Genetic diversity and relationships among different tomato varieties revealed by EST-SSR markers.
The genetic diversity and relationship of 42 tomato varieties sourced from different geographic regions was examined with EST-SSR markers. The genetic diversity was between 0.18 and 0.77, with a mean of 0.49; the polymorphic information content ranged from 0.17 to 0.74, with a mean of 0.45. This indicates a fairly high degree of diversity among these tomato varieties. Based on the cluster analysis using unweighted pair-group method with arithmetic average (UPGMA), all the tomato varieties fell into 5 groups, with no obvious geographical distribution characteristics despite their diverse sources. The principal component analysis (PCA) supported the clustering result; however, relationships among varieties were more complex in the PCA scatterplot than in the UPGMA dendrogram. This information about the genetic relationships between these tomato lines helps distinguish these 42 varieties and will be useful for tomato variety breeding and selection. We confirm that the EST-SSR marker system is useful for studying genetic diversity among tomato varieties. The high degree of polymorphism and the large number of bands obtained per assay shows that SSR is the most informative marker system for tomato genotyping for purposes of rights/protection and for the tomato industry in general. It is recommended that these varieties be subjected to identification using an SSR-based manual cultivar identification diagram strategy or other easy-to-use and referable methods so as to provide a complete set of information concerning genetic relationships and a readily usable means of identifying these varieties.


A shared haplotype for dentatorubropallidoluysian atrophy (DRPLA) in Italian families testifies of the recent introduction of the mutation.
To clarify the population history of dentatorubropallidoluysian atrophy (DRPLA) in Italy and to date back the introduction of the mutation, we reconstructed extended haplotypes flanking the CAG repeat in 10 patients of Italian ancestry, analyzing their similarity/dissimilarity as a function of distance from the CAG repeat. Our aim was to compare the hypothesis of a single, recent genealogy connecting all the observed haplotypes with the alternative hypothesis of multiple introductions by more distantly related haplotypes from outer sources. Polymorphic DNA markers were chosen to cover a region of 153 kb flanking the CAG repeat, that is, informative for dating the age of the DNA segment unaffected by recombination. In all patients, an expansion of the ATN1 CAG segment was confirmed residing onto the same narrow haplotype described to be associated with the CAG expansion in the Japanese and Portuguese populations. We also observed the disruption of the DRPLA haplotype at longer distances, on both sides of the CAG. Our results are compatible with a single founder in the last 600 years, most likely before the last 270 years. These estimates for the Sicilian population largely overlap a period in which the Japanese haplotype with the DRPLA mutation could have been introduced by the Portuguese maritime travelers.


CD4+ cell count at antiretroviral therapy initiation and economic restoration in rural Uganda.
To determine whether earlier initiation of antiretroviral therapy (ART) is associated with better economic outcomes. Prospective cohort study of HIV-positive patients on ART in rural Uganda. Patients initiating ART at a regional referral clinic in Uganda were enrolled in the Uganda AIDS Rural Treatment Outcomes study starting in 2005. Data on labor force participation and asset ownership were collected on a yearly basis, and CD4 cell counts were collected at pre-ART baseline. We fitted multivariable regression models to assess whether economic outcomes at baseline and in the 6 years following ART initiation varied by baseline CD4 cell count. Five hundred and five individuals, followed up to 6 years, formed the estimation sample. Participants initiating ART at CD4 cell count at least 200 cells/μl were 13 percentage points more likely to be working at baseline (P < 0.01, 95% confidence interval 0.06-0.21) than those initiating below this threshold. Those in the latter group achieved similar labor force participation rates within 1 year of initiating ART (P < 0.01 on the time indicators). Both groups had similar asset scores at baseline and demonstrated similar increases in asset scores over the 6 years of follow-up. ART helps participants initiating therapy at CD4 cell count below 200 cells/μl rejoin the labor force, though the findings for participants initiating with higher CD4 cell counts suggests that pretreatment declines in labor supply may be prevented altogether with earlier therapy. Baseline similarities in asset scores for those with early and advanced disease suggest that mechanisms other than morbidity may help drive the relationship between HIV infection and economic outcomes.


Role of sputum differential cell count in detecting airway inflammation in patients with chronic bronchial asthma or COPD.
Sputum may provide an alternative source of bronchial cells to investigate characteristics of airway inflammation and its functional correlates in patients with asthma or chronic obstructive pulmonary disease (COPD). Two groups of clinically stable patients were studied: a group of 43 patients with mild or moderate asthma and a group of 18 patients with COPD. Twenty normal subjects formed a control group. Sputum production was either spontaneous or induced with inhaled hypertonic saline for five minute periods for up to 20 minutes. The concentration of saline was increased at intervals of 10 minutes from 3% to 4%. Plugs from the lower respiratory tract were selected for differential counting in cytocentrifugation preparations. Bronchial provocation tests were performed by inhaling progressive concentrations of histamine from a DeVilbiss 646 nebuliser and the concentration of histamine which caused a 20% fall in the forced expiratory volume in one second (FEV1) was calculated (PC20FEV1). Neutrophils predominated in the sputum of subjects with COPD while eosinophils predominated in the sputum of those with chronic asthma. However, in 28% of asthmatic subjects an increased percentage of neutrophils was found. In asthmatic patients the differential count of eosinophils was inversely related to the FEV1, FEV1/VC, and bronchial hyperresponsiveness, and directly related to clinical scores. The cellular profile of sputum in normal subjects and in patients with asthma and COPD is different. The concentration of eosinophils in the sputum correlates with the severity of asthma.


Radioimmunotherapy for non-Hodgkin's lymphoma with yttrium 90 ibritumomab tiuxetan.
The increasing incidence of non-Hodgkin's lymphoma (NHL), coupled with the lack of optimal treatment options, has prompted the development of novel treatments. Of these, radioimmunotherapy is one of the most promising. Two of the radiolabeled monoclonal antibody therapies being studied in the treatment of NHL are yttrium 90 (90Y) ibritumomab tiuxetan and iodine 131 (131I) tositumomab. The radionuclides 90Y and 131I emit beta radiation; 131I also emits gamma radiation, thus requiring more elaborate precautionary measures to limit radiation exposure. The monoclonal antibody portions of the drugs target the CD20 surface antigen that is present on the majority of B-cell lymphomas, resulting in direct radiation to the targeted cells, as well as indirect targeting of adjacent cells (known as the crossfire effect). Clinical trials of 90Y ibritumomab tiuxetan in patients with NHL have produced promising results. The safe and effective use of radioimmunotherapy requires a multidisciplinary team approach in which nurses play a central role.


Archaebiotics: proposed therapeutic use of archaea to prevent trimethylaminuria and cardiovascular disease.
Trimethylamine (TMA) is produced by gut bacteria from dietary ingredients. In individuals with a hereditary defect in flavin-containing monooxygenase 3, bacterial TMA production is believed to contribute to the symptoms of trimethylaminuria (TMAU; fish-odor syndrome). Intestinal microbiota TMA metabolism may also modulate atherosclerosis risk by affecting trimethylamine oxide (TMAO) production levels. We propose that reducing TMA formation in the gut by converting it to an inert molecule could be used to prevent or limit these human diseases, while avoiding the major drawbacks of other clinical interventions. Reducing TMA levels by microbiological interventions could also be helpful in some vaginoses. Particular members of a recently discovered group of methanogens, that are variably present in the human gut, are unusual in being apparently restricted to utilizing only methyl compounds including TMA as substrates. We confirmed experimentally that one of these strains tested, Methanomassiliicoccus luminyensis B10, is able to deplete TMA, by reducing it with H 2 for methanogenesis. We therefore suggest that members of this archaeal lineage could be used as treatments for metabolic disorders.


The Moderating Roles of Resilience and Coping Strategy on Well-Being of Victimized Forensic Workers.
Previous research on workplace victimization has often disregarded forensic psychiatric populations and not yet been extended to the coronavirus pandemic. The present study expected the isolation of the government-issued lockdown to increase aggressive behavior in forensic patients, ultimately decreasing the general well-being of victimized forensic workers. Possible buffering protective factors (resilience and active coping) and enhancing risk factors (avoidant coping and passive coping) were investigated with the intention of optimizing the general well-being of at-risk forensic workers. The valid sample (N = 311) consisted of Dutch and Belgian forensic workers (74.6% females) with at least 9 hours of weekly patient contact, and with a mean age of 37.99 (SD = 12.20). Participants reported the number of violent incidents in the past 2 months, as well as completed a questionnaire battery including measures of well-being, resilience, and coping strategies. A significant increase of victimization during the lockdown compared to after it was lifted was found, however, the study did not find evidence to support that this negatively influenced the worker's general well-being. Active coping was found to be a significant moderator and protective factor for the general well-being of victimized forensic workers. In contrast, resilience, avoidant and passive coping were not significant moderators in this association. The present study has valuable clinical implications that could lead to preparatory and preventative measures for forensic workers at risk of being victimized. Future research may investigate constructs such as life satisfaction and post-traumatic growth, as well as be broadened into prison populations.


Update on Immunosuppression in Liver Transplantation.
The standard therapy for decompensated end-stage chronic liver disease of any etiology and acute fulminant hepatic failure is liver transplantation (LT). Advances in immunosuppressive therapy decreased the rates of acute and chronic rejections. Thus, graft and patient survivals have significantly improved. However, long-term adverse effects of prolonged use of immunosuppressive agents such as malignancies, opportunistic infections, metabolic disorders, and other organ toxicities have now become a major concern. Consequently, alternative approaches are needed to deescalate the customary drugs and their side effects. Therapy must be individualized and additional preventive measures should be taken by patients with particular risk factors or predisposed to certain adverse effects. Current opinion favors a combination of agents with different mechanism of actions and toxicity profiles. Corticosteroids are employed in immediate and early postoperative period. Although they have a pronounced side effect profile, calcineurin inhibitors (CNIs) are still the backbone of early and late phase immunosuppressive regimens because of their proved efficacy. Antimetabolites are frequent choices for steroid and/or CNI-sparing strategies. Studies also have established a role for mammalian target of rapamycin (mTOR) inhibitors in specific groups of recipients. Biologic agents are a hot topic of interest and made their way into current strategies for induction. Agents extrapolated from other transplantation or immunologic experience are being evaluated. Tasdogan BE, Ma M, Simsek C, et al. Update on Immunosuppression in Liver Transplantation. Euroasian J Hepato-Gastroenterol 2019;9(2):96-101.


