Patellar taping for patellofemoral pain: a systematic review and meta-analysis to evaluate clinical outcomes and biomechanical mechanisms.
Patellar taping is frequently used to treat patellofemoral pain (PFP). This systematic review and meta-analysis (1) evaluates the efficacy of patellar taping for patients with PFP, (2) compares the efficacy of various taping techniques and (3) identifies potential biomechanical mechanisms of action. The MEDLINE, CINAHL, SPORTSDiscus, Web of Science and Google Scholar databases were searched in January 2013 for studies evaluating the effects of patellar taping on pain and lower-limb biomechanics in individuals with PFP. Three independent reviewers assessed each paper for inclusion and two assessed for quality. Means and SDs were extracted from each included study to allow effect size calculations. Twenty studies were identified. There is moderate evidence that (1) tailored (customised to the patient to control lateral tilt, glide and spin) and untailored patellar taping provides immediate pain reduction of large and small effect, respectively and (2) tailored patellar taping promotes earlier onset of vastus medialis oblique (VMO) contraction (relative to vastus lateralis contraction). There is limited evidence that (1) tailored patellar taping combined with exercise provides superior pain reduction compared to exercise alone at 4 weeks, (2) untailored patellar taping added to exercise at 3-12 months has no benefit and (3) tailored patellar taping promotes increased internal knee extension moments. Tailoring patellar taping application (ie, to control lateral tilt, glide and spin) to optimise pain reduction is important for efficacy. Evaluation of tailored patellar taping beyond the immediate term is limited and should be a research priority. Possible mechanisms behind patellar taping efficacy include earlier VMO onset and improved knee function capacity (ie, ability to tolerate greater internal knee extension moments).


Disitamab vedotin in combination with immune checkpoint inhibitors for locally and locally advanced bladder urothelial carcinoma: a two-center's real-world study.
Objective: Our study aims to assess the effectiveness and safety profile of Disitamab Vedotin (DV, RC48-ADC), an innovative humanized anti-HER2 antibody conjugated with tubulin-disrupting antimitotic drug monomethyl auristatin E (MMAE) via a cleavable peptide linker. This treatment combined immune checkpoint inhibitors as part of the bladder sparing approach for selected patients suffering from locally and locally advanced bladder urothelial carcinoma. Patients and methods: We conducted a two-center, real-world study involving locally advanced urothelial carcinoma (UC) patients. Patients were classified based on HER2 expression (IHC 3+/2+/1+) or lack of HER2 expression (IHC 0). The primary endpoint was the objective response rate (ORR), assessed by the investigator following the criteria of RECIST V1.1. Secondary endpoints encompassed the pathological complete response rate (pCR), pathological partial response rate (pPR), and pathological stable disease (pSD), along with recurrence-free survival (RFS), the pathological downstaging rate, and the safety profile of the treatment. Results: In this study, nine patients were enrolled, with a median follow-up duration of 12.0 months. The overall confirmed ORR was 88.9%, Five patients achieved a complete response (CR), and three patients achieved a partial response (PR). The radiological complete response (rCR) aligned perfectly with pCR. The median radiological progression-free survival (rPFS) spanned 12.0 months (range from 8.0 to 17.0 months). One patient diagnosed with disease progression (PD) underwent a radical cystectomy. The pathological stage evolved from T2N0M0 to T3aN2M0, followed by adjuvant chemotherapy with a gemcitabine-cisplatin (GC) combination radiotherapy. At the 9-month follow-up, neither recurrence nor metastasis was observed. The rate and intensity of complications were manageable among these patients, with no evidence of grade 4 and 5 adverse events. Conclusion: The combination of DV and PD-1 demonstrated considerable activity in the objective response rate (ORR) in patients with HER2 IHC 0/1+/2+/3+ muscle-invasive bladder cancer (MIBC), along with the longest reported median radiological progression-free survival (rPFS) to date. With an extended duration of treatment, the safety profile of DV plus PD-1 was also confirmed to be manageable.


Activation of a plasma membrane-associated neutral sphingomyelinase and concomitant ceramide accumulation during IgG-dependent phagocytosis in human polymorphonuclear leukocytes.
The sphingomyelin cycle, which plays an important role in regulation of cell growth, differentiation, and apoptosis, involves the formation of ceramide by the action of a membrane-associated, Mg2+-dependent, neutral sphingomyelinase and/or a lysosomal acid sphingomyelinase. In human polymorphonuclear leukocytes (PMNs), ceramide production correlates with and plays a role in the regulation of functional responses such as oxidant release and Fcgamma receptor-mediated phagocytosis. To increase our understanding of the sphingomyelin cycle in human PMNs, the cellular location of neutral and acid sphingomyelinases was investigated in resting, formylmethionylleucylphenylalanine (FMLP)-activated, and FMLP-activated PMNs engaged in phagocytosis. In resting PMNs, a Mg2+-dependent, neutral sphingomyelinase was the predominant activity and was localized to the plasma membrane fractions along with the majority of ceramide. Upon FMLP-activation, there was a 1. 9-fold increase in this neutral, Mg2+-dependent sphingomyelinase activity, which increased to 2.7-fold subsequent to phagocytosis of IgG opsonized targets. This increase in sphingomyelinase activity was restricted to the plasma membrane fractions, which were also the site of increased ceramide levels. Phospholipase D (PLD) activity, which is a target of ceramide action and is required for phagocytosis, was also found primarily in the plasma membrane fractions of FMLP-activated and phagocytosing PMNs. Our findings indicate that in human PMNs engaged in phagocytosis, the sphingomyelin cycle is restricted to the plasma membrane where intracellular targets of ceramide action, such as PLD, are localized.


Health-related quality of life among breast cancer patients compared to cancer survivors and age-matched women in the general population in Vietnam.
This study compared the health-related quality of life (HRQoL) of breast cancer (BC) patients, survivors, and age-matched women from the general population in Vietnam to address the paucity of HRQoL research and contribute to the robust assessment of BC screening and care in Vietnam. The standardised EQ-5D-5L instrument was incorporated in an online survey and a hospital-based face-to-face survey, and together with data from the Vietnam EQ-5D-5L norms study. χ2 tests assessed EQ-5D health profile associations and a Tobit regression model investigated the association between overall health status (EQ-VAS/utility scores) and sociodemographic and clinical characteristics. A total of 309 participants (107 patients undergoing treatment and 202 survivors who had completed treatment) provided usable responses. The dimensions that affected mostly the HRQoL of women with BC were pain/discomfort and anxiety/depression. Current patients and survivors differed significantly regarding HRQoL dimensions of mobility, self-care, usual activities, and anxiety/depression. Their health utilities were 0.74 and 0.84, respectively, compared with 0.91 for age-matched Vietnamese women in the general population (p < 0.001). Treatment status (survivor vs patient), younger age, higher monthly household income, and higher education levels were associated with higher health utility. The results point to unmet needs in mental health support and well-being and for attention to be given to the development of a biopsychosocial system of cancer diagnosis, treatment, and care. The results will also inform future assessments of the comparative value for money of interventions intended to impact on breast cancer in Vietnam.


Modulation of expressivity in PDGFRB-related infantile myofibromatosis: a role for PTPRG?
Infantile myofibromatosis is a rare genetic disorder characterized by the development of benign tumors in the skin, muscle, bone, and viscera. The molecular pathogenesis is still incompletely known. An autosomal dominant form had been reported as causally related with mutations in the gene for platelet-derived growth factor receptor beta (PDGFRB). We report here two siblings with infantile myofibromatosis and with a PDGFRB mutation identified by exome sequence analysis. However, the unaffected mother also had the same PDGFRB mutation. We showed that both children had also inherited from their healthy father a heterozygous mutation in the gene for receptor protein tyrosine phosphatase gamma (PTPRG), an enzyme known to dephosphorylate PDGFRB. We suggest that in this family, the additional mutation in PTPRG may explain the full phenotypic penetrance in the siblings affected, in comparison with the unaffected mother.


Short-term outcomes of 59 dogs treated for ilial body fractures with locking or non-locking plates.
To determine the influence of plating systems on the clinical outcomes in dogs treated for ilial fractures. Retrospective study. Fifty-nine dogs (63 hemipelves). Radiographs and medical records of dogs with ilial fractures presented to Iowa State University between 2003 and 2019 were reviewed. After fracture reduction, fractures were fixed with a locking plate system (LPS) or non-locking plate system (NLS). Perioperative, long-term complications, and follow-up data were recorded. The frequency of implant failure and pelvic collapse were compared using a logistic and linear regression analysis, respectively. Where the univariate test was statistically significant, a multivariate analysis across categories was performed to identify statistically different categories. LPS and NLS implants were used in 25/63 and 38/63 hemipelves, respectively. Median follow-up time was 8 weeks (3-624 weeks). Implant failure occurred in 18/63 (29%) of fracture repairs, consisting of 17 with NLS and 1 with LPS. Revision surgery was recommended in five cases of implant failure, all with NLS. The probability of implant failure was higher when fractures were fixed with NLS (p = .0056). All other variables evaluated did not seem to influence outcome measures. The variable with the most influence on the outcomes of dogs treated for ilial fractures consisted of the fixation method (NLS vs. LPS). Fractures repaired with NLS were nearly 20 times more likely to fail than those repaired with LPS. Surgeons should consider repairing ilial body fractures in dogs with LPS to reduce the risk of short-term implant failure.


Limited, uneven progress in increasing racial and ethnic diversity of dental school graduates.
To assess racial and ethnic diversity of graduates of each dental school compared to the diversity of populations they draw from and to assess changes over time nationally and by school. We calculated diversity of graduates by school and nationally between 2010-2012 and 2017-2019 using the Integrated Post-secondary Education Data System (IPEDS) and compared the diversity of each state's college age population using data from the American Community Survey. We accounted for differences between in-state and out-of-state students attending public and private schools based on data from the American Dental Association's Survey of Dental Education Series. A diversity index (DI) was calculated for each school. A DI of 0.5 means that the representation of Black or Hispanic individuals among the graduates is half of their representation in the benchmark population. Among the 63 dental schools analyzed, only seven had a DI of greater than 0.5 for Black graduates (two of which were Historically Black Colleges and Universities) in 2017-2019. For Hispanic graduates, 20 schools had a DI above 0.5. Nationally, while the number of Black graduates increased between 2010-2012 and 2017-2019, the percentage decreased from 5.8% to 5.1%. The percentage of Hispanic graduates increased from 6.4% to 8.7%. Black and Hispanic individuals are underrepresented among dental school graduates. Increasing the diversity of the dental workforce could help address significant oral health disparities experienced by Black and Hispanic people. More needs to be done by the dental education community to increase racial and ethnic diversity of dental graduates.


