Vector-initiated transitive RNA interference in the filamentous fungus Aspergillus oryzae.
RNA interference (RNAi), modulates gene expression via cleavage of double-stranded RNA (dsRNA) by Dicer, producing 21-25 nucleotide silence-inducing RNAs (siRNAs). In association with Argonaute containing complexes, these siRNAs target sequence-specific degradation of the homologous single-stranded messenger RNA. In the majority of eukaryotes, degradation occurs within the boundaries of the dsRNA target. In Arabidopsis thaliana and Caenorhabditis elegans, gene silencing can also take place transitively, impacting transcripts from coding sequences that are adjacent to the intended target gene. Here we demonstrate effective transitive RNAi in the ascomycete Aspergillus oryzae. Fragments of 174 bp and 499 bp derived from the A. oryzae wA gene involved in spore color development, were inserted immediately upstream of an inverted repeat derived from the Escherichia coli gene encoding for Hygromycin Phosphotransferase B (hph), which provided a double-stranded hph RNAi trigger. Introduction of this construct into A. oryzae host cells produced transformants with spores that were lighter in color than those of wild type. Real-time RT-qPCR analysis demonstrated a direct correspondence of steady-state wA mRNA level to spore color. An A. oryzae strain deficient in RNA-dependent RNA Polymerase (RdRP) produced exclusively wild type colored spores when transformed with a wA transitive RNAi construct. Conversely, increased expression of RdRP enhanced the incidence of wA gene silencing via transitive RNAi.


Cost analysis of tubal anastomosis by laparoscopy and by laparotomy.
To compare the costs of tubal anastomosis performed by laparoscopy and by laparotomy. Cost analysis study using the hospital administrative database (Canadian Task Force classification II-2). University teaching hospital. Eighty-nine women. Intervention. Tubal anastomosis by laparoscopy (43) and by laparotomy (46). Tubal anastomosis took longer when performed by laparoscopy than by laparotomy; however, the total time patients spent in the operating room was similar. Women treated by laparoscopy spent more time in the recovery room. Labor costs for nurses in the operating room and recovery room costs were significantly higher in the laparoscopy group. Costs for operating room supplies were similar. Pharmacy costs and expenses in the ward were lower in the laparoscopy group. The mean total cost for laparoscopic tubal anastomosis was $861 +/- 137 and for laparotomy was $1348 +/- 188 (p <0.001). Laparoscopic tubal anastomosis costs less than tubal anastomosis by laparotomy.


Component X of mammalian pyruvate dehydrogenase complex: structural and functional relationship to the lipoate acetyltransferase (E2) component.
The lipoate acetyltransferase (E2, Mr 70,000) and protein X (Mr 51,000) subunits of the bovine pyruvate dehydrogenase multienzyme complex (PDC) core assembly are antigenically distinct polypeptides. However comparison of the N-terminal amino acid sequence of the E2 and X polypeptides reveals significant homology between the two components. Selective tryptic release of the 14C-labelled acetylated lipoyl domains of E2 and protein X from native PDC generates stable, radiolabelled 34 and 15 kDa fragments, respectively. Thus, in contrast to E2 which contains two tandemly-arranged lipoyl domains, protein X appears to contain only a single lipoyl domain located at its N-terminus.


Chemokine regulation of naïve T cell traffic in health and disease.
A central feature of the immune response is the precise spatio-temporal convergence of T cells and antigen presenting cells (APC) in particular microenvironments within secondary lymphoid organs (SLO). CCR7 and its ligands CCL19 and CCL21 have been identified as the gatekeepers for both naïve T lymphocytes and dendritic cells (DC) to these defined anatomical compartments. A new perception on the regulation of lymphocyte traffic in lymph nodes (LN) has come from observations that sphingosine-1-phosphate (S1P) receptor agonists affect T cell entry and exit from these organs. Recent developments in intravital microscopy (IVM) techniques reveal unexpected autonomous random motion of lymphocytes within secondary lymphoid tissues, and provoke questions about the mechanisms that guide their compartmental navigation.


A new species, new host records and life cycle data for lepocreadiids (Digenea) of pomacentrid fishes from the Great Barrier Reef, Australia.
A new species of lepocreadiid, Opechonoides opisthoporus n. sp., is described infecting 12 pomacentrid fish species from the Great Barrier Reef, Australia, with Abudefduf whitleyi Allen & Robertson as the type-host. This taxon differs from the only other known member of the genus, Opechonoides gure Yamaguti, 1940, in the sucker width ratio, cirrus-sac length, position of the testes, position of the pore of Laurer's canal, and relative post-testicular distance. The new species exhibits stenoxenic host-specificity, infecting pomacentrids from seven genera: Abudefduf Forsskål, Amphiprion Bloch & Schneider, Neoglyphidodon Allen, Neopomacentrus Allen, Plectroglyphidodon Fowler & Ball, Pomacentrus Lacépède and Stegastes Jenyns. Phylogenetic analyses of 28S rDNA sequence data demonstrate that O. opisthoporus n. sp. forms a strongly supported clade with Prodistomum orientale (Layman, 1930) Bray & Gibson, 1990. The life cycle of this new species is partly elucidated on the basis of ITS2 rDNA sequence data; intermediate hosts are shown to be three species of Ctenophora. New host records and molecular data are reported for Lepocreadium oyabitcha Machida, 1984 and Lepotrema amblyglyphidodonis Bray, Cutmore & Cribb, 2018, and new molecular data are provided for Lepotrema acanthochromidis Bray, Cutmore & Cribb, 2018 and Lepotrema adlardi (Bray, Cribb & Barker, 1993) Bray & Cribb, 1996. Novel cox1 mtDNA sequence data showed intraspecific geographical structuring between Heron Island and Lizard Island for L. acanthochromidis but not for L. adlardi or O. opisthoporus n. sp.


Clinical performance of CAD-CAM partial-coverage restorations: Experienced versus less-experienced operators.
Less-experienced operators have been shown to require additional training to achieve results similar to those of experienced operators. However, clinical data comparing the survival and success of ceramic restorations by experienced and less-experienced operators by using the computer-aided design and computer-aided manufacturing (CAD-CAM) technology are lacking. The purpose of this retrospective clinical study was to analyze and compare the clinical performance of CAD-CAM lithium disilicate restorations fabricated by less-experienced (predoctoral dental students) and experienced (dentists) operators. Patients who received an adhesively luted CAD-CAM lithium disilicate restoration between 2011 and 2019 were included in the study. Clinical performance was assessed by calibrated examiners by using World Dental Federation (FDI) criteria. Success and survival were calculated by the Kaplan-Meier method and statistically compared by log-rank tests and univariate Cox regression analyses. FDI criteria were compared by using Mann-Whitney-U tests (α=.05). Ninety-two restorations (students: n=65, dentists: n=27) were assessed (mean ±standard deviation time from insertion: 4.04 ±1.55 years). The survival rates after 2 years (students: 93.8%, mean annual failure rate [mAFR]: 3.1%; dentist: 96.3%, mAFR: 1.9%) and after 4 years (students: 87.3%, mAFR: 3.3%; dentists: 88.3%, mAFR: 3.1%) were not significantly different (P=.525). Also, success rates after 2 (students: 90.8%, mAFR: 4.7%; dentists: 92.6%, mAFR: 3.8%) and 4 years (students: 82.4%, mAFR: 4.7%; dentists: 76.1%, mAFR: 6.6%) were not significantly different (P=.778). FDI criteria were also not significantly different between less-experienced and experienced operators (P≥.110). Operator experience did not affect the short-term clinical performance of CAD-CAM lithium disilicate restorations.


Highly stable core-surface-crosslinked nanoparticles as cisplatin carriers for cancer chemotherapy.
Cisplatin is a potent anticancer drug with low solubility in water. A new type of highly stable polymer micelles, namely core-surface-crosslinked nanoparticles (SCNPs) made from amphiphilic brush copolymers, were evaluated as the carrier of cisplatin. Cisplatin could be loaded in the SCNPs with poly(epsilon-caprolactone) (PCL) cores and hydrophilic poly(ethylene glycol) (PEG) or poly[2-(N,N-dimethylamino)ethyl methacrylate] (PDMA) shells with high loading efficiency (approximately 90%). In vitro cellular uptake experiments indicated that both SCNPs could be easily taken up by SKOV-3 ovarian cancer cells. Both cell proliferation assay and IC50 measurements indicated that cisplatin encapsulated in the SCNPs had much enhanced cytotoxicity to the cancer cells compared to free cisplatin. The positive charges on the PCL/PDMA SCNPs promoted the cellular internalization of the nanoparticles, resulting in higher cytotoxicity of cisplatin in these SCNPs. The IC50 of the cisplatin encapsulated in PCL/PDMA SCNPs was as low as 0.01 microg/mL, lower than that of cisplatin in PCL/PEG SCNPs and free cisplatin.


Movement and retention of derelict fishing nets in Northwestern Hawaiian Island reefs.
Derelict fishing nets pose hazards to marine systems as they travel through the ocean or become ensnared on coral reefs. Understanding of the movement of nets within shallow atolls can help to optimize operations to protect these shallow reefs. In 2018, six derelict fishing nets at Manawai (Pearl and Hermes Reef) in the Northwestern Hawaiian Islands were tagged with satellite-transmitting buoys and tracked for three years. This study reveals that nets that enter the atoll from the northeast travel southwest towards the center of the atoll, and nets in the center can remain ensnared on the same reef for at least three years. This study shows that satellite buoys are a successful approach to tracking derelict net movement, and can inform future debris removal missions.


Interferon-gamma increased epithelial barrier function via upregulating claudin-7 expression in human submandibular gland duct epithelium.
Tight junctions (TJs) are necessary for salivary gland function and may serve as indicators of salivary gland epithelial dysfunction. IgG4-related disease (IgG4-RD) is a newly recognized fibro-inflammatory condition which disrupts the TJ associated epithelial barrier. The salivary glands are one of the most frequently involved organs in IgG4-RD, however, changes of the TJ associated epithelial barrier in salivary gland duct epithelium is poorly understood. Here, we investigated the regulation and function of TJs in human submandibular gland ductal epithelial cells (HSDECs) in normal and IgG4-RD. We examined submandibular gland (SMG) tissue from eight control individuals and 22 patients with IgG4-RD and established an HSDEC culture system. Immunohistochemistry, immunocytochemistry, western blotting, and measurement of transepithelial electrical resistance (TER) were performed. Claudin-4, claudin-7, occludin, and JAM-A were expressed at the apical side of the duct epithelium in submandibular gland (SMG) tissue and at the cell borders in HSDECs of normal and IgG4-RD. The expression and distribution of TJs in SMG tissue were not different in control individuals and patients with IgG4-RD in vivo and in vitro. Although interferon-gamma (IFNγ) generally disrupts the integrity and function of TJs, as manifested by decreased epithelial barrier function, IFNγ markedly increased the epithelial barrier function of HSDECs via upregulation of claudin-7 expression in HSDECs from patients with IgG4-RD. This is the first report showing an IFNγ-dependent increase in epithelial barrier function in the salivary gland duct epithelium. Our results provide insights into the functional significance of TJs in salivary gland duct epithelium in physiological and pathological conditions, including IgG4-RD.