Molecular basis for persistent hepatitis B virus infection in the liver after clearance of serum hepatitis B surface antigen.
Hepatitis B virus (HBV) DNA has been detected by polymerase chain reaction (PCR) in the liver of patients with resolved chronic HBV infection and sustained clearance of hepatitis B surface antigen (HBsAg) from serum. However, it is unknown whether the virus is transcriptionally active at this time or if the covalently closed circular (CCC) replicative intermediate of HBV DNA can still be detected. Therefore, hepatic nucleic acid extracts from seven patients who had cleared serum HBsAg were assessed by (PCR) for either reverse-transcribed HBV RNA, or an intact direct repeat region of the HBV genome indicative of the CCC replicative intermediate of HBV DNA. HBV transcripts were detected in four of seven patients in the study group, whereas an intact direct repeat region of the HBV genome was detected in three. Evidence for viral transcription and replication was more frequently detected in patients who had recently cleared serum HBsAg, but HBV RNA was also detected in one patient 5 years after HBsAg clearance, and an intact direct repeat region of HBV DNA was detected in another subject at nearly 4 years after resolution of disease. Therefore, hepatic HBV transcription may be associated with replicative intermediates of persistent HBV DNA in patients who have cleared HBsAg from serum, suggesting that, on occasion, HBV may not be in a latent state but undergoing low-level replication.


Secernin-1 as a novel prognostic biomarker candidate of synovial sarcoma revealed by proteomics.
We aimed to develop prognostic biomarkers for synovial sarcoma employing a proteomic approach. We examined the proteomic profile of synovial sarcoma using two-dimensional difference gel electrophoresis (2D-DIGE). We identified 20 protein spots whose intensity was statistically different (p<0.01) between a group of eight patients who were alive and continuously disease-free for over five years and a group of five patients who died of the disease within two years post diagnosis. Mass spectrometric protein identification demonstrated that these 20 spots corresponded to 17 distinct gene products. Three of the 20 spots corresponded to secernin-1 and had higher intensity in the good prognosis group. The prognostic performance of secernin-1 was further examined immunohistochemically in 45 synovial sarcoma cases. The 5-year survival rate was 77.6% and 21.8% for patients with secernin-1 positive and negative primary tumors respectively (p=0.0015). The metastasis-free survival was significantly higher in the patient group with high secernin-1 expression compared to that with low expression (p=0.0012). Uni- and multivariate analyses revealed that secernin-1 expression was a powerful prognostic factor compared to other clinico-pathological parameters examined. These results indicate that secernin-1 may be used as a biomarker to predict the overall and metastasis-free survival in synovial sarcoma patients.


Relationship between tumor location, size, and WHO grade in meningioma.
OBJECTIVE Prior studies have investigated preoperative risk factors for meningioma; however, no association has been shown between meningioma tumor size and tumor grade. The objective of this study was to investigate the relationship between tumor size and grade in a large single-center study of patients undergoing meningioma resection. METHODS A retrospective chart review of patients undergoing meningioma resection at the University of California, San Francisco, between 1985 and 2015 was performed. Patients with incomplete information, spinal meningiomas, multiple meningiomas, or WHO grade III meningiomas were excluded. The largest tumor dimension was used as a surrogate for tumor size. Univariate and multivariate logistic regression models were used to investigate the relationship between tumor grade and tumor size. A recursive partitioning analysis was performed to identify groups at higher risk for atypical (WHO grade II) meningioma. RESULTS Of the 1113 patients identified, 905 (81%) had a WHO grade I tumor and in 208 (19%) the tumors were WHO grade II. The median largest tumor dimension was 3.6 cm (range 0.2-13 cm). Tumors were distributed as follows: skull base (n = 573, 51%), convexity/falx/parasagittal (n = 431, 39%), and other (n = 109, 10%). On univariate regression, larger tumor size (p < 0.001), convexity/falx/parasagittal location (p < 0.001), and male sex (p < 0.001) were significant predictors of WHO grade II pathology. After controlling for interactions, multivariate regression found male sex (OR 1.74, 95% CI 1.25-2.43), size 3-6 cm (OR 1.69, 95% CI 1.08-2.66), size > 6 cm (OR 3.01, 95% CI 1.53-5.94), and convexity/falx/parasagittal location (OR 1.83, 95% CI 1.19-2.82) to be significantly associated with WHO grade II. Recursive partitioning analysis identified male patients with tumors > 3 cm as a high-risk group (32%) for WHO grade II meningioma. CONCLUSIONS Larger tumor size is associated with a greater likelihood of a meningioma being WHO grade II, independent of tumor location and male sex, which are known risk factors.


New alleles in calpastatin gene are associated with meat quality traits in pigs.
Suggestive QTL affecting raw firmness scores and average Instron force, tenderness, juiciness, and chewiness on cooked meat were mapped to pig chromosome 2 using a three-generation intercross between Berkshire and Yorkshire pigs. Based on its function and location, the calpastatin (CAST) gene was considered to be a good candidate for the observed effects. Several missense and silent mutations were identified in CAST and haplotypes covering most of the coding region were constructed and used for association analyses with meat quality traits. Results demonstrated that one CAST haplotype was significantly associated with lower Instron force and cooking loss and higher juiciness and, therefore, this haplotype is associated with higher eating quality. Some of the sequence variation identified may be associated with differences in phosphorylation of CAST by adenosine cyclic 3', 5'-monophosphate-dependent protein kinase and may in turn explain the meat quality phenotypic differences. The beneficial haplotype was present in all the commercial breeds tested and may provide significant improvements for the pig industry and consumers because it can be used in marker-assisted selection to produce naturally tender and juicy pork without additional processing steps.


Shear-driven segregation of dense granular mixtures in a split-bottom cell.
Shear-driven segregation of dense granular mixtures has been associated with a number of interesting pattern formation problems. We use experimental and computational split-bottom cells to isolate segregation effects associated with shear gradients from those associated with gravity. We find the effect of shear gradients much less dramatic than initial observations of segregation suggest. While a segregation pattern emerges in a circular split-bottom cell that appears coincident with the shear gradient, we find the pattern is orthogonal to the active segregation flux. We measure a toroidal convection roll that, in conjunction with gravity-driven segregation, is likely responsible for the dramatic horizontal segregation pattern. On the other hand, computational results from a parallel split-bottom cell indicate a subtle segregation flux associated with the shear gradient. The nature of the driving mechanism is unknown. A current predictive form of kinetic theory based on binary collisions dominating the particle dynamics predicts segregation in the opposite direction from observed trends. This indicates the direction of shear-driven segregation depends on the nature of the flow itself, collisional or frictional.


Multicenter, double-blind, placebo-controlled trial of terfenadine in seasonal allergic rhinitis and conjunctivitis.
A multiclinic, double-blind, parallel and controlled study was conducted in the 1982 spring pollen season to evaluate and compare the effects of terfenadine, 60 mg bid with those of chlorpheniramine 4 mg tid and placebo for a treatment period of seven days in patients with seasonal allergic rhinitis and conjunctivitis. Of a total of 397 patients enrolled in the seven study centers, 345 patients were accepted for evaluation of efficacy and 393 patients for safety. The results show that based on the physicians' assessment of the overall efficacy, terfenadine was significantly superior to placebo and comparable to chlorpheniramine in the relief of allergic symptoms, with moderate to complete relief being observed in 60% (68/113) of the terfenadine-treated patients, in 60% (71/119) of the chlorpheniramine-treated patients, and in 30% (34/119) of the placebo-treated patients. The daily evaluation of severity of symptoms by the patients show that the effect of terfenadine and chlorpheniramine was evident on the first day after entry, reached a peak on the second day after entry, and persisted thereafter. Side effects were minor and infrequent in all treatment groups. There was no statistically significant difference in the incidence of sedation between the terfenadine (7.6%) and placebo (2.4%) groups whereas the incidence of sedation with chlorpheniramine (19%) was significantly higher. In conclusion, terfenadine is as effective as chlorpheniramine in the treatment of symptoms of seasonal allergic rhinitis and conjunctivitis with an incidence of sedation not significantly different from that with placebo and significantly less than with chlorpheniramine.


C-terminal ECFP fusion impairs synaptotagmin 1 function: crowding out synaptotagmin 1.
To allow the monitoring of synaptotagmin 1 trafficking in vivo, we generated transgenic mice expressing a synaptotagmin 1-enhanced cyan fluorescent protein (ECFP) fusion protein under control of the Thy1 promoter. Transgenic synaptotagmin 1-ECFP is expressed throughout the brain where it localizes to synapses and marks synapses in vivo. However, when we crossed transgenic synaptotagmin 1-ECFP mice with synaptotagmin 1 knock-out mice, we detected no rescue of survival or function. Furthermore, viral overexpression of synaptotagmin 1-ECFP in synaptotagmin 1-deficient neurons failed to restore normal Ca2+-triggered release, whereas overexpression of wild type synaptotagmin 1 did so efficiently. To determine whether synaptotagmin 1-ECFP is non-functional because the ECFP-fusion interferes with its biochemical activities, we measured Ca2+-independent binding of synaptotagmin 1-ECFP to SNARE complexes, and Ca2+-dependent binding of synaptotagmin 1-ECFP to phospholipids and to itself. Although the apparent Ca2+ affinity of synaptotagmin 1-ECFP was decreased compared with wild type synaptotagmin 1, we observed no major changes in Ca2+-dependent or -independent activities, indicating that the non-functionality of the synaptotagmin 1-ECFP fusion protein was not because of inactivation of its biochemical properties. These data suggest that synaptotagmin 1-ECFP is suitable for monitoring synaptic vesicle traffic in vivo because the synaptotagmin 1-ECFP marks synaptic vesicles without participating in exocytosis. In addition, the data demonstrate that synaptotagmin 1 function requires a free C terminus, possibly because of spatial constraints at the release sites.


