Return to Physical Activity Following Flatfoot Reconstruction.
Progressive collapsing foot deformity (PCFD) is a debilitating condition encompassing several interrelated, progressive deformities requiring a combination of reconstructive procedures. Few studies investigate returns to activity following flatfoot reconstruction, and existing studies only examine 1 or 2 of the numerous procedures employed. This study aims to provide the first generalizable assessment of returns to sports and physical activity following reconstruction surgery in patients with flexible flatfoot deformity. Patients aged 18-60 years who underwent reconstructive surgery between February 16 and May 19 for symptomatic flexible-stage flatfoot deformity were identified by registry review. Eighty-two of 113 eligible patients (73%) were reached at a mean 2.9 years (range, 2.0-5.4) of follow-up with mean age at surgery of 48.9 years (range, 18-59). Returns to physical activity were evaluated with a sports-specific survey. Clinical outcomes were evaluated with Patient-Reported Outcomes Measurement Information System (PROMIS) scores. Patients reported participation in 21 specific sports and activities. One-fourth (25.6%) of patients (21/82) reported increased difficulty with physical activities postoperatively, 15.9% reported equal difficulty, and 58.5% (48/82) reported decreased difficulty. Median return times were 9-12 months for participation and 12-18 months to reach maximum preoperative participation levels. Improvements in Physical Function (P= .001), Pain Interference (P < .001), Pain Intensity (P <.001), and Global Physical Health (P = .004) were associated with increased satisfaction with respect to sports and physical activities. This study investigated participation in specific sports and physical activities following flatfoot reconstruction. Our findings suggest mixed outcomes, where many patients reported life-changing improvements but many also experienced prolonged pain and difficulty after surgery. Some patients reported increased difficulty or inability to return to their preoperative maximum level of participation, indicating that flatfoot reconstructions can lead to athletic limitations. Although flatfoot reconstruction can be a powerful tool to increase patients' capacity to engage in physical activity, in our cohort many patients had reduced physical activity outcomes. Level IV, retrospective case series.


[Effect of retS gene on biosynthesis of 2, 4-diacetyl-phloroglucinol in Pseudomonas fluorescens 2P24].
Regulator of exopolysaccharide and type III secretion (RetS) is a hybrid sensor kinase/response regulator protein located on bacterial membrane, and essential for expression of numerous genes. In Pseudomonas aeruginosa, RetS modulates the phosphorylation state of another kinase GacS via a direct interaction. The goal of this study is to study the effect of retS on the antibiotic 2,4-diacetyl-phloroglucinol (2,4-DAPG) production in P. fluorescens 2P24. Production of 2,4-DAPG in strain 2P24 and its mutants was quantified by HPLC. To determine the effect of RetS on the Gac/Rsm pathway, the promoters of the small RNA genes rsmX/rsmY/rsmZ and regulatory genes rsmA/rsmE in the Rsm pathway were fused with a promoterless lacZ, and the promoter activities were measured in 2P24 and the retS-deficient mutants. Genetic inactivation of the retS in strain 2P24 increased the production of an uncharacterized red pigment and the antibiotic 2,4-DAPG. RetS negatively regulated the transcription of the small RNAs RsmX and RsmZ. In the retS and gacS double mutant or the retS and gacA double mutant, all the phenotypic changes caused by the retS deletion were reversed to the level of gacS or gacA single gene mutant. In P. fluorescens 2P24, RetS negatively regulates the production of antibiotic 2,4-DAPG through the Gac/Rsm pathway.


[Hemosorption in the treatment of toxemia and septic conditions after severe mechanical trauma].
Under study was the significance of hemosorption in complex therapy of patients with complications resulting from a severe mechanical trauma. Lethality was found to decrease from 68% to 45%. The results obtained allow the use of hemosorption to be recommended for the treatment of endogenous intoxication in patients with complications of a severe mechanical trauma.


Effect of spectrum bias on the diagnostic accuracy of spectral-domain optical coherence tomography in glaucoma.
To evaluate the influence of a control group on the diagnostic accuracy of spectral-domain optical coherence tomography (SD-OCT) in early glaucoma. In a diagnostic, case-control study, 119 eyes of 60 normal subjects with no findings suspicious for glaucoma (control cohort 1); 76 eyes of 41 subjects referred by general ophthalmologists as glaucoma suspects based on optic disc morphology, but found by glaucoma experts to be normal but with physiological variations in their optic nerves (control cohort 2); and 65 eyes of 46 early-glaucoma patients (cases) underwent imaging of the optic nerve head (ONH), retinal nerve fiber layer (RNFL), and ganglion cell complex (GCC) by SD-OCT. Areas under the receiver operating characteristic curves (AUC) of ONH parameters discriminating glaucomatous eyes from normal eyes of control cohort 2 were significantly lesser (P < 0.001) than those discriminating glaucomatous eyes from normal eyes of control cohort 1. AUCs of RNFL parameters discriminating glaucomatous eyes from normal eyes of control cohorts 2 and 1 were comparable. Although the AUCs of GCC thickness parameters were comparable, AUCs of GCC focal and global loss volume in control cohort 2 (0.684 and 0.671. respectively) were significantly less (P < 0.05) than in control cohort 1 (0.881 and 0.841, respectively). The effectiveness of most SD-OCT parameters in detecting glaucoma significantly decreased when evaluated against a clinically relevant control group with suspicious-looking optic nerves compared with that against a control group consisting of normal subjects with no findings suspicious for glaucoma.


Using NGS-methylation profiling to understand the molecular pathogenesis of young MI patients who have subsequent cardiac events.
Globally, ischaemic heart disease is a major contributor to premature morbidity and mortality. A significant number of young Myocardial Infarction (MI) patients (aged <55 y) have subsequent cardiac events within a year of their index event. This study used Next Generation Sequencing (NGS) methylation to understand the pathogenesis in this subset of young MI patients, comparing them to a cohort of patients without recurrent events. Cases and controls were matched for age, gender, ethnicity, and comorbidities. Differential methylation analyses were performed on Reduced Representation Bisulphite Sequencing (RRBS) data. Across the group and within case-control pairs' variation were analysed. Pairwise comparisons across each matched case-control pair resulted in a list of genes that were consistently significantly differentially methylated between all 16 matched pairs. This gene list was input into pathway analysis databases. Of particular relevance to cardiac pathology the following pathways were identified as over-represented in the patients with recurrent events; cell adhesion, transcription regulation and cardiac electrical conduction, specifically relating to calcium channel activity. This study looked at methylation differences between two populations of young MI patients. There were significantly different methylation profiles between the two groups studied; key pathways were identified as specifically affected in the patients with recurrent cardiac events. Matched pairwise comparisons and detailed interpretations of DNA methylation data may help to elucidate complex pathogeneses within and between clinical subtypes. Further analysis will determine whether these epigenomic differences can be useful as predictive biomarkers of clinical progression.