Corynebacterium glutamicum WhcD interacts with WhiA to exert a regulatory effect on cell division genes.
Corynebacterium glutamicum WhcD plays an important regulatory role in cell division. Binding of WhcD to the promoter region of its target genes, such as ftsZ, was observed by electrophoretic mobility shift assays (EMSA) using purified fusion proteins; however, binding could only be observed in the presence of WhiA. Although WhcD alone did not bind to the DNA, it stimulated binding of WhiA to the promoter region of the cell division gene ftsZ. Binding of WhcD and WhiA to DNA did not occur in the presence of the oxidant diamide. Purified WhcD and WhiA physically interacted in vitro. The presence of diamide did not disrupt the WhcD-WhiA interaction but affected binding of WhiA to the promoter region of ftsZ. The GACAC motif and adjacent sequences were found to be important for binding of the WhcD-WhiA complex to the DNA. Collectively, our results suggest that WhcD enhances the WhiA DNA-binding activity by physically interacting with WhiA. In addition, loss of WhiA DNA-binding activity in the presence of an oxidant agent may suggest a role for this protein as a switch that controls cell division in cells under oxidative stress.


FDG-Avid But Pentixafor-Negative in EBV-Associated T-Cell Lymphoproliferative Disorders.
An 18-year-old woman with intermittent fever, pancytopenia, abnormal liver function, and enlarged lymph nodes and hepatosplenomegaly was clinically suspected as hemophagocytic lymphohistiocytosis. 18 F-FDG PET/CT showed increased metabolism in multiple lymph nodes, which were highly suggestive of lymphoma. No increased CXCR4 expression in lymph nodes was demonstrated on 68 Ga-pentixafor PET/CT. Subsequent right neck lymph node biopsy pathology revealed EBV-associated lymphoproliferative disorders. Our case shows that 68 Ga-pentixafor PET/CT may have potential value in differentiating EBV-associated lymphoproliferative disorders from lymphomas.


Association of β-Lactam Allergy Documentation and Prophylactic Antibiotic Use in Surgery: A National Cross-Sectional Study of Hospitalized Patients.
Alternative antibiotics for surgical prophylaxis are associated with increased adverse events and surgical site infection compared to cefazolin. In a sample of perioperative inpatients from 100 hospitals in the United States, cefazolin was 9-fold less likely to be used in patients with a documented β-lactam allergy whereas clindamycin was 45-fold more likely.


Long-term follow-up and prognostic factor analysis in clear cell adenocarcinoma of the ovary.
There were a few reports of a large number of patients with clear cell adenocarcinoma (CCA) of the ovary because of the low incidence of CCA. This study compared the clinical factors affecting long-term survival of patients with CCA to those with serous cystadenocarcinoma (SCA). One hundred and seventy-eight CCA and 311 SCA patients treated between 1987 and 2000 were retrospectively evaluated. Differences in survival rates were calculated using log-rank test and Cox's proportional hazards analysis was used to identify independent prognostic factors. The ratio of stage I was significantly higher than that of SCA. There was no significant difference of 8-year survival rate in each stage between CCA and SCA. However, the patients with stage IIIb or IIIc CCA showed significantly worse prognosis than those with SCA. Positive peritoneal or ascitic cytology, the presence of residual tumor, more than 100 ml ascites were demonstrated to have the significant impact on survival by univariate analysis. Multivariate analysis demonstrated that stage, more than 100 ml ascites, and the presence of residual tumor were significant prognostic factors of CCA overall survival. Distribution of stage and substage differed between CCA and SCA in this study. Thus, substaging is quite important for comparison of prognoses between histologies, and CCA showed poorer prognoses than serous adenocarcinoma in stages IIIb and IIIc.


Video-Assisted Thoracoscopic Sleeve Lobectomy for Centrally Located Non-small Cell Lung Cancer: A Meta-analysis.
Whether video-assisted thoracoscopic surgery (VATS) sleeve lobectomy could be an alternative to traditional thoracotomy sleeve lobectomy in treating centrally located non-small cell lung cancer (NSCLC) remains unclear. Therefore, we conducted the first meta-analysis to compare the effects of VATS sleeve lobectomy with thoracotomy sleeve lobectomy. We systematically searched relevant studies from Pubmed, Embase, and Web of Science on May 12, 2020. Data for analysis included short-term outcomes (blood loss, lymph node dissected, operation time, hospital stay, complications) and long-term outcomes (3-year overall survival (OS) and progression-free survival (PFS) rates). We calculated the weighted mean differences (WMDs) for continuous data and risk ratio (RR) for pooling categorical data. We finally included 5 retrospective cohort study consisting of 436 patients. VATS sleeve lobectomy yielded significantly less blood loss (WMD = -37.83; 95% confidence intervals (CIs) = [-58.56, -17.11]; P < 0.001) than thoracotomy sleeve lobectomy and comparable total number of dissected lymph node to thoracotomy sleeve lobectomy (WMD = - 0.07; 95%CI = [-1.14, 0.99]; P = 0.89). However, VATS sleeve lobectomy consumed significantly more operation time than thoracotomy sleeve lobectomy (WMD = 49.00; 95%CI = [14.67, 83.34]; P = 0.005). VATS sleeve lobectomy yielded significantly less postoperative hospital stay time than thoracotomy sleeve lobectomy (WMD = -1.68; 95%CI = [-2.98, -0.39]; P = 0.011) and comparable postoperative complication rate to thoracotomy sleeve lobectomy (RR = 0.84; 95%CI = [0.49, 1.44]; P = 0.52). Moreover, VATS sleeve lobectomy yielded comparable 3-year OS (RR = 1.08; 95%CI = [0.95, 1.22]; P = 0.23) and PFS (RR = 1.15; 95%CI = [0.96, 1.37]; P = 0.13) rates to thoracotomy sleeve lobectomy. No significant heterogeneities were observed. VATS sleeve lobectomy yielded less surgical trauma than thoracotomy sleeve lobectomy and improved postoperative recovery without compromising oncological prognosis. Even though VATS sleeve lobectomy may consume more operation time, it could be recommended as an alternative to thoracotomy sleeve lobectomy for treating centrally located NSCLC in carefully selected cases.


The NOTCH4 locus is associated with susceptibility to schizophrenia.
Linkage disequilibrium mapping of the MHC region in 80 British parent-offspring trios showed that NOTCH4 was highly associated with schizophrenia. The A-->G substitution in the promoter region and the (CTG)n repeat in exon 1 of NOTCH4 may be candidate sites conferring susceptibility to schizophrenia.


Low-density lipoprotein receptor genotypes modify the sera metabolome of patients with homozygous familial hypercholesterolemia.
Homozygous familial hypercholesterolemia (HoFH) is an extremely rare metabolism disorder usually caused by low-density lipoprotein receptor (LDLR) mutations. LDLR genotype is commonly known to determine blood concentrations of LDL cholesterol. However, effects of LDLR genotype on holistic metabolome remain unclear. Herein, we present metabolomic, genetic, and clinical datasets from a large multi-center panel of 142 patients with LDLR-mutated HoFH. We found that true homozygotes and compound heterozygotes showed few differences in clinical and metabolomic phenotypes. Compared with defective/defective mutation carriers, patients carrying one or two null mutation showed profound alterations in clinical laboratory lipids and serum cholesterol esters, lysophosphocholines, bile acids, and amino acids. Importantly, these altered metabolites are implicated in multiple biochemical reactions and associated with LDL cholesterol. This study extends the first map of different LDLR genotypes influencing the metabolome and suggests that the small-molecule metabolites serve as potential targets to mitigate the deleterious impact of LDLR mutations on HoFH.


Scanning electron microscopy of polypoid cystitis--a reversible lesion of the human bladder.
Scanning electron microscopy was performed on forceps biopsies from 13 patients with catheter-associated polypoid cystitis. Pleomorphic microvilli, which varied considerably in appearance, were found on surface cells of all cases with polypoid cystitis. Practically all of the characteristics of pleomorphic microvilli assigned earlier to bladder tumor cells in humans were identified on specimens from patients with polypoid cystitis. Pleomorphic microvilli were not observed in the bladder biopsies from control patients. The appearance of pleomorphic microvilli was consistent with reversible lesions of the human bladder and they are not specific markers of pre-neoplastic hyperplasia or malignancy.


A novel growth differentiation factor-9 (GDF-9) related factor is co-expressed with GDF-9 in mouse oocytes during folliculogenesis.
Growth differentiation factor-9 (GDF-9) is a transforming growth factor-b (TGF-b) family member which is expressed in the oocytes in mouse ovaries (McGrath, S.A., Esquela, A.F., Lee, S.J., 1995. Oocyte-specific expression of growth/differentiation factor-9. Mol. Endocrinol. 9, 131-136). GDF-9 is indispensable for normal folliculogenesis since female mice deficient for the GDF-9 gene are infertile due to an arrest of follicular growth at the primary follicle stage (Dong, J., Albertini, D.F., Nishimori, K., Kumar, T.R. , Lu, N., Matzuk, M.M., 1996. Growth differentiation factor-9 is required during early ovarian folliculogenesis. Nature 383, 531-535). We searched the GenBank Expressed Sequence Tag (EST) database with the mouse GDF-9 cDNA sequence, and identified from a mouse 2-cell embryo library an EST cDNA that encodes a putative member of the TGF-b superfamily, and named it as GDF-9B. Northern blot hybridization analyses of mouse ovaries revealed a single transcript of approximately 4.0 kilobases (kb) for GDF-9B and of 2.0 kb for GDF-9. We cloned by reverse transcription-polymerase chain reaction from mouse ovarian RNA a partial 821-base pair GDF-9B cDNA that spans the sequence encoding the putative mature region of GDF-9B. The COOH-terminal region of GDF-9B appears to be 53% homologous to GDF-9. Moreover, like GDF-9, GDF-9B lacks the cysteine residue needed for the covalent dimerization of several TGF-b family members. Using in situ hybridization analysis, we demonstrate that GDF-9B and GDF-9 mRNAs are co-localized in the oocyte. We also show that GDF-9B and GDF-9 genes are co-ordinately expressed during follicular development.


