in DLD-1 failed to restore WIF-1 expression suggesting that c-Myc is involved in initiating rather than maintaining WIF-1 epigenetic silencing. In a genome-wide screen, DNAJA4, TGFβ-induced and TRIM59 were repressed by c-Myc overexpression and DNA promoter hypermethylation. Our data reveal novel insights into c-Myc-mediated DNA methylation-dependent transcriptional silencing, a mechanism that might contribute to the dysregulation of Wnt signaling in cancer.


Preparation of nurses to meet the needs of an ethnically diverse society: educational implications.
In recognizing the multiethnic composition of contemporary society in the UK, this paper considers the implications for nurse education in respect of preparing practitioners who are capable of meeting the needs of an ethnically diverse population. It begins by considering briefly some contextual issues relating to the health needs of ethnic minorities and how they impact upon nursing practice. The literature clearly indicates some of the complexities that nurses face in attempting to deliver care which is responsive to the individual needs of patients in a multicultural context. The question then arises as to how nurse education can most effectively prepare future practitioners. Consideration is given to the curriculum content, methods and approaches that may be utilized together with different teaching and learning strategies. The case is made for the need for nurse educators to also consider the recruitment and support of students from ethnic minority groups, if nursing is to make progress in responding more appropriately to the needs of ethnic minority groups in the UK.


Learning from healthcare workers' experiences with personal protective equipment during the COVID-19 pandemic in Aotearoa/New Zealand: a thematic analysis and framework for future practice.
Safety and welfare are critical as pandemic-related demands on the healthcare workforce continue. Access to personal protective equipment (PPE) has been a central concern of healthcare workers throughout the COVID-19 pandemic. Against the backdrop of an already strained healthcare system, our study aimed to explore the experiences of healthcare workers with PPE during the first COVID-19 surge (February-June 2020) in Aotearoa/New Zealand (NZ). We also aimed to use these findings to present a strengths-based framework for supporting healthcare workers moving forward. Web-based, anonymous survey including qualitative open-text questions. Questions were both closed and open text, and recruitment was multimodal. We undertook inductive thematic analysis of the dataset as a whole to explore prominent values related to healthcare workers' experiences. October-November 2020 in New Zealand. 1411 healthcare workers who used PPE during surge one of the COVID-19 pandemic. We identified four interactive values as central to healthcare workers' experiences: transparency, trust, safety and respect. When healthcare workers cited positive experiences, trust and safety were perceived as present, with a sense of inclusion in the process of stock allocation and effective communication with managers. When trust was low, with concerns over personal safety, poor communication and lack of transparency resulted in perceived lack of respect and distress among respondents. Our proposed framework presents key recommendations to support the health workforce in terms of communication relating to PPE supply and distribution built on those four values. Healthcare worker experiences with PPE access has been likened to 'the canary in the coalmine' for existing health system challenges that have been exacerbated during the COVID-19 pandemic. The four key values identified could be used to improve healthcare worker experience in the future.


Performance statistics of a head-operated force-reflecting rehabilitation robot system.
Researchers in the rehabilitation engineering community have been designing and developing a variety of passive/active devices to help persons with limited upper extremity function to perform essential daily manipulations. Devices range from low-end tools such as head/mouth sticks to sophisticated robots using vision and speech input. While almost all of the high-end equipment developed to date relies on visual feedback alone to guide the user providing no tactile or proprioceptive cues, the "low-tech" head/mouth sticks deliver better "feel," because of the inherent force feedback through physical contact with the user's body. However, the disadvantage of a conventional head/mouth stick is that it can only function in a limited workspace and the performance is limited by the user's strength. It therefore seems reasonable to attempt to develop a system that exploits the advantages of the two approaches: the power and flexibility of robotic systems with the sensory feedback of a headstick. The system presented in this paper reflects the design philosophy stated above. This system contains a pair of master-slave robots with the master being operated by the user's head and the slave acting as a telestick. Described in this paper are the design, control strategies, implementation and performance evaluation of the head-controlled force-reflecting telestick system.


Emotional graphic cigarette warning labels reduce the electrophysiological brain response to smoking cues.
There is an ongoing public debate about the new graphic warning labels (GWLs) that the Food and Drug Administration (FDA) proposes to place on cigarette packs. Tobacco companies argued that the strongly emotional images FDA proposed to include in the GWLs encroached on their constitutional rights. The court ruled that FDA did not provide sufficient scientific evidence of compelling public interest in such encroachment. This study's objectives were to examine the effects of the GWLs on the electrophysiological and behavioral correlates of smoking addiction and to determine whether labels rated higher on the emotional reaction (ER) scale are associated with greater effects. We studied 25 non-treatment-seeking smokers. Event-related potentials (ERPs) were recorded while participants viewed a random sequence of paired images, in which visual smoking (Cues) or non-smoking (non-Cues) images were preceded by GWLs or neutral images. Participants reported their cigarette craving after viewing each pair. Dependent variables were magnitude of P300 ERPs and self-reported cigarette craving in response to Cues. We found that subjective craving response to Cues was significantly reduced by preceding GWLs, whereas the P300 amplitude response to Cues was reduced only by preceding GWLs rated high on the ER scale. In conclusion, our study provides experimental neuroscience evidence that weighs in on the ongoing public and legal debate about how to balance the constitutional and public health aspects of the FDA-proposed GWLs. The high toll of smoking-related illness and death adds urgency to the debate and prompts consideration of our findings while longitudinal studies of GWLs are underway.


Self- and co-regulation of physiological activity during mother-daughter interactions: The role of adolescent non-suicidal self-injury.
Non-suicidal self-injury (NSSI) is a significant public health concern that is thought to increase risk for future self-injurious behaviors, including suicide attempts. Notably, NSSI is especially prevalent among adolescents, which underscores a critical need to identify modifiable risk factors that could be targeted to reduce future risk. The current study examined self- and co-regulation of physiological responses during mother-daughter interactions in adolescent girls with and without a history of NSSI. Participants were 60 girls aged 13-17 with (n = 27) and without (n = 33) a history of NSSI and their mothers. Adolescents and their mothers completed positive and negative interaction tasks during which physiological reactivity was assessed via respiratory sinus arrhythmia (RSA). Using Actor-Partner Interdependence Modeling (APIM), we found that adolescents with an NSSI history demonstrated a higher RSA setpoint than adolescents without this history during the negative, but not positive, interaction task. In addition, there were differences in co-regulation during the negatively valenced interaction, such that mothers of daughters with NSSI were more reactive to fluctuations in their daughters' RSA than mothers of daughters without an NSSI history. These findings highlight intra- and interpersonal aspects of physiological dysregulation associated with NSSI that could provide promising targets of intervention to reduce future risk in adolescent girls.