Link Between Clinical Predictors of Heterotopic Ossification and Histological Analysis in Combat-Injured Service Members.
Heterotopic ossification (HO) is a debilitating condition that occurs following traumatic injury and may restrict range of motion and delay rehabilitation. The timing and efficacy of surgical resection have varied widely, and there is a gap in knowledge between clinical predictors of HO recurrence and histological analysis. Thirty-three service members seen at Walter Reed National Military Medical Center for symptomatic HO were enrolled in an institutional review board-approved study. Participants took oxytetracycline on four scheduled days prior to HO resection to determine the mineral apposition rate (bone growth rate). Detailed histological analyses included scanning electron microscopy with backscattered electron imaging and light microscopy. Data indicated that the mineral apposition rate of trauma-induced HO was approximately 1.7 μm/day at the time of operative intervention, which was 1.7 times higher than the rate in non-pathological human bone. The mineral apposition rate and postoperative alkaline phosphatase values were demonstrated to be positively and significantly related (ρ = 0.509, p = 0.026, n = 19). When the analysis was limited to patients with no more than a two-year period from injury to excision (thereby removing outliers who had a longer time period than their counterparts) and traumatic brain injury and nonsteroidal anti-inflammatory drugs (known correlates with HO development) were controlled for in the statistical analysis, the mineral apposition rate and recurrence severity were significantly related (ρ = -0.572, p = 0.041, n = 11). Data demonstrated a link between benchtop research and bedside care, with the mineral apposition rate elevated in patients with HO and correlated with recurrence severity; however, a larger sample size and more clinical factors are needed to refine this model. Prognostic Level II. See Instructions for Authors for a complete description of levels of evidence.


The induction of synthesis of L-type pyruvate kinase in cultured rat hepatocytes.
Hepatocytes were isolated from preweaned neonatal and adult rats and maintained in primary monolayer culture. Cells from preweaned newborns possessed no L-type pyruvate kinase, nor did they synthesize the enzyme. Incubation for 48-72 h in culture medium supplemented with 2 mM-fructose and 0.1 microM-insulin induced the synthesis of L-type pyruvate kinase, as judged by increased enzyme activity and the increased incorporation of [3H]leucine into immunoprecipitable L-type pyruvate kinase. Hepatocytes isolated from 48 h-starved adult rats incorporated less [3H]leucine into L-type pyruvate kinase than did cells isolated from high-carbohydrate-diet-fed rats. The rate of enzyme synthesis by cells from 48 h-starved rats was increased by the inclusion of fructose and insulin in the incubation medium, after a lag phase of 24-48 h. After 4 days in culture in the presence of fructose and insulin, hepatocytes from 48 h-starved rats synthesized L-type pyruvate kinase at similar rates to hepatocytes isolated from high-carbohydrate-diet-fed rats.


Is the Dinoflagellate Amoebophrya Really Missing an mtDNA?
Mitochondrial DNA (mtDNA) is a universal hallmark of aerobic eukaryotes. That is why the recent suggestion by John et al. (2019. An aerobic eukaryotic parasite with functional mitochondria that likely lacks a mitochondrial genome. Sci Adv. 5(4):eaav1110.) that the aerobic dinoflagellate Amoebophrya sp. strain AT5 (Syndiniales) lacks mtDNA was so remarkable. Here, by reanalyzing recently published genomic and transcriptomic data from three Amoebophrya strains, we provide evidence of a cryptic, highly reduced mtDNA in this clade. More work is needed before one can definitively say if Amoebophrya has or does not have an mtDNA, but for now, the data are pointing toward the existence of one. Ultimately, we urge caution when basing supposedly absent genomic features on single line evidences.


Dynamic evolution of a vesicle formed by comb-like block copolymer-tethered nanoparticles: a dissipative particle dynamics simulation study.
Vesicles are well-sealed capsules that can store or transport substances. Their dynamic evolution mechanisms are important to fulfill specific functions. In the present study, dissipative particle dynamics (DPD) simulation was employed to study the formation, fusion, and fission pathways of vesicles. Our results show that comb-like block copolymer-tethered nanoparticles can self-assemble into stable vesicles, which may be a good candidate for drug entrapment and controlled release of drug. The spontaneous fusion of this type of vesicles has been studied, whose mechanism is different from that of the vesicles formed from comb-like block copolymers. However, the fission mechanisms of the two types of vesicles are similar. To summarize, the introduction of nanoparticles alters the fusion mechanism, but causes no difference in the fission pathway of vesicles. The reason may be attributed to the nature of nanoparticles. The fusion and fission processes always experience the congregation of nanoparticles. The large and rigid nanoparticles inevitably result in changes of surface tension of the vesicle that is a key factor influencing the membrane dynamics.


Bithionol Potently Inhibits Human Soluble Adenylyl Cyclase through Binding to the Allosteric Activator Site.
The signaling molecule cAMP regulates functions ranging from bacterial transcription to mammalian memory. In mammals, cAMP is synthesized by nine transmembrane adenylyl cyclases (ACs) and one soluble AC (sAC). Despite similarities in their catalytic domains, these ACs differ in regulation. Transmembrane ACs respond to G proteins, whereas sAC is uniquely activated by bicarbonate. Via bicarbonate regulation, sAC acts as a physiological sensor for pH/bicarbonate/CO2, and it has been implicated as a therapeutic target, e.g. for diabetes, glaucoma, and a male contraceptive. Here we identify the bisphenols bithionol and hexachlorophene as potent, sAC-specific inhibitors. Inhibition appears mostly non-competitive with the substrate ATP, indicating that they act via an allosteric site. To analyze the interaction details, we solved a crystal structure of an sAC·bithionol complex. The structure reveals that the compounds are selective for sAC because they bind to the sAC-specific, allosteric binding site for the physiological activator bicarbonate. Structural comparison of the bithionol complex with apo-sAC and other sAC·ligand complexes along with mutagenesis experiments reveals an allosteric mechanism of inhibition; the compound induces rearrangements of substrate binding residues and of Arg(176), a trigger between the active site and allosteric site. Our results thus provide 1) novel insights into the communication between allosteric regulatory and active sites, 2) a novel mechanism for sAC inhibition, and 3) pharmacological compounds targeting this allosteric site and utilizing this mode of inhibition. These studies provide support for the future development of sAC-modulating drugs.


Tamoxifen therapy of metastatic breast cancer.
Inhibition of estrogen action at the target tissue level with the antiestrogen, tamoxifen, has proved highly successful in the treatment of hormone-responsive breast cancer. In randomized clinical trials involving postmenopausal patients, tamoxifen has been found to be as effective as high-dose estrogens, androgens, progestins, and the aromatase inhibitor, aminoglutethimide. Because of its remarkable lack of significant toxicity, tamoxifen is now considered the first endocrine treatment of choice of hormone-responsive postmenopausal breast cancer. Although effective in a significant fraction of premenopausal patients, tamoxifen cannot be considered a substitute for ovariectomy because it usually does not suppress menses and because response to castration may occur after progression during antiestrogen therapy. Current evidence suggests that there is no major advantage in combining tamoxifen with other endocrine therapies or chemotherapy. At present, it appears preferable to use different modalities of endocrine therapy sequentially in those patients with hormone-responsive cancers. Chemotherapy should be delayed until maximum benefit has been obtained from endocrine therapy.


African swine fever virus infection in the argasid host, Ornithodoros porcinus porcinus.
The pathogenesis of African swine fever virus (ASFV) infection in Ornithodoros porcinus porcinus was examined in nymphal ticks infected with the ASFV isolate Chiredzi/83/1. At times postinfection (p.i.) ranging from 6 h to 290 days, ticks or dissected tick tissues were titrated for virus and examined ultrastructurally for evidence of virus replication. The ASFV infection rate in ticks was 100% in these experiments, and virus infection was not associated with a significant increase in tick mortality. Initial ASFV replication occurred in phagocytic digestive cells of the midgut epithelium. Subsequent infection and replication of ASFV in undifferentiated midgut cells was observed at 15 days p.i. Generalization of virus infection from midgut to other tick tissues required 2 to 3 weeks and most likely involved virus movement across the basal lamina of the midgut into the hemocoel. Secondary sites of virus replication included hemocytes (type I and II), connective tissue, coxal gland, salivary gland, and reproductive tissue. Virus replication was not observed in the nervous tissue of the synganglion, Malpighian tubules, and muscle. Persistent infection, characterized by active virus replication, was observed for all involved tick tissues. After 91 days p.i., viral titers in salivary gland and reproductive tissue were consistently the highest detected. Successful tick-to-pig transmission of ASFV at 48 days p.i. correlated with high viral titers in salivary and coxal gland tissue and their secretions. A similar pattern of virus infection and persistence in O. porcinus porcinus was observed for three additional ASFV tick isolates in their associated ticks.


Isolation and characterization of novel multifunctional recombinant family 26 glycoside hydrolase from Mehsani buffalo rumen metagenome.
Rumen microbiota harbor a diverse set of carbohydrate-active enzymes (CAZymes), which play a crucial role in the degradation of a complex plant polysaccharide thereby providing metabolic energy to the host animals. Earlier, we reported CAZYme analysis from the buffalo rumen metagenome by high throughput shotgun sequencing. Among the various CAZymes, glycoside hydrolase family 26 (GH26) enzymes have a number of industrial applications including in paper, oil, biofuel, food, feed, pharmaceutical, coffee, and detergent industries. Here, we report isolation and characterization of GH26 enzyme from the buffalo rumen metagenome. A novel GH26 gene composed of 1,119 base pairs was successfully amplified using the gene-specific primers inferred based on the contig generated from metagenome sequence assembly and cloned in a pET32a (+) expression vector as an N-terminal histidine tag fusion protein. A novel GH26 protein from an unknown rumen microorganism shared a maximum of 68% identity with the Prevotella ruminicola 23 encoded carbohydrate esterase family 7 and 46% with Bacteroides sp. 2_1_33B encoded mannan endo-1, 4-β-mannosidase. The recombinant GH26-histidine tag fusion protein was expressed in Escherichia coli and purified using Ni-NTA affinity chromatography. The purified enzyme displayed multifunctional activities against various carbohydrate substrates including locust bean gum, beechwood xylan, pectin, and carboxymethyl cellulose suggesting mannanase, xylanase, pectin esterase, and endoglucanase activities, respectively.


The Stearoyl-CoA Desaturase-1 (Desat1) in Drosophila cooperated with Myc to Induce Autophagy and Growth, a Potential New Link to Tumor Survival.
Lipids are an important energy supply in our cells and can be stored or used to produce macromolecules during lipogenesis when cells experience nutrient starvation. Our proteomic analysis reveals that the Drosophila homologue of human Stearoyl-CoA desaturase-1 Desat1) is an indirect target of Myc in fat cells. Stearoyl-CoA desaturases are key enzymes in the synthesis of monounsaturated fatty acids critical for the formation of complex lipids such as triglycerides and phospholipids. Their function is fundamental for cellular physiology, however in tumors, overexpression of SCD-1 and SCD-5 has been found frequently associated with a poor prognosis. Another gene that is often upregulated in tumors is the proto-oncogene c-myc, where its overexpression or increased protein stability, favor cellular growth. Here, we report a potential link between Myc and Desat1 to control autophagy and growth. Using Drosophila, we found that expression of Desat1, in metabolic tissues like the fat body, in the gut and in epithelial cells, is necessary for Myc function to induce autophagy a cell eating mechanism important for energy production. In addition, we observed that reduction of Desat1 affects Myc ability to induce growth in epithelial cells. Our data also identify, in prostatic tumor cells, a significant correlation between the expression of Myc and SCD-1 proteins, suggesting the existence of a potential functional relationship between the activities of these proteins in sustaining tumor progression.