Development of an in vitro fibrin clot model to evaluate fibrinolytic agents for wound care application.
To describe an in vitro fibrin clot model that could reliably assess the fibrinolytic activity of enzymatic debriding agents for wound care application. A model of a fibrin clot was reconstructed in vitro by mixture of human fibrinogen and (alpha)-thrombin supplemented with factor XIII. These clots were then treated with enzymatic ointments. Fibrinolytic activity was investigated by measuring D-dimer levels, using an automated immunoturbidimetric Liatest D-dimer assay. Collagenase and papain-urea ointments demonstrated fibrinolytic activity which was macroscopically visible. Their effect was identical on the in vitro reconstructed fibrin clot and ex vivo collected wound fibrin clot; collagenase and papain-urea both induced a complete degradation and dissolution of both fibrin clots after 24 hours of treatment. This was associated with an increase in D-dimer concentration. This reconstructed fibrin clot in vitro model has the potential to predict the efficacy of fibrinolytic agents and therefore appears to be a suitable model for in vitro assays. This study was supported by a grant from URGO Laboratory.


Pathogenesis of renal dysfunction in chicks experimentally induced by avian nephritis virus.
Renal dysfunction in chicks infected with avian nephritis virus (ANV) at 1 day or 1 week of age was studied. Two chicks inoculated per os and one inoculated intraperitoneally, both infected at 1 day of age, died with visceral urate deposits within 10 to 13 dpi, and the surviving infected chicks showed a markedly reduced weight gain. In a chronological study, ANV was consistently isolated from the kidney irrespective of the route of infection or age of chick. Fluorescent antigen to ANV was also detected as cytoplasmic granules in the tubular cells, accompanied by necrosis of tubular cells. A high concentration of serum uric acid was detected 4 to 13 dpi in chicks infected at 1 day of age and was frequently coincident with detection of the tubular cell necrosis. These results suggest that the increased serum uric acid concentration is caused by viral damage of kidney tubular cells.


Expression and purification of monospecific and bispecific recombinant antibody fragments derived from antibodies that block the CD80/CD86-CD28 costimulatory pathway.
The development of recombinant techniques for rapid cloning, expression, and characterization of cDNAs encoding antibody (Ab) subunits has revolutionized the field of antibody engineering. By fusion to heterologous protein domains, chain shuffling, or inclusion of self-assembly motifs, novel molecules such as bispecific Abs can be generated that possess the subset of functional properties designed to fit the intended application. We describe the engineering of Ab fragments produced in bacteria for blocking the CD28-CD80/CD86 costimulatory interaction in order to induce tolerance against transplanted organs. We designed single-chain Fv antibodies, monospecific and bispecific diabodies, and a bispecific tetravalent antibody (BiTAb) molecule directed against the CD80 and/or CD86 costimulatory molecules. These recombinant Ab molecules were expressed in Escherichia coli, followed by purification and evaluation for specific interaction with their respective antigen in an enzyme-linked immunosorbent assay (ELISA). A specific sandwich ELISA confirmed the bispecificity of the bispecific diabodies and the BiTAb.


Integrating Medical Genetics Into Precision Oncology Practice in the Veterans Health Administration: The Time Is Now.
Increased access and utilization of tumor profiling of cancers in our veteran population uncovered a modest number of potentially pathogenic germline variants (PPGVs) that require genetics referral for follow-up evaluation and germline sequencing. Challenges identified specific to the veteran population include paucity of genetics providers, either at a veteran's VA facility or nearby non-VA facilities. We sought to investigate the number of veterans who would benefit from having such resources at both local and national levels. Annotated clinical reports of mutations identified by tumor-only profiling and medical records of veterans with solid tumors at the Veterans Administration Ann Arbor Healthcare System (VA AAHS) between 2015 and 2020 were reviewed. PPGVs were identified according to society recommendations (such as ESMO and American Board of Medical Genetics and Genomics), expert review, and/or previously published criteria. After the analysis of our local VA population, these same criteria were then applied to veterans in the National Precision Oncology Program (NPOP). Two hundred eight veterans underwent tumor profiling at the VA AAHS over the defined time period. This included 20 different primary tumor sites with over half (n = 130) being advanced cancer at diagnosis. Of these, 18 veterans (8.5%) had mutations suggestive of a PPGV. Applying these criteria to the larger NPOP database (n = 20,014), a similar percentage (6%) of PPGVs were identified. These results indicate a PPGV frequency (6%-9% of veterans) consistent with the prevalence of inherited cancer predisposition syndromes in the general population, underscoring the need for medical genetics as part of standard oncologic care for veterans. We explore current and future care delivery models to optimize incorporation of medical genetics and genetic counseling to best serve veterans needing such services.


Belief in pain.
There is a traditional view of pain as a conscious phenomenon which satisfies the following two principles at least: Pain is essentially a belief- or cognition-independent sensation, given for consciousness in an immediate way, and pain's unitary physical base is responsible for both its phenomenal or felt qualities and its functional, causal features. These are "The Raw Feels Principle" (P) and "The Unity of Pain Principle" (Q). Each is shown to be implausible. Evidence comes from recent pain research in a number of dimensions. A further argument against The Unity of Pain Principle is constructed by analogy with facts about blindsight. The consequences of rejecting P or Q are examined in light of (i) the traditional epistemological role of pain as exemplar of sensations thought to be foundational for knowledge, (ii) the prospects of a modularity theory of the pain system, (iii) recent accounts of pain among contemporary philosophers, (iv) a naturalist and realist conception of pain, and (v) views about the status of Folk Psychology.


Evidence for a KCl-Stimulated, Mg-ATPase on the Golgi of Corn Coleoptiles.
Membranes of corn (Zea mays, cv Trojan 929) coleoptiles were fractionated by sucrose density gradient centrifugation and the locations of organelles were determined using marker enzymes and electron microscopy. Latent IDPase (or UDPase) was selected as the Golgi marker and UDPG-sterol glucosyl transferase was selected as the plasma membrane (PM) marker, because they were clearly separable from markers for the other organelles. Golgi-rich and PM-rich fractions were studied in relation to their ATPase activities. The pH optimum of the KCl, Mg(2+)-ATPase of the PM-rich fraction from a step gradient was 6.0 to 6.5, while the Golgi-rich fraction had peaks at pH 6.0 to 6.5 and pH 7.5. It is hypothesized that the peak at pH 6.0 to 6.5 for the Golgi-rich fraction is due to PM-contamination, while the peak at pH 7.5 represents the activity of a Golgi ATPase. To reduce PM contamination, Golgi-rich fractions obtained from step or rate-zonal gradients were recentrifuged isopycnically on linear sucrose gradients. The distribution of KCl, Mg(2+)-ATPase activity was measured at pH 6.5 and 7.5. The pH 6.5 ATPase was coincident with UDPG-sterol glucosyl transferase, a PM marker, while the pH 7.5 ATPase overlapped with latent UDPase, a Golgi marker. These results provide strong evidence for a KCl, Mg(2+)-ATPase, active at pH 7.5, associated with the Golgi membranes of corn coleoptiles.