Prostate cancer screening: Attitudes and practices of family physicians in Ontario.
: The utility of prostate cancer screening is controversial. We sought to determine whether Ontario's family physicians believe it is beneficial and to characterize their screening protocols. : A survey was developed with input from urologists, family physicians and the Ontario Medical Association's Section on General and Family Practice. Questions covered three domains: (1) demographics, (2) beliefs about screening utility and (3) screening practices. All 7302 family physicians in Ontario were invited by email to complete the online survey. : A total of 969 physicians completed the survey; 955 (52.0% male, 48.0% female) were included. Most (80.97%) use prostate-specific antigen (PSA) and digital rectal examination (DRE) for screening; 9.4% use DRE alone and 7.15% PSA. Of the respondents, 8.3% do not offer prostate cancer screening. Most physicians begin offering screening at age 50 (72.9%) and stop at ages 70 or 80 (68.4%); 17.9% offer lifelong screening. In response to the statement "screening with DRE provides a survival benefit," 37.6% and 32.6 agreed and disagreed, respectively. For "screening with PSA provides a survival benefit," 43.3% agreed and 31.0% disagreed. For the statement "the benefits of prostate cancer screening outweigh the risks," 51.4% agreed and 22.0% disagreed. : Although 91.7% of respondents offer prostate cancer screening, they are divided over its utility. Only 51.4% were convinced that the benefits outweighed the harms. There is significant variability between physicians' screening protocols. A limitation of this study is the possibility of selection bias. Nevertheless, this is the largest sample of Ontario family physicians ever surveyed about prostate cancer screening and highlights divergent physician practices and a need for more conclusive evidence on screening utility.


Fiber-optic pulsed photothermal radiometry for fast surface-temperature measurements.
Temperature measurement based on pulsed photothermal radiometry is described. In this technique a body is irradiated by a laser pulse and its temperature is inferred from the shape of the emitted photothermal-signal curve. A prototypical system based on a pulsed CO(2) laser, an IR detector, and IR-transmitting silver halide optical fibers was constructed and used to evaluate the feasibility of this technique. An important feature of the technique is that changes in sample emissivity or geometric factors do not introduce errors in the temperature determination. Theory, simulation, and experimental results are given and discussed.


Training and technical assistance to enhance capacity building between prevention research centers and their partners.
The Centers for Disease Control and Prevention has administered the Prevention Research Centers Program since 1986. We quantified the number and reach of training programs across all centers, determined whether the centers' outcomes varied by characteristics of the academic institution, and explored potential benefits of training and technical assistance for academic researchers and community partners. We characterized how these activities enhanced capacity building within Prevention Research Centers and the community. The program office collected quantitative information on training across all 33 centers via its Internet-based system from April through December 2007. Qualitative data were collected from April through May 2007. We selected 9 centers each for 2 separate, semistructured, telephone interviews, 1 on training and 1 on technical assistance. Across 24 centers, 4,777 people were trained in 99 training programs in fiscal year 2007 (October 1, 2006-September 30, 2007). Nearly 30% of people trained were community members or agency representatives. Training and technical assistance activities provided opportunities to enhance community partners' capacity in areas such as conducting needs assessments and writing grants and to improve the centers' capacity for cultural competency. Both qualitative and quantitative data demonstrated that training and technical assistance activities can foster capacity building and provide a reciprocal venue to support researchers' and the community's research interests. Future evaluation could assess community and public health partners' perception of centers' training programs and technical assistance.


Quantum dots in molecular detection of disease.
The unique photophysical properties of semiconductor quantum dots (QDs) have made them ideal for use as spectral labels and luminescent probes. In this review, applications are presented in which QDs function as active participants in nanoscale biosensor assemblies, where replacing traditional molecular fluorophores results in improved assay performance. Specific focus is on disease detection with applications including multiplexed target detection, mutation detection by coincidence analysis and QD-based FRET reporters for miRNA detection and DNA methylation analysis.


Intraoperative Regional Cerebral Oxygenation During Pediatric Thoracoscopic Surgery: A Systematic Review.
Background: Ventilating a pediatric patient during thoracoscopy is challenging. Few studies have highlighted the impact of capnothorax in children by measuring regional cerebral oxygen saturation (rcSO2) with near infrared spectroscopy. In this systematic review, we aimed to summarize the data from relevant studies and assess whether thoracoscopy in children is associated with intraoperative pathological cerebral desaturation. Methods: The authors systematically searched four databases for relevant studies on the measurement of rcSO2 during pediatric thoracoscopic procedures. The primary outcome was the proportion of patients with pathological desaturation, that is, >20% decline in the intraoperative rcSO2. Risk of bias among the included studies was estimated using the Newcastle-Ottawa scale. Results: The systematic search resulted in 776 articles, of which 7 studies were included in the analysis. In total, 88 patients (99 procedures) with an age ranging from 0 days to 8.1 years were included. Of these, 43 (49%) patients were neonates. The included cohort had esophageal atresia and tracheoesophageal fistula (n = 26), long-gap esophageal atresia (n = 5), congenital diaphragmatic hernia (n = 14), and congenital pulmonary airway malformations and other conditions needing lung resection (n = 43). Of the total 99 procedures, pathological desaturation was noticed in 13 (13.1%, 95% confidence interval 7.2-21.4) of them. Upon quality assessment, most of the studies were weaker in the selection and comparability domains. Conclusion: In this review, pathological cerebral desaturation was noticed in 13.1% of the pediatric thoracoscopic procedures. However, due to limited methodological quality of the included studies, further randomized multicentric studies comparing rcSO2 in open versus thoracoscopic surgeries are needed to derive definitive conclusions.


Chronic administration of nalmefene leads to increased food intake and body weight gain in mice.
Nalmefene is an orally available opioid receptor antagonist that has been shown to suppress appetite in humans, but its effects on chronic food intake and body weight remain unclear. Here, we report that chronic (21-day) oral administration of nalmefene at 2 or 10 mg/kg/day in diet-induced obese (DIO) mice led to significant increases (9-11%) in cumulative food intake. Mice in the nalmefene-treated groups also gained body weight at a rate faster than the control. Body composition analysis showed that the extra body weight gains in the treated animals were mostly due to increased fat accumulation. Since acute nalmefene treatment showed a trend toward a decrease rather than an increase in food intake, it is possible that the orexigenic effect of chronic oral administration of nalmefene was caused by pharmacologically active metabolites rather than the drug itself. Our results argue against the potential use of nalmefene for treating human obesity.


Isolation and properties of highly purified Halobacterium cutirubrum deoxyribonucleic acid-dependent ribonucleic acid polymerase.
1. The subunits alpha and beta of Halobacterium cutirubrum DNA-dependent RNA polymerase have been purified to electrophoretic homogeneity. Both have mol.wt. 18000 and they are required in equimolar amounts for optimum activity. 2. The instability of the complete enzyme, alphabeta, in the absence of salt is due to the rapid inactivation of the beta subunit in these conditions. 3. Nearest-neighbour analysis of the product formed on poly[d(A-T)] as template shows that the enzyme copies the latter accurately. 4. The enzyme initiates new chains with purine nucleoside triphosphates exclusively. 5. The product obtained in the standard assay conditions contains some high mol.wt. (>16S) material, but consists primarily of short chains, of average length 70-80 nucleotide units. 6. The template specificity of the complete enzyme has been studied at high and low ionic strength. Its extreme dependence on salt concentration is unrelated to the gross overall base composition of the DNA used. 7. T(7) DNA is transcribed asymmetrically and the enzyme selectively copies the T(7) ;early' genes. 8. Preliminary amino acid analyses of alpha and beta subunits show that their overall content of acidic, basic and neutral amino acids does not differ appreciably from that of Escherichia coli RNA polymerase.