Eco-corona formation on the nanomaterials in the aquatic systems lessens their toxic impact: A comprehensive review.
Recent studies have shown that nanosized materials including plastics as a major cause of concern in the aquatic ecosystem. Fortunately, in the aquatic environment, the surface of the materials is often colonized by exudates of aquatic microorganisms (biofilm), where these materials are attached and surrounded by a secreted matrix with a sticky layer. The significance of these biofilms on the existence and beneficial implications of these pollutants has been studied in recent decades. Here we develop the concept of these pollutants as a complex matrix of polymers to which Extracellular Polymeric Substances (EPS) binds to form eco-corona modifying its density and surface charge of these particles. This review critically integrates the outstanding properties and functions of algal EPS in the aquatic environment and their dynamic interactions of early colonization on the surface of these pollutants, the impact of biofilm formation on stability, reactivity and, toxicity from the current literature. Due to the modifications of the environmental processes, EPS can have an impact on the toxicity thus special attention is focused on their behavior to decrease the toxicity of the pollutants in the aquatic environment. Although there has been an increasing number of researches in this area, further progress is needed to explore the extent to which ecological processes could be impacted, including the modifications in the behavior of aquatic pollutants. Thus, this review provides a recent perspective into the mechanisms of how eco-corona formation mitigates the toxicity of nanomaterials prevalent in aquatic ecosystems.


Ethanol tolerance measured electrophysiologically in hippocampal slices and not in neuromuscular junctions from chronically ethanol-fed rats.
The electrophysiologically measured effects of perfused ethanol were examined in neuromuscular junctions and hippocampal slices removed concurrently from naive and ethanol-tolerant rats. At the neuromuscular junction, ethanol augmented the frequency height, and half-width of spontaneous miniature endplate potentials (Mepp). No tolerance was noted to these effects in junctions from ethanol-fed animals. Ethanol depressed the stratum radiatum evoked hippocampal CA1 population spike. Clearcut tolerance to this effect was noted in slices from chronically ethanol-fed animals.


Combination TL01 ultraviolet B phototherapy and topical calcipotriol for psoriasis: a prospective randomized placebo-controlled clinical trial.
Previous studies have demonstrated the ultraviolet (UV)-sparing effect of combining topical calcipotriol with broadband UVB in the treatment of psoriasis. To determine if the combination of narrowband TL01 UVB phototherapy and topical calcipotriol produces the same UVB-sparing effect. This was a randomized, placebo-controlled, blinded clinical trial. Fifty psoriasis patients were recruited, 25 of whom were randomized into the active group who received TL01 phototherapy together with twice-daily application of calcipotriol cream 50 microg g(-1). The control group received TL01 phototherapy and twice-daily application of a topical emollient as placebo. TL01 phototherapy was given three times per week starting at 70% minimal erythema dose with 20% increments as tolerated for up to approximately 20 sessions. Patients were assessed using the Psoriasis Area and Severity Index (PASI) and Psoriasis Disability Index (PDI). They were evaluated at treatment sessions 8, 14 and 20, and followed up at 5 and 10 weeks post-treatment. Statistical analysis was performed using a two-tailed t-test. There were no significant differences in demographic characteristics and baseline PASI and PDI scores between the two groups. The mean PASI score declined significantly (P < 0.01) for both groups after treatment. The difference in mean PASI score reduction from baseline between the two groups was only significant during the first eight sessions, with a net reduction of 3.6 (95% confidence interval 1.0-6.2, P = 0.008) in the active group relative to the control group. The mean PDI score declined significantly (P < 0.05) for both groups, but there was no statistical difference in mean PDI score reduction between the two groups (P = 0.8) at the end of treatment. The mean cumulative UVB dose for the active group was significantly lower (P < 0.02) at 16 204 mJ cm-2 compared with 21 082 mJ cm-2 for the control group. We conclude that combining TL01 phototherapy with topical calcipotriol cream has a UVB-sparing effect.


Long-term monitoring of treatment with recombinant human growth hormone by serial determinations of type III procollagen-related antigens in serum.
Inasmuch as recombinant human growth hormone is now more generally available for the treatment of different types of short stature, there is a need for better short-term indicators of treatment success. In healthy children, serum concentrations of antigens related to the aminoterminal propeptide of type III procollagen (P-III-NP) closely follow the growth velocity curve. P-III-NP was measured longitudinally in 20 children with growth hormone deficiency during 6 months of human growth hormone substitution therapy. Two different radioimmunoassay systems were used; one recognizes predominantly the intact propeptide showing a lesser affinity to a smaller monomeric peptide (RIAgnost assay), while the other assay detects both forms equally (FAB assay). These results were compared to the growth response [median 5.6 (0.4 to 13.9) cm in 6 months] and to other established growth correlated parameters (somatomedin C, alkaline phosphatase). A relatively better growth response correlated significantly with high pretreatment P-III-NP (RIAgnost assay) values (r = 0.56) and delayed bone age (r = -0.70). A combination of these parameters in multiple regression analysis increased the cumulative prediction value to above 60% (r2 = 0.61). On the other hand, P-III-NP (FAB assay) values proved to be best in monitoring treatment, correlating with the individual growth rate during the first 3 months (r = 0.40; p less than 0.05), during the consecutive 3 months (r = 0.66; p less than 0.001), and during the total 6-month period (r = 0.46; p less than 0.05). All other parameters showed associations to growth only during some treatment periods.(ABSTRACT TRUNCATED AT 250 WORDS)


Nebivolol/Hydrochlorothiazide : a new fixed-dose combination for effective simplified antihypertensive therapy.
Hypertension is a major independent risk factor for cardiovascular (CV) morbidity and mortality and is combined with additional CV risk factors, such as diabetes mellitus, dyslipidaemia, heart disease, metabolic syndrome, smoking and obesity in many hypertensive individuals. The prevention of CV disease with antihypertensive therapy has been widely demonstrated, with a 30-40% reduction in stroke and a 20% reduction in coronary events in patients receiving antihypertensive drugs compared with untreated hypertensive individuals. Even small decreases in blood pressure (BP) correlate with a significantly lower incidence of CV events; this is independent of drug class used, but dependent on the extent of BP reduction achieved. All antihypertensive drugs lower BP to a similar extent and are suitable for long-term therapy. Treatment guidelines recommend that antihypertensive treatment should be initiated early to reach a target BP value of ≤140/90 mmHg, or lower in high-risk patients, in order to maximize the long-term benefits in CV mortality and morbidity reduction. However, the proportion of patients responding to monotherapy is generally low; therefore, first-line therapy with a combination of antihypertensive drugs is recommended in non-responders and those with multiple risk factors or subclinical organ damage. The combination of β-blocker/thiazide diuretic is frequently used in clinical practice and as reference treatment in clinical trials. Nebivolol, a third-generation cardioselective vasodilatory β-blocker, shows an additive effect in reducing systolic and diastolic BP when combined with hydrochlorothiazide (HCTZ). In monotherapy non-responders, the combination improves response and BP normalization rates compared with monotherapy. Since pharmacokinetic studies have shown that fixed-dose nebivolol-HCTZ therapy is bioequivalent to the two agents administered concomitantly, this combination is useful in monotherapy non-responders and for those who require rapid BP control to prevent end-organ damage. In addition, a simplified regimen improves patient compliance - a major obstacle to achieving target BP levels. In addition to providing enhanced BP reduction and control, nebivolol-HCTZ is well tolerated, with a similar incidence of adverse events to that observed with either monotherapy, and a neutral impact on lipid and glucose metabolism.


The carcinogenic effect of cigarette smoke. The effect of cigarette smoke on human gastric mucosal cells in organ culture.
The carcinogenic effect of cigarette smoke on certain organs is well established epidemiologically, but the evidence in relation to gastric carcinoma is inconclusive. The present study reports the effect of cigarette smoke condensate (CSC) on gastric mucosa grown in organ culture and its effect is compared with that of the known gastric carcinogen, N-methyl-N1-nitro-N-nitrosoguanidine (NG). Viability of the cells was assessed by the uptake of labelled glucose and glycine. It was found that, as gauged by lactate and beta-glucuronidase production into the ambient fluid, CSC produced a response typical of malignancy that did not differ from that with NG.


Outcomes with microsurgery of common peroneal nerve lesions.
The purpose of this retrospective study is to present our results with peroneal nerve lesions, to examine the relative significance of various factors, to assess their effect on outcome, and to establish guidelines of treatment for the microsurgical management of these difficult lesions. Over a 33-year period, a total of 62 patients were treated at McGill University and the Microsurgical Research Center, Eastern Virginia Medical School. The clinical records of all patients treated for peroneal nerve lesions were reviewed for retrospective analysis. Of 62 patients, 35 had microneurolysis and nerve decompression of the common peroneal nerve (CPn) as the only surgical procedure while 27 required reconstruction with nerve grafting. Postoperatively muscle power was graded from M + 4 to M - 5 in 27 patients, from M - 4 to M4 in 26 patients, from M - 3 to M + 3 in 8 patients, and from M - 2 to M + 2 in 1 patient. The behavioral video data showed a mean preoperative ankle dorsiflexion of 6.79° ± 5.6 and postoperative ankle dorsiflexion of 37.9° ± 9.3. Overall, excellent functional results were achieved in 27 of 62 patients (43%) with peroneal palsy who underwent microsurgical reconstruction, and good results were observed in 25 patients (40%). Despite previous widespread pessimism, the surgical repair of CPn lesion is worthwhile, yielding good to excellent results in the majority of patients, after a careful preoperative consultation, establishment of a sound strategy of reconstruction and using aggressive and atraumatic microsurgery.


Transcaval Access for Large Bore Devices.
Examine the latest data and techniques regarding transcaval access and closure. Transcaval access was proven to be a feasible and a translatable skill in a 100 patient open-label prospective study. No late complications from fistulas occurred and of all patients alive at 1 year, one fistula remained open. Transcaval is a viable access route for large bore devices. With adequate planning, bleeding and vascular complications are minimal. It should be integrated into the rubric of transcatheter large bore access.