Mechanism of proteinuria. IV. Effect of renin on hemoglobin excretion.
1. Administration of renin to the rat causes an increase in the glomerular permeability to hemoglobin and a decrease in tubular absorption of this substance as measured by functional methods. 2. Morphologic examination in the rat and in the rabbit reveals an increased concentration of hemoglobin in the tubular lumina, confirmatory evidence of increased permeability. However, there is no significant effect upon the formation of hemoglobin droplets within the renal cells. 3. By analogy with the events observed in neutral red "secretion," it is suggested that hemoglobin is absorbed by both a "direct" and an "indirect" mechanism and that renin administration affects or depresses the "direct" mechanism only.


Musculoskeletal disorders among nurses compared with two other occupational groups.
There is a high incidence of low back pain (LBP) among nurses. However, few longitudinal studies have investigated musculoskeletal disorders (MSDs) at other anatomical sites in nurses. To describe the cumulative incidence and persistence/recurrence of MSDs of the low back, neck, shoulder, elbow, wrist/hand and knee among New Zealand nurses, to investigate the impact of MSDs on work and functional tasks and to compare findings for nurses with those in postal workers and office workers. Participants completed a postal survey at baseline and again 1 year later. Information was collected about MSDs in the previous 1 month and 12 months and about the ability to attend work, undertake work duties and perform functional tasks. Among nurses, the low back was the site with the highest cumulative incidence and highest prevalence of persistent/recurrent, work-disabling and functional-task-disabling pain. Work-disabling LBP was more prevalent among nurses and postal workers than office workers (P < 0.001). Nurses had a substantial prevalence of work-disabling shoulder pain (10%) and functional-task-disabling knee (19%) and wrist/hand pain (16%). With the exception of the elbow, each occupational group had a high prevalence of persistent/recurrent MSDs at all anatomical sites. LBP continues to have a substantial impact among nurses. Other less commonly considered MSDs, such as shoulder, wrist/hand and knee pain, also made work or functional tasks difficult, suggesting that primary and secondary prevention efforts should consider MSDs at other anatomical sites as well as the low back.


Molecular structure and evolutionary origin of human cardiac muscle actin gene.
Two recombinant phages that contain cardiac muscle actin gene were isolated from a human DNA library and their structures were determined. Restriction analysis indicates that both clones carry the same EcoRI 13-kilobase fragment where the coding sequence is mapped. The cloned DNA hybridized with polyadenylylated RNA from human fibroblasts, which directs the synthesis of cytoplasmic beta- and gamma-actin in vitro. However, sequence determination of the cloned DNA showed that the entire coding sequence perfectly matched the amino acid sequence of cardiac muscle actin. The initiation codon is followed by a cysteine codon that is not found at the amino-terminal site of any actin isoform, suggesting the necessity of post-translational processing for in vivo actin synthesis. There are five introns interrupting exons at codons 41/42, 150, 204, 267, and 327/328. Surprisingly, these intron locations are exactly the same as those of the rat skeletal muscle actin gene but different from those of nonmuscle beta-actin gene. Nucleotide sequences of all exon/intron boundaries agree with the G-T/A-G rule (G-T at the 5' and A-G at the 3' termini of each intron). The 3'-untranslated sequence has no homology to that of nonmuscle beta- or gamma-actin gene, but Southern blot hybridization has shown that this region has considerable homology to that of one of the other actin genes. These results indicate that the recombinant phages, which we have isolated, contain cardiac muscle actin gene and that cardiac muscle actin gene and skeletal muscle actin genes are derived from their ancestor gene at a relatively recent time in evolutionary development.


L-Phenylalanine inhibition of muscle pyruvate kinase.
The allosteric inhibition of M1-type pyruvate kinase from rabbit skeletal muscle by phenylalanine is reciprocally dependent on Mg2+ and phosphoenolpyruvate concentrations. At pH 8, phenylalanine acts as a competitive inhibitor with respect to Mg2+ and phosphoenolpyruvate, and vice versa. Phenylalanine introduces sigmoidicity into the dependence of the reaction velocity on [Mg2+]. In vitro kinetic analysis indicates that phenylalanine inhibition of muscle pyruvate kinase is unlikely to have regulatory significance in vivo.


[Determination of heterophilic antibodies in infants: apropos of nine cases].
The most frequent and known disease produced by Epstein-Barr virus is infectious mononucleosis. It is usually diagnosed after the age of two in pediatric practice based on the characteristic hematologic picture and Paul-Bunnell test. In infants there is rarely communications of this cases in the literature and, moreover, Paul-Bunnell test is commonly negative. We have collected nine infants below two years old with mononucleosis syndrome characteristic hematologic picture and with heterophil antibodies positive reaction in all of them. Clinical and laboratory finding, and it likely relation to the EBV infection are discussed.


Modes and consequences of thrombin's interaction with fibrin.
Thrombin mediates the balance between coagulant and fibrinolytic forces and has numerous cellular effects. This intricate balance is maintained by biochemical mechanisms that regulate thrombin activity. Disruption of this balance could lead to bleeding or thrombosis. Once thrombin is generated, two major mechanisms regulate its activity. By binding fibrin, thrombin's activity is localized to the thrombus, a process that limits its systemic procoagulant effects. Thrombin that escapes into the circulation is efficiently inactivated by plasma inhibitors, such as antithrombin, or is sequestered by thrombomodulin on the endothelium. Although thrombin's interaction with fibrin limits its systemic effects, fibrin-bound thrombin resists inactivation and can produce a local procoagulant stimulus that triggers thrombus growth. Direct thrombin inhibitors were developed, at least in part, to target fibrin-bound thrombin. These agents are finding their niche for the prevention and treatment of venous and arterial thrombosis. The mechanisms by which thrombin binds fibrin are reviewed in this paper. As well, the potential pathological consequences of thrombin's interaction with fibrin are discussed.


Predictive value of the Ages and Stages Questionnaire® for school performance and school intervention in late preterm- and term-born children.
The new health supervision guidelines emphasize the importance of surveillance or a formal developmental screening test at 4 years, one of the most used tests is Ages & Stages Questionnaire. Nevertheless, there is still not enough evidence whether these tools will be useful to predict future school performance (SP). The objectives of this study were to evaluate the Ages & Stages Questionnaire 3rd edition 48-month interval (ASQ3-48) translated to Spanish for predicting the need for school intervention (SI) and poor SP at 8-9 years of age, in late preterm infants (LPIs) and term-born infants (Terms) and to compare the prevalence of SI and poor SP in the two groups. Data were collected from a cohort of 75 LPIs and 58 Terms assessed with ASQ3-48 and with a further assessment of academic results at 8-9 years, through a standardized school test of the Education Department of Catalonia. SI data were obtained through a parent report. Area under the curve (AUC), sensitivity, specificity and predictive values were calculated, and logistic regression analysis was used. The prevalence of poor SP was 12.8%, without statistically significant differences between LPIs and Terms. LPIs had higher SI than Terms (29.3% vs. 10.3%, P = 0.001). AUC for poor SP was 0.73 and for receiving SI was 0.56 without differences between the two groups. The sensitivity of the ASQ3-48 for poor SP was 41%, for specificity 92%, and for receiving SI 14% and 89%, respectively. Poor SP was related to having positive screening in the ASQ3-48 (OR 6.5 [95% CI, 1.9-22.2]) while having received SI was related to late prematurity (OR 3.6 [95% CI, 1.3-9.6]). The ASQ3-48 shows acceptable predictive properties for poor SP but not for receiving SI. No differences were found in SP between LPI and Term cohorts, but LPIs are likelier to require SI.


Use of eCompliance, an innovative biometric system for monitoring of tuberculosis treatment in rural Uganda.
Directly observed therapy short-course (DOTS) requires direct observation of tuberculosis (TB) patients and manual recording of doses taken. Programmatically, manual tracking is both time-consuming and prone to human error. Our project in western Uganda assessed the impact on TB treatment outcomes of a comprehensive patient support program including eCompliance, a biometric medical record device, with the aim of increasing TB patient retention. Through an observational study of 142 patients, DOTS outcomes of patients in the intervention group were compared with two control groups. Descriptive statistical comparisons, case-cohort analysis, and difference in change over time were used to assess the impact. Intervention patients had a higher cure rate than all other patients (55.6% versus 28.3% [P < 0.01]) and the odds of having a "cured" outcome were 3.17 higher (P < 0.05). The intervention group had a statistically significantly lower odds of having a negative outcome (0% versus.17% [P < 0.01]) than patients from the control groups. Additionally, the intervention group had a lost to follow-up rate lower than all other groups (0% versus 7%) that was trending on significant. In resource-limited settings, implementing comprehensive DOTS including eCompliance may reduce the occurrence of negative DOTS outcomes for patients.


Heparin-immobilized pluronic/PVA composite microparticles for the sustained delivery of ionic drug.
Heparin-immobilized Pluronic (F-68)/Polyvinylalcohol (PVA) composite microparticles were designed and characterized for the sustained drug delivery of ionic drug. Venlafaxine, antidepressant medication, was used as a model drug. For the efficient loading of ionic drug, heparin was immobilized into F-68/PVA composite microparticles. Differential scanning calorimetry (DSC) was used to understand the intra/intermolecular interactions in the heparin-immobilized F-68/PVA composite gels containing model drug. For the application as a sustained drug delivery system, the loading amount and release pattern of loaded drug were measured using high performance liquid chromatography (HPLC).