Alterations in visual cortical activation and connectivity with prefrontal cortex during working memory updating in major depressive disorder.
The present study examined the impacts of major depressive disorder (MDD) on visual and prefrontal cortical activity as well as their connectivity during visual working memory updating and related them to the core clinical features of the disorder. Impairment in working memory updating is typically associated with the retention of irrelevant negative information which can lead to persistent depressive mood and abnormal affect. However, performance deficits have been observed in MDD on tasks involving little or no demand on emotion processing, suggesting dysfunctions may also occur at the more basic level of information processing. Yet, it is unclear how various regions in the visual working memory circuit contribute to behavioral changes in MDD. We acquired functional magnetic resonance imaging data from 18 unmedicated participants with MDD and 21 age-matched healthy controls (CTL) while they performed a visual delayed recognition task with neutral faces and scenes as task stimuli. Selective working memory updating was manipulated by inserting a cue in the delay period to indicate which one or both of the two memorized stimuli (a face and a scene) would remain relevant for the recognition test. Our results revealed several key findings. Relative to the CTL group, the MDD group showed weaker postcue activations in visual association areas during selective maintenance of face and scene working memory. Across the MDD subjects, greater rumination and depressive symptoms were associated with more persistent activation and connectivity related to no-longer-relevant task information. Classification of postcue spatial activation patterns of the scene-related areas was also less consistent in the MDD subjects compared to the healthy controls. Such abnormalities appeared to result from a lack of updating effects in postcue functional connectivity between prefrontal and scene-related areas in the MDD group. In sum, disrupted working memory updating in MDD was revealed by alterations in activity patterns of the visual association areas, their connectivity with the prefrontal cortex, and their relationship with core clinical characteristics. These results highlight the role of information updating deficits in the cognitive control and symptomatology of depression.


Prospective study of histoplasmosis in patients infected with human immunodeficiency virus: incidence, risk factors, and pathophysiology.
Histoplasmosis is a common opportunistic infection in patients with human immunodeficiency virus (HIV) infection who reside in areas where Histoplasma capsulatum is endemic. We undertook a prospective study of a cohort of 304 HIV-Infected patients in Kansas City from October 1990 through March 1993 to define the incidence-specific risk factors, and pathophysiology of histoplasmosis. The annual incidence of histoplasmosis was 4.7%; 74% of the patients with histoplasmosis were symptomatic (all of whom had disseminated disease). A history of exposure to chicken coops, a positive baseline serology for complement-fixing antibodies to Histoplasma mycelium antigen, and a baseline CD4+ lymphocyte count of < 150/microL were associated with an increased risk for histoplasmosis. Histoplasmin reactivity and the presence of pulmonary calcifications were not useful markers for patients at high risk. Symptomatic infection occurred in 9.9% of patients with evidence of prior exposure to H. capsulatum, in 4.0% of patients without documented prior exposure, and in 3.0% of patients who were anergic; these findings suggest that the pathophysiology of histoplasmosis in patients with AIDS involves reactivation of latent infection in some cases and dissemination of exogenously acquired infection in other cases.


DNA methylation mediates persistent epileptiform activity in vitro and in vivo.
Epilepsy is a chronic brain disorder involving recurring seizures often precipitated by an earlier neuronal insult. The mechanisms that link the transient neuronal insult to the lasting state of epilepsy are unknown. Here we tested the possible role of DNA methylation in mediating long-term induction of epileptiform activity by transient kainic acid exposure using in vitro and in vivo rodent models. We analyzed changes in the gria2 gene, which encodes for the GluA2 subunit of the ionotropic glutamate, alpha-amino-3-hydroxy-5-methyl-4-isoxazole proprionic acid receptor and is well documented to play a role in epilepsy. We show that kainic acid exposure for two hours to mouse hippocampal slices triggers methylation of a 5' regulatory region of the gria2 gene. Increase in methylation persists one week after removal of the drug, with concurrent suppression of gria2 mRNA expression levels. The degree of kainic acid-induced hypermethylation of gria2 5' region varies between individual slices and correlates with the changes in excitability induced by kainic acid. In a rat in vivo model of post kainic acid-induced epilepsy, we show similar hypermethylation of the 5' region of gria2. Inter-individual variations in gria2 methylation, correlate with the frequency and intensity of seizures among epileptic rats. Luciferase reporter assays support a regulatory role for methylation of gria2 5' region. Inhibition of DNA methylation by RG108 blocked kainic acid-induced hypermethylation of gria2 5' region in hippocampal slice cultures and bursting activity. Our results suggest that DNA methylation of such genes as gria2 mediates persistent epileptiform activity and inter-individual differences in the epileptic response to neuronal insult and that pharmacological agents that block DNA methylation inhibit epileptiform activity raising the prospect of DNA methylation inhibitors in epilepsy therapeutics.


Exercise-induced translocation of skeletal muscle glucose transporters.
Skeletal muscle contractile activity results in increased rates of glucose transport that are associated with an increase in the number and activity of plasma membrane glucose transporters. In the current study it was determined whether exercise causes a translocation of glucose transporters from an intracellular pool to the plasma membrane and whether exercise and insulin stimulate the same glucose transporter protein. Plasma membrane glucose transporter number, measured by cytochalasin B binding, increased from 10.1 +/- 0.73 to 15.0 +/- 1.4 pmol/mg protein (P less than 0.01) in muscle of exercised rats, whereas microsomal membrane transporters decreased significantly from 6.0 +/- 0.7 to 4.2 +/- 0.4 pmol/mg protein (P less than 0.05). Western blot analysis using the monoclonal antibody mAb 1F8 (specific for GLUT-4) demonstrated a 45% increase in plasma membrane GLUT-4 from exercised skeletal muscle compared with controls, whereas microsomal membranes from the exercised muscle had a concomitant 25% decrease in GLUT-4 protein. These data suggest that exercise recruits transporters to the plasma membrane from an intracellular microsomal pool, similar to the translocation of transporters that occurs with insulin stimulation. Furthermore, both exercise and insulin stimulate the translocation of GLUT-4 in skeletal muscle, while GLUT-1 is not altered.


Effect of natural starters used for sourdough bread in Morocco on phytate biodegradation.
Phytase activity was studied in natural sourdough bread starters to determine physicochemical characteristics (phytic acid hydrolysis, dough rising capacity and pH) in the flour and during sourdough fermentation. Fermentation microorganisms (yeasts and lactic acid bacteria) were also characterized. Results showed a decrease of phytic acid in sourdoughs started with traditional starters, and wide variation in phytase activity. Microorganism counts were high at the end of fermentation, indicating higher fermenting activity of the starters. Yeast populations showed wide variation and lactic acid bacteria had high counts in the fermentation. Phytase activity was demonstrated in starter cultures made of lactic acid bacteria and yeast isolates, the most interesting of which were Saccharomyces cerevisiae combined with Lactobacillus plantarum and Leuconostoc mesenteroides.