Successful treatment of nonocclusive mesenteric ischemia in a reconstructed jejunum after esophagectomy and remnant gastric tube resection: a case report.
Nonocclusive mesenteric ischemia (NOMI), an ischemic bowel disease without a disruption of the mesenteric blood flow or strangulation of the mesentery or intestine, may cause a lethal clinical course. We report a very rare case of jejunal necrosis caused by NOMI in the pedicled mesentery of the reconstructed jejunum after remnant gastric tube resection for heterochronous gastric tube cancer after esophagectomy. An 80-year-old man visited our department with chief complaints of fever and appetite loss after 4 months from gastric tube resection and digestive reconstruction with pedicled jejunum. Contrast-enhanced computed tomography (CT) revealed impaired blood flow without torsion of the mesentery, severe wall thickness, and micro-penetration in the reconstructed jejunum and combined pyothorax in the right thoracic cavity. Esophagogastroduodenoscopy demonstrated extensive mucosal necrosis confined to the jejunum, which was elevated in the thoracic cavity. The jejunal necrosis due to NOMI occurring in the reconstructed jejunum was suspected, and lifesaving small bowel resection with right thoracotomy was considered necessary. However, radical operation with right thoracotomy was considered to be excessively invasive and not valid due to the patient's poor physical status, advanced age, and presence of left adrenal metastasis from the remnant gastric cancer. Therefore, we selected the conservative treatment with fasting, transnasal drainage, and administration of antibiotics due to the patient's intention. CT-guided right thoracic drainage for the intrathoracic abscess was needed 10 days after starting treatment and the inflammatory response rapidly improved. Follow-up CT and esophagogastroduodenoscopy revealed the improvement in the ischemic changes in jejunal mucosa without perforation. Intake was initiated at 20 days after symptom onset, and the patient was discharged at 40 hospital days without any complications and sequelae. To the best of our knowledge, this is the first case of NOMI occurring in the reconstructed jejunum after remnant gastric tube resection that was successfully treated with a conservative treatment. For NOMI, it is important to make appropriate diagnosis based on imaging findings and perform proper assessment of the patient's condition. Conservative treatments may be also useful depending on the patient's condition.


Temporal Associations of Screen Time and Anxiety Symptoms Among Adolescents.
There are indications that screen time is associated with symptoms of anxiety in adolescents, but from a longitudinal perspective, the magnitude, specificity, and temporal precedence in this association are relatively under-explored. To address this gap, over the course of 4 years, we annually surveyed adolescents assessing their usage of various types of screen time per day and symptoms of anxiety.


Comparative effectiveness of care coordination interventions in the emergency department: a systematic review.
To conduct a systematic review on the effectiveness of emergency department (ED)-based care coordination interventions. We reviewed any randomized controlled trial or quasi-experimental study indexed in MEDLINE, CINAHL, Web of Science, Cochrane, or Scopus that evaluated the effectiveness of ED-based care coordination interventions. To be included, interventions had to incorporate information from previous visits, provide educational services on continuing care, provide post-ED treatment plans, or transfer information to continuing care providers. Studies had to quantify information transfer or report ED revisits, hospitalizations, or follow-up rates. Randomized controlled trial quality was assessed with the Jadad score. Of 23 included articles, 14 were randomized controlled trials and 9 were quasi-experimental studies. Randomized controlled trial quality ranged from 2 to 3 on a 5-point scale. The majority of the studies (17) were conducted at a single center. Of nineteen studies that developed post-ED plans, 12 were effective in improving follow-up rates or reducing repeated ED visits. Four studies found paradoxically higher ED visit rates. Of 4 that used educational services for continuing care, 2 were effective. Of the 2 evaluating information transfer, 1 was effective. One study assessed incorporating information from other sites and found higher rates of information transfer, but utilization was not studied. The majority of ED-based care coordination interventions focus on interfacing with outpatient providers, and about two thirds have been effective in increasing follow-up rates or reducing repeated ED utilization. Other types of interventions have shown similar effectiveness, but fewer have been studied.


Reducing total mercury and methylmercury accumulation in rice grains through water management and deliberate selection of rice cultivars.
Rice consumption has been identified as a major route of methylmercury (MeHg) exposure in some areas of inland China. We investigated two potential mitigation methods (water management and deliberate selection of rice cultivars) to reduce the amount of total mercury (Hg) and MeHg within the grain. Rice grown aerobically had markedly reduced total Hg and MeHg concentrations as well as a much lower proportion of MeHg in the grain. Remarkably, there were considerable variations in the total Hg and MeHg concentrations as well as the proportion of MeHg in the grain among the 24 cultivars grown in the same paddy soil. The Hg tolerance index (expressed as % mean of control root growth) also varied substantially among the different cultivars. Furthermore, negative correlations were found between the total Hg and MeHg concentrations (P<0.05) of grain and the proportion of MeHg in the grain (P<0.01).


Effect of exercise on ventricular diastolic time in coronary artery disease.
To evaluate diastolic time during uninterrupted upright exercise, 28 normal volunteers (group 1) and 12 men with coronary artery disease (group 2) were studied by ear densitography. Electromechanical systole-heart rate and diastolic time-heart rate regression equations during upright exercise were obtained from group 1. Electromechanical systole-heart rate had an inverse linear relation (electromechanical systole = 480 - 1.4 heart rate) and diastolic time-heart rate had an inverse nonlinear relation (diastolic time = 1206e-0.02 heart rate). Although there were no significant differences in electromechanical systole and diastolic time at 1 minute of exercise between patients with and without CAD, at peak exercise prolongation of electromechanical systole and consequent shortening of diastolic time in patients with CAD were observed. This disproportionate shortening of diastole with lengthening of systole at peak exercise tends to decrease myocardial perfusion and, hence, oxygen supply, while increasing myocardial oxygen demand, contributing to aggravation of ischemia in patients with CAD.


Impact of positive peri-stent vascular remodeling after sirolimus-eluting and paclitaxel-eluting stent implantation on 5-year clinical outcomes: intravascular ultrasound analysis from the Poststent Optimal Stent Expansion Trial multicenter randomized trial.
Positive peri-stent vascular remodeling (PPVR) after drug-eluting stent (DES) implantation is an important mechanism of late-acquired stent malapposition (LASM). A total of 226 patients (sirolimus-eluting stent [SES], n=105; paclitaxel-eluting stent [PES], n=121) from the Poststent Optimal Stent Expansion Trial who underwent a post-intervention and 9-month follow-up intravascular ultrasound were followed clinically for 5 years. PPVR was arbitrarily defined as a >10% increase in the external elastic membrane volume index at follow-up. PPVR and LASM occurred more frequently with SESs than with PESs. The 5-year rate of major adverse cardiac events was lower with SES than with PES (10.7% vs. 23.2%, P=0.002). The late and very late stent thrombosis (ST) rate was similar between the 2 DES types, but it was higher in patients with PPVR than in those without PPVR (8.8% vs. 1.3%, P=0.009) regardless of the DES type. Early discontinuation (<1 year) of dual antiplatelet therapy (DAPT; hazard ratio [HR], 24.14; 95% confidence interval [CI]: 4.90-118.87; P<0.001), PPVR (HR, 14.94; 95%CI: 1.85-120.46; P=0.011), LASM (HR, 8.01; 95%CI: 1.93-33.16; P=0.004), and stent length (HR, 1.14; 95%CI: 0.98-1.32 per mm; P=0.078) were associated with increased risk of late and very late ST. PPVR and LASM development after DES implantation, along with early discontinuation of DAPT and longer stent length, are important risk factors of late and very late ST.


Ultrastructure of juvenile hormone-induced degenerating flight muscles in a bark beetle, Ips paraconfusus.
Topical application of 5 microgram of a juvenile hormone analogue (JHA), ZR-615, to female callow adults of Ips paraconfusus induced degeneration of the dorsoventral flight muscles. Within 24 h after JHA-treatment the diameter of the myofibrils was reduced to almost half due to the lysis of the peripheral myofilaments. Mitochondria showed conspicuous degenerative changes like swelling, dissolution of the matrix or presence in the matrix of dense filamentous material or myelin-like figures. Degeneration of the mitochondria seemed to take place inside isolation membranes derived from sarcoplasmic reticulum. A number of granular osmiophilic bodies appeared in the sarcoplasm. Three days after JHA-treatment the muscles were very thin and sheath-like. Most of the mitochondria had already degenerated. The dense sarcoplasm contained numerous crystalline bodies. The granular dense bodies were also more frequent. The myofibrils were comprised of only occasional small bundles of myofilaments. The tubules of the T system enclosed an amorphous material. The nuclei and the tracheal system remained intact but they were crowded due to the decreased volume of the muscle. In some specimens degeneration of the myofibrils and mitochondria was completed by the third day. No sign of degeneration was observed in the flight muscles of acetone treated control insects.


Coagulant toxicity and effectiveness in a slaughterhouse wastewater treatment plant.
Attempts were made in this study to examine the toxicity of polymer/alum addition to the aeration tank effluent prior to sludge flotation as practiced in a slaughterhouse wastewater treatment plant. Based on the Microtox toxicity assay, alum at concentrations 100-200 mg/L was found to slightly increase the toxicity level of slaughterhouse wastewater effluent. However, at higher concentrations (300-1000 mg/L), significant residual chronic toxicity remained in all slaughterhouse wastewater effluents, independent of the treatment process. Polymer, on the other hand, removed organics and solids, but polymer effluents are more toxic than alum at extremely low concentration. Results indicated that alum and polymer caused inhibitory effects to the system at soluble concentrations of approximately 400 and 60 mg/L and above, respectively. The data also indicated that the solids collected in both tests (polymer/alum) were much more toxic than those from the effluents. Sediment samples from the polymer tests were the most toxic. Furthermore, the effluent toxicity of the coagulants was dramatically more significant when used after the settlement of solids than when used in mixed liquor. In addition, strong correlations were observed between the observed toxicity for a series of supernatants and the coagulant concentrations of alum/polymer processes, and between supernatants and solids collected in both tests.


Excessive workload, uncertain career opportunities and lack of funding are important barriers to recruiting and retaining primary care medical researchers: a qualitative interview study.
Research activity in primary care has been steadily increasing, but is still insufficient and more researchers are needed. Many initiatives have been launched to recruit and retain primary care researchers, but only little is known about barriers and facilitators to a research career in primary care. To examine barriers and facilitators to recruiting and retaining primary care medical researchers. Semi-structured interviews with 33 primary care medical researchers, all medical doctors. We used a phenomenological approach to analysing the interviews. Important barriers to pursuing a research career in primary care were heavy workload, isolation at work, short-term funding and low salary. Important facilitators to attracting and retaining primary care researchers were the desire and opportunity to improve primary care, the flexible working conditions, the career opportunities, including the possibility of combining university-based research with clinical work and a friendly and competent research environment. Better strategies for recruiting and retaining researchers are a prerequisite for the development of primary care, and in future the main emphasis should be on working conditions, networking and mentoring. Studies including those primary care physicians who have chosen not to do research are highly needed.


Preparation of colloidal CdSe and CdS/CdSe nanoparticles from sodium selenosulfate in aqueous polymers solutions.
Cadmium selenide nanoparticles formation at the interaction between CdCl2 and Na2SeSO3 in aqueous solutions of sodium polyphosphate and gelatin has been studied. Structural and optical properties of CdSe nanoparticles have been characterized. It has been shown that the temperature and the ratio of reagents concentrations are the basic parameters, controlling the size of CdSe nanoparticles. Photocatalytic activity of CdS nanoparticles in Na2SeSO3 reduction has been found and investigated; structural and optical properties of binary CdS/CdSe nanoparticles have been characterized. This photoreaction, when carried out in the presence of CdCl2, results in the formation of composite CdS/CdSe nanoparticles. It has been shown that slow interaction of adsorbed selenosulfate with surface-trapped CdS conduction band electrons is the limiting stage of the photocatalytic reaction.