Acne--natural history, facts and myths.
This article describes the epidemiology and natural history of acne and discusses the common aetiological myths and beliefs surrounding acne. Acne is a polymorphic disease with noninflammatory and inflammatory aspects and a wide spectrum of severity. The severe inflammatory patterns tend to be those that most frequently scar and therefore require early intervention. The pattern of disease, relative severity, distribution, and patient's social circumstances will influence appropriate management. However, effective management also includes patient education about acne, its natural history, and dispelling acne myths.


Direct Reconstruction of CT-based Attenuation Correction Images for PET with Cluster-Based Penalties.
Extremely low-dose CT acquisitions for the purpose of PET attenuation correction will have a high level of noise and biasing artifacts due to factors such as photon starvation. This work explores a priori knowledge appropriate for CT iterative image reconstruction for PET attenuation correction. We investigate the maximum a posteriori (MAP) framework with cluster-based, multinomial priors for the direct reconstruction of the PET attenuation map. The objective function for direct iterative attenuation map reconstruction was modeled as a Poisson log-likelihood with prior terms consisting of quadratic (Q) and mixture (M) distributions. The attenuation map is assumed to have values in 4 clusters: air+background, lung, soft tissue, and bone. Under this assumption, the MP was a mixture probability density function consisting of one exponential and three Gaussian distributions. The relative proportion of each cluster was jointly estimated during each voxel update of direct iterative coordinate decent (dICD) method. Noise-free data were generated from NCAT phantom and Poisson noise was added. Reconstruction with FBP (ramp filter) was performed on the noise-free (ground truth) and noisy data. For the noisy data, dICD reconstruction was performed with the combination of different prior strength parameters (β and γ) of Q- and M-penalties. The combined quadratic and mixture penalties reduces the RMSE by 18.7% compared to post-smoothed iterative reconstruction and only 0.7% compared to quadratic alone. For direct PET attenuation map reconstruction from ultra-low dose CT acquisitions, the combination of quadratic and mixture priors offers regularization of both variance and bias and is a potential method to derive attenuation maps with negligible patient dose. However, the small improvement in quantitative accuracy relative to the substantial increase in algorithm complexity does not currently justify the use of mixture-based PET attenuation priors for reconstruction of CT images for PET attenuation correction.


Delivering the Same Intervention to Hispanic/Latinos With Pre-diabetes and Diabetes. Early Evidence of Success in a Longitudinal Mixed Method Study.
Despite nationwide efforts to address the diabetes epidemic and reduce prevalence disparities, higher rates persist among the poor, especially those with limited literacy. Currently, individuals with abnormal glycemia who have pre-diabetes and diabetes qualify for different programs. However, evidence suggests that, for low-income Hispanic/Latinos, offering a single intervention to all those with abnormal glycemia may provide a more culturally acceptable and effective approach. Our objective was to explore the feasibility of such an intervention led by community health workers (CHWs) among low-income Hispanic/Latinos with diabetes and at risk for diabetes. Using a quasi-experimental mixed method design, we assessed weight, glycosylated hemoglobin, diabetes knowledge, and behavior changes of Hispanic/Latinos participants with pre-diabetes and diabetes living in Southern California. Biometric measurements, blood tests, and surveys were collected at baseline and 3 months post-intervention. Interviews and focus group discussions provided qualitative data. Although the program was less costly, results exceeded those reported for low-income H/L attending the National Diabetes Prevention Program and did not differ between pre-diabetes and diabetes groups. Instead, including individuals at different stages of the dysglycemic spectrum seemed to have enhanced the intervention. Physician referral and attendance of family/friends were associated with better outcomes. Our findings indicate that a joint prevention/self-management intervention led by CHWs for low-income Hispanic/Latinos with diabetes and with pre-diabetes is feasible and cost-effective, providing results that could help reduce the success gap. Incorporating suggestions and replicating this study on a larger scale could help determine whether or not results are reproducible.


Mitotic regulators govern progress through steps in the centrosome duplication cycle.
Centrosome duplication is marked by discrete changes in centriole structure that occur in lockstep with cell cycle transitions. We show that mitotic regulators govern steps in centriole replication in Drosophila embryos. Cdc25(string), the expression of which initiates mitosis, is required for completion of daughter centriole assembly. Cdc20(fizzy), which is required for the metaphase-anaphase transition, is required for timely disengagement of mother and daughter centrioles. Stabilization of mitotic cyclins, which prevents exit from mitosis, blocks assembly of new daughter centrioles. Common regulation of the nuclear and centrosome cycles by mitotic regulators may ensure precise duplication of the centrosome.


Undiagnosed bleeding states and medical treatment.
Relatively little attention has been placed in mild hemophilia in the orthopedic literature even though there are probably an equal number of patients with mild as compared to classic manifestations. Mild hemophilia and similar related conditions frequently are undiagnosed and may represent major problems and if associated with surgery, catastrophic results. Patients with milder bleeding diatheses may not have a diagnosis made until late in life. Mild hemophilia and hemophiloid conditions are defined and typical findings are given. Other causes of intra or postoperative hemorrhagic states are mentioned. Three case reports of mild Factor VIII and Factor XI deficiency, undiagnosed prior to the author's medical treatment, are presented. Knowledge of the typical historical findings, diagnostic methods, and proper treatment of patients seeking medical treatment with mild hemophilia, hemophiloid and similar conditions explaining hemorrhage following trauma and intra and postoperatively will help to prevent undesirable results as well as severe complications.