[Inhibiting effect of S-adenosyl-L-homocysteine and its structural analogs on the process of enzymatic methylation of tRNA].
The action of S-adenosyl-l-homocysteine (S-Ado-Hcy), its four structural analogues S-Ino-Hcy, S-Guo-Hcy, S-Urd-Hcy, S-Cyd-Hcy and the five corresponding sulfoxides on tRNA methylases has been investigated. The data obtained in the study of overall incorporation of 14CH3-groups into an unfractioned tRNA preparation suggested that both the affinity of the inhibitors tested for various methylases and the type of inhibition were different. The experiments performed with unfractioned tRNA preparation permit to get an idea of the average inhibitory potency of each of the compounds. The study of their action on individual tRNA methylases by means of fractionation of minor components produced demonstrated that the affinity of the inhibitors tested for various methylases was really different. Thus, S-Ado-Hcy, S-Ino-Hcy and S-Urd-Hcy practically do not inhibit m1A methylase but have the highest affinity for m5C methylase. In an experiment with tRNAPhe which is a substrate for a single, namely m5C methylase, the type of inhibition of this methylase by S-Cyd-Hcy was revealed; it was found to be non-competitive with respect to S-Ado-Met, and the S-Cyd-Hcy concentration reducing the methylation by 50 percent was 1.2-10(-4) M.


Inhibitory effect of PPARγ on NLRP3 inflammasome activation.
Rationale: Stimulation of the NLRP3 inflammasome by metabolic byproducts is known to result in inflammatory responses and metabolic diseases. However, how the host controls aberrant NLRP3 inflammasome activation remains unclear. PPARγ, a known regulator of energy metabolism, plays an anti-inflammatory role through the inhibition of NF-κB activation and additionally attenuates NLRP3-dependent IL-1β and IL-18 production. Therefore, we hypothesized that PPARγ serves as an endogenous modulator that attenuates NLRP3 inflammasome activation in macrophages. Methods: Mouse peritoneal macrophages with exposure to a PPARγ agonist at different stages and the NLRP3 inflammasome-reconstituted system in HEK293T cells were used to investigate the additional anti-inflammatory effect of PPARγ on NLRP3 inflammasome regulation. Circulating mononuclear cells of obese patients with weight-loss surgery were used to identify the in vivo correlation between PPARγ and the NLRP3 inflammasome. Results: Exposure to the PPARγ agonist, rosiglitazone, during the second signal of NLRP3 inflammasome activation attenuated caspase-1 and IL-1β maturation. Moreover, PPARγ interfered with NLRP3 inflammasome formation by decreasing NLRP3-ASC and NLRP3-NLRP3 interactions as well as NLRP3-dependent ASC oligomerization, which is mediated through interaction between the PPARγ DNA-binding domain and the nucleotide-binding and leucine-rich repeat domains of NLRP3. Furthermore, PPARγ was required to limit metabolic damage-associated molecular pattern-induced NLRP3 inflammasome activation in mouse macrophages. Finally, the mature caspase-1/PPARγ ratio was reduced in circulating mononuclear cells of obese patients after weight-loss surgery, which we define as an "NLRP3 accelerating index". Conclusions: These results revealed an additional anti-inflammatory role for PPARγ in suppressing NLRP3 inflammasome activation through interaction with NLRP3. Thus, our study highlights that PPARγ agonism may be a therapeutic option for targeting NLRP3-related metabolic diseases.


Evaluation of Factors Associated with Anxiety and Depression in Chinese Visiting Scholars in the United States During the COVID-19 Pandemic Assessed by Online Questionnaires.
BACKGROUND This study aimed to use online questionnaires to evaluate the factors associated with anxiety and depression in Chinese visiting scholars in the United States during the COVID-19 pandemic. MATERIAL AND METHODS Using a cross-sectional design, 311 Chinese scholars visiting 41 states in the United States were interviewed on 20 and 21 April 2020 through WeChat using the Patient Health Questionnaire-9 (PHQ-9) and the Generalized Anxiety Disorder-7 (GAD-7) questionnaire. RESULTS Of these 311 visiting scholars, 69 (22.2%) reported no symptoms of anxiety or depression, whereas 63 (20.3%) reported severe anxiety and 67 (21.5%) reported severe depression. Risk of anxiety was 93% higher in visiting scholars with than without accompanying parents in the US (odds ratio [OR], 1.93; 95% confidence interval [CI], 1.01-3.68) and was 1.72-fold (95% CI, 1.04-2.84) higher in those experiencing stress about family members with COVID-19. Stresses about personal security and return to China on schedule were associated with 1.73-fold (95% CI, 1.03-2.92) and 3.00-fold (95% CI, 1.51-5.95) higher risks of anxiety, respectively. Risks of depression were 1.86-fold (95% CI, 1.14-3.05), 1.84-fold (95% CI, 1.10-3.07), and 3.45-fold (95% CI, 1.72-6.92) higher in visiting Chinese scholars who were than were not experiencing stresses about financial support, personal security and return to China on schedule, respectively. CONCLUSIONS Chinese scholars visiting the United States during the COVID-19 pandemic experienced severe psychological distress. Surveys that include larger numbers of visiting scholars are warranted.


Causal effects in clinical and epidemiological studies via potential outcomes: concepts and analytical approaches.
A central problem in public health studies is how to make inferences about the causal effects of treatments or agents. In this article we review an approach to making such inferences via potential outcomes. In this approach, the causal effect is defined as a comparison of results from two or more alternative treatments, with only one of the results actually observed. We discuss the application of this approach to a number of data collection designs and associated problems commonly encountered in clinical research and epidemiology. Topics considered include the fundamental role of the assignment mechanism, in particular the importance of randomization as an unconfounded method of assignment; randomization-based and model-based methods of statistical inference for causal effects; methods for handling noncompliance and missing data; and methods for limiting bias in the analysis of observational data, including propensity score matching and sensitivity analysis.


Human Schwann cells retain essential phenotype characteristics after immortalization.
Schwann cells (SCs) play an important role in the pathogenesis of peripheral nerve diseases and represent a potential target for development of therapies. However, use of primary human SCs (hSCs) for in vitro models is limited because these cells are difficult to prepare and maintain in high yield and purity under common cell culture conditions. To circumvent this obstacle, we immortalized primary human fetal SCs using the SV40 large T-antigen and human telomerase reverse transcriptase expression vectors. After cloning, selection, and purification, we evaluated several immortalized SC lines for their ability to express extracellular matrix (ECM) molecules and myelinate embryonic rat sensory axons. In addition, we established a gene expression profile and explored their sensitivity to oxidative stress in a simple in vitro assay. Immortalized hSC clones expressed common glial markers and a broad variety of growth factors, receptors, and ECM molecules as determined by immunocytochemistry, microarray, and quantitative reverse transcription-polymerase chain reaction. In neuron-SC co-cultures, these cells were able to myelinate rat dorsal root ganglia neurons, although their effectiveness was lower in comparison to primary rat SCs. In toxicity assays, immortalized hSCs remain susceptible to oxidative stress induced by H(2)O(2). This study shows that, using specific immortalization techniques, it is possible to establish hSC lines that retain characteristics of typical primary hSCs. These cells are particularly useful for drug screening and studies aimed at disease mechanisms involving SCs.


Effects of cyanobacteria Synechocystis spp. in the host-parasite model Crassostrea gasar-Perkinsus marinus.
Perkinsosis is a disease caused by protozoan parasites from the Perkinsus genus. In Brazil, two species, P. beihaiensis and P. marinus, are frequently found infecting native oysters (Crassostrea gasar and C. rhizophorae) from cultured and wild populations in several states of the Northeast region. The impacts of this disease in bivalves from Brazil, as well as the interactions with environmental factors, are poorly studied. In the present work, we evaluated the in vitro effects of the cyanobacteria Synechocystis spp. on trophozoites of P. marinus and haemocytes of C. gasar. Four cyanobacteria strains isolated from the Northeast Brazilian coast were used as whole cultures (WCs) and extracellular products (ECPs). Trophozoites of P. marinus were exposed for short (4h) and long (48h and 7days, the latter only for ECPs) periods, while haemocytes were exposed for a short period (4h). Cellular and immune parameters, i.e. cell viability, cell count, reactive oxygen species production (ROS) and phagocytosis of inert (latex beads) and biological particles (zymosan and trophozoites of P. marinus) were measured by flow cytometry. The viability of P. marinus trophozoites was improved in response to WCs of Synechocystis spp., which could be a beneficial effect of the cyanobacteria providing nutrients and reducing reactive oxygen species. Long-term exposure of trophozoites to ECPs of cyanobacteria did not modify in vitro cell proliferation nor viability. In contrast, C. gasar haemocytes showed a reduction in cell viability when exposed to WCs, but not to ECPs. However, ROS production was not altered. Haemocyte ability to engulf latex particles was reduced when exposed mainly to ECPs of cyanobacteria; while neither the WCs nor the ECPs modified phagocytosis of the biological particles, zymosan and P. marinus. Our results suggest a negative effect of cyanobacteria from the Synechocystis genus on host immune cells, in contrast to a more beneficial effect on the parasite cell, which could together disrupt the balance of the host-parasite interaction and make oysters more susceptible to P. marinus as well as opportunistic infections.


Dual Fatty Acid Elongase Complex Interactions in Arabidopsis.
Very long chain fatty acids (VLCFAs) are involved in plant development and particularly in several cellular processes such as membrane trafficking, cell division and cell differentiation. However, the precise role of VLCFAs in these different cellular processes is still poorly understood in plants. In order to identify new factors associated with the biosynthesis or function of VLCFAs, a yeast multicopy suppressor screen was carried out in a yeast mutant strain defective for fatty acid elongation. Loss of function of the elongase 3 hydroxyacyl-CoA dehydratase PHS1 in yeast and PASTICCINO2 in plants prevents growth and induces cytokinesis defects. PROTEIN TYROSIN PHOSPHATASE-LIKE (PTPLA) previously characterized as an inactive dehydratase was able to restore yeast phs1 growth and VLCFAs elongation but not the plant pas2-1 defects. PTPLA interacted with elongase subunits in the Endoplasmic Reticulum (ER) and its absence induced the accumulation of 3-hydroxyacyl-CoA as expected from a dehydratase involved in fatty acid (FA) elongation. However, loss of PTPLA function increased VLCFA levels, an effect that was dependent on the presence of PAS2 indicating that PTPLA activity repressed FA elongation. The two dehydratases have specific expression profiles in the root with PAS2, mostly restricted to the endodermis, while PTPLA was confined in the vascular tissue and pericycle cells. Comparative ectopic expression of PTPLA and PAS2 in their respective domains confirmed the existence of two independent elongase complexes based on PAS2 or PTPLA dehydratase that are functionally interacting.