Chronic mild stress alters the somatostatin receptors in the rat brain.
The involvement of somatostatin (SST) and its receptors in the pathophysiology of depression and stress has been evidenced by numerous studies. The purpose of the present study was to find whether chronic mild stress (CMS), an animal model of depression, affects the SST receptors in the rat brain and pituitary, as well as the level of SST in plasma. In CMS model, rats were subjected to 2 weeks of stress and behaviorally characterized using the sucrose consumption test into differently reacting groups based on their response to stress, i.e., stress-reactive (anhedonic), stress-non-reactive (resilient), and invert-reactive rats (characterized by excessive sucrose intake). We measured specific binding of [125I]Tyr3-Octreotide, expression of mRNA encoding sst2R receptors in the rat brains, expression of SST and its receptors in rat pituitary, and the level of SST in the plasma. The obtained results show decreases in binding of [125I]Tyr3-Octreotide in most of rat brain regions upon CMS and no significant differences between three stressed groups of animals, except for significant up-regulation of sst2 receptor in medial habenula (MHb) in the stress-reactive group. In the same group of animals, significant increase in plasma SST level was observed. There are two particularly sensitive sites distinguishing the response to stress in CMS model. In the brain, it is MHb, while on the periphery this predictor is SST level in plasma. These changes may broaden an understanding of the mechanisms involved in the stress response and point to the intriguing role of MHb.


Musculoskeletal changes in women with spinal cord injury: a twin study.
Bone mineral density (BMD), bone geometry, and muscle cross-sectional area (CSA) were investigated in two sets of monozygotic female twins, where one of each pair had sustained a spinal cord injury (SCI). Twin pair 1 (TP1) and twin pair 2 (TP2) were 32 and 47 yr old and were 7 and greater than 20 yr postinjury, respectively. BMD was measured using dual-energy X-ray absorptiometry. Computed tomography was used to measure volumetric BMD, bone geometry, and muscle CSA of the thigh and calf. For the SCI twin of TP1, BMDs of hip, distal femur, proximal tibia, and spine were 59.5, 46.6, 53.1, and 93.3%, respectively, of values of the noninjured twin. For TP2, corresponding values in the SCI twin were 36.2, 35.9, 39.2, and 62.2%, respectively, of the non-SCI twin. Average muscle CSAs of the SCI twins were 31.2+/-2.3% and 31.0+/-6.1% of the values for their non-SCI twin, indicating that muscle CSAs were reduced by 70%. Among the SCI twins, volumetric BMDs were 83.4+/-2.4% and 87.0+/-3.45% of the non-SCI twins' values. Moments of inertia in the SCI twins ranged from 62.4 to 97.9% of the non-SCI twins' values at mid-femur and calf. This study reveals that in addition to the declines in muscle CSA and BMD, important changes in bone geometry occur in women after SCI.


Mesenchymal stromal cell potency to treat acute kidney injury increased by ultrasound-activated interferon-γ/interleukin-10 axis.
Mesenchymal stromal cell (MSC) therapies combined with renal pulsed focused ultrasound (pFUS) pretreatment increase MSC homing and improve cisplatin-induced acute kidney injury (AKI) better than MSC alone. However, mechanisms underlying improved outcomes remain unknown. We hypothesize pFUS up-regulates renal interferon-γ (IFNγ) and stimulates MSC to produce interleukin-10 (IL-10) after migrating to kidneys. To demonstrate initially, MSC cultured with IFNγ up-regulated IL-10. More MSC-derived IL-10 was detected in kidneys when IFNγ-stimulated MSC were infused and they improved AKI better than unstimulated MSC. Next, IFNγ-knockout mice with AKI received pFUS+MSC, but MSC-derived IL-10 expression and AKI were similar to using MSC alone. AKI in wild-type mice receiving pFUS and IL-10-deficient MSC was also unimproved compared to administering IL-10-deficient MSC alone. Indoleamine 2,3-dioxygenase (IDO), an anti-inflammatory enzyme up-regulated in MSC by IFNγ, was up-regulated during AKI, but was not further elevated in MSC from pFUS-treated kidneys, suggesting that IDO is not involved in improved AKI healing by pFUS+MSC. These data suggest IFNγ is up-regulated by pFUS and after i.v.-infused MSC home to pFUS-treated kidneys, IFNγ stimulates additional IL-10 production by MSC to improve AKI. Analogous mechanisms of ultrasound-treated tissue microenvironments stimulating therapeutic MSC may exist in other pathologies where adjuvant ultrasound techniques are successful.


Impact of thymidine phosphorylase and CD163 expression on prognosis in stage II colorectal cancer.
Tumor-associated macrophages (TAM) are known to facilitate colorectal cancer (CRC) growth. High macrophage infiltration in thymidine phosphorylase (TYMP) expressing CRC may correspond to poor prognosis. The prognostic impact of the expression CD163, a receptor associated with TAM, and TYMP in stroma, respectively, tumor tissue is not yet established. The aim of this study was to identify the potential associations between TYMP and CD163 expression levels and relapse-free survival (RFS) of patients with stage II CRC, and if microdissection is of importance. Stage II CRC patients, radically resected with relapse (n = 104), were matched to patients with a 5-year relapse-free follow-up (n = 206). Gene expression of TYMP and CD163 was analyzed in snap-frozen tumor tissues and in microdissected formalin-fixed tumor tissues separated into tumor epithelium and stroma. TYMP expression was high in poorly differentiated tumors, right-sided CRC, and tumors with high microsatellite instability CD163-expressing macrophages near tumor epithelial cells had high expression in poorly differentiated and T4 tumors. High TYMP expression in tumor epithelial cells was in the multivariate analyses associated with shorter relapse-free survival (hazard ratio 1.66; 95% confidence interval: 1.09-2.56; p < 0.05). TYMP expression in tumor epithelial cells was associated with RFS and emphasizes the need for tissue microdissection. Additional studies are needed to establish whether TYMP and CD163 could add clinically relevant information to identify high-risk stage II patients that could benefit from adjuvant chemotherapy.


Activation of a chondrocyte volume-sensitive Cl(-) conductance prior to macroscopic cartilage lesion formation in the rabbit knee anterior cruciate ligament transection osteoarthritis model.
The anterior cruciate ligament transection (ACLT) rabbit osteoarthritis (OA) model confers permanent knee instability and induces joint degeneration. The degeneration process is complex, but includes chondrocyte apoptosis and OA-like loss of cartilage integrity. Previously, we reported that activation of a volume-sensitive Cl(-) current (ICl,vol) can mediate cell shrinkage and apoptosis in rabbit articular chondrocytes. Our objective was therefore to investigate whether ICl,vol was activated in the early stages of the rabbit ACLT OA model. Adult Rabbits underwent unilateral ACLT and contralateral arthrotomy (sham) surgery. Rabbits were euthanized at 2 or 4 weeks. Samples were analyzed histologically and with assays of cell volume, apoptosis and electrophysiological characterization of ICl,vol. At 2 and 4 weeks post ACLT cartilage appeared histologically normal, nevertheless cell swelling and caspase 3/7 activity were both significantly increased compared to sham controls. In cell-volume experiments, exposure of chondrocytes to hypotonic solution led to a greater increase in cell size in ACLT compared to controls. Caspase-3/7 activity, an indicator of apoptosis, was elevated in both ACLT 2wk and 4wk. Whole-cell currents were recorded with patch clamp of chondrocytes in iso-osmotic and hypo-osmotic external solutions under conditions where Na(+), K(+) and Ca(2+) currents were minimized. ACLT treatment resulted in a large increase in hypotonic-activated chloride conductance. Changes in chondrocyte ion channels take place prior to the onset of apparent cartilage loss in the ACLT rabbit model of OA. Further studies are needed to investigate if pharmacological inhibition of ICl,vol decreases progression of OA in animal models.


Neuropeptides and the blood-brain barrier in goldfish.
The general activity level of a goldfish is easily monitored by placing it in water to a depth of 2.5 cm in an aquarium on top of an activity meter. With this system, goldfish were administered a 5 microliter (80 microgram/kg) intracranial (IC) or intraperitoneal (IP) injection of one of 21 compounds and tested for general activity. The results indicated that activity decreased significantly over time and the peptides differentially decreased activity, with the greatest alterations in activity produced by two new enkephalin analogs: D-Ala2, F5Phe4-enkephalin-NH2 and Nalpha Nepsilon-bis-(D-Ala2-enkephalin)-Lys-NH2. Overall, decreased activity began approximately 3 min after an IC injections and 6 min after an IP injection. The longer latency after IP injections may indicate the time required for the substance, either in its original or fragmented form, to reach and cross the blood-brain barrier and makes a primary peripheral effect unlikely. Most of the peptides or possibly their metabolites appeared to enter the brain since no significant difference in activity existed after IC and IP injections, with both producing reliable decreases from the control. In summary, peptides can exert behavioral effects after both IC and IP administration in goldfish.


IoT Applications with 5G Connectivity in Medical Tourism Sector Management: Third-Party Service Scenarios.
5G is the next generation of mobile communication technology. Current generation of wireless technologies is being evolved toward 5G for better serving end users and transforming our society. Supported by 5G cloud technology, personal devices will extend their capabilities to various applications, supporting smart life. They will have significant role in health, medical tourism, security, safety, and social life applications. The next wave of mobile communication is to mobilize and automate industries and industry processes via Machine-Type Communication (MTC) and Internet of Things (IoT). The current key performance indicators for the 5G infrastructure for the fully connected society are sufficient to satisfy most of the technical requirements in the healthcare sector. Thus, 5G can be considered as a door opener for new possibilities and use cases, many of which are as yet unknown. In this paper we present heterogeneous use cases in medical tourism sector, based on 5G infrastructure technologies and third-party cloud services.


Greater trochanter osteotomy as a component of cementless total hip replacement: Five cases in four dogs.
To describe the surgical technique and report the long-term outcome of greater trochanteric osteotomy (GTO) as a component of cementless total hip replacement (THR) in dogs with severe medialization of the greater trochanter or chronic craniodorsal hip luxation. Short case series. Four dogs treated with five THRs. Data collected from medical records included signalment, indication for THR, duration of clinical signs, body weight, pre- and post-operative radiographic assessment, surgical templating, osteotomy technique, THR implant selection, surgical time, complications, and long-term clinical and radiographic follow-up. Clinical outcomes were determined based on in-hospital history, orthopedic examination, and radiographic evaluation. All five surgical procedures resulted in satisfactory long-term clinical results at follow-up a median of 48.2 months (range, 34-56 months) after THR. There were no minor complications and one major complication. One dog experienced post-operative luxation unrelated to the GTO and was successfully treated with a cup revision. GTO was effective in facilitating cementless THR in dogs with either severe medialization of the greater trochanter or chronic craniodorsal luxation.