The Potency of Diarylamine Radical-Trapping Antioxidants as Inhibitors of Ferroptosis Underscores the Role of Autoxidation in the Mechanism of Cell Death.
Two aromatic amines (ferrostatin-1 and liproxstatin-1) were recently identified from high-throughput screening efforts to uncover potent inhibitors of ferroptosis, the necrotic-like cell death induced by inhibition of glutathione peroxidase 4 (GPX4), deletion of the corresponding gpx4 gene, or starvation of GPX4 of its reducing cosubstrate, glutathione (GSH). We have since demonstrated that these two aromatic amines are highly effective radical-trapping antioxidants (RTAs) in lipid bilayers, suggesting that they subvert ferroptosis by inhibiting lipid peroxidation (autoxidation) and, thus, that this process drives the execution of ferroptosis. Herein, we show that diarylamine RTAs used to protect petroleum-derived products from autoxidation can be potent inhibitors of ferroptosis. The diarylamines investigated include representative examples of additives to engine oils, greases and rubber (4,4'-dialkyldiphenylamines), core structures of dyes and pharmaceuticals (phenoxazines and phenothiazines), and aza-analogues of these three classes of compounds that we have recently shown can be modified to achieve much greater reactivity. We find that regardless of how ferroptosis is induced (GPX4 inhibition, gpx4 deletion or GSH depletion), compounds which possess good RTA activity in organic solution (kinh > 105 M-1 s-1) and lipid bilayers (kinh > 104 M-1 s-1) are generally potent inhibitors of ferroptosis (in mouse embryonic fibroblasts). Likewise, structural analogs that do not possess RTA activity are devoid of antiferroptotic activity. These results further support the argument that lipid peroxidation (autoxidation) plays a major role in the mechanism of cell death induced by either GPX4 inhibition, gpx4 deletion, or GSH depletion. Moreover, it offers clear direction that ongoing medicinal chemistry efforts on liproxstatin and ferrostatin derivatives, which have been proposed as lead compounds for the treatment and/or prevention of ischemia/reperfusion injury, renal failure, and neurodegeneration, can be widened to include other aminic RTAs. To aid in these efforts, some relevant structure-reactivity relationships are discussed.


[Prospective multi-center study of one-step nucleic acid amplification assay for the detection of sentinel lymph nodes metastases in breast cancer patients].
To evaluate the roles of Sysmex RD100i one-step nucleic acid amplification (OSNA) assay in the intraoperative assessments of breast cancer sentinel lymph nodes (SLNs). A total of 552 consecutive prospective patients were enrolled from five centers nationwide from February to December 2010. And SLNs were sliced into alternating 2 mm blocks. The odd blocks were tested by the OSNA assay intraoperatively and the even ones assessed by postoperative histology. In addition, intraoperative histological assessments were performed on the even blocks of 211 patients by frozen section and all blocks by touch imprint cytology. A total of 1188 SLNs were removed. The mean turnaround time of the assay was 37.3 min. There was no significant difference of turnaround time at each center (P = 0.074). As compared to postoperative histology, the overall performance of the assay had an accuracy of 91.4% (1086/1188), a sensitivity of 83.7% (159/190) and a specificity of 92.9% (927/998). The sensitivity of the assay was higher than frozen section (77.6% (59/76) vs 69.7% (53/76), P = 0.286) and was significantly higher than touch imprint cytology (83.6% (158/189) vs 76.2% (144/189), P = 0.044). For nodes with micro-metastases, the sensitivity of the assay was higher than frozen section (8/17 vs 4/17, P = 0.289) and was significantly higher than touch imprint cytology (62.5% (30/48) vs 35.4% (17/48), P = 0.007). As an accurate and rapid intraoperative assay for assessing breast SLNs, the OSNA assay may replace frozen section and touch imprint cytology for clinical applications.


Arousal responses of the newborn piglet to airways obstruction and rebreathing during normothermia and hyperthermia.
Most experimental studies of arousal to respiratory stressors have been performed under neutral thermal conditions. The present study focussed on arousal responses with the added challenge of heat stress. The subjects were two groups of 10 sedated piglets aged 4-6 days and 2-6 days respectively. Respiratory stimuli (partial and total airways obstruction (AO) or rebreathing (RB) expired gases) were applied during non-rapid-eye-movement (NREM) sleep. Measurements of heart rate, SaO2, intra-pleural pressure or inspired CO2 and O2 were recorded during tests in normothermia and hyperthermia. Hyperthermia significantly shortened the time to arousal. Thus, arousal times from partial and total AO and from RB were: in normothermia, 40.3 +/- 5.8, 9.2 +/- 0.7 and 94.9 +/- 14.7 s respectively; in hyperthermia, 17.8 +/- 3.7, 7.6 +/- 0.9 and 68.5 +/- 9.1 s respectively. Cardio-respiratory variables at arousal were similar in both thermal states. Seven non-sedated piglets were challenged with RB stimuli during normothermia only to determine the influence of sleep state on arousal. Arousal in REM sleep was delayed approximately 2-3-fold (NREM, 41.2 +/- 8.2 s; REM, 88.5 +/- 18.1 s), occurring at a lower SaO2 and higher inspired CO2. It is concluded that arousal from NREM sleep under adverse conditions of hyperthermia shortens the arousal time from asphyxial stimuli induced by AO and RB with no change in the arousal threshold.


Directive support, nondirective support, and health behaviors in a community sample.
Social support is associated with improved psychological functioning, physical health, and health-promoting behaviors. However, research suggests that health outcomes might depend upon the type rather than the amount of support provided to recipients. This study assessed the relationship among nondirective and directive support, and health behaviors (i.e., physical activity, fruit and vegetable intake, alcohol use) in a community sample of 304 adults. Results revealed that nondirective support was related to greater fruit and vegetable intake and lower alcohol use; directive support was not associated with these behaviors. For physical activity, a similar trend emerged, but support variables did not explain any variance above that explained by demographic variables. These findings offer a unique contribution to the literature, as they suggest that certain types of supportive behaviors are important in encouraging positive health behaviors.


Texture analysis of 18F-FDG PET/CT to predict tumour response and prognosis of patients with esophageal cancer treated by chemoradiotherapy.
This retrospective study was done to examine whether the heterogeneity in primary tumour F-18-fluorodeoxyglucose (18F-FDG) distribution can predict tumour response and prognosis of patients with esophageal cancer treated by chemoradiotherapy (CRT). The enrolled 52 patients with esophageal cancer underwent 18F-FDG-PET/CT studies before CRT. SUVmax, SUVmean, metabolic tumour volume (MTV, SUV ≥ 2.5), total lesion glycolysis (TLG) and six heterogeneity parameters assessed by texture analysis were obtained. Patients were classified as responders or non-responders according to Response Evaluation Criteria in Solid Tumors. Progression-free survival (PFS) and overall survival (OS) were calculated by the Kaplan-Meier method. Prognostic significance was assessed by Cox proportional hazards analysis. Thirty four non-responders showed significantly higher MTV (p = 0.006), TLG (p = 0.007), intensity variability (IV; p = 0.003) and size-zone variability (SZV; p = 0.004) than 18 responders. The positive and negative predictive values for non-responders were 77 % and 69 % in MTV, 76 % and 100 % in TLG, 78 % and 67 % in IV and 78 % and 82 % in SZV, respectively. Although PFS and OS were significantly shorter in patients with high MTV (PFS, p = 0.018; OS, p = 0.014), TLG (PFS, p = 0.009; OS, p = 0.025), IV (PFS, p = 0.013; OS, p = 0.007) and SZV (PFS, p = 0.010; OS, p = 0.007) at univariate analysis, none of them was an independent factor, while lymph node status, stage and tumour response status were independent factors at multivariate analysis. Texture features IV and SZV, and volumetric parameters MTV and TLG can predict tumour response, but all of them have limited value in prediction of prognosis of patients with esophageal cancer treated by CRT.