Yersinia outer proteins (YOPS) E, K and N are antigenic but non-protective compared to V antigen, in a murine model of bubonic plague.
The pathogenic Yersiniae produce a range of virulence proteins, encoded by a 70 kb plasmid, which are essential for infection, and also form part of a contact-dependent virulence mechanism. One of these proteins, V antigen, has been shown to confer a high level of protection against parenteral infection with Y. pestis in murine models, and is considered to be a protective antigen. In this study, the protective efficacy of V antigen has been compared in the same model with that of other proteins (YopE, YopK and YopN), which are part of the contact-dependent virulence mechanism. Mice immunised with two intraperitoneal doses of V antigen or each of the Yops, administered with either Alhydrogel or interleukin-12, produced high antigen-specific serum IgG titres. As shown in previous studies, V+Alhydrogel was fully protective, and 5/5 mice survived a subcutaneous challenge with 90 or 9x10(3) LD50's of Y. pestis GB. In addition, these preliminary studies also showed that V+IL-12 was partially protective: 4/5 or 3/5 mice survived a challenge with 90 or 9x10(3) LD50's, respectively. In contrast, none of the mice immunised with the Yops survived the challenges, and there was no significant delay in the mean time to death compared to mice receiving a control protein. These results show that using two different vaccine regimens, Yops E, K and N, failed to elicit protective immune responses in a murine model of plague, whereas under the same conditions, V antigen was fully or partially protective.


PPP1CC2 can form a kinase/phosphatase complex with the testis-specific proteins TSSK1 and TSKS in the mouse testis.
The mouse protein phosphatase gene Ppp1cc is essential for male fertility, with mutants displaying a failure in spermatogenesis including a widespread loss of post-meiotic germ cells and abnormalities in the mitochondrial sheath. This phenotype is hypothesized to be responsible for the loss of the testis-specific isoform PPP1CC2. To identify PPP1CC2-interacting proteins with a function in spermatogenesis, we carried out GST pull-down assays in mouse testis lysates. Amongst the identified candidate interactors was the testis-specific protein kinase TSSK1, which is also essential for male fertility. Subsequent interaction experiments confirmed the capability of PPP1CC2 to form a complex with TSSK1 mediated by the direct interaction of each with the kinase substrate protein TSKS. Interaction between PPP1CC2 and TSKS is mediated through an RVxF docking motif on the TSKS surface. Phosphoproteomic analysis of the mouse testis identified a novel serine phosphorylation site within the TSKS RVxF motif that appears to negatively regulate binding to PPP1CC2. Immunohistochemical analysis of TSSK1 and TSKS in the Ppp1cc mutant testis showed reduced accumulation to distinct cytoplasmic foci and other abnormalities in their distribution consistent with the loss of germ cells and seminiferous tubule disorganization observed in the Ppp1cc mutant phenotype. A comparison of Ppp1cc and Tssk1/2 knockout phenotypes via electron microscopy revealed similar abnormalities in the morphology of the mitochondrial sheath. These data demonstrate a novel kinase/phosphatase complex in the testis that could play a critical role in the completion of spermatogenesis.


Spectrum of mutation and frequency of allelic deletion of the p53 gene in ovarian cancer.
The p53 gene encodes a nuclear phosphoprotein present in low levels in normal human cells. The wild-type form of this protein functions to restrain inappropriate cellular proliferation. Approximately one half of human epithelial ovarian cancers have mutations in the p53 gene and overexpress the mutant protein product. Deletion of one allele of the p53 gene also frequently occurs in these cancers. We sought to define the spectrum of mutations in the p53 gene in epithelial ovarian cancer with respect to both the specific codons involved and the type of mutations observed. We also examined the frequency of allelic deletion of the p53 gene in cancers containing p53 gene mutations. Tissue samples from the epithelial ovarian cancers of 62 patients were obtained during initial laparotomy. Histologic examination was done to ensure that the experimental samples used in this study contained more than 75% cancer cells. Total RNA was extracted from these samples and separately from matched control noncancerous regions of the surgical specimen or white blood cells. The purified RNAs were reverse transcribed to generate cDNA copies of exons 4-10 of the p53 gene. Two rounds of polymerase chain reaction (PCR) were conducted to produce enough template for DNA sequence analysis of the regions of interest within the p53 gene. Dideoxy sequencing of at least two independent productions of each amplified DNA template was done to confirm the validity of the mutations found. Allelic deletions were identified by PCR and gel electrophoretic techniques to examine three polymorphisms within the p53 gene in cancer-normal DNA pairs. We identified 45 mutations in exons 5-8 of the p53 gene, where mutations frequently have been found in other cancer types. An additional mutation was identified in exon 4. Overall, 72% of the mutations were transitions, 24% transversions, and 4% microdeletions. Allelic deletion of the other p53 allele was seen in 67% of ovarian cancers in which a p53 mutation was present. Germ-line p53 mutations were not found in any patients whose cancers had p53 mutations. Like p53 mutations in other types of human cancers, those in epithelial ovarian cancers are diverse and occur frequently in exons 5-8. The predominance of transition mutations suggests that p53 mutations in ovarian cancer arise because of spontaneous errors in DNA synthesis and repair rather than the direct interaction of carcinogens with DNA. These molecular data are consistent with data from epidemiologic studies that have failed to demonstrate a convincing relationship between exposure to environmental carcinogens and the development of ovarian cancer.


Oxidative modifications of mitochondrial complex II are associated with insulin resistance of visceral fat in obesity.
Obesity, particularly visceral adiposity, has been linked to mitochondrial dysfunction and increased oxidative stress, which have been suggested as mechanisms of insulin resistance. The mechanism(s) behind this remains incompletely understood. In this study, we hypothesized that mitochondrial complex II dysfunction plays a role in impaired insulin sensitivity in visceral adipose tissue of subjects with obesity. We obtained subcutaneous and visceral adipose tissue biopsies from 43 subjects with obesity (body mass index ≥ 30 kg/m2) during planned bariatric surgery. Compared with subcutaneous adipose tissue, visceral adipose tissue exhibited decreased complex II activity, which was restored with the reducing agent dithiothreitol (5 mM) ( P < 0.01). A biotin switch assay identified that cysteine oxidative posttranslational modifications (OPTM) in complex II subunit A (succinate dehydrogenase A) were increased in visceral vs. subcutaneous fat ( P < 0.05). Insulin treatment (100 nM) stimulated complex II activity in subcutaneous fat ( P < 0.05). In contrast, insulin treatment of visceral fat led to a decrease in complex II activity ( P < 0.01), which was restored with addition of the mitochondria-specific oxidant scavenger mito-TEMPO (10 µM). In a cohort of 10 subjects with severe obesity, surgical weight loss decreased OPTM and restored complex II activity, exclusively in the visceral depot. Mitochondrial complex II may be an unrecognized and novel mediator of insulin resistance associated with visceral adiposity. The activity of complex II is improved by weight loss, which may contribute to metabolic improvements associated with bariatric surgery.


Histochemical localization of hydrolytic enzymes during corneal graft reaction.
We studied hydrolytic enzymes in the graft bed during the corneal graft reaction. Two enzymes, beta-glucuronidase and acid phosphatase, were used as the markers of lysosomal hydrolytic enzymes. These were traced histochemically at various times before and after the onset of the graft reaction in a corneal xenograft model. Results showed a relationship between the degree of graft opacity, cellular infiltration and enzyme localization. It appeared that during the early stages of graft reaction, the granulocytes, and during the late stages, the agranulocytes were the main source of the enzymes.


Validating 4 Staging Systems for Cutaneous Squamous Cell Carcinoma Using Population-Based Data: A Nested Case-Control Study.
Cutaneous squamous cell carcinoma (cSCC) has metastatic potential, but the prognostic value of current staging systems in nonselected patients is uncertain. To assess the effect of risk factors for metastasis and to evaluate validity and usefulness of 4 staging systems for cSCC using population-based data. This was a nationwide, population-based, nested case-control study. The Cancer Registry of Norway, which receives compulsory data on all cancers in the Norwegian population of approximately 5.2 million inhabitants. All patients diagnosed as having a primary invasive cSCC in the period 2000 to 2004 (n = 6721) were identified. Of these, 112 patients were diagnosed with metastasis within 5 years. As control patients, 112 patients with cSCC without metastases, matched for sex and age at diagnosis, were identified by random. Clinical data and biopsy specimens of primary cSCC were collected for all 224 patients. The biopsies were reexamined histologically by an experienced pathologist using well-established criteria for cSCC, yielding 103 patients with metastasis (cases) and 81 cSCC without metastasis (controls). The ability of 4 cSCC staging systems (ie, the American Joint Committee on Cancer, 7th edition [AJCC 7] staging system, the staging system used by Breuninger et al, the Brigham and Women's Hospital [BWH] staging system, and the AJCC, 8th edition [AJCC 8] staging systems) to identify patients who developed metastasis with 5 years of follow-up. External validation was performed by logistic regression, discrimination (sensitivity, specificity, proportion correctly classified, concordance index), and calibration (R2, Hosmer-Lemeshow test, plots) statistics. Of 6721 patients; 3674 (54.7%) were men and 3047 (45.3%) were women, with a mean age at diagnosis of 78 years. Of the 103 patients with cSCC diagnosed with metastasis within 5 years, 60 [58.3%] were men, and mean [SD] age 72.7 [13.5] years. Of the 81 patients with cSCC without metastasis, 51 [63.0%] were men, and mean [SD] age 74.6 [11.7] 15 years. The staging systems distinguished poorly to moderately between patients who developed metastasis and those who did not. The ability to cluster patients with similar outcomes within the same staging category was low, particularly when using the AJCC 7 system. Using the AJCC 7 system, the risk of metastasis for T2 patients was 3-fold, compared with T1 patients (OR, 2.96; 95% CI, 1.43-6.15). In the system used by Breuninger et al, high-risk categories for diameter and tumor thickness and the BWH system's T2b category collected relatively homogeneous groups. In the systems used by Breuninger et al and Brigham-Women's Hospital, risk of metastasis was significantly elevated with increasing stage or risk category. Using the system by Breuninger et al, the risk of metastasis was less than 3-fold for tumors in the high-risk category of the combined variable (OR, 2.72; 95% CI, 1.29-5.74). The BWH system gave ORs for metastasis at 4.6 (95% CI, 2.23-9.49) and 21.31 (95% CI, 6.07-74.88) for the T2a and T2b categories, respectively. Using population-based data, 4 current staging systems for cSCC were unsatisfactory in identifying nonselected cSCC patients at high risk for metastasis. The system used by Breuninger gave the best results.