Potential for seed-mediated gene flow in agroecosystems from transgenic safflower (Carthamus tinctorius L.) intended for plant molecular farming.
Safflower has been transformed for field scale molecular farming of high-value proteins including several pharmaceuticals. Viable safflower seed remaining in the soil seed bank after harvest could facilitate seed and pollen-mediated gene flow. Seeds may germinate in subsequent years and volunteer plants may flower and potentially outcross with commodity safflower and/or produce seed. Seeds from volunteers could become admixed with conventional crops at harvest, and/or replenish the seed bank. Seed in following crops could be transported locally and internationally and facilitate gene flow in locations where regulatory thresholds and public acceptance differ from Canada. Seed-mediated gene flow was examined in three studies. Safflower seed loss and viability following harvest of commercial fields of a non-transgenic cultivar were determined. We assessed seed longevity of transgenic and non-transgenic safflower, on the soil surface and buried at two depths. Finally, we surveyed commercial safflower fields at different sites and measured density and growth stage of safflower volunteers, in other crops the following year and documented volunteer survival and viable seed production. Total seed loss at harvest in commercial fields, ranged from 231 to 1,069 seeds m(-2) and the number of viable seeds ranged from 81 to 518 seeds m(-2). Safflower has a relatively short longevity in the seed bank and no viable seeds were found after 2 years. Based on the seed burial studies it is predicted that winter conditions would reduce safflower seed viability on the soil surface by >50%, leaving between 40 and 260 viable seeds m(-2). The density of safflower volunteers emerging in the early spring of the following year ranged from 3 to 11 seedlings m(-2). Safflower volunteers did not survive in fields under chemical fallow, but in some cereal fields small numbers of volunteers did survive and generate viable seed. Results will be used to make recommendations for best management practices to reduce seed-mediated gene flow from commercial production of plant molecular farming with safflower.


Metagenomics in animal gastrointestinal ecosystem: Potential biotechnological prospects.
Microbial metagenomics---the applications of the genomics suit of technologies to nonculturable microorganisms, is coming of age. These approaches can be used for the screening and identification of nonculturable gastrointestinal (GI) microflora for assessing and exploiting them in nutrition and the health of the host. Advances in technologies designed to access this wealth of genetic information through environmental nucleic acids extraction and analysis have provided the means of overcoming the limitations of conventional culture-dependent microbial genetic exploitation. The molecular techniques and bioinformatics tools will result in reliable insights into the animals' GI microbial structure and activity of the livestock gut microbes in relation to functional interactions, temporal and spatial relationships among different microbial consortia and dietary ingredients. Further developments and applications of these methods promise to provide the opportunity to link distribution and identity of various GI microbes in their natural habitats, and explore their use for promoting livestock health and industrial development.


Modification of the cell surface expression of histocompatibility antigens induced by the neurotoxin 2,5 hexanedione.
Class I histocompatibility antigens (HLA) are expressed on the surface of almost all nucleated mammalian cells; the expression of this surface antigenic molecule may be changed or abrogated by several factors. In this paper, a modification in HLA expression in a human carcinoma cell line following exposure to the neurotoxicant 2,5 hexanedione is reported. This compound is known to produce a wide spectrum of subcellular pathological events; in this study, we describe an effect on the surface and cytoplasmic distribution of both light and heavy subunits of HLA antigens, demonstrated by immunocytochemical and immunoelectron microscopy techniques. Human carcinoma cells, which under normal growing conditions express the HLA, abrogate the surface expression of this glycoprotein after exposure to 2,5 hexanedione and an intracytoplasmic accumulation seems to occur. Several possibilities are discussed, such as an effect of the toxicant on the transport of the nascent glycoprotein.


The development of the relationship between dorsal root afferents and motoneurons in the larval bullfrog spinal cord.
The relationship of dorsal root afferents to motoneuron somata and dendrites was studied by labelling dorsal and ventral roots of the tadpole lumbar enlargement with HRP at different stages of hindlimb development. Procedures were used which allowed for sequential light and electron microscopic analysis to determine whether close appositions between labelled elements represented synaptic contacts. Lateral motor column (LMC) motoneuron dendrites grow first into the lateral funiculus, and later begin arborizing within the spinal gray, concurrent with the arrival of developing dorsal root afferent fibers. Mature-appearing synaptic contacts between dorsal root afferents and motoneuron dendrites are established first on distal dendrites, and are observed on progressively more proximal dendrites as hindlimb development proceeds. Migrating motoneurons were also labelled in some animals. Distinct dorsal and ventral migratory pathways were noted; cells migrating dorsally were contacted by developing dorsal root afferents. Migrating motoneurons were associated with radially oriented processes, and were often closely apposed to other cells. The coincident development of dorsal root projections and the motoneuron dendrites which these fibers innervate in the adult, as well as the interaction between these two systems during cell migration, suggest that these two systems may be interdependent in establishing their normal relationship during development.


Daphnia magna can tolerate short-term starvation without major changes in lipid metabolism.
Daphnia magna is a common crustacean that is adapted to brief spells of fasting. Lipids are naturally a major component of their diet and are stored as energy reserves. However, there has been some controversy in the literature on the extent to which dietary lipids are used directly for complex lipid formation in Daphnia. We examined lipid metabolism in D. magna by labeling the animals using [1-14C]acetate and then followed the turnover of radiolabeled lipids during a pulse chase. Daphnia were either fed or maintained without food during the chase period. The decrease in radioactivity during the chase was relatively unaffected by feeding, although there were some differences in the distribution of radioactivity between lipid classes or individual FA. The polar lipids, which were four times better labeled than nonpolar lipids, contained the most radioactivity in the zwitterionic phosphoglycerides, PE and PC. Under the experimental conditions, the turnover of the polar membrane lipids was unaffected by feeding. Within nonpolar lipids, TAG accounted for up to about 80% of the label, followed by DAG. Overall, our data show that D. magna is capable of high rates of lipid radiolabeling de novo and, in addition, is able to use--and indeed may be dependent on--some dietary components such as the PUFA linoleate and alpha-linolenate. The results also clearly show that Daphnia is able to tolerate brief spells of fasting (24 h) with very little change to its lipid metabolism.


Comparison of morbidity associated with laparoscopic myomectomy and hysterectomy for the treatment of uterine leiomyomas.
To compare short-term morbidity and quality of life after laparoscopic hysterectomy (LH) and laparoscopic myomectomy (LM) for the treatment of symptomatic uterine leiomyomas. We performed a prospective, observational study of women who were eligible for both surgical procedures. After informed consent was obtained, each participant was asked to complete the SF-12v2 Health Survey before surgery and to repeat it seven days and 28 days after surgery. Data on short-term morbidities, such as operative time, blood loss, length of hospital stay, and surgical complications, were collected by an obstetrician-gynaecologist. Women who underwent LH were compared by non-parametric statistical analyses with those who underwent LM. Sixty-one women were recruited between January 1 and December 31, 2008, including 40 who underwent LM and 21 LH. Women who underwent LH were older, had higher parity, and were less likely to have infertility than those who chose LM. Median LH operative time of 223 minutes (IQR 214 to 241) was slightly longer than for LM (188 minutes, IQR 154 to 239; P = 0.02). However, we found no difference between the two groups in terms of SF-12v2 fluctuation, blood loss, hospital stay, and short-term complications. Laparoscopic myomectomy is a viable alternative to laparoscopic hysterectomy for women with symptomatic leiomyomas who want conservative surgery. The procedures have similar morbidity and impact on quality of life.


Primary Immune Deficiency Treatment Consortium (PIDTC) update.
The Primary Immune Deficiency Treatment Consortium (PIDTC) is a collaboration of 41 North American centers studying therapy for rare primary immune deficiency diseases (PIDs), including severe combined immune deficiency (SCID), Wiskott-Aldrich syndrome (WAS), and chronic granulomatous disease (CGD). An additional 3 European centers have partnered with the PIDTC to study CGD. Natural history protocols of the PIDTC analyze outcomes of treatment for rare PIDs in multicenter longitudinal retrospective, prospective, and cross-sectional studies. Since 2009, participating centers have enrolled more than 800 subjects on PIDTC protocols for SCID, and enrollment in the studies on WAS and CGD is underway. Four pilot projects have been funded, and 12 junior investigators have received fellowship awards. Important publications of the consortium describe the outcomes of hematopoietic cell transplantation for SCID during 2000-2009, diagnostic criteria for SCID, and the pilot project of newborn screening for SCID in the Navajo Nation. The PIDTC Annual Scientific Workshops provide an opportunity to strengthen collaborations with junior investigators, patient advocacy groups, and international colleagues. Funded by the National Institute of Allergy and Infectious Diseases and the Office of Rare Diseases Research, National Center for Advancing Translational Sciences, the PIDTC has recently received renewal for another 5 years. Here we review accomplishments of the group, projects underway, highlights of recent workshops, and challenges for the future.


Genetic rearrangements in Pseudomonas amygdali pathovar aesculi shape coronatine plasmids.
Plant pathogenic Pseudomonas species use multiple classes of toxins and virulence factors during host infection. The genes encoding these pathogenicity factors are often located on plasmids and other mobile genetic elements, suggesting that they are acquired through horizontal gene transfer to confer an evolutionary advantage for successful adaptation to host infection. However, the genetic rearrangements that have led to mobilization of the pathogenicity genes are not fully understood. In this study, we have sequenced and analyzed the complete genome sequences of four Pseudomonas amygdali pv. aesculi (Pae), which infect European horse chestnut trees (Aesculus hippocastanum) and belong to phylogroup 3 of the P. syringae species complex. The four investigated genomes contain six groups of plasmids that all encode pathogenicity factors. Effector genes were found to be mostly associated with insertion sequence elements, suggesting that virulence genes are generally mobilized and potentially undergo horizontal gene transfer after transfer to a conjugative plasmid. We show that the biosynthetic gene cluster encoding the phytotoxin coronatine was recently transferred from a chromosomal location to a mobilizable plasmid that subsequently formed a co-integrate with a conjugative plasmid.