Efficacy of sorafenib monotherapy versus sorafenib-based loco-regional treatments in advanced hepatocellular carcinoma.
Although sorafenib is accepted as the standard of care in advanced hepatocellular carcinoma (HCC), its therapeutic benefit is marginal. Here, we aimed to compare the efficacy and safety of sorafenib monotherapy (S-M) and sorafenib-based loco-regional treatments (S-LRTs) in advanced HCC. From 2007 to 2012, 290 patients with advanced HCC (Barcelona Clinic Liver Cancer stage C) with S-M (n = 226) or S-LRTs (n = 64) were reviewed retrospectively. Survival outcomes and treatment-related toxicities between two groups were analyzed. Variables related to tumor burden and liver function were similar between the groups (all P > 0.05). Within the entire population, the S-LRTs group had both longer median overall survival (OS) (8.5 vs 5.5 months, P = 0.001) and progression-free survival (PFS) (5.3 vs 3.0 months, P = 0.002) than the S-M group. Furthermore, the S-LRTs group had longer Os than the S-M group in a subgroup with neither extrahepatic spread (EHS) nor regional nodal involvement (RNI) (18.0 vs 7.8 months, P = 0.019) and in a subgroup with EHS and/or RNI (8.3 vs 4.8 months, P = 0.028). In addition, the S-LRTs group had longer PFS than the S-M group in the subgroup with neither EHS nor RNI (9.6 vs 3.2 months, P = 0.027). Related toxicity was similar between two groups. Combined use of sorafenib and LRTs may provide better treatment outcomes without significantly increasing treatment-related toxicities, even in patients with EHS and/or RNI. Therefore, addition of active LRTs might be considered, if feasible.


c-Jun-like immunoreactivity in apoptosis is the result of a crossreaction with neoantigenic sites exposed by caspase-3-mediated proteolysis.
Previous reports in various cells and species have shown that apoptotic cells are specifically and strongly labeled by certain c-Jun/N-terminal antibodies, such as c-Jun/sc45. This kind of immunoreactivity is confined to the cytoplasm. It is not due to c-Jun but appears to be related to c-Jun-like neoepitopes generated during apoptosis. This study was planned to gain further information about c-Jun-like immunostaining during apoptosis and to evaluate these antibodies as possible tools for characterizing cell death. Most of the experiments were performed in chick embryo spinal cord. When the apoptotic c-Jun-like immunoreactivity and caspase-3 immunostaining patterns were compared, we found that both antibodies immunostained the same dying cells in a similar pattern. In contrast to TUNEL staining, which reveals a positive reaction in both apoptotic and necrotic dying cells, active caspase-3 and c-Jun/sc45 antibodies are more selective because they stained only apoptotic cells. When cytosolic extracts from normal tissues were digested in vitro with caspase-3, c-Jun/sc45 immunoreactivity was strongly induced in several proteins, as demonstrated by Western blotting. Similar results were found when normal tissue sections were treated with caspase-3. Our results show that c-Jun/sc45 antibodies react with neoepitopes generated from cell proteins cleaved by activated caspases during apoptosis. We conclude that c-Jun/sc45 antibodies may be useful for detecting apoptosis. They can even be used in archival paraffin-embedded tissue samples.


Salicylates inhibit lipopolysaccharide-induced transcriptional activation of the tissue factor gene in human monocytic cells.
Binding of plasma Factor VII/VIIa to the tissue factor (TF) receptor initiates the coagulation protease cascades. TF expression by circulating monocytes is associated with thrombotic and inflammatory complications in a variety of diseases. Transcriptional activation of the human TF gene in monocytic cells exposed to bacterial lipopolysaccharide (LPS) is mediated by binding of c-Rel/p65 heterodimers to a kappa B site in the TF promoter. Here, we report that a family of anti-inflammatory agents, known as the salicylates, inhibited LPS induction of TF activity and TF gene transcription in human monocytes and monocytic THP-1 cells at clinically relevant doses. Furthermore, sodium salicylate blocked the LPS-induced proteolytic degradation of I kappa B alpha, which prevented the nuclear translocation of c-Rel/p65 heterodimers. In contrast, two other nonsteroidal anti-inflammatory drugs, ibuprofen and indomethacin, did not inhibit LPS induction of the TF gene. These results indicated that salicylates inhibited LPS induction of TF gene transcription in monocytic cells by preventing nuclear translocation of c-Rel/p65 heterodimers. The clinical benefits of salicylates in the treatment of several diseases, including atherosclerosis and rheumatoid arthritis, may be related to their ability to reduce monocyte gene expression.


Outcomes of Vitrectomy in Pediatric Retinal Detachment with Proliferative Vitreoretinopathy.
To report outcomes of pars plana vitrectomy (PPV) in pediatric retinal detachment (RD) with proliferative vitreoretinopathy (PVR), complications, factors influencing the final anatomical and functional results. Retrospective consecutive case series of 14 eyes. Average postoperative follow-up period was 34 months. Mean age of patients was 10 years; eleven patients (79%) were males. The most common etiology was trauma (57%), the second-myopia (36%) and one case of uveitis (7%). At the day of presentation, the best-corrected visual acuity (BCVA) was worse than hand motion (50%); macula was detached in 86% of cases. Simultaneous PPV and phacoemulsification with intraocular lens (IOL) implantation were performed in 12 cases (86%). The most common endotamponade during PPV was silicone oil (93%). Anatomic reattachment was accomplished in 86% of cases. Final BCVA was equal or better than 0.1 in 50% of patients. The postoperative complications were found in 5 eyes (36%). Complete PPV was allowed for anatomically reattached retina and preserved vision in pediatric complex RD with PVR. However, visual outcomes were not satisfactory. Preserving vision in children with RD is of great importance for their future motor and intellectual development. This trial is registered with ClinicalTrials.gov Identifier: NCT03208205.


Anthropometric measurements and body proportions among Chinese children.
There are significant racial differences in body proportions. Such data are not readily available for Chinese children. This article reports a cross section study of body proportions of 2193 Hong Kong Chinese children, ages 4 to 16 years, with equal gender distribution. Standing height, sitting height, and arm span were measured with standard equipment and methodology. Lower segment height was calculated as the difference between standing height and sitting height. Statistical analysis of the results showed a high linear correlation of the standing height with arm span, sitting height, and lower segment height, with a correlation coefficient ranging from 0.965 to 0.983 for both genders. When expressed as the ratio of standing height to arm span, the value was relatively constant and changed only linearly from 1.03 to 1 in girls and 1.03 to 0.98 in boys ages 4 to 16 years. The ratio of sitting height to lower segment height varied from a mean of 1.4 to 1.14 in boys and 1.36 to 1.18 in girls ages 4 to 16 years. The Chinese children were found to have a proportional limb segmental length relative to the trunk that differed significantly from the proportionally longer limbs in whites and blacks.