Recent advances on aqueous solubility prediction.
Aqueous solubility is one of the major physiochemical properties to be optimized in drug discovery. It is related to absorption and distribution in the ADME-Tox (Absorption, Distribution, Metabolism, Excretion, and Toxicity). Aqueous solubility and membrane permeability are the two key factors that affect a drug's oral bioavailability. Because of the importance of aqueous solubility, a lot of efforts have been spent on developing reliable models to predict this physiochemical property. Although some progress has been made and a lot of models have been constructed, it is concluded that accurate and reliable aqueous models targeted to predict solubility of drug-like molecules, have not emerged based on the outcome of an aqueous solubility prediction campaign sponsored by Goodman et al. In this review paper, we provide a snapshot of the latest development in the field. The challenges of developing high quality aqueous solubility models as well as the strategies of surmounting those challenges have been discussed. We conclude that the biggest challenge of modeling aqueous solubility is to collect more high quality, unskewed and drug-relevant solubility data which are sufficient diverse to cover most the chemical space of drugs. The second challenge is to develop good descriptors to account for the lattice energy of solvation. In order to develop accurate and predictable in silico solubility models, the key is to collect a sufficient number of high quality experimental data and the suspicious data must be verified. In addition, the molecular descriptors must be relevant to the energies in the solvation process (the lattice energy for crystal packing, the energy of forming cavity in solvent, and the solvation energy), and the models must be carefully cross-validated and evaluated using the external data sets.


Environmental factors influencing the risk of autism.
Autism is a developmental disability with age of onset in childhood (under 3 years old), which is characterized by definite impairments in social interactions, abnormalities in speech, and stereotyped pattern of behaviors. Due to the progress of autism in recent decades, a wide range of studies have been done to identify the etiological factors of autism. It has been found that genetic and environmental factors are both involved in autism pathogenesis. Hence, in this review article, a set of environmental factors involved in the occurrence of autism has been collected, and finally, some practical recommendations for reduction of the risk of this devastating disease in children are represented.


Ancient recruitment by chromists of green algal genes encoding enzymes for carotenoid biosynthesis.
Chromist algae (stramenopiles, cryptophytes, and haptophytes) are major contributors to marine primary productivity. These eukaryotes acquired their plastid via secondary endosymbiosis, whereby an early-diverging red alga was engulfed by a protist and the plastid was retained and its associated nuclear-encoded genes were transferred to the host genome. Current data suggest, however, that chromists are paraphyletic; therefore, it remains unclear whether their plastids trace back to a single secondary endosymbiosis or, alternatively, this organelle has resulted from multiple independent events in the different chromist lineages. Both scenarios, however, predict that plastid-targeted, nucleus-encoded chromist proteins should be most closely related to their red algal homologs. Here we analyzed the biosynthetic pathway of carotenoids that are essential components of all photosynthetic eukaryotes and find a mosaic evolutionary origin of these enzymes in chromists. Surprisingly, about one-third (5/16) of the proteins are most closely related to green algal homologs with three branching within or sister to the early-diverging Prasinophyceae. This phylogenetic association is corroborated by shared diagnostic indels and the syntenic arrangement of a specific gene pair involved in the photoprotective xanthophyll cycle. The combined data suggest that the prasinophyte genes may have been acquired before the ancient split of stramenopiles, haptophytes, cryptophytes, and putatively also dinoflagellates. The latter point is supported by the observed monophyly of alveolates and stramenopiles in most molecular trees. One possible explanation for our results is that the green genes are remnants of a cryptic endosymbiosis that occurred early in chromalveolate evolution; that is, prior to the postulated split of stramenopiles, alveolates, haptophytes, and cryptophytes. The subsequent red algal capture would have led to the loss or replacement of most green genes via intracellular gene transfer from the new endosymbiont. We argue that the prasinophyte genes were retained because they enhance photosynthetic performance in chromalveolates, thus extending the niches available to these organisms. The alternate explanation of green gene origin via serial endosymbiotic or horizontal gene transfers is also plausible, but the latter would require the independent origins of the same five genes in some or all the different chromalveolate lineages.


Development of a portable 3D ultrasound imaging system for musculoskeletal tissues.
3D ultrasound is a promising imaging modality for clinical diagnosis and treatment monitoring. Its cost is relatively low in comparison with CT and MRI, no intensive training and radiation protection is required for its operation, and its hardware is movable and can potentially be portable. In this study, we developed a portable freehand 3D ultrasound imaging system for the assessment of musculoskeletal body parts. A portable ultrasound scanner was used to obtain real-time B-mode ultrasound images of musculoskeletal tissues and an electromagnetic spatial sensor was fixed on the ultrasound probe to acquire the position and orientation of the images. The images were digitized with a video digitization device and displayed with its orientation and position synchronized in real-time with the data obtained by the spatial sensor. A program was developed for volume reconstruction, visualization, segmentation and measurement using Visual C++ and Visualization toolkits (VTK) software. A 2D Gaussian filter and a Median filter were implemented to improve the quality of the B-scan images collected by the portable ultrasound scanner. An improved distance-weighted grid-mapping algorithm was proposed for volume reconstruction. Temporal calibrations were conducted to correct the delay between the collections of images and spatial data. Spatial calibrations were performed using a cross-wire phantom. The system accuracy was validated by one cylinder and two cuboid phantoms made of silicone. The average errors for distance measurement in three orthogonal directions in comparison with micrometer measurement were 0.06+/-0.39, -0.27+/-0.27, and 0.33+/-0.39 mm, respectively. The average error for volume measurement was -0.18%+/-5.44% for the three phantoms. The system has been successfully used to obtain the volume images of a fetus phantom, the fingers and forearms of human subjects. For a typical volume with 126 x 103 x 109 voxels, the 3D image could be reconstructed from 258 B-scans (640 x 480 pixels) within one minute using a portable PC with Pentium IV 2.4 GHz CPU and 512 MB memories. It is believed that such a portable volume imaging system will have many applications in the assessment of musculoskeletal tissues because of its easy accessibility.


Mental health and psychosocial characteristics in adolescent obesity: a population-based case-control study.
In this population-based study we compared self-esteem, social background, social and academic competence, behavioural problems and lifestyle in 58 obese adolescents (BMI > or =99.6th percentile or > or =30 kg/m2), aged 14-18 y, with 58 sex- and age-matched controls of normal weight. The instruments used were: I Think I Am, Youth Self Report and a lifestyle questionnaire. The obese group was on average, 40 kg heavier than the controls. The obese individuals rated themselves significantly lower in physical characteristics, but in all other aspects of self-esteem, mental health and social and academic competence there were no differences between the two groups. There were significant socioeconomic differences, with more obese adolescents living with only one parent and with the mothers in the obese group having, in general, lower education than those in the control group. This study confirms previous observations that obesity is associated with special socioeconomic conditions in youth, but that obese adolescents do not differ from their normal-weight peers in other aspects of mental health.


Incidence of outbreak-associated COVID-19 cases by industry in Ontario, Canada, 1 April 2020-31 March 2021.
The objective of our study was to estimate the rate of workplace outbreak-associated cases of COVID-19 by industry in labour market participants aged 15-69 years who reported working the majority of hours outside the home in Ontario, Canada. We conducted a population-based cross-sectional study of COVID-19 workplace outbreaks and associated cases reported in Ontario between 1 April 2020 and 31 March 2021. All outbreaks were manually classified into two-digit North American Industry Classification System codes. We obtained monthly denominator estimates from the Statistics Canada Labour Force Survey to estimate the incidence of outbreak-associated cases per 100 000 000 hours among individuals who reported the majority of hours were worked outside the home. We performed this analysis across industries and in three distinct time periods. Overall, 12% of cases were attributed to workplace outbreaks among working-age adults across our study period. While incidence varied across the time periods, the five industries with the highest incidence rates across our study period were agriculture, healthcare and social assistance, food manufacturing, educational services, and transportation and warehousing. Certain industries have consistently increased the incidence of COVID-19 over the course of the pandemic. These results may assist in ongoing efforts to reduce transmission of COVID-19 by prioritising resources, as well as industry-specific guidance, vaccination and public health messaging.


Questionnaires to Assess Facilitators and Barriers of Early Mobilization in Critically Ill Patients; Which One to Choose? A Systematic Review.
Implementing and performing early mobilization is a complex process requiring multidisciplinary input and cooperation. To gain insight in its facilitators and barriers, various surveys have been developed. A systematic review was conducted, to identify the psychometric properties, feasibility and suitability of questionnaires to assess facilitators and barriers of early mobilization in critically ill patients. Data were extracted regarding a.o. definition of early mobilization, development, psychometric properties, content and themes, question format. The search identified 537 publications of which 13 unique questionnaires were included. The questionnaires showed wide variation in extensiveness of development. Only six questionnaires actually assessed validity and reliability. Which questionnaire to choose depends on the aim of its use, required level of detail and specifics of the ICU, though three questionnaires were recommended as their definition of early mobilization covered a broad range of activities, including nursing related mobility activities. International consensus on what constitutes early mobilization is desirable.


Effect of chitosan on the biological properties of sweet basil (Ocimum basilicum L.).
The effect of the treatment of chitosan at various concentrations (0.01%, 0.05%, 0.1%, 0.5%, and 1%) upon sweet basil (Ocimum basilicum L.) before seeding and transplanting was investigated in aspects of the amount of phenolic and terpenic compounds, antioxidant activity, and growth of the basil, as well as the phenylalanine ammonia lyase (PAL) activity. The total amount of the phenolic and terpenic compounds increased after the chitosan treatment. Especially, the amounts of rosmarinic acid (RA) and eugenol increased 2.5 times and 2 times, respectively, by 0.1% and 0.5% chitosan treatment. Due to the significant induction of phenolic compounds, especially RA, the corresponding antioxidant activity assayed by the DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging test increased at least 3.5-fold. Also, the activity of PAL, a key regulatory enzyme for the phenylpropanoid pathway, increased 32 times by 0.5% chitosan solution. Moreover, after the elicitor chitosan treatment, the growth in terms of the weight and height of the sweet basil significantly increased about 17% and 12%, respectively. Our study demonstrates that an elicitor such as chitosan can effectively induce phytochemicals in plants, which might be another alternative and effective means instead of genetic modification.