Experimental and computational studies on a protonated 2-pyridinyl moiety and its switchable effect for the design of thermolytic devices.
1D and 2D NMR investigations as well as computational studies, including static quantum-mechanics calculations, density function theory formalism, and classical molecular dynamics, were applied to determine the protonation sites in the thermolabile protecting group (TPG) containing a 2-pyridynyl moiety within its structure. This protecting group has three possible sites for protonation: an azomethine (pyridinic) atom (N1), 2-aminoethanol residue (N2), and 4-amino substituent (N4). Our investigations showed that the protonation mainly occurs on the N1 atom. Such protonation seems to be a major inhibitory factor in the thermal removal of 2-pyridynyl TPG by the "chemical switch" approach and decreases the aromaticity of the pyridine ring. We also discussed possible participation of N2 nitrogen in irreversible intramolecular cyclization under acidic conditions.


Cash transfer programs and child health and family economic outcomes: a systematic review.
Family income is an important determinant of child and parental health. In Canada, cash transfer programs to families with children have existed since 1945. This systematic review aimed to examine the association between cash transfer programs to families with children and health outcomes in Canadian children (ages 0 to 18) as well as family economic outcomes. We reviewed academic and grey literature published up to November 2021. Additional studies were identified through reference review. We included any study that examined children 0-18 years old and/or their parents, took place in Canada and reported Canada-specific data, and reported child, youth and/or parental health outcomes, as well as family economic outcomes. Risk of bias was assessed by two reviewers using a modified Newcastle-Ottawa Scale. Our search yielded 23 studies meeting the inclusion criteria out of 7052 identified. Eight studies in total measured child health outcomes, including birth outcomes, child overall health, and developmental and behavioural outcomes, and four directly addressed parental health, including mental health, injuries, and obesity. Most studies reported generally positive associations, though some findings were specific to certain subgroups. Some studies also examined fertility and labour force participation outcomes, which described varying effects. Cash transfer programs to families with children in Canada are associated with better child and parental health outcomes. Additional research is needed to evaluate the mechanisms of effects, and to identify which types and levels of government transfers are most effective, and target populations, to optimize the positive effects of these benefits.


Orthopaedic Gene Therapy: Twenty-Five Years On.
Orthopaedics pioneered the expansion of gene therapy beyond its traditional scope of diseases that are caused by rare single-gene defects. Orthopaedic applications of gene therapy are most developed in the areas of arthritis and regenerative medicine, but several additional possibilities exist. Invossa, an ex vivo gene therapeutic for osteoarthritis, was approved in South Korea in 2017, but its approval was retracted in 2019 and remains under appeal; a Phase-III clinical trial of Invossa has restarted in the U.S. There are several additional clinical trials for osteoarthritis and rheumatoid arthritis that could lead to approved gene therapeutics for arthritis. Bone-healing and cartilage repair are additional areas that are attracting considerable research; intervertebral disc degeneration and the healing of ligaments, tendons, and menisci are other applications of interest. Orthopaedic tumors, genetic diseases, and aseptic loosening are additional potential targets. If successful, these endeavors will expand the scope of gene therapy from providing expensive medicines for a few patients to providing affordable medicines for many.


Drug interactions: a study and evaluation of their incidence in Victoria.
This study attempted to quantify the incidence of drug interactions in general practice prescribing in Victoria. We used a computer program to search 428 prescriptions and associated medication records for possible interactions. We made 2445 drug/drug comparisons; the computer program generated 37 possible interaction alerts, none of which warranted a change in therapy. This type of computer program, unless extremely sophisticated, can only serve as a primary alert; a health professional should then check the validity of interactions it identifies.


[Fluorescence angiography of the anterior eye segment in the evaluation of impaired microcirculation within the diagnosis of severity of eye burns].
We used fluorescence angiography to study the vascular microcirculation in the anterior eye segment of 14 patients with severe eye bums. The severity of eye burns is not only related with an impaired blood circulation in the conjunctiva, it also depends on a microcirculation status of episcleral vessels. Different degrees of permeability of fluorescein through the conjunctival or episcleral vessels make it possible to specify circulation disorders in both cases. Whether the impaired microcirculation in the anterior eye segment is transient or not can be established through a second fluorescence angiography made during days 7-8 from the burn trauma. The severity of eye burn depends directly on the sizes of ischemic regions of the sclera.


Public Awareness, Prevalence and Potential Determinants of Bovine Tuberculosis in Selected Districts of Gamo Zone, Southern Ethiopia.
Bovine tuberculosis (bTB) is infectious chronic disease of animals mainly caused by bacillus Mycobacterium bovis. It is known endemic disease of cattle in Ethiopia. In the current study sites, there is little information on bovine tuberculosis. Thus, this study was aimed to assess public awareness; and estimate prevalence and potential a risk factors of bovine tuberculosis. A cross-sectional study was conducted, November 2018 to May 2019, on dairy cattle randomly selected from smallholder farms of Arba Minch Zuria and Chencha districts. Comparative intradermal tuberculin test (CIDT) was used in the diagnosis of bTB in dairy animals. CIDT was administered to 221 dairy cattle. Questionnaire survey was performed on 110 smallholders to assess risk factors accountable for the event of bTB in human population. 8.2% animal level prevalence of bTB was documented in the study sites. Dairy cattle with age of 4 to 8 eight years were 34% more likely to be affected by bTB less than 4 years age cattle (OR=1.34). However, factors like housing, body condition, herd size and parity were not significantly related with the event of bTB. The assessment of bTB awareness was shown 29.7% participants knew that cattle could be infected, and 13.4% of the respondents believed that it can spread from animal to human and vice versa. Besides, 66.1% of participants had habit for consumption of raw milk and soured milk products. Moreover, respondent's attitude about zoonoses and consequences of bTB was risky. Present study discloses that bTB is animal health problem affecting dairy cattle in Gamo zone, Southern Ethiopia. Based on the current findings, public awareness creation, bovine tuberculosis test and segregation strategy should be applied to minimize the public health hazards and risk factors for bovine origin tuberculosis.


Atypical sensors for direct and rapid neuronal detection of bacterial pathogens.
Bacterial infection can threaten the normal biological functions of a host, often leading to a disease. Hosts have developed complex immune systems to cope with the danger. Preceding the elimination of pathogens, selective recognition of the non-self invaders is necessary. At the forefront of the body's defenses are the innate immune cells, which are equipped with particular sensor molecules that can detect common exterior patterns of invading pathogens and their secreting toxins as well as with phagocytic machinery. Inflammatory mediators and cytokines released from these innate immune cells and infected tissues can boost the inflammatory cascade and further recruit adaptive immune cells to maximize the elimination and resolution. The nervous system also seems to interact with this process, mostly known to be affected by the inflammatory mediators through the binding of neuronal receptors, consequently activating neural circuits that tune the local and systemic inflammatory states. Recent research has suggested new contact points: direct interactions of sensory neurons with pathogens. Latest findings demonstrated that the sensory neurons not only share pattern recognition mechanisms with innate immune cells, but also utilize endogenous and exogenous electrogenic components for bacterial pathogen detection, by which the electrical firing prompts faster information flow than what could be achieved when the immune system is solely involved. As a result, rapid pain generation and active accommodation of the immune status occur. Here we introduced the sensory neuron-specific detector molecules for directly responding to bacterial pathogens and their signaling mechanisms. We also discussed extended issues that need to be explored in the future.