Can malaria parasite pathogenesis be prevented by treatment with tumor necrosis factor-alpha?
We consider a model incorporating the influence of innate and adaptive immune responses on malaria pathogenesis. By calculating the model reproduction number for a special representation of cytokine interaction, we have shown that the cytokine tumour necrosis factor-α can be administered to inhibit malaria infection. We have also found that if the cytokine F ∗ and a generic drug of efficacy ε are administered as dual therapy then clearance of the parasite can be achieved even for a generic drug of low efficacy. Our study is recommending administration of dual therapy as a strategy to prevent parasites from developing resistance to malaria treatment drugs.


[A case of atypical iris corneal endothelial syndrome].
Iridocorneal endothelial syndrome is a rare ophthalmic disease, most of which are unilateral and common in women. The rate of misdiagnosis and missed diagnosis is relatively high due to its various clinical manifestations. In this case, the patient presented uncontrollable high intraocular pressure, corneal edema leading to difficult observation of corneal endothelium morphology, and accompanied by a small amount of iris neovascularization. No clearly diagnosis was made before glaucoma surgery. Further examination was performed after corneal clearance, and the final diagnosis was iris corneal endothelial syndrome (Chandler syndrome).


Cell division-associated expression of an epitope, KH17, on early developing thymocytes.
A newly established monoclonal antibody, KH17, detects a unique epitope temporarily expressed on early developing CD3-thymocytes confined to a cycling stage. KH17 is detectable on a part of CD4-CD8-,CD4-CD8+, and CD4+CD8+ cells, but not on CD4+CD8- thymocytes. By four-color flow cytometry analysis using KH17, we were able to define the heterogeneity of immature CD4-CD8- thymocytes by the expression of KH17 and IL-2R. In Thy-1-congeneic bone marrow chimeras, the appearance of KH17-IL-2R+ thymocytes preceded the increase of KH17+IL-2R- cells. The antibody could also divide CD3-CD4-CD8+ cells into two subpopulations, KH17+ and KH17-, which showed a continuum. In the fetal thymus there was a rapid and dramatic increase of KH17-CD4+CD8+ thymocytes concomitant with a decrease of KH17+CD4-CD8+ thymocytes in later gestation days. KH17 is not expressed on resting peripheral T cells, but is expressed on a large proportion of Con A-activated blastic spleen cells. The KH17 molecules precipitated from Con A-activated spleen cells were 55 and 75 kd polypeptides, but different from IL-2R subunits.


Haematological changes in the garden lizard, Calotes nemoricola due to the defensive secretion of the grasshopper, Poecilocerus pictus.
Defensive secretion of the Grasshopper, Poecilocerus pictus produced significant changes in the blood components of the predator lizard, C. nemoricola. The red blood cell (RBC) count, haemoglobin (Hb) concentration, packed cell volume (PCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC) and white blood cell (WBC) count decreased greatly, whereas mean corpuscular volume (MCV) showed significant increase following the administration of lethal dose of defensive secretion. These changes however were found to be reversible toward control levels in the lizards injected with sublethal dose. Disrupted erythropoeitic activity in lizards by the cardenolides in the defensive secretion was suggested.


[Polymorphism of POU1F1 gene and PRL gene and their combined effects on milk performance traits in Chinese Holstein cattle].
Three novel SNPs were found by DNA sequencing, PCR-RFLP and CRS-PCR methods were used for genotyping in 979 Chinese Holstein cattle. One SNP, G1178C, was identified in exon 2 of POU1F1 gene. Two novel SNPs, A906G and A1134G, were identified in 5'-flanking regulatory region (5'-UTR) of PRL gene. The association between polymorphisms of the two genes and milk performance traits were analyzed with PROC GLM of SAS. The results showed that GC genotype at 1178 locus of POU1F1 gene was advantageous for milk yield, milk protein yield, and milk fat yield. AG genotype at 906 locus was advantageous for milk yield. There was no significant difference between 1134 locus and milk performance traits of 5'-UTR of PRL gene. Analysis of genotype combination effect on milk production traits showed that the effect of combined genotype was not simple sum of single genotypes and the effects of gene pyramiding seemed to be more important in molecular breeding.


Local and Interregional Neurochemical Associations Measured by Magnetic Resonance Spectroscopy for Studying Brain Functions and Psychiatric Disorders.
Magnetic resonance spectroscopy (MRS) studies have found significant correlations among neurometabolites (e.g., between glutamate and GABA) across individual subjects and altered correlations in neuropsychiatric disorders. In this article, we discuss neurochemical associations among several major neurometabolites which underpin these observations by MRS. We also illustrate the role of spectral editing in eliminating unwanted correlations caused by spectral overlapping. Finally, we describe the prospects of mapping macroscopic neurochemical associations across the brain and characterizing excitation-inhibition balance of neural networks using glutamate- and GABA-editing MRS imaging.


Multilayered assemblies composed of brilliant yellow and poly(allylamine) for an optical pH sensor.
Multilayered thin films containing poly(allylamine) (PAA) and brilliant yellow (BY) were prepared on a quartz slide by a layer-by-layer (LBL) deposition technique. The UV-visible spectra of the PAA/BY films were sufficiently changed depending upon the pH value of the solution in which the film was immersed. The response of the PAA/BY films was very fast (within a second) upon pH change from 9.0 to 5.0, while the response time was ca. 100 s upon pH change from 5.0 to 9.0.


New diagnostic approach to intracystic lesions of the breast by fiberoptic ductoscopy.
Intracystic tumors of the breast are uncommon and, at the time of ultrasonography and aspiration cytology, it is difficult to distinguish cancer from a benign tumor. The Fiberoptic Ductoscopy System (FDS) is an emerging technique allowing direct visual access to the ductal system of the breast. FDS was inserted through the cannulae into the cavity and we observed the intracystic tumors (3 intracystic papillomas and 2 intracytsic papillary carcinomas). The appearance of the malignant tumors was irregular, rough-shaped and they tended to bleed. On the contrary, benign tumors had smooth surfaces without bleeding. Cytological findings showed malignant cells in one out of two breast cancer patients. In addition, in the immunohistochemical study of resected tumor tissues from 5 patients, we observed positive reactions with anti-ErbB-2 antibody in 2 intracystic papillary carcinomas. In contrast, none of the histologically confirmed benign lesions (3 intracystic papillomas) gave positive results. In conclusion, the use of FDS as a non-invasive technique may provide valuable information.