Development of a zoomorphic test specimen for constancy testing on digital X-ray systems in veterinary radiology.
Technical failures and incorrect usage of digital X-ray systems may lead to a decreasing image quality, artefacts and a higher dose exposure of staff and patients. Although there are no regulations regarding constancy testing in veterinary radiology all operators are required to avoid unnecessary exposure. The aim of this study was to develop a reasonably inexpensive zoomorphic 3D-printed test specimen for constancy testing that allows the detection of changing image quality by visual analysis. Primarily, a calibration curve of the attenuation factor of the 3D-printing material (ZP150) was determined. MATLAB converted every pixel value of a thorax X-ray image of a Beagle dog into an equivalent thickness of printing material. The thickness distribution was printed using a 3D-printer. This printed test specimen was additionally provided with five thin aluminium discs to simulate lung nodules. To evaluate the usability for constancy testing 12 X-ray images of the test specimen were made. Two images (reference and control) were taken with the minimum dose in order to obtain images suitable for diagnosis purposes. Eight images were taken with a dose differing 30-140% from the reference dose by varying current-time product (mAs) or tube voltage (kVp). Two images were taken with the same parameters as the reference image but edited with different image processing. Six veterinarians (general practitioners) evaluated ten chosen structures in the X-ray images in a Visual Grading Analysis and scored the image quality of these structures for every image in comparison to the reference image. A Visual Grading Analysis Score was calculated and statistically analysed. A higher current-time product led to a negligibly better evaluation of the X-ray image. The lower the current-time product the worse the X-ray images were scored. Likewise, both increasing and decreasing of the tube voltage led to lower scores. A zoomorphic test specimen can be used for constancy testing of digital X-ray systems in veterinary medicine. Especially a lower dose can be recognised due to deviation in the image quality when compared to the reference image. The 3D-printed test specimen is less expensive than test equipment used in human medicine.


Alcohol and hepatitis C virus--interactions in immune dysfunctions and liver damage.
Hepatitis C virus infection affects 170 million people worldwide, and the majority of individuals exposed to HCV develop chronic hepatitis leading to progressive liver damage, cirrhosis, and hepatocellular cancer. The natural history of HCV infection is influenced by genetic and environmental factors of which chronic alcohol use is an independent risk factor for cirrhosis in HCV-infected individuals. Both the hepatitis C virus and alcohol damage the liver and result in immune alterations contributing to both decreased viral clearance and liver injury. This review will capture the major components of the interactions between alcohol and HCV infection to provide better understanding for the molecular basis of the dangerous combination of alcohol use and HCV infection. Common targets of HCV and alcohol involve innate immune recognition and dendritic cells, the critical cell type in antigen presentation and antiviral immunity. In addition, both alcohol and HCV affect intracellular processes critical for hepatocyte and immune cell functions including mitochondrial and proteasomal activation. Finally, both chronic alcohol use and hepatitis C virus infection increase the risk of hepatocellular cancer. The common molecular mechanisms underlying the pathological interactions between alcohol and HCV include the modulation of cytokine production, lipopolysaccharide (LPS)-TLR4 signaling, and reactive oxygen species (ROS) production. LPS-induced chronic inflammation is not only a major cause of progressive liver injury and fibrosis, but it can also contribute to modification of the tissue environment and stem cells to promote hepatocellular cancer development. Alteration of these processes by alcohol and HCV produces an environment of impaired antiviral immune response, greater hepatocellular injury, and activation of cell proliferation and dedifferentiation.


Disparate effects of deficient expression of connexin43 on atrial and ventricular conduction: evidence for chamber-specific molecular determinants of conduction.
Myocardial conduction depends on intercellular transfer of current at gap junctions. Atrial myocytes express three different gap junction channel proteins-connexin43 (Cx43), connexin45 (Cx45), and connexin40 (Cx40)-- whereas ventricular myocytes express only Cx43 and Cx45. However, the physiological roles of individual connexins are unknown. We have previously shown that mice heterozygous for a null mutation in the gene encoding Cx43 (Cx43(+/-) mice) express 50% of the normal amount of Cx43 in ventricular myocardium and exhibit marked slowing of ventricular conduction. To determine whether atrial conduction is affected in Cx43(+/-) mice, we measured atrial conduction velocity in isolated hearts, performed detailed ECG and electrophysiological studies in intact animals, and determined the amount of cardiac connexins in atrial and ventricular tissue. Ventricular conduction velocity was reduced by 38% in Cx43(+/-) mice compared with wild-types, but atrial conduction velocity in the same hearts was normal. QRS duration was significantly greater in Cx43(+/-) mice than in wild-types, but P-wave duration and amplitude did not differ. Atrial expression of Cx43 was reduced by 50%. These results indicate that Cx43 is a principal conductor of intercellular current in the ventricle because ventricular conduction is significantly slowed when Cx43 content is reduced by only 50%. In contrast, a similar reduction in Cx43 content in atrial muscle has no effect on atrial conduction, suggesting that Cx40 (which is expressed in atrial but not ventricular myocytes) is a major electrical coupling protein in atrial muscle. Thus, Cx43 and Cx40 may be chamber-specific determinants of myocardial conduction.


Validating the association of Oxford classification and renal function deterioration among Taiwanese individuals with Immunoglobulin A nephropathy.
Validation of the Oxford classification (MEST and MEST-C) for Immunoglobulin A nephropathy (IgAN) in the Taiwanese population is lacking. Our study aimed to validate this classification and assess individual lesion impact. We conducted a retrospective cohort study at Taichung Veterans General Hospital, Taiwan (Jan 2011-Jul 2023). Composite renal outcomes were evaluated using clinical conditions and estimated glomerular filtration rate (eGFR). We used Kaplan-Meier, univariable/multivariable logistic regression and ROC curves. Subgroup analysis considered eGFR < or ≥ 30.0 ml/min/1.73 m2. In 366 renal biopsies, serum creatinine was 1.34 mg/dl, eGFR 53.8 ml/min/1.73 m2, urine protein-creatinine ratio 1159 mg/g. T1/T2 lesions had lowest baseline eGFR (39.6/11.5 ml/min/1.73 m2), correlating with poorest renal survival (median survival 54.7/34.4 months). Univariable analysis linked all individual variables to worse renal outcomes. Multivariable analysis (MEST/MEST-C) showed only T1/T2 linked to worse outcomes. T score had highest predictive power (AUC 0.728, sensitivity 60.2%, specificity 83.6%), with MEST having high AUC (0.758). No extra predictive power was seen transitioning MEST to MEST-C. Subgroup analysis (eGFR < 30.0 ml/min/1.73 m2) associated C1 with improved renal outcomes (odds ratio 0.14, 95% CI 0.03-0.65). T lesion correlated with worse outcomes across subgroups. The T lesion consistently correlated with worse renal outcomes across all groups and baseline statuses. Integrating the C lesion into the transition from MEST to MEST-C did not enhance predictive power. Importantly, the C1 lesion was linked to improved renal outcomes in the eGFR < 30.0 ml/min/1.73 m2 subgroup, likely due to treatment effects.


Risk profiles for metabolic syndrome and its transition patterns for the elderly in Beijing, 1992-2009.
There have been few reports on the development of metabolic disorders, especially when they are considered as a cluster. The purpose of this study was to describe risk profiles for metabolic syndrome (MetS) in elderly dwellers in Beijing, and to find their transition patterns over time. Data were derived from Beijing longitudinal study of aging, a community-based cohort study hosted by Xuanwu hospital. There were 3,257 elderly people aged 55 years or over recruited in 1992. MetS was assessed for the years 1992, 2000, and 2009. Finally, 363 subjects with complete information for components of MetS in the three years were included in the study. The criteria of MetS recommended by the joint interim statement criteria were adopted. Latent transition analysis was used to calculate the transition probabilities between adjacent visits. A risk typology consisting of four time-invariant groups was detected based on the components of MetS for all subjects. Low MetS risk group, BP risk group, BP-HDL risk group, and BP-FPG-TG risk group were found. The probability of staying at the same status was higher at the two intervals across 18 years. Four latent groups were extracted based on three assessments for the components of MetS, together with their transition patterns. Findings suggested various trajectories for MetS components. Different combinations of intervention strategy might be needed for MetS risk groups.


Cognitive specificity in internalizing and externalizing problems in community and clinic-referred children.
Examined whether cognitive features in Beck's (1967) model of depression and his cognitive content-specificity hypothesis (Beck, Brown, Steer, Eidelson, & Riskind, 1987) are broadband specific features that distinguish internalizing problems from externalizing problems. From a community (n = 389; 196 boys, 193 girls; age M = 9.98, SD = 1.52) and clinic (n = 82; 55 boys, 27 girls; age M = 10.42, SD = 2.91) sample, 4 groups were defined on behavioral measures and then examined on cognitive measures: externalizing only (n = 33), internalizing only (n = 41), comorbid externalizing and internalizing (n = 53), and control (neither externalizing nor internalizing; n = 35). Both the internalizing only and comorbid groups reported significantly more cognitive disturbances (negative cognitive triad, cognitive processing distortions, and "depressive" and "anxious" thought content) than both of the externalizing only and control groups in both the community and clinic samples. The results were not related to either overall level of psychopathology (as reported by mothers) or social desirability in children's reporting. These findings suggest that the cognitive features in Beck's model distinguish internalizing from externalizing problems and add to a growing literature that calls for further conceptual refinement of cognitive models for understanding both narrow-band and broadband syndromes.


Pinto bean consumption reduces biomarkers for heart disease risk.
To determine effects of daily intake of 1/2 cup pinto beans, black-eyed peas or carrots (placebo) on risk factors for coronary heart disease (CHD) and diabetes mellitus (DM) in free-living, mildly insulin resistant adults over an 8 week period. Randomized, crossover 3x3 block design. Sixteen participants (7 men, 9 women) received each treatment for eight-weeks with two-week washouts. Fasting blood samples collected at beginning and end of periods were analyzed for total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol, triacylglycerols, high-sensitivity C-reactive protein, insulin, glucose, and hemoglobin A1c. A significant treatment-by-time effect impacted serum TC (p = 0.026) and LDL (p = 0.033) after eight weeks. Paired t-tests indicated that pinto beans were responsible for this effect (p = 0.003; p = 0.008). Mean change of serum TC for pinto bean, black-eyed pea and placebo were -19 +/- 5, 2.5 +/- 6, and 1 +/- 5 mg/dL, respectively (p = 0.011). Mean change of serum LDL-C for pinto bean, black-eyed pea and placebo were -14 +/- 4, 4 +/- 5, and 1 +/- 4 mg/dL, in that order (p = 0.013). Pinto beans differed significantly from placebo (p = 0.021). No significant differences were seen with other blood concentrations across the 3 treatment periods. Pinto bean intake should be encouraged to lower serum TC and LDL-C, thereby reducing risk for CHD.