Role of CD8 T cells in rat IgE responses.
The role of the cytokines interleukin-4 and interferon-gamma in the regulation of IgE responses in the mouse and man have focused on the role of CD4 T cells. In the rat, antigen-specific CD8 T cells, generated following inhalation of antigen, have been shown to be capable of suppressing IgE responses. Repeated intraperitoneal injections of 1 ng ricin and 1 microgram antigen established a long-lived IgE response in both low- and high-IgE responder rat strains (Wistar and Brown Norway). The duration of the IgE antibody response was 204 and 248 days, respectively. Total IgE levels rose from 30 +/- 20 to 39,000 +/- 7,500 ng/ml in the Wistar rat and from 120 +/- 100 to 47,000 +/- 8,000 ng/ml in the Brown Norway rat. An even greater (10(4)-fold) increase was seen in antigen-specific IgE antibody levels. Ricin alone had no effect and concomitant or prior stimulation with antigen was required. The proportion of CD4+ and CD8+ cells present in the spleen at the peak of the IgE response was markedly increased compared with animals given ricin or antigen alone. Furthermore, CD8 T cells were approximately 100 times more sensitive to ricin than CD4 T cells. These data suggest that enhancement of IgE responses in ricin-treated animals results from the selective deletion of T cells which suppress IgE and are of the CD8 phenotype.


Structure-function relationships of soybean proglycinins at subunit levels.
Glycinin consists of five kinds of subunits, group I (A1aB1b, A1bB2, and A2B1a) and group II (A3B4 and A5A4B3). cDNAs for individual subunits were cloned by reverse transcription-polymerase chain reaction method and expressed in Escherichia coli using pET vector. The recombinant proglycinins were purified by ammonium sulfate fractionation and column chromatography in the form of homotrimers. Physicochemical properties such as molecular dimensions, solubility, surface hydrophobicity, thermal stability, and emulsifying ability of individual proglycinins were studied. Molecular dimensions were proportional to molecular size for all proglycinins except A2B1a. Solubility was intrinsic to each proglycinin. At the ionic strength of 0.5, all proglycinins except A1aB1b showed a very low solubility at acidic pH, but A1aB1b was soluble to higher than 60%. At ionic strength 0.08, all proglycinins exhibited isoelectric precipitation, although A2B1a and A1bB2 were not completely insoluble. The order of emulsifying ability (A1bB2 < A2B1a < A5A4B3 < A3B4 < or = A1aB1b) was not of the same for surface hydrophobicity (A5A4B3 < A1aB1b < or = A3B4 < A1bB2 < A2B1a) and thermal stability (A1bB2 << A2B1a < or = A5A4B3 < A3B4 < or = A1aB1b).


Square-wave voltammetric sensing of Lawsone (2-hydroxy-1,4-naphthoquinone) based on the enhancement effect of cationic surfactant on anodically pretreated boron-doped diamond electrode.
In this reported work, an anodically pretreated boron-doped diamond (BDD) electrode was used for the inexpensive, simple and quick detection of a natural dye, lawsone. Lawsone had a well-defined, irreversible and diffusion-controlled oxidation peak at approximately +0.19 V in phosphate buffer solution (PBS, 0.1 M, pH 2.5) using cyclic voltammetry (CV). The oxidation peak heights of lawsone were significantly increased in PBS using the cationic surfactant cetyltrimethylammonium bromide (CTAB). Under optimized experimental conditions, the calibration curve was linear over a concentration range of 0.1-5.0 μM with detection limit of 0.029 μM in 0.1 M PBS (pH 2.5) containing 0.1 mM CTAB by using square-wave voltammetry (SWV). To evaluate the practical applicability of the BDD electrode, it was used for the quantification of lawsone in commercial henna, a natural dye made from the leaves of the henna plant.


Transcriptomic dissection of sexual differences in Bemisia tabaci, an invasive agricultural pest worldwide.
Sex difference involving chromosomes and gene expression has been extensively documented. In this study, the gender difference in the sweetpotato whitefly Bemisia tabaci was investigated using Illumina-based transcriptomic analysis. Gender-based RNAseq data produced 27 Gb reads, and subsequent de novo assembly generated 93,948 transcripts with a N50 of 1,853 bp. A total of 1,351 differentially expressed genes were identified between male and female B. tabaci, and majority of them were female-biased. Pathway and GO enrichment experiments exhibited a gender-specific expression, including enriched translation in females, and enhanced structural constituent of cuticle in male whiteflies. In addition, a putative transformer2 gene (tra2) was cloned, and the structural feature and expression profile of tra2 were investigated. Sexually dimorphic transcriptome is an uncharted territory for the agricultural insect pests. Molecular understanding of sex determination in B. tabaci, an emerging invasive insect pest worldwide, will provide potential molecular target(s) for genetic pest control alternatives.


The incidence and associations of malignancy in a large cohort of patients with biopsy-determined idiopathic inflammatory myositis.
The South Australian (SA) myositis database has registered all patients with biopsy-proven inflammatory myositis in SA from 1980 to 2009. We determined the incidence and associations of malignancy in myositis by linking this database with the SA cancer registry. Standardized incidence ratios (SIR) for malignancy were determined using the total SA population over the same time period, stratified by age and gender. The SIR for cancer in the myositis population (n = 373) was 1.39, p = 0.047. There was a trend towards an increased SIR in dermatomyositis but no increased risk of malignancy in polymyositis or inclusion body myositis. Malignancies of the lung and prostate were the commonest and 28 % of malignancies occurred within one year of IIM diagnosis. The odds of developing cancer were significantly raised in the presence of a shawl sign, male gender, and in patients with overlap syndrome or rheumatoid arthritis whilst myalgia was a significant protective factor. HLA-A28 allele was overrepresented in patients with malignancy (11 vs 2 %, p = 0.006). Patients in SA with myositis are at modestly increased risk for malignancy. We report clinical and genetic risk factors allowing the identification of patients at greatest risk for malignancy.


Dysmetabolism and Sleep Fragmentation in Obstructive Sleep Apnea Patients Run Independently of High Caffeine Consumption.
Daytime hypersomnolence, the prime feature of obstructive sleep apnea (OSA), frequently leads to high coffee consumption. Nevertheless, some clinicians ask for patients' caffeine avoidance. Caffeinated drinks are sometimes associated with more severe OSA. However, these effects are not consensual. Here we investigated the effect of caffeine consumption on sleep architecture and apnea/hypopnea index in OSA. Also, the impact of caffeine on variables related with dysmetabolism, dyslipidemia, and sympathetic nervous system (SNS) dysfunction were investigated. A total of 65 patients diagnosed with OSA and 32 without OSA were included after given written informed consent. Polysomnographic studies were performed. Blood was collected to quantify caffeine and its metabolites in plasma and biochemical parameters. 24 h urine samples were collected for catecholamines measurement. Statistical analyses were performed by SPSS: (1) non-parametric Mann-Whitney test to compare variables between controls and OSA; (2) multivariate logistic regression testing the effect of caffeine on sets of variables in the 2 groups; and (3) Spearmans' correlation between caffeine levels and comorbidities in patients with OSA. As expected OSA development is associated with dyslipidemia, dysmetabolism, SNS dysfunction, and sleep fragmentation. There was also a significant increase in plasma caffeine levels in the OSA group. However, the higher consumption of caffeine by OSA patients do not alter any of these associations. These results showed that there is no apparent rationale for caffeine avoidance in chronic consumers with OSA.