DNA ploidy of colorectal carcinoma by tumour site, gender and history of noncolorectal malignancies.
DNA ploidy was analysed by flow cytometer from frozen samples of 205 colorectal carcinomas. Sixty-two percent of the tumours had an abnormal DNA stemline. Forty-eight percent of carcinomas in the right colon, 62% of carcinomas in the left colon and 74% of carcinomas in the rectum were aneuploid (p = 0.007). Sixty-nine percent of tumours in males and 55% of tumours in females were aneuploid (p = 0.029). The difference in frequency of aneuploidy between females and males was greatest in tumours of the right colon, where 37% of the tumours in females and 62% of the tumours in males were aneuploid (p = 0.047). The percentage of diploid colorectal carcinomas was higher (55%) in patients with a history of noncolorectal malignancy than in others (34%, p = 0.031). These results suggest that flow cytometry may be helpful in understanding the development of colorectal carcinomas.


Altered expression of the xenobiotic transporter P-glycoprotein in liver and liver tumours of mummichog Fundulus heteroclitus from a creosote-contaminated environment.
P-glycoproteins (Pgps) are involved in efflux of xenobiotics from drug-resistant cell lines and tumours, and in excretion of toxicants from normal tissues. Recently, investigators have proposed that Pgp activity contributes to resistance or tolerance of certain aquatic species to pollutants. In the present study using immunoblot and immunohistochemical techniques, we found elevation of Pgp in liver and liver tumours of creosote-resistant mummichog from a contaminated site in the Elizabeth River, Virginia. Immunoblots of mummichog liver extracts showed an immunoreactive band at 170 kDa and indicated two- to three-fold elevation of Pgp in livers of resistant fish relative to those from a reference site. Laboratory exposures of reference site fish to a model PAH (3-methylcholanthrene), however, produced no increase in liver Pgp levels as measured by immunoblot. Normal mummichog liver sections showed specific immunohistochemical staining for Pgp on the canalicular surface of hepatocytes. In the majority of hepatic neoplasms we observed a high level of over-expression and altered patterns of Pgp expression. However we did not observe Pgp over-expression in early proliferative lesions. Elevation of Pgp in livers and liver tumoursof these resistant mummichog may contribute to their survival in a heavily contaminated environment.


Assessment of the radionuclide composition of "hot particles" sampled in the Chernobyl nuclear power plant fourth reactor unit.
Fuel-containing materials sampled from within the Chernobyl Nuclear Power Plant (ChNPP) Unit 4 Confinement Shelter were spectroscopically studied for gamma and alpha content. Isotopic ratios for cesium, europium, plutonium, americium, and curium were identified, and the fuel burn-up in these samples was determined. A systematic deviation in the burn-up values based on the cesium isotopes in comparison with other radionuclides was observed. The studies conducted were the first ever performed to demonstrate the presence of significant quantities of 242Cm and 243Cm. It was determined that there was a systematic underestimation of activities of transuranic radionuclides in fuel samples from inside of the ChNPP Confinement Shelter, starting from 241Am (and going higher) in comparison with the theoretical calculations.


Nonvasomotor influence of sodium nitroprusside on arteriolar remote response to methacholine.
Vascular communication functions to facilitate blood distribution within tissues and can be demonstrated as conducted vasomotor responses. This study was designed to determine if local application of sodium nitroprusside (SNP) would affect arteriolar function at remote sites. In the cheek pouch of anesthetized hamsters, local application of SNP and nifedipine caused arteriolar dilation only at the site of application (7.1 +/- 0.5 and 7.4 +/- 0.6 microm), but not at remote sites. The application of SNP enhanced subsequent remote, nitric oxide (NO)-independent dilation in response to methacholine, which was applied at a site upstream from the SNP application site (6.7 +/- 0.7 versus 4.5 +/- 0.7 microm for methacholine alone). This potentiating effect was also observed following application of 3-morpholinosydnonimine, but not following nifedipine. This nonvasomotor influence of SNP was not affected by N(omega)-nitro-L-arginine (L-NA), tetrodotoxin (TTX), Gap 27 peptide or halothane. Attenuated local dilation in response to methacholine by L-NA could be partially recovered following downstream application of SNP, suggesting that SNP-induced potentiation was associated with enhanced vasodilatory signals at the methacholine application site. Thus, our results suggest that SNP induces nonvasomotor signals in arterioles to affect the network distribution of blood flow. Intrinsic NO, TTX-sensitive Na(+) channels and gap junctional communication do not seem to play a major role in the conduction of the nonvasomotor signals.


First-line bevacizumab in combination with chemotherapy for HER2-negative metastatic breast cancer: pooled and subgroup analyses of data from 2447 patients.
Bevacizumab has consistently demonstrated improved progression-free survival (PFS) and response rate when combined with first-line chemotherapy for HER2-negative metastatic breast cancer (mBC). However, the lack of a significant overall survival (OS) difference continues to attract debate, and identification of patients deriving greatest benefit from bevacizumab remains elusive. Individual patient data from three randomised phase III trials in the first-line HER2-negative mBC setting were analysed, focusing specifically on efficacy in poor-prognosis patients. The meta-analysis (n = 2447) demonstrated a PFS hazard ratio (HR) of 0.64 (95% confidence interval [CI] 0.57-0.71; median 9.2 months with bevacizumab versus 6.7 months with non-bevacizumab therapy) and response rate of 49% versus 32%, respectively. The OS HR was 0.97 (95% CI 0.86-1.08); median 26.7 versus 26.4 months, respectively. In patients with triple-negative mBC, the HRs for PFS and OS were 0.63 (95% CI 0.52-0.76) and 0.96 (95% CI 0.79-1.16), respectively. Median PFS was 8.1 months with bevacizumab versus 5.4 months with chemotherapy alone, median OS was 18.9 versus 17.5 months, respectively, and 1-year OS rates were 71% versus 65%. Bevacizumab improves efficacy, including 1-year OS rates, both overall and in subgroups of poor-prognosis patients with limited treatment options.


Th1 dominance in the immune response to live Salmonella typhimurium requires bacterial invasiveness but not persistence.
Factors responsible for the predictable generation of Th1 or Th2 immune responses to microorganisms in vivo are not well characterized, although the ability of antigen presenting cells (APC) to provide co-stimulation, the kinetics of MHC-peptide ligand generation as well as the cytokine environment are all considered important factors for the differential Th1/Th2 priming of T cells. Our earlier findings of an IFN-gamma-dominant, Th1-type response to live Salmonella typhimurium (Stm) and a Th2-type response to killed Stm suggested that persistence of viable bacteria might be an important factor in the generation of IFN-gamma-dominant responses. Using genetically susceptible and resistant strains of mice to limit bacterial replication and persistence in vivo, we show that mice of the lty(r) genotype, capable of a 10-fold better clearance of Stm, mount an IFN-gamma-dominant immune response following immunization with live Stm similar to that in the lty(s) strain. Further, metabolically defective mutants of Stm, aroA and purA, when used in the live form, also elicit IFN-gamma-dominant immune responses similar to the wild-type Stm strain despite their inability to proliferate in vivo. While a laboratory strain of Escherichia coli, which is antigenically cross-reactive but non-invasive, elicits hardly any IFN-gamma in immune responses, an invasive strain of E. coil induces an IFN-gamma-dominant response. These data together indicate that, while entry of bacteria into macrophages is likely to be critical for the generation of IFN-gamma-dominant immune responses, their persistence is not.