Apoptosis of bystander T cells induced by human immunodeficiency virus type 1 with increased envelope/receptor affinity and coreceptor binding site exposure.
Apoptosis of uninfected bystander CD4(+) T cells contributes to T-cell depletion during human immunodeficiency virus type 1 (HIV-1) pathogenesis. The viral and host mechanisms that lead to bystander apoptosis are not well understood. To investigate properties of the viral envelope glycoproteins (Env proteins) that influence the ability of HIV-1 to induce bystander apoptosis, we used molecularly cloned viruses that differ only in specific amino acids in Env. The ability of these strains to induce bystander apoptosis was tested in herpesvirus saimiri-immortalized primary CD4(+) T cells (CD4/HVS), which resemble activated primary T cells. Changes in Env that increase affinity for CD4 or CCR5 or increase coreceptor binding site exposure enhanced the capacity of HIV-1 to induce bystander apoptosis following viral infection or exposure to nonreplicating virions. Apoptosis induced by HIV-1 virions was inhibited by CD4, CXCR4, and CCR5 antibodies or by the CXCR4 inhibitor AMD3100, but not the fusion inhibitor T20. HIV-1 virions with mutant Envs that bind CXCR4 but are defective for CD4 binding or membrane fusion induced apoptosis, whereas CXCR4 binding-defective mutants did not. These results demonstrate that HIV-1 virions induce apoptosis through a CXCR4- or CCR5-dependent pathway that does not require Env/CD4 signaling or membrane fusion and suggest that HIV-1 variants with increased envelope/receptor affinity or coreceptor binding site exposure may promote T-cell depletion in vivo by accelerating bystander cell death.


Pathways in gut-brain communication: evidence for distinct gut-to-brain and brain-to-gut syndromes.
The rich interconnectedness between gut and brain is increasingly being identified. This article reviews the evidence for brain-gut and gut-brain syndromes, particularly recent epidemiological evidence, and animal studies demonstrating bi-directionality at the formative stage of development. Narrative literature review with selection for relevance and quality. Population surveys show a strong correlation between anxiety, depression, and functional gastrointestinal disorders, contradicting early suspicions that the high prevalence of anxiety and depression in the clinic was mainly due to neurotic health seeking behaviour. Five and 12 year follow-up shows that psychological distress can predict later onset of a functional gastrointestinal disorder and vice versa. Brain-gut pathways include the autonomic nervous system, hypothalamic-pituitary-adrenal axis including corticotrophin releasing factor directly acting on the gut. Gut-brain pathways include ascending pain pathways, cytokines including tumour necrosis factor alpha in response to bacterial translocation and inflammation, 5-hydroxytryptamine secretion by entero-endocrine cells and psychoactive chemicals of bacterial origin which may enter the blood stream, such as gamma-aminobutyric acid, fatty acids and 5-hydroxytryptamine precursors. The ability to control rodent temperament and HPA responsiveness with early modification of gastrointestinal flora, and the effects of early stress on the barrier function of the gastrointestinal tract and flora, suggests an ability of both systems to prime each other in early life for later problems. This hypothesis seems to be supported by a possible protective effect of a probiotic strain of bacteria in a model of early rat psychological trauma. Psychological treatments are known to improve functional gastrointestinal disorders, the next wave of research may involve preventative microbiological gut based treatments for primary psychological presentations, both to treat the presenting complaint and inoculate against later functional gastrointestinal disorders.


Real-time Tracking of Handheld Object from Color or Depth Images.
Object tracking during rehabilitation could help a therapist to evaluate a patient's movement and progress. Hence, we present an image-based method for real-time tracking of handheld objects due to its ease of use and availability of color or depth cameras. We use an efficient projective point correspondence method and generalize the use of precomputed spare viewpoint information to allow real-time tracking of a rigid object. The method runs at more than 30 fps on a CPU while achieving submillimeter accuracy on synthetic datasets and robust tracking on a semi-synthetic dataset.Clinical relevance Real-time, accurate, and robust tracking of an object using an image-based method is a promising tool for rehabilitation applications as it is practical for clinical settings.


The Development and Validation of a Subjective Assessment Tool for the Hip in the Athletic Population.
No validated functional assessments are available that are designed specifically to evaluate the performance and function of the athletic hip. Subsections of some validated outcome assessments address recreation, but a full assessment dedicated to athletic hip function does not exist. Current hip scoring systems may not be sensitive to subtle changes in performance and function in an athletic, younger population. The patient-athlete subjective scoring system developed in this study will be validated, reliable, and responsive in the evaluation of hip function in the athlete. Cross-sectional study; Level of evidence, 3. Based on the results of a pilot questionnaire administered to 18 athletic individuals, a final 10-item questionnaire was developed. Two hundred fifty competitive athletes from multiple sports completed the final questionnaire and 3 previously validated hip outcome assessments. Each athlete was self-assigned to 1 of 3 injury categories: (1) playing without hip/groin trouble; (2) playing, but with hip/groin trouble; and (3) not playing due to hip/groin trouble. The injury categories contained 196, 40, and 14 athletes, respectively. Correlations between the assessment scores and injury categories were measured. Responsiveness testing was performed in an additional group of 24 injured athletes, and their scores before and after intervention were compared. The Kerlan-Jobe Orthopaedic Clinic (KJOC) Athletic Hip Score showed high correlation with the modified Harris Hip Score, the Nonarthritic Hip Score, and the International Hip Outcome Tool. The new score stratified athletes by injury category, demonstrated responsiveness and accuracy, and varied appropriately with improvements in injury category after treatment of injuries. The new KJOC Athletic Hip Score is valid, reliable, and responsive for evaluation of the hip in an athletic population. The results support its use for the functional assessment of the hip in future studies.


Evaluating Surveillance Methods for Arboviral Vectors of La Crosse Virus and West Nile Virus of Southern Appalachia.
To monitor mosquito-borne diseases, public health departments conduct mosquito and pathogen surveillance. Our objective was to evaluate mosquito monitoring methods for collecting La Crosse virus (LACV) and West Nile virus (WNV) vectors (Aedes and Culex mosquitoes, respectively) in southern Appalachia. Centers for Disease Control and Prevention (CDC) light traps baited with carbon dioxide (CO(2)), CDC light traps baited with CO(2) and BG lure, BG-Sentinel traps baited with CO(2), gravid traps baited with oak (Quercus)-water infusion, and resting traps were compared in eastern Tennessee in 2013. Traps operated at 8 different urban sites throughout Knox County were randomly assigned to and rotated among 6 plots within each site. Results were specific for each vector; the BG-Sentinel trap was the best method for Aedes triseriatus, the CDC trap baited with CO(2) and BG lure was the best method for Ae. albopictus, and the gravid trap was the best method for Ae. japonicus. Culex erraticus collections varied by week and trapping method, indicating no single method was best, but the questing traps collected more mosquitoes. There was no significant trapping difference for Cx. pipiens complex in this region using the methods tested. The results suggest using a combination of trapping methods when sampling for LACV and/or WNV mosquito vectors in southern Appalachia. Effective trapping methods are necessary to enable accurate surveillance, improve control methods, enhance understanding of dispersal, and use for early detection of vectors and pathogens.


Two quasi-stable lead(ii) hydrides at ambient temperature.
Simple reactions of their terphenyl lead bromide precursors with DIBAL-H in diethyl ether solution at ca. -78 °C leads to the isolation of the hydrides {Pb(μ-H)ArPri4}2 (ArPri4 = C6H3-2,6-(C6H3-2,6-Pri2)2) (1) and {Pb(μ-H)ArMe6}2 (ArMe6 = C6H3-2,6-(C6H2-2,4,6-Me3)2) (2) in good yield (60-80%). The isolated solids are stable at up to 5 °C for several weeks but are thermally labile in solution. Hydride 1 decomposes to the diplumbyne ArPri4PbPbArPri4, while 2 decomposes to the plumbylene Pb(ArMe6)2. The decomposition of 1 was determined to be zero order with a rate constant of ca. 2.0 × 10-5 M min-1 at 298 K.


VALIDITY OF FIELD TESTS TO ESTIMATE CARDIORESPIRATORY FITNESS IN CHILDREN AND ADOLESCENTS: A SYSTEMATIC REVIEW.
To systematically review the literature to verify the validity of field-tests to evaluate cardiorespiratory fitness (CRF) in children and adolescents. The electronic search was conducted in the databases: Medline (PubMed), SPORTDiscus, Scopus, Web of Science, in addition to the Latin American databases LILACS and SciELO. The search comprised the period from the inception of each database until February 2015, in English and Portuguese. All stages of the process were performed in accordance with the PRISMA flow diagram. After confirming the inclusion criteria, eligibility, and quality of the studies, 43 studies were analyzed in full; 38 obtained through the searches in the electronic databases, and 5 through private libraries, and references from other articles. Of the total studies, only 13 were considered high quality according to the adopted criteria. The most commonly investigated test in the literature was the 20-meter shuttle run (SR-20 m), accounting for 23 studies, followed by tests of distances between 550 meters and 1 mile, in 9 studies, timed tests of 6, 9, and 12 minutes, also 9 studies, and finally bench protocols and new test proposals represented in 7 studies. The SR-20-m test seems to be the most appropriate to evaluate the CRF of young people with the equation of Barnett, recommended to estimate VO2 peak. As an alternative for evaluating CRF, the 1-mile test is indicated with the equation proposed by Cureton for estimating VO2 peak.


Establishment of a novel cell line (TS-2) of pre-B acute lymphoblastic leukemia with a t(1;19) not involving the E2A gene.
The t(1;19)(q23;p13) translocation involving the E2A gene on chromosome 19p13.3 is a nonrandom translocation that is often seen in childhood pre-B-cell acute lymphoblastic leukemia (ALL). However, recent studies have demonstrated the presence of immunophenotypic and molecular heterogeneity among patients with the cytogenetically identical chromosome translocation. Here we report a novel pre-B ALL cell line, TS-2, with t(1;19) translocation not involving the E2A gene. The breakpoint of t(1;19) in TS-2 was demonstrated to be at 19p13.3, a region indistinguishable from the locus of the E2A gene, by cytogenetic study and fluorescence in situ hybridization. However, rearrangement of the E2A gene was not detected in TS-2 by Southern blot analysis. Moreover, the expressions of PBX1 or E2A/PBX1 fusion genes were not detected by an extensive study with Northern blot analysis and reverse transcription-polymerase chain reaction. These findings suggest that TS-2 may have a genetic abnormality involving uncharacterized gene(s) at 19p13.3 distinct from the E2A gene and, therefore, may be useful for investigating the heterogeneity of molecular pathogenesis in leukemias with t(1;19)(q23;p13) translocation.