PON-P2: prediction method for fast and reliable identification of harmful variants.
More reliable and faster prediction methods are needed to interpret enormous amounts of data generated by sequencing and genome projects. We have developed a new computational tool, PON-P2, for classification of amino acid substitutions in human proteins. The method is a machine learning-based classifier and groups the variants into pathogenic, neutral and unknown classes, on the basis of random forest probability score. PON-P2 is trained using pathogenic and neutral variants obtained from VariBench, a database for benchmark variation datasets. PON-P2 utilizes information about evolutionary conservation of sequences, physical and biochemical properties of amino acids, GO annotations and if available, functional annotations of variation sites. Extensive feature selection was performed to identify 8 informative features among altogether 622 features. PON-P2 consistently showed superior performance in comparison to existing state-of-the-art tools. In 10-fold cross-validation test, its accuracy and MCC are 0.90 and 0.80, respectively, and in the independent test, they are 0.86 and 0.71, respectively. The coverage of PON-P2 is 61.7% in the 10-fold cross-validation and 62.1% in the test dataset. PON-P2 is a powerful tool for screening harmful variants and for ranking and prioritizing experimental characterization. It is very fast making it capable of analyzing large variant datasets. PON-P2 is freely available at http://structure.bmc.lu.se/PON-P2/.


Sudden loading during a dynamic lifting task: a simulation study.
It is believed that nurses risk the development of back pain as a consequence of sudden loadings during tasks in which they are handling patients. Forward dynamics simulations of sudden loads (applied to the arms) during dynamic lifting tasks were performed on a two-dimensional whole-body model. Loads were in the range of -80 kg to 80 kg, with the initial load being 20 kg. Loading the arm downwards with less than that which equals a mass of 20 kg did not change the compressive forces on the spine when compared to a normal lifting motion with a 20 kg mass in the hands. However when larger loads (40 kg to 80 kg extra in the hands) were simulated, the compressive forces exceeded 13,000 N (above 3400 N is generally considered a risk factor). Loading upwards led to a decrease in the compressive forces but to a larger backwards velocity at the end of the movement. In the present study, it was possible to simulate a fast lifting motion. The results showed that when loading the arms downwards with a force that equals 40 kg or more, the spine was severely compressed. When loading in the opposite direction (unloading), the spine was not compressed more than during a normal lifting motion. In practical terms, this indicates that if a nursing aide tries to catch a patient who is falling, large compressive forces are applied to the spine.


Zinc fingers--folds for many occasions.
Zinc finger domains (ZnFs) are common, relatively small protein motifs that fold around one or more zinc ions. In addition to their role as a DNA-binding module, ZnFs have recently been shown to mediate protein:protein and protein:lipid interactions. This small zinc-ligating domain, often found in clusters containing fingers with different binding specificities, can facilitate multiple, often independent intermolecular interactions between nucleic acids and proteins. Classical ZnFs, typified by TFIIIA, ligate zinc via pairs of cysteine and histidine residues but there are at least 14 different classes of Zn fingers, which differ in the nature and arrangement of their zinc-binding residues. Some GATA-type ZnFs can bind to both DNA and a variety of other proteins. Thus proteins with multiple GATA-type fingers can play a complex role in regulating transcription through the interplay of these different binding selectivities and affinities. Other ZnFs have more specific functions, such as DNA-binding ZnFs in the nuclear hormone receptor proteins and small-molecule-binding ZnFs in protein kinase C. Some classes of ZnFs appear to act exclusively in protein-only interactions. These include the RING family of ZnFs that are involved in ubiquitination processes and in the assembly of large protein complexes, LIM, TAZ, and PHD domains. We review the similarities and differences in structure and functions of different ZnF classes and highlight the versatility of this fold.


[Photodynamic therapy in adult-onset foveomacular vitelliform dystrophy].
We present the case of a patient who underwent photodynamic therapy (PDT) for adult-onset foveomacular vitelliform dystrophy (AFVD) at another clinic, having been misdiagnosed as having a choroidal neovascular membrane. Two years post-treatment, the patient had severe central visual loss and showed angiographic signs of choroidal hypoperfusion. AFVD is a rare clinical entity, usually having a fair visual prognosis, although some cases may show episodes of severe visual loss during their clinical course. Optical coherence tomography (OCT) is valuable in revealing the distinct images of AFVD from the features of a choroidal neovascular membrane. PDT using verteporfin may have negative long term effects if used in patients with AFVD.


Temporal trends in sperm count: a systematic review and meta-regression analysis.
Reported declines in sperm counts remain controversial today and recent trends are unknown. A definitive meta-analysis is critical given the predictive value of sperm count for fertility, morbidity and mortality. To provide a systematic review and meta-regression analysis of recent trends in sperm counts as measured by sperm concentration (SC) and total sperm count (TSC), and their modification by fertility and geographic group. PubMed/MEDLINE and EMBASE were searched for English language studies of human SC published in 1981-2013. Following a predefined protocol 7518 abstracts were screened and 2510 full articles reporting primary data on SC were reviewed. A total of 244 estimates of SC and TSC from 185 studies of 42 935 men who provided semen samples in 1973-2011 were extracted for meta-regression analysis, as well as information on years of sample collection and covariates [fertility group ('Unselected by fertility' versus 'Fertile'), geographic group ('Western', including North America, Europe Australia and New Zealand versus 'Other', including South America, Asia and Africa), age, ejaculation abstinence time, semen collection method, method of measuring SC and semen volume, exclusion criteria and indicators of completeness of covariate data]. The slopes of SC and TSC were estimated as functions of sample collection year using both simple linear regression and weighted meta-regression models and the latter were adjusted for pre-determined covariates and modification by fertility and geographic group. Assumptions were examined using multiple sensitivity analyses and nonlinear models. SC declined significantly between 1973 and 2011 (slope in unadjusted simple regression models -0.70 million/ml/year; 95% CI: -0.72 to -0.69; P < 0.001; slope in adjusted meta-regression models = -0.64; -1.06 to -0.22; P = 0.003). The slopes in the meta-regression model were modified by fertility (P for interaction = 0.064) and geographic group (P for interaction = 0.027). There was a significant decline in SC between 1973 and 2011 among Unselected Western (-1.38; -2.02 to -0.74; P < 0.001) and among Fertile Western (-0.68; -1.31 to -0.05; P = 0.033), while no significant trends were seen among Unselected Other and Fertile Other. Among Unselected Western studies, the mean SC declined, on average, 1.4% per year with an overall decline of 52.4% between 1973 and 2011. Trends for TSC and SC were similar, with a steep decline among Unselected Western (-5.33 million/year, -7.56 to -3.11; P < 0.001), corresponding to an average decline in mean TSC of 1.6% per year and overall decline of 59.3%. Results changed minimally in multiple sensitivity analyses, and there was no statistical support for the use of a nonlinear model. In a model restricted to data post-1995, the slope both for SC and TSC among Unselected Western was similar to that for the entire period (-2.06 million/ml, -3.38 to -0.74; P = 0.004 and -8.12 million, -13.73 to -2.51, P = 0.006, respectively). This comprehensive meta-regression analysis reports a significant decline in sperm counts (as measured by SC and TSC) between 1973 and 2011, driven by a 50-60% decline among men unselected by fertility from North America, Europe, Australia and New Zealand. Because of the significant public health implications of these results, research on the causes of this continuing decline is urgently needed.