Childhood cancer epidemiology in the Czech Republic (1994-2016).
The knowledge of cancer burden in the population, its time trends and the possibility of international comparison is an important starting point for cancer control programmes. Our study aimed to evaluate trends in childhood cancer epidemiology of patients aged 0-14 years in the period 1994-2016 in the Czech Republic. Data on childhood cancers have been obtained from the Czech National Cancer Registry. These data were validated using the clinical database of childhood cancer patients and combined with data from death certificates. Incidence and mortality trends were assessed by the joinpoint regression method. The life tables method was used to calculate the overall age-standardised five-year survival. The incidence trend was stable; the age-standardised (world) cancer incidence - ASR (W) - was 173.7 per 1 million children in the period 1994-2016. However, there was apparent significant decrease in mortality: ASR (W) dropped from 58.1 per 1 million children in 1994 to 21.4 per 1 million children in 2016. The overall five-year survival increased over time by 10 %. Statistically significant improvements in survival were observed in patients with lymphoid leukaemia, astrocytomas, neuroblastomas, osteosarcomas and rhabdomyosarcomas. Such a relevant increase in survival rates, and therefore also a decrease in mortality rates in the Czech Republic, is most likely due to improvements in diagnostic and treatment methods since the 1990s, which were facilitated by the concentration of childhood cancer patients in children's cancer centres.


Semi-automated colony-forming unit counting for biosafety level 3 laboratories.
Biosafety level 3 decontamination precautions motivate measuring microbial colonies using consumable photography instead of expensive automated plate counters or smartphones, and assaying drug treatments-with multiple concentrations per treatment, replicates, and controls-produces hundreds of images. Here, we present a protocol for semi-automated image analysis by hand-tuning three parameters. The parameters control for non-uniform colony growth and artifacts such as lid condensation, reflections, and plating streaks. We describe steps to prepare images, tune parameters, and plot dose-response relationships. For complete details on the use and execution of this protocol, please refer to Larkins-Ford et al.1.


A Novel Ultrasound-Guided Technique for Hysteroscopic Adhesiolysis in High-Risk Patients.
Hysteroscopy is a common gynecologic surgical procedure. Certain diagnoses, notably intrauterine adhesions and cervical stenosis, make hysteroscopy more complicated because of an increased likelihood of complications. Three patients, 1 with cervical stenosis and 2 with Asherman syndrome, underwent ultrasound (US)-guided adhesiolysis. Access to the uterine cavity was obtained by either direct balloon-aided dilation or the US-guided Seldinger technique, followed by balloon-aided dilation to enter the endometrial cavity and disrupt intrauterine/intracervical adhesions. In this case series, we describe a novel approach of using US-guided balloon dilation to safely and effectively treat intrauterine adhesions and to decrease the risk of perforation.


In vitro non-enzymatic glycation and formation of browning products in the bovine lens alpha-crystallin.
Calf lens LMW alpha-crystallin was glycated by incubating with various sugars (glucose, glucose-6-P and ribose) for 21 days. All sugars induced disulfide formation, but ribose also produced higher molecular weight cross-linked species. The ribocated protein turned yellow in color and had a strong blue fluorescence (Ex/Em = 370/450 nm) typical for a browning product. The chromophore of the browning product showed a new circular dichroism (CD) band at 320-330 nm. Conformational study indicated that the browning reaction destablized protein and may play a significant role in protein aggregation and insolubilization.


Continuum models in neurobiology and information processing.
Continuum models have been used with considerable success for single neurons but have been neglected in the study of neuronal populations. In more popular discrete neuronal network models, the geometric details of the neuronal centers are usually neglected. We here give a continuum nonlinear dynamical model and an approximate model which admits the possibility of ascertaining the roles of the various connectivity patterns from center to center in the central nervous system. Frequency transfer characteristics are used to incorporate the nonlinear dynamics of single neurons. Simple examples are evaluated both analytically and numerically and the results presented graphically.


Determination of the position of the Qi.- quinone binding site from the protein surface of the cytochrome bc1 complex in Rhodobacter capsulates chromatophores.
The technique of distance measurement, utilizing spin relaxation enhancement by an external probe, has been extended to the study of intrinsic semiquinone radicals through the use of holmium-EDTA complexes and continuous wave electron paramagnetic resonance spectroscopy. This technique has been used to determine the distance of the semiquinone anion, Qi (also designated as Qn.- or Qc.-), from the surface of the ubiquinone cytochrome c oxidoreductase, consisting of only three subunits, in membrane particles from Rhodobacter capsulates. The location of the semiquinone anion is 6-10 A from the N side protein, establishing that there are two separate quinone reaction sites, i.e., 'Qi' and 'Qo', within this complex on opposite sides of the membrane. The results are discussed in relation to reported ENDOR, EPR, and optical studies of the mitochondrial counterpart.


Sensory temporal processing in adults with early hearing loss.
This study examined tactile and visual temporal processing in adults with early loss of hearing. The tactile task consisted of punctate stimulations that were delivered to one or both hands by a mechanical tactile stimulator. Pairs of light emitting diodes were presented on a display for visual stimulation. Responses consisted of YES or NO judgments as to whether the onset of the pairs of stimuli was perceived simultaneously or non-simultaneously. Tactile and visual temporal thresholds were significantly higher for the deaf group when compared to controls. In contrast to controls, tactile and visual temporal thresholds for the deaf group did not differ when presentation locations were examined. Overall findings of this study support the notion that temporal processing is compromised following early deafness regardless of the spatial location in which the stimuli are presented.


Potentiation of lethality and increase in body temperature by combined use of d-methamphetamine and morphine in mice.
Lethality and change in body temperature in mice were examined after subcutaneous injection of d-methamphetamine and morphine alone or in combination. The LD50 values for methamphetamine and morphine were calculated to be 95 and 670 mg/kg body wt., respectively. When a non-lethal dose of morphine (300 mg/kg) was administered with various doses of methamphetamine, the LD50 for methamphetamine was reduced to 5 mg/kg, indicating a marked potentiation of toxicity by combined use of both drugs. Injection of 5 mg/kg of methamphetamine produced slight hyperthermia, while 300 mg/kg of morphine decreased the body temperature of mice. However, when both drugs were used concomitantly, a marked increase in body temperature was observed. Hyperthermia was also observed when the dose of morphine was reduced to 50 mg/kg. It is postulated that hyperthermia is probably one of the contributory factors in the potentiated toxicity by combined use of morphine and methamphetamine.


Geomagnetic activity, humidity, temperature and headache: is there any correlation?
Meterological factors influence several biological functions. Geomagnetic activity (GMA) can be considered a trigger factor of migraine attacks. We studied the possible relationship between 40 migraine patients and some meteorological factors: humidity, temperature and geomagnetic activity in particular. All frequency changes of geomagnetic activity, temperature and humidity values are recorded daily. The study was performed from March to June 1988 over a geographically small area in order to avoid climatic and environmental influences. Our results indicate a significant correlation between geomagnetic activity and migraine attack frequency. Controversial opinions concerning the modalities of data collection and the possible relationships between environment and headache raise the need of further studies.


Dissipation of Energy by Dry Granular Matter in a Rotating Cylinder.
We study experimentally the dissipation of energy in a rotating cylinder which is partially filled by granular material. We consider the range of angular velocity corresponding to continous and stationary flow of the granulate. In this regime, the stationary state depends on the angular velocity and on the filling mass. For a wide interval of filling levels we find a universal behavior of the driving torque required to sustain the stationary state as a function of the angular velocity. The result may be of relevance to industrial applications, e.g. to understand the power consumption of ball mills or rotary kilns and also for damping applications where mechanical energy has to be dissipated in a controlled way.


Gene therapy for ventricular tachyarrhythmias.
Cardiac arrest is the leading cause of death in the United States and other developed countries. Ventricular tachyarrhythmias are the most prominent cause of cardiac arrest, and patients with structural heart disease are at increased risk for these abnormal heart rhythms. Drug and device therapies have important limitations that make them inadequate to meet this challenge. We and others have proposed development of arrhythmia gene therapy as an alternative to current treatment methods. In this review, I discuss the basic mechanisms of ventricular arrhythmias and summarize the literature on the use of gene therapy for ventricular tachyarrhythmias.


Fabrication of Tβ4-Exosome-releasing artificial stem cells for myocardial infarction therapy by improving coronary collateralization.
Currently, stem cell transplantations in cardiac repair are limited owing to disadvantages, such as immunological rejection and poor cell viability. Although direct injection of exosomes can have a curative effect similar to that of stem cell transplantation, high clearance hinders its application in clinical practice. Previous reports suggested that induction of coronary collateralization can be a desired method of adjunctive therapy for someone who had missed the optimal operation time to attenuate myocardial ischemia. In this study, to mimic the paracrine and biological activity of stem cells, we developed artificial stem cells that can continuously release Tβ4-exosomes (Tβ4-ASCs) by encapsulating specific exosomes within microspheres using microfluidics technology. The results show that Tβ4-ASCs can greatly promote coronary collateralization in the periphery of the myocardial infarcted area, and its therapeutic effect is superior to that of directly injecting the exosomes. In addition, to better understand how it works, we demonstrated that the Tβ4-ASC-derived exosomes can enhance the angiogenic capacity of coronary endothelial cells (CAECs) via the miR-17-5p/PHD3/Hif-1α pathway. In brief, as artificial stem cells, Tβ4-ASCs can constantly release functional exosomes and stimulate the formation of collateral circulation after myocardial infarction, providing a feasible and alternative method for clinical revascularization.


Metagenomic discovery of novel enzymes and biosurfactants in a slaughterhouse biofilm microbial community.
DNA derived from environmental samples is a rich source of novel bioactive molecules. The choice of the habitat to be sampled predefines the properties of the biomolecules to be discovered due to the physiological adaptation of the microbial community to the prevailing environmental conditions. We have constructed a metagenomic library in Escherichia coli DH10b with environmental DNA (eDNA) isolated from the microbial community of a slaughterhouse drain biofilm consisting mainly of species from the family Flavobacteriaceae. By functional screening of this library we have identified several lipases, proteases and two clones (SA343 and SA354) with biosurfactant and hemolytic activities. Sequence analysis of the respective eDNA fragments and subsequent structure homology modelling identified genes encoding putative N-acyl amino acid synthases with a unique two-domain organisation. The produced biosurfactants were identified by NMR spectroscopy as N-acyltyrosines with N-myristoyltyrosine as the predominant species. Critical micelle concentration and reduction of surface tension were similar to those of chemically synthesised N-myristoyltyrosine. Furthermore, we showed that the newly isolated N-acyltyrosines exhibit antibiotic activity against various bacteria. This is the first report describing the successful application of functional high-throughput screening assays for the identification of biosurfactant producing clones within a metagenomic library.