Anti-vibrio and immune-enhancing activity of medicinal plants in shrimp: A comprehensive review.
Disease epidemics in shrimp aquaculture increase apace with the development of aquaculture systems throughout the world. The disease caused by Vibrio spp. (vibriosis) is considered the most devastating, which has made it the most feared bacterial disease in the shrimp sector. In aquaculture, several strategies have already been applied to control Vibrio strains, including chemicals, probiotics, antibiotics, natural products from plants, including plant oils; hence, there has been considerable attention for using plants in shrimp aquaculture to provide sustainable, eco-friendly and safe compounds, such as alkaloids, saponins, terpenoids and flavonoids for replacing chemical compounds and antibiotics in current aquaculture. Medicinal plants may also have immunostimulating activity, increase growth and resistance in shrimps. The present paper aims to review the inhibition of Vibrio spp. in shrimp by medicinal plants, using both in vitro or/and in vivo techniques. Several medicinal plants appear capable of inhibiting growth of Vibrio pathogens outside living shrimp or in the body of shrimp, through enhancing growth and immune capacity when shrimps are fed or injected with them. In the current review Gracilaria spp. (Gracilariaceae family) and Sargassum spp. (family Sargassaceae) have been used most for in vitro and in vivo experiments. Among the terrestrial plants, Eucalyptus camaldulensis, Psidium guajava, Rhodomyrtus tomentosa, and Syzygium cumini (Myrtaceae family) had significant activity against Vibrio.


UK and Ireland Joint Advisory Group (JAG) consensus statements for training and certification in diagnostic endoscopic ultrasound (EUS).
International endoscopy societies vary in their approach for credentialing individuals in endoscopic ultrasound (EUS) to enable independent practice; however, there is no consensus in this or its implementation. In 2019, the Joint Advisory Group on GI Endoscopy (JAG) commissioned a working group to examine the evidence relating to this process for EUS. The aim of this was to develop evidence-based recommendations for EUS training and certification in the UK. Under the oversight of the JAG quality assurance team, a modified Delphi process was conducted which included major stakeholders from the UK and Ireland. A formal literature review was made, initial questions for study were proposed and recommendations for training and certification in EUS were formulated after a rigorous assessment using the Grading of Recommendation Assessment, Development and Evaluation tool and subjected to electronic voting to identify accepted statements. These were peer reviewed by JAG and relevant stakeholder societies before consensus on the final EUS certification pathway was achieved. 39 initial questions were proposed of which 33 were deemed worthy of assessment and finally formed the key recommendations. The statements covered four key domains, such as: definition of competence (13 statements), acquisition of competence (10), assessment of competence (5) and postcertification mentorship (5). Key recommendations include: (1) minimum of 250 hands-on cases before an assessment for competency can be made, (2) attendance at the JAG basic EUS course, (3) completing a minimum of one formative direct observation of procedural skills (DOPS) every 10 cases to allow the learning curve in EUS training to be adequately studied, (4) competent performance in summative DOPS assessments and (5) a period of mentorship over a 12-month period is recommended as minimum to support and mentor new service providers. An evidence-based certification pathway has been commissioned by JAG to support and quality assure EUS training. This will form the basis to improve quality of training and safety standards in EUS in the UK and Ireland.


Ayahuasca may help to improve self-compassion and self-criticism capacities.
Ayahuasca is a psychedelic brew that originated in the Amazon basin. The psychological effects of this drug are becoming better understood due to the growing research interest in identifying new potential therapeutic agents for the treatment of emotion dysregulation and other disorders. Previous studies suggest that ayahuasca enhances mindfulness-related capacities (decentering, non-judging, non-reacting and acceptance) and emotion regulation. The aim of the present exploratory study was to determine the effects of ayahuasca on self-compassion in a community sample. We administered validated questionnaires (the Self-Compassion Scale-Short Form and Forms of Self-Criticism and Self-Reassurance) to evaluate pre-post changes in self-compassion and self-criticism/self-reassurance in 45 volunteers (27 women; 60%) before and after (≤24 h) an ayahuasca ceremony. Most participants (n = 29; 67.4%) had previously used ayahuasca. Ayahuasca resulted in significant improvements, with medium to large effect sizes (η2 = 0.184-0.276), in measures of self-compassion (p < 0.05), self-criticism (p < 0.01) and self-reassurance (p < 0.01). The findings of this study suggest that ayahuasca promotes well-being and self-compassion, which could have a therapeutic effect on individuals with negative affect and other psychopathological conditions. Large, controlled studies are needed to confirm these findings.


Oscillation suppression and chimera states in time-varying networks.
Complex network theory has offered a powerful platform for the study of several natural dynamic scenarios, based on the synergy between the interaction topology and the dynamics of its constituents. With research in network theory being developed so fast, it has become extremely necessary to move from simple network topologies to more sophisticated and realistic descriptions of the connectivity patterns. In this context, there is a significant amount of recent works that have emerged with enormous evidence establishing the time-varying nature of the connections among the constituents in a large number of physical, biological, and social systems. The recent review article by Ghosh et al. [Phys. Rep. 949, 1-63 (2022)] demonstrates the significance of the analysis of collective dynamics arising in temporal networks. Specifically, the authors put forward a detailed excerpt of results on the origin and stability of synchronization in time-varying networked systems. However, among the complex collective dynamical behaviors, the study of the phenomenon of oscillation suppression and that of other diverse aspects of synchronization are also considered to be central to our perception of the dynamical processes over networks. Through this review, we discuss the principal findings from the research studies dedicated to the exploration of the two collective states, namely, oscillation suppression and chimera on top of time-varying networks of both static and mobile nodes. We delineate how temporality in interactions can suppress oscillation and induce chimeric patterns in networked dynamical systems, from effective analytical approaches to computational aspects, which is described while addressing these two phenomena. We further sketch promising directions for future research on these emerging collective behaviors in time-varying networks.


Cancer Tissue Engineering: A Novel 3D Polystyrene Scaffold for In Vitro Isolation and Amplification of Lymphoma Cancer Cells from Heterogeneous Cell Mixtures.
Isolation and amplification of primary lymphoma cells in vitro setting is technically and biologically challenging task. To optimize culture environment and mimic in vivo conditions, lymphoma cell lines were used as a test case and were grown in 3-dimension (3D) using a novel 3D tissue culture polystyrene scaffold with neonatal stromal cells to represent a lymphoma microenvironment. In this model, the cell proliferation was enhanced more than 200-fold or 20,000% neoplastic surplus in 7 days when less than 1% lymphoma cells were cocultured with 100-fold excess of neonatal stroma cells, representing 3.2-fold higher proliferative rate than 2D coculture model. The lymphoma cells grew and aggregated to form clusters during 3D coculture and did not maintained the parental phenotype to grow in single-cell suspension. The cluster size was over 5-fold bigger in the 3D coculture by day 4 than 2D coculture system and contained less than 0.00001% of neonatal fibroblast trace. This preliminary data indicate that novel 3D scaffold geometry and coculturing environment can be customized to amplify primary cancer cells from blood or tissues related to hematological cancer and subsequently used for personalized drug screening procedures.


Inhibition of 7,12-dimethylbenz(a)anthracene-induced mammary carcinogenesis by retinyl acetate.
The administration of 2.5 mg retinyl acetate daily in the diet to female Sprague-Dawley rats beginning 7 days after the intragastric instillation of either 2.5, 5, or 15 mg 7,12-dimethylbenz(a)anthracene (CMBA) resulted in a reduction in the incidence of benign mammary tumors of 37, 30, and 31%, respectively. An equally significant reduction in the number of tumors was also evident. Although no difference was noted in the percentage incidence of mammary adenocarcinomas between the placebo and 2.5 mg retinyl acetate-treated groups at the 2.5-mg DMBA level, the percentage incidence was reduced by 52 and 39% in these groups at the 5- and 15-mg DMBA dose. Furthermore, the number of adenocarcinomas was also significantly reduced. Although both the percentage incidence and number of tumors were reduced by treatment with 1 mg retinyl acetate, these differences were not statistically significant. Liver histology and liver function tests of rats of the retinyl acetate groups did not differ from that of the control group. Similarly, the estrus cycle of treated animals did not differ from that of control rats. These data indicate that relatively large doses of retinyl acetate significantly inhibit the development of DMBA-induced mammary adenocarcinomas and benign tumors. Furthermore, the suppression of mammary tumorigenesis is apparently not the result of an alteration in either the metabolism of DMBA or estrogen nor to an inhibition of tumor growth resulting from retinyl acetate toxicity. The inhibitory effect of retinyl acetate may be related to the effect of retinoids on epithelial cell differentiation and/or reversal of carcinogen-induced anaplasia.


Comparison of toxicity associated with early morning versus late afternoon radiotherapy in patients with head-and-neck cancer: a prospective randomized trial of the National Cancer Institute of Canada Clinical Trials Group (HN3).
Based on our demonstration of a circadian rhythm in the human oral mucosa cell cycle, with most cells in the G(1) phase in the morning and M phase at night, we hypothesized that morning radiotherapy (RT) would lead to less oral mucositis than afternoon RT. A total of 216 patients were randomized to morning (8-10 AM) vs. afternoon (4-6 PM) RT and stratified by radiation dose, smoking status, and center. Patients receiving primary or postoperative RT alone were eligible. Oral mucositis was scored using the Radiation Therapy Oncology Group (RTOG) criteria and a validated scoring system. Of 205 evaluable patients, 52.9% vs. 62.4% developed RTOG Grade 3 or greater mucositis after morning vs. afternoon RT, respectively (p = 0.17). Morning RT was also associated with significantly less weight loss after 5 months (p = 0.024). In a subgroup of 111 patients treated to a dose of 66-70 Gy in 33-35 fractions, exploratory analyses revealed a significant reduction in Grade 3 or greater mucositis with morning RT (44.6% vs. 67.3%, p = 0.022) and a longer interval to the development of Grade 3 or greater mucositis (median, >7.9 vs. 5.6 weeks, p = 0.033). In 53 patients, who smoked during therapy, a significant reduction was found in Grade 3 or greater mucositis with morning RT (42.9% vs. 76%, p = 0.025). In this proof of principle study, morning RT was associated with significantly less weight loss after 5 months and an apparent reduction in oral mucositis in a subset of patients receiving >/=66 Gy and in patients who smoked during therapy.