Lithium enhancement of central 5-HT transmission induced by 5-HT precursors.
The effects of acute and chronic lithium chloride administration on synaptic transmission between bulbospinal norepinephrine (NE) or 5-hydroxy-tryptamine (5-HT) pathways and sympathetic preganglionic neurons were tested in unanesthetized, spinal cats. Discharges recorded from sympathetic preganglionic white rami were evoked by stimulation of spinal reflex pathways or descending excitatory pathways in the cervical spinal cord. Acute lithium administration (2 meq/kg) produced insignificant depression of the reflex pathway but markedly depressed transmission through the intraspinal pathway, an effect that was prevented by depletion or blockage of 5-HT. These observations and the failure of lithium to alter the typical effects of L-dopa on both pathways indicate that lithium does not affect transmission through the excitatory NE pathway. L-Tryptophan (1,0 mg/kg) alone produced little or no depression of either pathway, but 3--4 hr after lithium, this dose of L-tryptophan gradually depressed transmission through both pathways by about 20%. After chronic lithium pretreatment (1 meq/kg twice a day for 3 days), L-tryptophan rapidly depressed transmission through spinal reflex and intraspinal pathways by 40% and 50% respectively. Chronic lithium pretreatment also more than doubled the depression of transmission through both pathways produced by 30 mg/kg of 5-HTP. The average of plasma lithium levels 8--10 hr after the last chronic dose was 1.5 meq/liter. These results support the proposal that lithium increases the uptake of L-tryptophan and 5-HTP by central 5-HT terminals and thereby enhances 5-HT synthesis which is reflected in increased transmission at central 5-HT synapses.


Health-promoting lifestyle behaviors of Spanish-speaking Hispanic adults.
The study purpose was to determine (a) health-promoting lifestyle behaviors of Spanish-speaking Hispanics, (b) differences in these behaviors by demographics and acculturation, and (c) the extent to which perceived health status, demographics, and acculturation explain health-promoting lifestyles. The Spanish-language Health-Promoting Lifestyle Profile II (HPLPII) and other instruments were completed by a convenience sample of 545 Hispanic adults recruited in several midwestern communities. Of the six behavioral dimension scores of the HPLPII, scores were lowest for physical activity and highest for spiritual growth. They differed by age, gender, employment status, marital status, and acculturation. Perceived health status, demographics, and acculturation explained 12% of the variance in overall health-promoting lifestyle. These findings can help guide culturally appropriate interventions designed to improve this population's overall health.


Prevalence and association of early onset severe hyperbilirubinemia in newborn in the East China region: Retrospective medical record analyses.
Research on the prevalence and association of hyperbilirubinemia is controversial because of different cultures, demographics, and clinical conditions. The etiology of hyperbilirubinemia is affected by the environment and other factors in the newborn. The World Health Organization recommended a 1-day hospital stay after uncomplicated delivery, jaundice assessment before discharge, and screening on 3rd and 7th days after birth for hyperbilirubinemia. However, the implementation of these recommendations is difficult in China. The objective of this study was to evaluate the prevalence and association of early onset severe hyperbilirubinemia in newborns in East China. Retrospective medical record analyses for 250 cesarean sections or vaginal deliveries, ≥2 kg body weight, and negative for Hepatitis B surface antigen by birth newborns were performed. A biochemical analyzer, quantitative assay, and quantitative polymerase chain reaction were used to evaluate total serum bilirubin, glucose-6-phosphate dehydrogenase (G6PD) deficiency, and gene variant phenotyping, respectively. A total in 33 (13%) newborns were reported with early onset severe hyperbilirubinemia (according to the American Academy of Pediatrics, total serum bilirubin ≥ 342 μmol/L within 6 hours of birth). All newborns with severe hyperbilirubinemia were hospitalized and underwent phototherapy. The mothers of all newborns had a gestational age ≥ 35 weeks. Hospitalization included artificial feeding, and breastfeeding was rare (P < .0001). ABO incompatibility ("O" blood type for mother and either "A" or "AB" or "B" blood type for newborn, P = .0411), G6PD deficiency (G6PD/6-phosphogluconate dehydrogenase ≤ 1.0 in quantitative assay, P = .0422), Rh incompatibility (the mother's blood type was Rh negative and newborn blood type was Rh positive, P = .0416), fewer genotype rs4149056 frequencies (P = .0452), higher genotype rs2306283 frequencies (P = .0461), and higher genotype rs1805173 frequencies (P = .0471) were independent parameter for early onset severe hyperbilirubinemia of newborns. The prevalence of early onset severe hyperbilirubinemia in Chinese newborns is 13% in the East China region. Blood incompatibility, G6PD deficiency, fewer genotype rs4149056 frequencies, higher genotype rs2306283 frequencies, and higher genotype rs1805173 frequencies were independent predictors of early onset severe hyperbilirubinemia among newborns in the East China region (Level of Evidence: IV; Technical Efficacy: Stage 5).


Genotype to phenotype mapping still needs underpinning by research in metabolism and enzymology.
The article 'Evidence that the metabolite repair enzyme NAD(P)HX epimerase has a moonlighting function' by Niehaus et al. published in this issue illustrates a number of the problems that still arise when attempting to translate genotypes to phenotypes, such as for interpreting mutant phenotypes or building genome-scale metabolic models. In this case, the mutation concerned appears to map to an enzyme in one of the little-known but essential metabolite repair pathways that have been discovered in recent years. However, the bioinformatic and experimental evidence presented suggests that the annotated enzyme activity of the mutated gene product, whilst correct, accounts neither for the phenotype nor for the chromosomal and transcriptional associations of the gene. The bioinformatic and metabolomic evidence presented points to an additional but important role for the gene product in pyridoxal phosphate homoeostasis, thus adding the enzyme to the expanding list of those with a 'moonlighting function'.


Decreased wound neutrophils and indiscrete margination in the pathogenesis of wound infection.
To assess the pathogenesis of increased susceptibility to infection and septic death in a rat model, neutrophils (PMNs) in the wound, circulating PMNs, and their in vivo activity were evaluated after 30% and 60% burns. Eight hours after injury there were twice as many PMNs in the wounds of rats that sustained 30% compared with 60% burns. There was no difference between these two groups in the number of circulating PMNs at 2, 4, 6, and 8 hours after injury. In vivo evaluation of PMN response to infusion of F-Met-Leu-Phe revealed that circulating PMNs were more sensitive 4 hours after 60% burns compared with sham burns. At this time PMNs were found to be less sensitive to zymosan-activated serum infusion after 30% burns compared with sham burns. However, the PMNs in rats with 30% burns were more sensitive to this stimulus than were PMNs in rats with 60% injuries. Thus rats with greater injury, known to be more susceptible to wound infection, have fewer PMNs in their wounds 8 hours after injury. This is preceded by an increased sensitivity of PMNs in vivo to bacterial chemotactic factor and a relative increase in sensitivity to wound factors. This unusual finding implicates indiscrete margination as a factor in the pathogenesis of infection.


Formative research to develop a mass media campaign to increase physical activity and nutrition in a multiethnic state.
Poor nutrition and physical inactivity are the second leading causes of preventable morbidity and mortality in the United States. Mass media campaigns have tremendous promise for reaching large segments of the population to influence these behaviors. There is still insufficient evidence in the literature, however, to recommend mass marketing campaigns for physical activity and nutrition. Successful mass media campaigns should have a formative research base that includes conducting preproduction research with the target audience, using theory as a conceptual foundation of the campaign, segmenting the audience into meaningful subgroups, and using a message approach that is targeted to and likely will be effective with the audience segment. In this study, these formative research steps were addressed to develop a mass media campaign based on the Theory of Planned Behavior to increase physical activity and fruit and vegetable consumption in 35-55-year-old adults in the state of Hawaii. For the walking campaign, our results identified time, a control belief, as the major barrier. For fruits and vegetable, the data suggested social norm (if others around me ate them) and control (if they were available). These data then were used to develop a mass media campaign based on these principals.


The inheritance of spontaneous amyloidosis development in mice: a model for hereditary threshold metabolic disorders.
To study the inheritance of spontaneous amyloidosis development in mice, we crossed the LLC strain (with a high incidence) with the A/J strain (with a low incidence) and with the HLC strain (with a zero incidence of amyloidosis). We produced the F1 and a backcross generation to each parental strain in each cross. Examination of the spleen, liver, and kidneys of each mouse for the presence of amyloid was made between age 15 and 18 months. The data fit neither a dominant nor a recessive single gene hypothesis for the development of amyloidosis. In consideration of amyloidosis as a hereditary threshold character, we developed an additive gene model. Subsequently, a test for fitness of the observed percentages to the expected percentages in the backcross generations was made according to this model. The observed percentages of amyloidosis in the spleens, livers, and individual mice agreed with the expected percentages in the LLC X A/J crosses but not in the LLC X HLC crosses. Therefore, we conclude that for the development of amyloidosis, a difference exists at one locus between LLC and A/J and at more than one locus between LLC and HLC. The probability of development of amyloidosis in an individual depends on the effects of the genes at these loci. For hereditary metabolic disorders that cannot be explained by either a single dominant or recessive gene hypothesis, this genetic model may be useful to test whether the development of the disease is due to additive effects of genes.


A comparative study between staple line reinforcement during laparoscopic sleeve gastrectomy and no reinforcement: an Egyptian experience.
Laparoscopic sleeve gastrectomy (LSG) has become an increasingly popular bariatric procedure. LSG still conveys some risks, including early staple line complications such as bleeding and leaks. It has been proposed that staple line complications can be reduced by staple line reinforcement (SLR). This study aimed to compare the short-term efficacy and safety of the SLR during LSG by oversewing versus no SLR in an Egyptian cohort over a period of 11 years. This is a retrospective study that analyzed data from patients undergoing LSG by the same surgeon over a period of 11 years. The patients' early postoperative complications were compared according to performing SLR. The SLR group showed significantly longer surgery time (p = 0.021) and a lower rate of postoperative bleeding (p = 0.027). All leakage cases occurred in the non-SLR group (0.7% vs. 0.0%) without statistical significance (p = 0.212). The two mortality cases occurred in the non-SLR group. The LOS was comparable in the two groups (p = 0.289). This study confirms the short-term benefits of SLR by oversewing during LSG in terms of a lower incidence of 30-day morbidity, particularly bleeding, and lower rates of reoperation, with a clinically questionable longer operation time.


Molecular basis of antibiotic self-resistance in a bee larvae pathogen.
Paenibacillus larvae, the causative agent of the devastating honey-bee disease American Foulbrood, produces the cationic polyketide-peptide hybrid paenilamicin that displays antibacterial and antifungal activity. Its biosynthetic gene cluster contains a gene coding for the N-acetyltransferase PamZ. We show that PamZ acts as self-resistance factor in Paenibacillus larvae by deactivation of paenilamicin. Using tandem mass spectrometry, nuclear magnetic resonance spectroscopy and synthetic diastereomers, we identified the N-terminal amino group of the agmatinamic acid as the N-acetylation site. These findings highlight the pharmacophore region of paenilamicin, which we very recently identified as a ribosome inhibitor. Here, we further determined the crystal structure of PamZ:acetyl-CoA complex at 1.34 Å resolution. An unusual tandem-domain architecture provides a well-defined substrate-binding groove decorated with negatively-charged residues to specifically attract the cationic paenilamicin. Our results will help to understand the mode of action of paenilamicin and its role in pathogenicity of Paenibacillus larvae to fight American Foulbrood.