[The validity and sensitivity to change of the Spanish self-administered version of the chronic respiratory questionnaire (CRQ-SAS)].
The interviewer-administered chronic respiratory questionnaire (CRQ-IA) is widely used and has demonstrated excellent properties for measuring health-related quality of life (HRQL) in patients with chronic obstructive pulmonary disease (COPD). However, the self-administered version (CRQ-SAS) in Spanish has not been validated. The aim of this trial was to evaluate the validity and the sensitivity of the Spanish version of the CRQ-SAS in patients with COPD. We randomized 40 patients with COPD (33 treated with pulmonary rehabilitation and 7 with liquid oxygen therapy) to one of the two methods of administration of CRQ (SAS vs. IA) both before and 8 weeks after the treatment. In addition, patients completed the SF-36 questionnaire, pulmonary function tests, and six-minute walk test. The CRQ-SAS demonstrated good longitudinal construct validity on all domains with a range of correlations, for the change scores, between 0.46 (P=.05) and 0.71 (P=.01). Regarding sensitivity to change, we observed a minimal clinically significant change in most domains (fatigue 0.71 [P=.02], emotional factor 0.62 [P=.04], control of the disease 0.83 [P=.06]). The Spanish version of CRQ-SAS is valid for evaluating HRQL in COPD patients. The correlations of the CRQ-SAS with other tools provide construct validity and show good sensitivity to change.


Human immunodeficiency virus type 1 mutants that escape neutralization by human monoclonal antibody IgG1b12. off.
IgG1b12, a human monoclonal antibody (MAb) to an epitope overlapping the CD4-binding site on gp120, has broad and potent neutralizing activity against most primary human immunodeficiency virus type 1 (HIV-1) isolates. To assess whether and how escape mutants resistant to IgG1b12 can be generated, we cultured primary HIV-1 strain JRCSF in its presence. An escape mutant emerged which was approximately 100-fold more resistant to neutralization by IgG1b12. Both virion-associated and solubilized gp120 from this variant had a reduced affinity for IgG1b12, and sequencing of its env gene showed that amino acid substitutions had occurred at three positions within gp120. Two (D164N and D182N) were located in V2, and one (P365L) was in C3. By site-directed mutagenesis, we demonstrated that the D182N and P365L mutations, but not D164N, contribute to the IgG1b12-resistant phenotype. However, the former two substitutions, individually or in combination, hinder the replication of the neutralization-resistant virus. Introduction of the D164N substitution into the P365L variant results in a nonviable virus (D164N/P365L). In contrast, addition of D164N to the D182N or D182N/P365L mutant partially restored replicative function to near wild-type levels. Furthermore, we found that all of the IgG1b12-resistant mutant viruses remained sensitive to other human MAbs, such as 2G12 and 2F5, and to the CD4-IgG molecule, except that the P365L-containing mutant was slightly resistant to CD4-IgG. These results suggest that escape from IgG1b12 neutralization is due to a local rather than a global modification of the gp120 structure. Our findings have implications for the therapeutic and prophylactic applications of antibodies for HIV-1 infection.


Melatonin, bone regulation and the ubiquitin-proteasome connection: A review.
Recently, investigators have shown that ubiquitin-proteasome-mediated protein degradation is critical in regulating the balance between bone formation and bone resorption. The major signal transduction pathways regulating bone formation are the RANK/NF-κB pathway and the Wnt/β-catenin pathway. These signal transduction pathways regulate the activity of mature osteoblasts and osteoclasts. In addition, the Wnt/β-catenin pathway is one of the major signaling pathways in the differentiation of osteoblasts. The ubiquitin ligases that are reported to be of major significance in regulating these pathways are the ubiquitin SCF(B-TrCP) ligase (which regulates activation of NF-κB via degradation of IkBα in osteoclasts, and regulates bone transcription factors via degradation of β-catenin), the Keap-Cul3-Rbx1 ligase (which regulates degradation of IkB kinase, Nrf2, and the antiapoptotic factor Bcl-2), and Smurf1. Also of significance in regulating osteoclastogenesis is the deubiquitinase, CYLD (cylindramatosis protein), which facilitates the separation of NF-κB from IkBα. The degradation of CYLD is also under the regulation of SCF(B-TrCP). Proteasome inhibitors influence the activity of mature osteoblasts and osteoclasts, but also modulate the differentiation of precursor cells into osteoblasts. Preclinical studies show that melatonin also influences bone metabolism by stimulating bone growth and inhibiting osteoclast activity. These actions of melatonin could be interpreted as being mediated by the ubiquitin ligases SCF(B-TrCP) and Keap-Cul3-Rbx, or as an inhibitory effect on proteasomes. Clinical trials of the use of melatonin in the treatment of bone disease, including multiple myeloma, using both continuous and intermittent modes of administration, are warranted.


[Prospective evaluation of the risk factors, etiology and the antimicrobial susceptibilities of the isolates in nosocomial bacteremic patients].
This study was aimed to assess the frequency, risk factors and etiology of bacteremia and antimicrobial susceptibilities of strains isolated from adult bacteremic patients hospitalized in Kocaeli University Hospital, Turkey during a 12-month period. Blood specimens obtained from a total of 2602 patients after at least 48 hours of hospitalization, were cultured in aerobic BACTEC 9120 and BACTEC 9050 (Becton-Dickinson, USA) automatized systems, and the identification and antibiotic susceptibilities of bacteria were performed by VITEK 1 (bioMerieux, France) system. Blood cultures of a total of 385 (14.7%) patients yielded bacterial growth; the contamination rate being 9.1% (238/2602) and nosocomial bacteria rate being 5.6% (147/2602). The distribution of the 147 (81 female, 67 male) bacteremic patients were as follows: 45% internal ward, 28% surgical ward and 27% intensive care unit. A total of 148 patients (81 female, 67 male) with no growth in their blood cultures were randomly chosen as the control group. The pathogens isolated from nosocomial bacteremias revealed 101 gram-positive bacteria (68.7%), 39 gram- negative bacilli (26.5%) and seven Candida species (4.7%). The most frequent gram-positive bacteria were staphylococci 84/101; 83.1%) and gram-negative bacteria were E. coli (14/39; 35.8%). Coagulase negative staphylococci (CNS) were isolated from 79% of the cultures evaluated as contaminants. Methicillin resistance was determined in 61.6% of S. aureus and 68.1% of CNS. Extended spectrum beta-lactamases were detected in 64% of E. coli, 62% of Pseudomonas spp., 47% of Klebsiella spp. and 24% of Acinetobacter spp. and resistance rates to ceftazidime and beta-lactam/beta-lactamase inhibitor combinations in these strains were 39-67%. The most effective drug group was found to be carbapenems with 3-26% resistance rates. The evalution of risk factors between patient and control groups revealed that mechanical ventilation (p < 0.05) and interventional catheters (except for intravenous catheters) (p < 0.05) were the most frequent sources of bacteremias. The most frequent focus of infection were the lungs (29.9%), followed by abdominal and catheter originated infections (23.8% each). The high rate of antibiotic resistance determined in the nosocomial bacterial isolates in our hospital indicated that antibiotic use policies in our setting should be taken under control. The high rate of contamination in blood cultures emphasize that more attention should paid for continous education about collection of blood culture specimens.