Thyrotropin-releasing hormone-stimulated [3H]inositol metabolism in GH3 pituitary tumor cells. Studies with lithium.
GH3 pituitary tumor cells were labeled to isotopic equilibrium with [3H]inositol. Thyrotropin-releasing hormone (TRH), which has been shown to stimulate inositol phospholipid metabolism in these cells, enhanced the accumulation of [3H]inositol-derived radioactivity in the cell's acid-soluble fraction. Separation of the [3H]inositol metabolites by ion-exchange chromatography revealed that TRH induced a rapid rise in the cellular content of [3H]inositol mono-, bis-, and trisphosphate. The latter two metabolites accumulated in a multiphasic manner with an initial peak 5-10 sec after TRH addition. This was followed by a short-lived decline and a secondary rise which left the metabolite levels elevated for at least 50 min. The GH3 cell [3H]inositol monophosphate and [3H] inositol content also rose in response to TRH, but the latter accumulated with a considerably slower time course than the phosphorylated derivatives. None of these responses could be mimicked by the calcium ionophore A23187. Incubation of GH3 cells with TRH in the presence of lithium led to an enhanced accumulation of [3H]inositol monophosphate and, to a lesser extent, of [3H]inositol bis- and trisphosphate. This accumulation rose in a linear fashion with time for at least 20 min, by which point 50% of the [3H]inositol-containing phospholipids had been depleted. When lithium was added 30 min after TRH, [3H]inositol monophosphate accumulated at the same rate as was found when TRH and lithium were added together, indicating that the TRH-induced phospholipid response in GH3 cells does not desensitize. Under normal conditions, approximately equal amounts of the three [3H]inositol phosphates were formed within 5 sec of TRH addition. However, when TRH was added to cells grown chronically in lithium-containing medium, or to cells incubating at a subphysiological temperature (25 degrees), greater than 90% of the metabolites formed were the bis- or trisphosphates. This indicates that the primary event stimulated by TRH is the breakdown by phospholipase C of phosphatidylinositol 4,5-bisphosphate and, perhaps also, of phosphatidylinositol 4-phosphate.


Removing a Bent Femoral Intramedullary Nail Cost Effectively: A Case Report.
A bent intramedullary (IM) nail becomes challenging and technically demanding to the orthopaedic surgeon for nail extraction. A broken nail can be easily removed through the fracture site. However, a bent nail has to be broken before it can be removed. Several studies and case reports outline the strategies and techniques for removing a bent IM nail. However, there is a paucity of guidelines and standard protocol describing the best and inexpensive strategy. We report a case where two years following surgery for intramedullary nailing of the right femur, the IM mail was bent following secondary trauma. We used a technique based on the principles of an ability to fully cut the nail and extract it in two pieces by using a Jumbo cutter which is available in the orthopaedic armamentarium. This technique is simple yet economical, with the likelihood of causing less soft tissue damage and thermal necrosis.


Sequence specificity of the post-translational proteolytic cleavage of vicilin, a seed storage protein of pea (Pisum sativum L.).
Amino acid sequence data from vicilin of pea (Pisum sativum L.) were compared with predicted sequences from complementary DNA species. The sites of potential post-translational proteolytic cleavage of vicilin precursor polypeptides were located in polar regions of the polypeptide, at acidic or amide residues. Proteolysis did not take place in precursors containing a functionally distinct sequence: neutral residue-hydrophobic residue-basic residue at the cleavage site. Differences between the genomic sequences encoding vicilin thus specify proteolytic cleavage of vicilin precursor polypeptides.


Conditional Knockout of GLT-1 in Neurons Leads to Alterations in Aspartate Homeostasis and Synaptic Mitochondrial Metabolism in Striatum and Hippocampus.
Expression of the glutamate transporter GLT-1 in neurons has been shown to be important for synaptic mitochondrial function in the cerebral cortex. Here we determined whether neuronal GLT-1 plays a similar role in the hippocampus and striatum, using conditional GLT-1 knockout mice in which GLT-1 was inactivated in neurons by expression of synapsin-Cre (synGLT-1 KO). Ex vivo 13C-labelling using [1,2-13C]acetate, representing astrocytic metabolism, yielded increased [4,5-13C]glutamate levels, suggesting increased astrocyte-neuron glutamine transfer, in the striatum but not in the hippocampus of the synGLT-1 KO. Moreover, aspartate concentrations were reduced - 38% compared to controls in the hippocampus and the striatum of the synGLT-1 KO. Mitochondria isolated from the hippocampus of synGLT-1 KO mice exhibited a lower oxygen consumption rate in the presence of oligomycin A, indicative of a decreased proton leak across the mitochondrial membrane, whereas the ATP production rate was unchanged. Electron microscopy revealed reduced mitochondrial inter-cristae distance within excitatory synaptic terminals in the hippocampus and striatum of the synGLT-1 KO. Finally, dilution of 13C-labelling originating from [U-13C]glucose, caused by metabolism of unlabelled glutamate, was reduced in hippocampal synGLT-1 KO synaptosomes, suggesting that neuronal GLT-1 provides glutamate for synaptic tricarboxylic acid cycle metabolism. Collectively, these data demonstrate an important role of neuronal expression of GLT-1 in synaptic mitochondrial metabolism in the forebrain.


Faeces, FACS, and functional assays--preparation of Isospora suis oocyst antigen and representative controls for immunoassays.
Highly purified antigen and appropriate controls are essential for antigen-specific immunoassays. In the case of Isospora suis, the causative agent of neonatal porcine coccidiosis, the only current source of antigen is oocysts isolated from faeces. The aim of this study was to develop a procedure for high-grade purification of I. suis oocysts from piglet faeces to obtain both antigen and representative controls suitable for in vitro re-stimulation of lymphocytes. This was achieved by use of filtration, density-gradient centrifugation and fluorescence-activated cell sorting (FACS). The feasibility for immunological studies was demonstrated with IFN-gamma ELISPOT assays after in vitro re-stimulation of lymphocytes from previously infected swine using the obtained antigen. The developed method allowed the production of highly purified antigen and representative controls from faeces with an oocyst recovery rate of 14%. Regarding the application of the obtained material it could be shown that lymphocytes from I. suis-infected pigs react in an antigen-specific manner in terms of an in vitro recall response by the production of IFN-gamma. This demonstrates the suitability of the developed method for the production of antigen and controls for sensitive immunological readout systems. Moreover, the detected specific IFN-gamma response encourages further functional studies on the cellular immune response to I. suis.


A core outcome domain set to assess cutaneous neurofibromas related to neurofibromatosis type 1 in clinical trials.
Cutaneous neurofibromas (cNF) are considered one of the highest burdens of neurofibromatosis type 1 (NF1). To date, no medical treatment can cure cNF or prevent their development. In that context, there is an urgent need to prepare and standardize the methodology of future trials targeting cNF. The objective was to develop a core outcome domain set suitable for all clinical trials targeting NF1-associated cNF. The validated approach of this work consisted of a three-phase methodology: i) generating the domains (systematic review of literature (SRL) and qualitative studies), ii) agreeing (three-round international e-Delphi consensus process, working groups), and iii) voting. i) The SLR and the qualitative studies (three types of focus groups and a French e-survey with 234 participants) resulted in a preliminary list of 31 candidate items and their corresponding definitions. ii) A total of 229 individuals from 29 countries participated in the first round of the e-Delphi: 71 patients, relatives or representatives (31.0%), 130 health-care professionals (HCP, 56.8%) and 28 researchers, representatives of a drug regulatory authority, industry or pharmaceutical company representatives or journal editors (12.2%). The overall participation rate was 74%. Between rounds 2 and 3, international workshops were held to better understand the disagreement amongst stakeholders. This phase led to the identification of 19 items as outcome sub-domains. iii) The items were fused to create 4 outcome domains ("clinical assessment", "daily life impact", "patient satisfaction" and "perception of health") and prioritized. The 7 items which did not reach consensus were decided to be marked for the research agenda. The final core outcome domain set reached 100% of the votes of the steering committee members. Although numerous outcomes can be explored in studies related to cNF in NF1, the present study offers 4 outcome domains that should be reported in all trial studies, agreed upon by international patients, relatives and representatives of patients, HCP, researchers, representatives of drug regulatory authorities or pharmaceutical companies, and journal editors. The next step will include the development of a set of core outcome measurement instruments to further standardize how these outcomes should be assessed.


Psychological comorbidity in children with chronic conditions.
The incidence and prevalence of chronic conditions in children are increasing along with a concomitant rise in the number of psychological problems they experience. When psychological comorbidity exists, the child is at greater risk for numerous problems. This article will define psychological comorbidity and discuss its incidence, presentation, associated factors, and interventions.


Genetics of Endometriosis.
Endometriosis is a complex, puzzling disease in women of reproductive age. The exact cause of the disease is unknown; however, different genetic and environmental factors contribute to the evolution of the disorder. Further investigation has led to the discovery of inheritance as a complex genetic trait. Candidate genes were identified to isolate regions of genes that affect disease risk. Additional linkage studies have been performed to map specific genes along the entire genome. Recent advances to determine the genetic component of endometriosis include genome-wide association studies. This chapter focuses on different approaches to identify the genetic links of endometriosis and its pathogenesis.


Age- and race-dependence of the fibroglandular breast density analyzed on 3D MRI.
The purpose of this study was to evaluate the age- and race-dependence of the breast fibroglandular tissue density based on three-dimensional breast MRI. The normal breasts of 321 consecutive patients including Caucasians, Asians, and Hispanics were studied. The subjects were separated into three age groups: Younger than 45, between 45 and 55, and older than 55. Computer algorithms based on body landmarks were used to segment the breast, and fuzzy c-means algorithm was used to segment the fibroglandular tissue. Linear regression analysis was applied to compare mean differences among different age groups and race/ethnicity groups. The obtained parameters were not normally distributed, and the transformed data, natural log (ln) for the fibroglandular tissue volume, and the square root for the percent density were used for statistical analysis. On the average, the transformed fibroglandular tissue volume and percent density decreased significantly with age. Racial differences in mean transformed percent density were found among women older than 45, but not among women younger than 45. Mean percent density was higher in Asians compared to Caucasians and Hispanics; the difference remained significant after adjustment for age, but not significant after adjusted for both age and breast volume. There was no significant difference in the density between the Caucasians and the Hispanics. The results analyzed using the MRI-based method show age- and race-dependence, which is consistent with literature using mammography-based methods.