Auditory stimulation of opera music induced prolongation of murine cardiac allograft survival and maintained generation of regulatory CD4+CD25+ cells.
Interactions between the immune response and brain functions such as olfactory, auditory, and visual sensations are likely. This study investigated the effect of sounds on alloimmune responses in a murine model of cardiac allograft transplantation. Naïve CBA mice (H2k) underwent transplantation of a C57BL/6 (B6, H2b) heart and were exposed to one of three types of music--opera (La Traviata), classical (Mozart), and New Age (Enya)--or one of six different single sound frequencies, for 7 days. Additionally, we prepared two groups of CBA recipients with tympanic membrane perforation exposed to opera for 7 days and CBA recipients exposed to opera for 7 days before transplantation (pre-treatment). An adoptive transfer study was performed to determine whether regulatory cells were generated in allograft recipients. Immunohistochemical, cell-proliferation, cytokine, and flow cytometry assessments were also performed. CBA recipients of a B6 cardiac graft that were exposed to opera music and Mozart had significantly prolonged allograft survival (median survival times [MSTs], 26.5 and 20 days, respectively), whereas those exposed to a single sound frequency (100, 500, 1000, 5000, 10,000, or 20,000 Hz) or Enya did not (MSTs, 7.5, 8, 9, 8, 7.5, 8.5 and 11 days, respectively). Untreated, CBA mice with tympanic membrane perforations and CBA recipients exposed to opera for 7 days before transplantation (pre-treatment) rejected B6 cardiac grafts acutely (MSTs, 7, 8 and 8 days, respectively). Adoptive transfer of whole splenocytes, CD4+ cells, or CD4+CD25+ cells from opera-exposed primary allograft recipients resulted in significantly prolonged allograft survival in naive secondary recipients (MSTs, 36, 68, and > 100 days, respectively). Proliferation of splenocytes, interleukin (IL)-2 and interferon (IFN)-γ production was suppressed in opera-exposed mice, and production of IL-4 and IL-10 from opera-exposed transplant recipients increased compared to that from splenocytes of untreated recipients. Flow cytometry studies showed an increased CD4+CD25+ Forkhead box P3 (Foxp3)+ cell population in splenocytes from those mice. Our findings indicate that exposure to opera music, such as La traviata, could affect such aspects of the peripheral immune response as generation of regulatory CD4+CD25+ cells and up-regulation of anti-inflammatory cytokines, resulting in prolonged allograft survival.


Prevalence and risk factors of anemia in children.
To identify the prevalence and factors associated with anemia in children attending Municipal Early Childhood Education Day Care Center (Centros Municipais de Educação Infantil [CMEI]) nurseries in Colombo-PR. Analytical, cross-sectional study with a representative sample of 334 children obtained by stratified cluster sampling, with random selection of 26 nurseries. Data collection was conducted through interviews with parents, assessment of iron intake by direct food weighing, and hemoglobin measurement using the finger-stick test. Bivariate association tests were performed followed by multiple logistic regression adjustment. The prevalence of anemia was 34.7%. Factors associated with anemia were: maternal age younger than 28 years old (p=0.03), male children (p=0.02), children younger than 24 months (p=0.01), and children who did not consume iron food sources (meat+beans+dark green leafy vegetables) (p=0.02). There was no association between anemia and iron food intake in CMEI. However, iron intake was well below the recommended levels according to the National Education Development Fund resolution, higher prevalence of anemia was observed in children whose intake of iron, heme iron, and nonheme iron was below the median. In terms of public health, the prevalence of anemia is characterized as a moderate problem in the studied population and demonstrates the need for coordination of interdisciplinary actions for its reduction in CMEI nurseries.


Macrophage-based therapeutic strategies in regenerative medicine.
Mounting evidence suggests that therapeutic cell and drug delivery strategies designed to actively harness the regenerative potential of the inflammatory response have great potential in regenerative medicine. In particular, macrophages have emerged as a primary target because of their critical roles in regulating multiple phases of tissue repair through their unique ability to rapidly shift phenotypes. Herein, we review macrophage-based therapies, focusing on the translational potential for cell delivery of ex vivo-activated macrophages and delivery of molecules and biomaterials to modulate accumulation and phenotype of endogenous macrophages. We also review current obstacles to progress in translating basic findings to therapeutic applications, including the need for improved understanding of context-dependent macrophage functions and the myriad factors that regulate macrophage phenotype; potential species-specific differences (e.g. humans versus mice); quality control issues; and the lack of standardized procedures and nomenclature for characterizing macrophages. Looking forward, the inherent plasticity of macrophages represents a daunting challenge for harnessing these cells in regenerative medicine therapies but also great opportunity for improving patient outcomes in a variety of pathological conditions.


Peptide analysis of mammalian decomposition fluid in relation to the post-mortem interval.
We report the results of a semi-quantitative peptide analysis of decomposition fluid under field-based conditions in the absence of a soil matrix. Sixteen domestic pig (Sus scrofa domesticus) cadavers were used to model human decomposition in trials conducted in the summer and winter months in Western Australia. Physical characteristics were recorded and targeted peptide components of decomposition fluid were analysed using high performance liquid chromatography-triple quadrupole mass spectrometry. Principal component analysis identified 29 peptides, originating from haemoglobin subunits alpha and beta, creatine kinase, beta-enolase and lactate dehydrogenase, that contributed to differences in the mean peak areas of samples collected during the early period of decomposition (days 6-12 and day 2 in winter and summer, respectively) and during the later period (days 24-34 and days 8-10 in winter and summer, respectively). Fold changes for 8 peptides between these periods were significantly different. Three peptides derived from haemoglobin subunit beta, one from beta-enolase and two from lactate dehydrogenase displayed consistent trends, in that a notable increase in mean peak area was followed by a marked decrease in both the summer and winter samples. When temperature was accounted for, these trends occurred at different time points in summer and winter, indicating that factors other than temperature had impacted the rate of degradation of the proteins involved. The single peptides derived from haemoglobin subunit alpha and creatine kinase displayed consistent increases in mean peak area for the summer samples, suggesting that temperature played the most significant role in their degradation. Further analyses revealed that 7 peptides (one originating from haemoglobin subunit alpha, three from haemoglobin subunit beta and three from lactate dehydrogenase) displayed consistent trends that could be correlated with total body score and with the early stages of decomposition. The consistent trends (mean peak area versus time) for peptides derived from several proteins during decomposition trials conducted under different temperature regimes further emphasised the potential of peptide analysis in time since death estimation.


Extrarenal Manifestations: Polycystic Liver Disease and Its Complications.
The liver is the commonest site of involvement outside of the kidney in autosomal dominant polycystic kidney disease. Most individuals with polycystic liver disease are asymptomatic and require no therapeutic interventions, but a small number of affected individuals who experience symptomatic polycystic liver disease develop medical complications as a result of massive enlargement of cyst number and size and hepatic parenchyma and its subsequent associated complications. This can lead to deterioration in overall health and quality of life, increasing morbidity and mortality. In this review, we will touch upon disease pathogenesis, prevalence, and complications and discuss recent advances in surgical and medical management.


Animal-assisted therapies for autism.
Animal-assisted therapies have been increasingly used as part of treatment for a range of emotional-behavioral conditions and have more recently been incorporated into treatment for children with symptoms associated with autism spectrum disorder. Autism spectrum disorder affects one in 36 children and early interventions can be very effective. The aims of this paper are to: 1) provide background into animal-assisted therapies including a breakdown on the subtypes of therapies, 2) report broad summaries of research outcomes across various domains: social-emotional, physical, quality of life, behavioral skills, and adaptive skills, and 3) summarize key takeaways for pediatric practitioners when supporting families of youth with autism spectrum disorder who are interested in participating in animal-assisted therapy. This paper is not a systematic review or meta-analysis as several rigorous review studies have been published already. This paper is an overview of the background and research for pediatric practitioners when advising families on treatment options. Implications for future research are covered.


Recent advances in analysis of new psychoactive substances by means of liquid chromatography coupled with low-resolution tandem mass spectrometry.
The number of methods for the analysis of new psychoactive substances (NPS) is continually increasing, and there is no indication that this trend will change in the near future. The constantly growing market of "designer drugs" makes it necessary to develop new methods of their analysis. The aim of this review is to present the multi-component methods of detection and identification of NPS using low-resolution tandem mass spectrometry coupled with liquid chromatography. For this purpose, 36 articles were selected by applying strictly defined search criteria. Due to the large differences in the matrices and physicochemical properties of the analytes, the described research methods are diverse. These differences are visible in sample preparation methods, chromatographic columns, mobile phases, gradients, or additives to mobile phases used. This work collects and organizes the existing information on the subject of NPS screening analysis methods and will be helpful to forensic scientists working on this topic.


The impact of viral and host elements on HIV fitness and disease progression.
Twenty-five years after the emergence of HIV onto the global scene, multiple advancements have been made in the understanding of HIV pathology. Thanks to the development of antiretroviral therapies, growing numbers of individuals with HIV infection experience slowed or halted acceleration to AIDS. Despite this, new HIV infections and AIDS-related morbidity and mortality are still common in the highly active antiretroviral therapy era. Recently, we and others have identified viral replicative fitness as a major determinant of HIV disease progression, which could have a major impact in the clinical setting. Therefore, in this review, we will discuss host and viral factors that affect viral fitness and its relationship on HIV pathogenesis.