Knockout of microRNA-21 attenuates alcoholic hepatitis through the VHL/NF-κB signaling pathway in hepatic stellate cells.
microRNA-21 (miRNA) is one of the most abundant miRNAs in chronic liver injuries including alcoholic liver injury. Previous studies have demonstrated that miR-21 plays a role in inflammation in the liver and functions in hepatic stellate cells (HSCs), which reside in the perisinusoidal space between sinusoidal endothelial cells and hepatocytes and regulate sinusoidal circulation. HSCs integrate cytokine-mediated inflammatory responses in the sinusoids and relay them to the liver parenchyma. Here, we showed that the activation of Von Hippel-Lindau (VHL) expression, by miR-21 knockout in vivo and anti-miR-21 or VHL overexpression in vitro, suppressed the production of proinflammatory cytokines, such as interleukin (IL)-6, monocyte chemoattractant protein-1, and IL-1β, in human HSCs during alcoholic liver injury. Sequence and functional analyses confirmed that miR-21 directly targeted the 3'-untranslated region of VHL. Immunofluorescence and real-time PCR analysis revealed that miR-21 depletion blocked NF-κB activation in human HSCs both in cultured HSCs as well as HSCs isolated from alcohol-related liver disease mice liver by laser capture microdissection. We also showed that conditioned medium from anti-miR-21-transfected HSCs suppressed human monocyte-derived THP-1 cell migration. Taken together, our study indicates that depletion of miR-21 may downregulate cytokine production in HSCs and macrophage chemotaxis during alcoholic liver injury and that the targeting of miR-21 may have therapeutic potential for preventing the progression of alcoholic liver diseases. NEW & NOTEWORTHY This study demonstrates that silencing microRNA-21 can inhibit cytokine production and inflammatory responses in human hepatic stellate cells during alcoholic liver injury and that the targeting of microR-21 in hepatic stellate cells may have therapeutic potential for prevention and treatment of alcoholic liver diseases.


AIDS pathogenesis: from models to viral dynamics in patients.
The course of HIV infection usually follows a pattern in which the immune system is initially able to limit viral replication, but ultimately fails. It is proposed that disease progression is a consequence of viral replication and evolution within the host, the immune response providing the selection pressure for increasing diversity. When the number of mutants rises above a certain threshold level, the immune system can no longer downregulate all the viral variants simultaneously and symptomatic disease results. Recent studies of the dynamics and kinetics of HIV infection have indicated that 30% of the free virus population in the plasma is replenished each day, a very high turnover rate. After intervention with nevirapine, resistant virus was observed to have replaced wild-type virus in the plasma within 2-4 weeks. Even in late-stage patients, the immune system has the capacity to regenerate large numbers of CD4 cells, but this ability does not continue indefinitely. This implies that to reverse immunodeficiency the principal objective of treatment in the management of HIV infection should be inhibition of viral replication and thus prevention of lymphocyte destruction.


Characteristics and Outcomes of Fasting Orders Among Medical Inpatients.
While many hospitalized patients have orders to fast in preparation for interventions, the extent to which these orders are necessary or adhere to evidence-based durations is unknown. In this study, we analyzed the length, indication, and associated outcomes of nil per os (NPO) orders for general medicine patients at an academic institution in the United States, and compared them to the best available evidence for recommended length of NPO. Of 924 NPO orders assessed, the indicated intervention was not performed for 183 (19.8%) orders, largely due to a change in plan (75/183, 41.0%) or scheduling barriers (43/183, 23.5%). When analyzed by indication, the median duration of NPO orders ranged from 8.3 hours for kidney ultrasound to 13.9 hours for upper endoscopy. For some indications, the literature suggested NPO orders may be unnecessary. Furthermore, in indications for which NPO was deemed necessary in the literature, the duration of most NPO orders was much longer than minimally required. These results suggest the need for establishing more robust practice guidelines or institutional protocols for NPO orders. Journal of Hospital Medicine 2017;12:36-39.


Social mindfulness and prosociality vary across the globe.
Humans are social animals, but not everyone will be mindful of others to the same extent. Individual differences have been found, but would social mindfulness also be shaped by one's location in the world? Expecting cross-national differences to exist, we examined if and how social mindfulness differs across countries. At little to no material cost, social mindfulness typically entails small acts of attention or kindness. Even though fairly common, such low-cost cooperation has received little empirical attention. Measuring social mindfulness across 31 samples from industrialized countries and regions (n = 8,354), we found considerable variation. Among selected country-level variables, greater social mindfulness was most strongly associated with countries' better general performance on environmental protection. Together, our findings contribute to the literature on prosociality by targeting the kind of everyday cooperation that is more focused on communicating benevolence than on providing material benefits.


Direct evidence of iNOS-mediated in vivo free radical production and protein oxidation in acetone-induced ketosis.
Diabetic patients frequently encounter ketosis that is characterized by the breakdown of lipids with the consequent accumulation of ketone bodies. Several studies have demonstrated that reactive species are likely to induce tissue damage in diabetes, but the role of the ketone bodies in the process has not been fully investigated. In this study, electron paramagnetic resonance (EPR) spectroscopy combined with novel spin-trapping and immunological techniques has been used to investigate in vivo free radical formation in a murine model of acetone-induced ketosis. A six-line EPR spectrum consistent with the alpha-(4-pyridyl-1-oxide)-N-t-butylnitrone radical adduct of a carbon-centered lipid-derived radical was detected in the liver extracts. To investigate the possible enzymatic source of these radicals, inducible nitric oxide synthase (iNOS) and NADPH oxidase knockout mice were used. Free radical production was unchanged in the NADPH oxidase knockout but much decreased in the iNOS knockout mice, suggesting a role for iNOS in free radical production. Longer-term exposure to acetone revealed iNOS overexpression in the liver together with protein radical formation, which was detected by confocal microscopy and a novel immunospin-trapping method. Immunohistochemical analysis revealed enhanced lipid peroxidation and protein oxidation as a consequence of persistent free radical generation after 21 days of acetone treatment in control and NADPH oxidase knockout but not in iNOS knockout mice. Taken together, our data demonstrate that acetone administration, a model of ketosis, can lead to protein oxidation and lipid peroxidation through a free radical-dependent mechanism driven mainly by iNOS overexpression.