Histopathologic and biochemical liver test abnormalities in chronic asymptomatic or oligosymptomatic alcoholics: a review.
To review the medical literature regarding the histopathologic and biochemical liver test abnormalities in chronic asymptomatic or oligosymptomatic alcoholics. Review of articles in the MEDLINE and LILACS databases regarding serum levels and prevalence of alterations in aspartate-aminotransferase, alanine-aminotransferase, alkaline phosphatase, and total bilirubin, in relation to liver histopathology, with or without discrimination of types of histopathologic alteration. Global mean prevalence rates of aspartate-aminotransferase and alanine-aminotransferase alterations were 86.3% and 51.1%; in cases with steatosis they were 79.1% and 38.5%; and in cases of hepatitis, 90.1% and 58%. In all studies, prevalence rates of aspartate-aminotransferase alterations were significantly higher with lower variability than those of alanine-aminotransferase. Mean aspartate-aminotransferase levels were higher than 2N (N is the upper normal limit of the method employed) in all cases with hepatitis histopathology, while those of alanine-aminotransferase were 1.48N, in the same cases. Prevalence of alkaline phosphatase and total bilirubin abnormalities were 74.5% and 74.9% globally; in cases of steatosis, they were 70.9% and 67.9%; and in cases of hepatitis, 75.9% and 77.7%. Mean alkaline phosphatase levels were above the upper normal limit in all cases, but those of total bilirubin were above normal in 4 of 7 hepatitis studies. Prevalence of aspartate-aminotransferase alteration was consistently related to presence of histopathologic abnormalities; an enzyme level higher than 2N suggests the diagnosis of alcoholic hepatitis.


Burden of Mental, Neurological, Substance Use Disorders and Self-Harm in North America: A Comparative Epidemiology of Canada, Mexico, and the United States.
To estimate the burden of mental, neurological, substance use disorders and self-harm (MNSS) in Canada, Mexico, and the United States. We extracted 2017 data from the Global Burden of Disease online database. Based on a previously developed framework to classify and aggregate the burden of specific disorders and symptoms, we reestimated the MNSS burden to include suicide, alcohol use, drug use, specific neurological, and painful somatic symptom disorders. We analyzed age-sex-specific patterns within and between countries. The MNSS burden is the largest of all disorder groupings. It is lowest in Mexico, intermediate in Canada, and highest in the United States. Exceptions are alcohol use, bipolar, conduct disorders, and epilepsy, which are highest in Mexico; and painful somatic syndromes and headaches, which are highest in Canada. The burden of drug use disorders in the United States is twice the burden in Canada, and 7 times the burden in Mexico. MNSS become the most burdensome of all disorder groups by age 10, staying at the top until age 60, and show a distinct pattern across the lifetime. The top three MNSS disorders for men are a combination of substance use disorders and self-harm (United States), with the addition of painful somatic syndromes (Canada), and headaches (Mexico). For women, the top three are headaches and depression (all countries), drug use (United States), neurocognitive disorders (Mexico), and painful somatic syndromes (Canada). MNSS are the most burdensome disease grouping and should be prioritized for funding in Canada, Mexico, and the United States.


Bedtime, sleep pattern, and incident cardiovascular disease in middle-aged and older Chinese adults: The dongfeng-tongji cohort study.
To investigate the associations of bedtime and a low-risk sleep pattern with incident cardiovascular disease (CVD). A total of 31,500 retirees were included from the Dongfeng-Tongji cohort in 2008-2010 and 2013. Sleep information was collected by questionnaires. CVD events were identified through the health care system until December 31, 2018. Cox proportional hazards regression models were performed to estimate hazard ratios (HRs) and 95% confidence intervals (CIs). During an average follow-up of 7.2 years, 8324 cases of incident CVD, including 6557 coronary heart disease (CHD) and 1767 stroke, were documented. U-shaped associations of bedtime with the risks of incident CVD and stroke were observed. Compared with bedtime between 10:01 p.m.-11:00 p.m., the HR (95% CI) for CVD was 1.10 (1.01-1.20) for ≤9:00 p.m., 1.07 (1.01-1.13) for 9:01 p.m.-10:00 p.m., and 1.32 (1.11-1.58) for >12:00 a.m., respectively, mainly driven by stroke risk (22%, 14%, and 70% higher for ≤9:00 p.m., 9:01 p.m.-10:00 p.m., and >12:00 a.m., respectively). The number of low-risk sleep factors, namely bedtime between 10:01 p.m.-12:00 a.m., sleep duration of 7-< 8 h/night, good/fair sleep quality, and midday napping ≤60 min, exhibited dose-dependent relationships with CVD, CHD, and stroke risks. Participants with 4 low-risk sleep factors had a respective 24%, 21%, and 30% lower risk of CVD, CHD, and stroke than those with 0-1 low-risk sleep factor. Individuals with early or late bedtimes had a higher CVD risk, especially stroke. Having low-risk sleep habits is associated with lower CVD risks.


Global and regional left ventricular function in patients undergoing transcatheter closure of secundum atrial septal defect.
This study sought to evaluate global and regional left ventricular (LV) function before and early after device closure of atrial septal defects (ASDs) in patients with normal pulmonary pressure. Global LV diastolic function was unaffected by ASD closure. An improvement in global LV systolic function at rest resulted in an increase in stroke volume at rest. Nevertheless, total cardiac output did not change after the procedure, because of a decrease in heart rate at rest counterbalancing the increase in stroke volume. Thus, lateral and inferior LV regional systolic function were preserved after device implantation. Moreover, no changes in regional LV diastolic function were highlighted during the study.


Correlations of glucose metabolism and structural damage to language function in aphasia.
Studies of aphasic patients using [18F]fluorodeoxyglucose positron computed tomographs have shown areas of metabolic depression in the left hemisphere larger than the area of infarction noted on CT. To evaluate these metabolic differences in relationship to language abnormalities, 11 patients had metabolic scans, CT, and were administered the Boston Diagnostic Aphasia Battery and the Porch Index of Communicative Ability. Correlation analyses were computed between metabolic, CT, and language data. CT demonstrated reliable correlations of speaking, oral reading, and repetition to Wernicke' area, consistent with current anatomic language models, while metabolic data from areas posterior, inferior, and superior to the traditional Wernicke's area and the head of the caudate nucleus also had reliable correlations with aphasic language function. The utilization of both structural and metabolic brain measures may improve our understanding of the anatomy of language as related to aphasia.


Stress-induced analgesia prevents the development of the tonic, late phase of pain produced by subcutaneous formalin.
Subcutaneous injection of formalin produces a biphasic pain response: a transient early phase followed by a tonic late phase. It has recently been suggested that development of the late phase depends upon the presence of the early one. In support of this suggestion, we now demonstrate that blocking the early phase by stress-induced analgesia prevents development of the late phase, whereas the same stressor given after the first phase does not. Both phases are manifested when stress-induced analgesia is blocked by the N-methyl-D-aspartate (NMDA) or opiate antagonists, MK-801 and naloxone.


Correction to: Fis1 depletion in osteoarthritis impairs chondrocyte survival and peroxisomal and lysosomal function.
In Fig. 1C, the wrong si-control image was used by mistake.


Overexpression of the halophyte Kalidium foliatum H⁺-pyrophosphatase gene confers salt and drought tolerance in Arabidopsis thaliana.
According to sequences of H(+)-pyrophosphatase genes from GenBank, a new H(+)-pyrophosphatase gene (KfVP1) from the halophyte Kalidium foliatum, a very salt-tolerant shrub that is highly succulent, was obtained by using reverse transcription PCR and rapid amplification of cDNA ends methods. The obtained KfVP1 cDNA contained a 2295 bp ORF and a 242 bp 3'-untranslated region. It encoded 764 amino acids with a calculated molecular mass of 79.78 kDa. The deduced amino acid sequence showed high identity to those of H(+)-PPase of some Chenopodiaceae plant species. Semi-quantitative PCR results revealed that transcription of KfVP1 in K. foliatum was induced by NaCl, ABA and PEG stress. Transgenic lines of A. thaliana with 35S::KfVP1 were generated. Three transgenic lines grew more vigorous than the wild type (ecotype Col-0) under salt and drought stress. Moreover, the transgenic plants accumulated more Na(+) in the leaves compared to wild type plants. These results demonstrated that KfVP1 from K. foliatum may be a functional tonoplast H(+)-pyrophosphatase in contributing to salt and drought tolerance.


Radical hysterectomy and tailored postoperative radiation therapy in the management of bulky stage 1B cervical cancer.
Ninety-two patients with Stage IB cervical cancers having a diameter equal to or greater than 4.0 cm were treated with radical surgery. Thirty-two patients received postoperative radiotherapy because of operative findings suggestive of high risk of pelvic recurrence. All 32 irradiated patients were treated with a standard pelvic field. Four patients also received paraaortic radiotherapy, and ten received intravaginal brachytherapy. Postoperative complications were seen in five patients (two nonirradiated, three irradiated). Projected 5-year survival is 79% (71% 5-year survival in irradiated patients, and 83% 5-year survival in nonirradiated patients). Preoperative evaluation of tumor volume was not found to reliably predict histologic high risk factors such as depth of stromal invasion, risk of lymph node metastases, or presence of extracervical/uterine involvement. A primary surgical approach in this group of patients with large-diameter Stage IB cervical cancers allows definition of those patients who might benefit from a combined surgical and radiotherapeutic approach to treatment based on findings at operation.


Patient selection for mechanical circulatory support.
Heart failure continues to be associated with high morbidity and mortality. Over the past decade, left ventricular assist devices have been shown to improve the survival and quality of life. However, it is quite clear that very sick patients do not do as well, and it is therefore imperative to select patients at the proper time. In this article, we review both the indications for considering left ventricular assist device therapy as well as discuss the considerations and therapy that should be done pre-operatively to possibly improve post-insertion outcomes.


Hydroxycholesterol substitution in ionizable lipid nanoparticles for mRNA delivery to T cells.
Delivery of nucleic acids, such as mRNA, to immune cells has become a major focus in the past decade with ionizable lipid nanoparticles (LNPs) emerging as a clinically-validated delivery platform. LNPs-typically composed of ionizable lipids, cholesterol, phospholipids, and polyethylene glycol lipids -have been designed and optimized for a variety of applications including cancer therapies, vaccines, and gene editing. However, LNPs have only recently been investigated for delivery to T cells, which has various therapeutic applications including the engineering of T cell immunotherapies. While several LNP formulations have been evaluated for mRNA delivery, recent work has demonstrated that the utilization of cholesterol analogs may enhance mRNA delivery. Other studies have shown that cholesterols modified with hydroxyl groups can alter endocytic recycling mechanisms. Here, we engineered a library of LNPs incorporating hydroxycholesterols to evaluate their impact on mRNA delivery to T cells by leveraging endosomal trafficking mechanisms. Substitution of 25% and 50% 7α-hydroxycholesterol for cholesterol in LNPs enhanced mRNA delivery to primary human T cells ex vivo by 1.8-fold and 2.0-fold, respectively. Investigation of endosomal trafficking revealed that these modifications also increase late endosome production and reduce the presence of recycling endosomes. These results suggest that hydroxyl modification of cholesterol molecules incorporated into LNP formulations provides a mechanism for improving delivery of nucleic acid cargo to T cells for a range of immunotherapy applications.