Superior Canal Dehiscence Syndrome Affecting 3 Families.
Superior canal dehiscence syndrome (SCDS) is an increasingly recognized cause of hearing loss and vestibular symptoms, but the etiology of this condition remains unknown. To describe 7 cases of SCDS across 3 families. This retrospective case series included 7 patients from 3 different families treated at a neurotology clinic at a tertiary academic medical center from 2010 to 2014. Patients were referred by other otolaryngologists or were self-referred. Each patient demonstrated unilateral or bilateral SCDS or near dehiscence. Clinical evaluation involved body mass index calculation, audiometry, cervical vestibular evoked myogenic potential testing, electrocochleography, and multiplanar computed tomographic (CT) scan of the temporal bones. Zygosity testing was performed on twin siblings. The diagnosis of SCDS was made if bone was absent over the superior semicircular canal on 2 consecutive CT images, in addition to 1 physiologic sign consistent with labyrinthine dehiscence. Near dehiscence was defined as absent bone on only 1 CT image but with symptoms and at least 1 physiologic sign of labyrinthine dehiscence. A total of 7 patients (5 female and 2 male; age range, 8-49 years) from 3 families underwent evaluation. Family A consisted of 3 adult first-degree relatives, of whom 2 were diagnosed with SCDS and 1 with near dehiscence. Family B included a mother and her child, both of whom were diagnosed with unilateral SCDS. Family C consisted of adult monozygotic twins, each of whom was diagnosed with unilateral SCDS. For all cases, dehiscence was located at the arcuate eminence. Obesity alone did not explain the occurrence of SCDS because 5 of the 7 cases had a body mass index (calculated as weight in kilograms divided by height in meters squared) less than 30.0. Superior canal dehiscence syndrome is a rare, often unrecognized condition. This report of 3 multiplex families with SCDS provides evidence in support of a potential genetic contribution to the etiology. Symptomatic first-degree relatives of patients diagnosed with SCDS should be offered evaluation to improve detection of this disorder.


Leptin receptor-mediated regulation of cholinergic neurotransmitter phenotype in cells of central nervous system origin.
Leptin is an adipocyte-secreted hormone that regulates body weight and exerts effects on hematopoiesis, reproduction, and immunity. The leptin receptor (OBR) shares sequence similarity and signaling capabilities with receptors for cytokines of the ciliary neurotrophic factor (CNTF) family. Our previous finding that CNTF and leptin exert similar anti-obesity effects and activate common neuronal signaling pathways, prompted us to investigate whether leptin may share with CNTF the ability to regulate the expression of specific neuronal genes. To this end, we established a cell line, derived from the murine septal cholinergic neuronal cell line SN-56, which stably expresses OBR. In this cell line, termed SN-56/OBR, leptin induces STAT transcription factor activation and STAT-dependent reporter gene expression in a manner similar to that of CNTF. Furthermore, in SN-56/OBR cells both CNTF and leptin produce changes in neurotransmitter and neuropeptide phenotype characteristic of cholinergic neurons, such as an increase in choline acetyltransferase and vasoactive intestinal polypeptide, and a decrease in neuropeptide Y expression. SN-56/OBR cells thus constitute an interesting new model system to investigate leptin action in cells of central nervous system origin. Possible physiological implications of OBR's intrinsic ability to regulate cholinergic phenotypic markers are discussed.


Shortest-path constraints for 3D multiobject semiautomatic segmentation via clustering and Graph Cut.
We derive shortest-path constraints from graph models of structure adjacency relations and introduce them in a joint centroidal Voronoi image clustering and Graph Cut multiobject semiautomatic segmentation framework. The vicinity prior model thus defined is a piecewise-constant model incurring multiple levels of penalization capturing the spatial configuration of structures in multiobject segmentation. Qualitative and quantitative analyses and comparison with a Potts prior-based approach and our previous contribution on synthetic, simulated, and real medical images show that the vicinity prior allows for the correct segmentation of distinct structures having identical intensity profiles and improves the precision of segmentation boundary placement while being fairly robust to clustering resolution. The clustering approach we take to simplify images prior to segmentation strikes a good balance between boundary adaptivity and cluster compactness criteria furthermore allowing to control the trade-off. Compared with a direct application of segmentation on voxels, the clustering step improves the overall runtime and memory footprint of the segmentation process up to an order of magnitude without compromising the quality of the result.


Trimethyllysine, a trimethylamine N-oxide precursor, provides near- and long-term prognostic value in patients presenting with acute coronary syndromes.
Trimethyllysine (TML) serves as a nutrient precursor of the gut microbiota-derived metabolite trimethylamine N-oxide (TMAO) and is associated with incident cardiovascular (CV) events in stable subjects. We examined the relationship between plasma TML levels and incident CV events in patients presenting with acute coronary syndromes (ACS). Plasma levels of TML were quantified in two independent cohorts using mass spectrometry, and its relationship with CV events was investigated. In a Cleveland Cohort (N = 530), comprised of patients presenting to the emergency department with chest pain and suspected ACS, TML was associated with major adverse cardiac events (MACE, myocardial infarction, stroke, need for revascularization, or all-cause mortality) over both 30 days [3rd tertile (T3), adjusted odds ratio (OR) 1.77, 95% confidence interval (CI) 1.04-3.01; P < 0.05] and 6 months (T3, adjusted OR 1.95, 95% CI 1.15-3.32; P < 0.05) of follow-up independent of traditional CV risk factors and indices of renal function. Elevated TML levels were also associated with incident long-term (7-year) all-cause mortality [T3, adjusted hazard ratio (HR) 2.52, 95% CI 1.50-4.24; P < 0.001], and MACE even amongst patients persistently negative for cardiac Troponin T at presentation (e.g. 30-day MACE, T3, adjusted OR 4.49, 95% CI 2.06-9.79; P < 0.001). Trimethyllysine in combination with TMAO showed additive significance for near- and long-term CV events, including patients with 'negative' high-sensitivity Troponin T levels. In a multicentre Swiss Cohort (N = 1683) comprised of ACS patients, similar associations between TML and incident 1-year adverse cardiac risks were observed (e.g. mortality, adjusted T3 HR 2.74, 95% CI 1.28-5.85; P < 0.05; and MACE, adjusted T3 HR 1.55, 95% CI 1.04-2.31; P < 0.05). Plasma TML levels, alone and together with TMAO, are associated with both near- and long-term CV events in patients with chest pain and ACS.


Risks and prevention of contamination of beef carcasses during the slaughter process in the United States of America.
A brief general description of the beef cattle slaughter process used in the United States of America is presented. Considerable variation in prevalence of microbial contamination at beef processing establishments has been reported. Primary factors which may explain this variation include: a) pathogen load on, or in, live animals entering the establishment; b) sanitation practices used by the establishment. Pathogen load on, or in, live animals depends on environmental exposure to contaminants, the health of the animal, and on situations stressful to the animal which may increase microbial shedding, such as disrupted access to feed and water, transport, handling and contact with other animals. Processing stations and slaughter practices which are potential sources of carcass contamination are listed, and points on carcasses where contamination is most likely to occur are noted. Contamination risk-reduction measures within an establishment include carcass treatments with anti-microbial agents, trimming, washing, steaming, chilling and gamma irradiation. Several good sanitation practices are described. Microbial monitoring is a necessary step for determining whether sanitation practices are efficacious, but the usefulness of microbial monitoring depends on the microbial tests selected, sampling procedures, frequency of sampling, the rapidity of receiving test results and consistent and accurate record-keeping and analysis. Federal inspection to ensure that processing establishments comply with new Federal regulations regarding the implementation of standard sanitation procedures and microbial testing should result in greater vigilance of good sanitation practices by establishments.


The Oxime Derivatives of 1-R-1H-Naphtho[2,3-d][1,2,3]triazole-4,9-dione 2-oxides: Synthesis and Properties.
To synthesize a novel chemotype based on the naphthoquinone scaffold with retained cytotoxicity and provisionally low intracellular oxidation potential. Derivatives of naphthoquinone, although potent anticancer agents, can exert heart toxicity due to generation of free oxygen species. In this study, we modified the scaffold by replacing one carbonyl group with the oxime moiety. Interestingly, only one carbonyl group in 1-R-1H-naphtho[2,3-d][1,2,3]triazole-4,9-dione 2-oxides reacted with hydroxylamine. The spatial structure was determined by X-ray analysis. New compounds were tested for the ability to form stable complexes with double stranded DNA by spectroscopy and molecular docking and to induce death of tumor cell lines and non-malignant counterparts. The resulting 1-R-1H-naphtho[2,3-d][1,2,3]triazole-4,9-dione 4-oxime 2-oxides were further acylated to produce a series of 1-R-1H-naphtho[2,3-d][1,2,3]triazole-4,9-dione 4-(O-acyloxime) 2-oxides. Newly synthesized compounds demonstrated a higher (in submicromolar or low micromolar range) cytotoxic potency against human colon and breast adenocarcinoma cell lines than to non-malignant skin fibroblasts. Spectroscopic measurements revealed that, unlike other classes of quinone derivatives, new naphthotriazoledione oxides did not form stable complexes with double stranded DNA regardless of their fitting to the DNA minor groove (as determined by molecular modeling). Thus, our chemical modifications yielded a new chemotype with good cytotoxic properties and yet-to-be-identified intracellular target(s).


Reduction of congenital nystagmus in a patient after smoking cannabis.
Smoking cannabis has been described to reduce acquired pendular nystagmus in MS, but its effect on congenital nystagmus is not known. To report the effect of smoking cannabis in a case of congenital nystagmus. A 19-year-old male with congenital horizontal nystagmus presented to the clinic after smoking 10 mg of cannabis. He claimed that the main reason for smoking cannabis was to improve his vision. At the next clinic appointment, he had not smoked cannabis for 3weeks. Full ophthalmologic examination and eye movement recordings were performed at each visit. Visual acuity improved by 3 logMar lines in the left eye and by 2 logMar lines in the right eye after smoking cannabis. The nystagmus intensities were reduced by 30% in primary position and 44%, 11%, 10% and 40% at 20-degree eccentricity to the right, left, elevation and depression, respectively, after smoking cannabis. Cannabis may be beneficial in the treatment of congenital idiopathic nystagmus (CIN). Further research to clarify the safety and efficacy of cannabis in patients with CIN, administered for example by capsules or spray, would be important.


Lipids associated with bovine kidney glomerular basement membranes.
1. After incubation of bovine glomeruli with D-[U-14C]glucose, about 21% of the total radioactivity is found in lipid extracts of glomerular basement membranes. 2. The concentration of lipids in glomerular basement membranes (4.3% of dry wt.) is lower than in the residual glomerular particles (10.8% of dry wt.). The concentrations of neutral lipids (13.9%), phospholipids (46.7%) and cholesterol (37.9%) in the total lipid extract of the glomerular basement membranes, however, differ from those in the residual glomerular particles (15.6, 54.0 and 30.9% respectively). Though residual glomerular particles show a higher lipid content, the radioactivity in this fraction only amounts to 38% of that found in the glomerular basement membranes. 3. The specific radioactivity of total glomerular basement-membrane lipids (12 600 d.p.m./mg) is about 4 times as high as that of the glomerular basement membranes. The specific radioactivities of the individual lipid components, however, differ. The highest values are found for phosphatidylcholine and triacylglycerols. The largest proportion of the radioactivity is found in the glycerol of the glycerides. The radioactivity in the fatty acids is much less and does not differ significantly in the various classes of lipids. 4. G.1.c. of methyl esters of the fatty acids does not reveal a clear difference between the fatty acid compositions of glomerular basement membranes and residual glomerular particles. 5. Treatment of glomerular basement-membrane preparations with ultrasound, the generally used procedure for glomerular basement-membrane preparations, drastically decreases the lipid content of glomerular basement membranes. 6. It is concluded that lipids are associated with the basement membranes. Further, the comparatively high radioactive labelling suggests that glomerular basement-membrane lipids may be an interesting class of substances for further pathological studies.


Functional and volumetric assessment of liver segments after portal vein embolization: Differences in hypertrophy response.
Patients considered for liver resection with insufficient volume or function of the future remnant liver are candidates for portal vein embolization to allow safe resection. The aim of this study is to analyze the volumetric and functional responses after portal vein embolization and to evaluate predictors of the hypertrophy response. All patients who underwent portal vein embolization before liver resection 2006-2017 were included. Patients who did not undergo computed tomography-volumetry and functional assessment with technetium-99m mebrofenin hepatobiliary scintigraphy before and after portal vein embolization were excluded. The functional and volumetric response rates were calculated. Multiple regression analysis was conducted to examine the relationship between the hypertrophy response and potential predictors. A total of 90 patients underwent portal vein embolization of the right liver. After 3 weeks, there was a significant increase in both volumetric and functional share of the future remnant liver (both P < .01). The increase in functional share exceeded the increase in volumetric share (P < .01). The median functional contribution of segment 4 after portal vein embolization was 41.5% (31.7%-48.7%) of the nonembolized lobe. Preoperative chemotherapy was not a significant predictor of the increase in function or volume. Compared with benign lesions, malignant diseases were significant negative predictors of the functional response. A total of 3 weeks after portal vein embolization, the functional response exceeded that of the volumetric response, meaning that the waiting time to resection potentially can be decreased. Segment 4 had a significant share of both volume and function, enabling surgical strategies only leaving segment 4 as a monosegment. Neoadjuvant chemotherapy had no negative influence on the hypertrophy response.


Management of Chemotherapy for Stage II Nasopharyngeal Carcinoma in the Intensity-Modulated Radiotherapy Era: A Review.
Nasopharyngeal carcinoma is an endemic disease with a high prevalence in Southeast Asia, Mediterranean countries, and Northern Africa. With substantial advances in screening and diagnosis, increasingly more early-stage (stage I~II) patients are being diagnosed. The undebated treatment modality for stage I patients is radiotherapy alone. However, controversies exist for patients with stage II disease, mostly revolving around the management of chemotherapy. However, the use of intensity-modulated radiotherapy for the treatment of nasopharyngeal carcinoma has increased recently, which has drastically improved survival outcomes. Thus, many oncologists have considered omitting chemotherapy for stage II patients in the intensity-modulated radiotherapy era. Unfortunately, prospective studies comparing concurrent radio-chemotherapy with intensity-modulated radiotherapy alone are limited. Notably, stage II nasopharyngeal carcinoma consists of three subgroups, among which stage T2N1M0 disease is unique and potentially warrants additional treatment including chemotherapy. Additionally, molecular biology techniques are advancing at an incredible speed. Instead of adopting a one-size-fits-all recommendation, exploring potential predictive biomarkers to select patients who are likely to derive benefit from chemotherapy is a better choice. In this review, we summarize the data from studies and reviews regarding chemotherapy for stage II nasopharyngeal carcinoma in the intensity-modulated radiotherapy era and discuss chemotherapy utility. Eventually, we conclude that IMRT alone may be sufficient for stage II nasopharyngeal carcinoma, but this needs to be verified by prospective studies in the near future, the evidence collected thus far suggests that concurrent chemo-radiotherapy without induction or adjuvant chemotherapy is yet to be necessary for patients with stage II disease.


Mycobacterium ulcerans Experimental Dormancy.
Whether Mycobacterium ulcerans, the etiological agent of Buruli ulcer in numerous tropical countries, would exist in a dormant state as reported for closely related Mycobacterium species, has not been established. Six M. ulcerans strains were exposed to a progressive depletion in oxygen for 2 months, using the Wayne model of dormancy previously described for M. tuberculosis, and further examined by microscopy after staining of dynamic, dormant, and dead mycobacteria (DDD staining), microcalorimetry and subculture in the presence of dead and replicative M. ulcerans as controls. Mycobacterium ulcerans CU001 strain died during the progressive oxygen depletion and four of five remaining strains exhibited Nile red-stained intracellular lipid droplets and a 14- to 20-day regrowth when exposed to ambient air, consistent with dormancy. A fifth M. ulcerans 19423 strain stained negative in DDD staining and slowly regrew in 27 days. Three tested M. ulcerans strains yielded microcalorimetric pattern similar to that of the negative (dead) homologous controls, differing from that of the homologous positive (replicative) controls. The relevance of these experimental observations, suggesting a previously unreported dormancy state of M. ulcerans, warrants further investigations in the natural ecological niches where M. ulcerans thrive as well as in Buruli ulcer lesions.


Automated, Vision-Based Goniometry and Range of Motion Calculation in Individuals With Suspected Ehlers-Danlos Syndromes/Generalized Hypermobility Spectrum Disorders: A Comparison of Pose-Estimation Libraries to Goniometric Measurements.
Generalized joint hypermobility (GJH) often leads clinicians to suspect a diagnosis of Ehlers Danlos Syndrome (EDS), but it can be difficult to objectively assess. Video-based goniometry has been proposed to objectively estimate joint range of motion in hyperextended joints. As part of an exam of joint hypermobility at a specialized EDS clinic, a mobile phone was used to record short videos of 97 adults (89 female, 35.0 ± 9.9 years old) undergoing assessment of the elbows, knees, shoulders, ankles, and fifth fingers. Five body keypoint pose-estimation libraries (AlphaPose, Detectron, MediaPipe-Body, MoveNet - Thunder, OpenPose) and two hand keypoint pose-estimation libraries (AlphaPose, MediaPipe-Hands) were used to geometrically calculate the maximum angle of hyperextension or hyperflexion of each joint. A custom domain-specific model with a MobileNet-v2 backbone finetuned on data collected as part of this study was also evaluated for the fifth finger movement. Spearman's correlation was used to analyze the angles calculated from the tracked joint positions, the angles calculated from manually annotated keypoints, and the angles measured using a goniometer. Moderate correlations between the angles estimated using pose-tracked keypoints and the goniometer measurements were identified for the elbow (rho =.722; Detectron), knee (rho =.608; MoveNet - Thunder), shoulder (rho =.632; MoveNet - Thunder), and fifth finger (rho =.786; custom model) movements. The angles estimated from keypoints predicted by open-source libraries at the ankles were not significantly correlated with the goniometer measurements. Manually annotated angles at the elbows, knees, shoulders, and fifth fingers were moderately to strongly correlated to goniometer measurements but were weakly correlated for the ankles. There was not one pose-estimation library which performed best across all joints, so the library of choice must be selected separately for each joint of interest. This work evaluates several pose-estimation models as part of a vision-based system for estimating joint angles in individuals with suspected joint hypermobility. Future applications of the proposed system could facilitate objective assessment and screening of individuals referred to specialized EDS clinics.


Er-doped photonic crystal fiber amplifier with 70 W of output power.
We report for the first time, to the best of our knowledge, on a large mode area Er-doped photonic crystal fiber laser system. The fiber core had a diameter of 40 μm and NA of <0.04 leading to a mode field diameter of about 31 μm around 1550 nm wavelength. More than 70 W of single-frequency output power at 1556 nm could be extracted using a master-oscillator power-amplifier scheme. Near diffraction limited output beam quality has been verified by measuring the TEM(00) content with a nonconfocal scanning ring cavity.


Double-blind study of oral gamma-vinyl GABA in the treatment of dystonia.
Six patients with different forms of dystonia were treated with gamma-vinyl GABA, a specific enzyme-activated inhibitor of GABA-transaminase, in a double-blind, placebo-controlled crossover study. gamma-Vinyl GABA therapy, 2 g daily for 2 weeks, was compared with placebo by weekly assessments. There were no consistent changes in three evaluation scores. Agents that augment CNS GABA are unlikely to benefit patients with generalized dystonia.


Opioid Management in CKD.
Patients with chronic kidney disease (CKD) experience a high pain and symptom burden. Concurrently, opioid prescription and use in patients with CKD continues to increase, leading to concern for opioid-related risks. Nephrologists increasingly face challenging clinical situations requiring further evaluation and treatment of pain, for which opioid use may be indicated. However, nephrologists are not commonly trained in pain management and may find it difficult to compile the necessary information and tools to effectively assess and treat potentially multidimensional pain. In these situations, they may benefit from using an evidence-based stepwise approach proposed in this article. We address current approaches to opioid use for pain management in CKD and offer a stepwise approach to individualized opioid assessment, focusing on kidney-specific concerns. This includes thorough evaluation of the pain experience, opioid use history, and treatment goals. We subsequently discuss considerations when initiating opioid therapy, strategies to reduce opioid-related risks, and recommended best practices for opioid stewardship in CKD. Using this sequential approach to opioid management, nephrologists can thereby gain a broad overview of key patient considerations, the foundation for understanding implications of opioid use, and a patient-tailored plan for opioid therapy.


Calibration of ac and dc magnetometers with a Dy2O3 standard.
The ac susceptibility and magnetization curves of a glued Dy(2)O(3) powder sample are measured by an ac susceptometer and a dc superconducting quantum interference device magnetometer, both of which have been calibrated previously. It is shown that the magnetic moment of the paramagnetic sample as a function of field and temperature may be accurately expressed by a combination of the Curie-Weiss law and the Langevin function at T > 45 K with three adjusting parameters, so that the dc magnetization curves and the magnitude and phase of ac susceptibility at different values of dc bias field measured by any magnetometer can be calibrated by using Dy(2)O(3) as a standard. The expressions are empirical and cannot be justified in the entire field and temperature range by existing theories of paramagnetism. Below 10 K, indication of approaching a possible phase transition is found. It is shown that pure Dy(2)O(3) powder may be used as a primary standard, with susceptibility [13.28(T + 17)](-1) emu/Oe/g at T > 50 K and H < 10 kOe, in consistency with the Curie-Weiss law and the quantum mechanical theory of paramagnetism.