Causal association evaluation of diabetes with Alzheimer's disease and genetic analysis of antidiabetic drugs against Alzheimer's disease.
Despite accumulating epidemiological studies support that diabetes increases the risk of Alzheimer's disease (AD), the causal associations between diabetes and AD remain inconclusive. The present study aimed to explore: i) whether diabetes is causally related to the increased risk of AD; ii) and if so, which diabetes-related physiological parameter is associated with AD; iii) why diabetes drugs can be used as candidates for the treatment of AD. Two-sample Mendelian randomization (2SMR) was employed to perform the analysis. Firstly, the 2SMR analysis provided a suggestive association between genetically predicted type 1 diabetes (T1D) and a slightly increased AD risk (OR = 1.04, 95% CI = [1.01, 1.06]), and type 2 diabetes (T2D) showed a much stronger association with AD risk (OR = 1.34, 95% CI = [1.05, 1.70]). Secondly, further 2SMR analysis revealed that diabetes-related physiological parameters like fasting blood glucose and total cholesterol levels might have a detrimental role in the development of AD. Thirdly, we obtained 74 antidiabetic drugs and identified SNPs to proxy the targets of antidiabetic drugs. 2SMR analysis indicated the expression of three target genes, ETFDH, GANC, and MGAM, were associated with the increased risk of AD, while CPE could be a protective factor for AD. Besides, further PPI network found that GANC interacted with MGAM, and further interacted with CD33, a strong genetic locus related to AD. In conclusion, the present study provides evidence of a causal association between diabetes and increased risk of AD, and also useful genetic clues for drug development.


Autophagy Contributes to the Rapamycin-Induced Improvement of Otitis Media.
Otitis media (OM) is a pervasive disease that involves hearing loss and severe complications. In our previous study, we successfully established a mouse model of human OM using Tlr2tm1Kir (TLR2-/-) mice with middle ear (ME) inoculation of streptococcal peptidoglycan-polysaccharide (PGPS). In this study, we found that hearing loss and OM infections in OM mice were significantly alleviated after treatment with rapamycin (RPM), a widely used mechanistic target of RPM complex 1 (mTORC1) inhibitor and autophagy inducer. First of all, we tested the activity of mTORC1 by evaluating p-S6, Raptor, and mTOR protein expression. The data suggested that the protein expression level of p-S6, Raptor and mTOR are decreased in TLR2-/- mice after the injection of PGPS. Furthermore, our data showed that both the autophagosome protein LC3-II, Beclin-1, ATG7, and autophagy substrate protein p62 accumulated at higher levels in mice with OM than in OM-negative mice. The expression of lysosomal-associated proteins LAMP1, Cathepsin B, and Cathepsin D increased in the OM mice compared with OM-negative mice. Rab7 and Syntaxin 17, which is necessary for the fusion of autophagosomes with lysosomes, are reduced in the OM mice. In addition, data also described that the protein expression level of p-S6, mTOR and Raptor are lower than PGPS group after RPM treatment. The accumulation of LC3-II, Beclin-1, and ATG7 are decreased, and the expression of Rab7 and Syntaxin 17 are increased significantly after RPM treatment. Our results suggest that autophagy impairment is involved in PGPS-induced OM and that RPM improves OM at least partly by relieving autophagy impairment. Modulating autophagic activity by RPM may be a possible effective treatment strategy for OM.


Kinematic cross-correlation induces sensory integration across separate objects.
In a basic cursor-control task, the perceived positions of the hand and the cursor are biased towards each other. We recently found that this phenomenon conforms to the reliability-based weighting mechanism of optimal multisensory integration. This indicates that optimal integration is not restricted to sensory signals originating from a single source, as is the prevailing view, but that it also applies to separate objects that are connected by a kinematic relation (i.e. hand and cursor). In the current study, we examined which aspects of the kinematic relation are crucial for eliciting the sensory integration: (i) the cross-correlation between kinematic variables of the hand and cursor trajectories, and/or (ii) an internal model of the hand-cursor kinematic transformation. Participants made out-and-back movements from the centre of a semicircular workspace to its boundary, after which they judged the position where either their hand or the cursor hit the boundary. We analysed the position biases and found that the integration was strong in a condition with high kinematic correlations (a straight hand trajectory was mapped to a straight cursor trajectory), that it was significantly reduced for reduced kinematic correlations (a straight hand trajectory was transformed into a curved cursor trajectory) and that it was not affected by the inability to acquire an internal model of the kinematic transformation (i.e. by the trial-to-trial variability of the cursor curvature). These findings support the idea that correlations play a crucial role in multisensory integration irrespective of the number of sensory sources involved.


Oral pigmented lesions: a retrospective analysis from Brazil.
Pigmented lesions are uncommon in the oral mucosa, and studies investigating the incidence and types of these lesions are desired to improve the diagnostic knowledge of clinicians. The aim of this study was to analyze the distribution of oral pigmented lesions in a Brazilian population. A retrospective descriptive cross-sectional study was performed. Oral pigmented lesions were retrieved from the files of two oral and maxillofacial pathology services from Brazil over a 45-year period (1974-2019). The clinical data and the diagnoses of each case were retrieved and included in a Microsoft Excel® database. From 77.074 lesions diagnosed in this period, 761 (0.99%) represented pigmented lesions of the oral mucosa, including 351 (46.1%) melanocytic and 410 (53.9%) non-melanocytic lesions, with a higher incidence in females (73.2%) between the fourth and seventh decades of life. Amalgam tattoo (53.6%) represented the most common lesion, followed by melanotic macule (18.3%) and racial pigmentation (10.8%). Other pigmented lesions included nevus (9.9%), post-inflammatory pigmentation (3%), melanoma (2.1%), melanoacanthoma (1.4%), smoker's melanosis (0.4%), drug-induced pigmentation (0.3%), and melanotic neuroectodermal tumor of infancy (0.1%). The buccal mucosa was the most commonly affected site (25.2%), followed by the alveolar ridge (14.5%), and gingiva (11.8%). The current findings were similar to previous studies with minor differences due methodology and characteristics of the services from where lesions were retrieved. The knowledge of these data may contribute to a better understanding of oral pigmented lesions and assist clinicians to better recognize and manage them.


Investigation of ligand binding to the multidrug resistance protein EmrE by isothermal titration calorimetry.
Escherichia coli multidrug resistance protein E (EmrE) is an integral membrane protein spanning the inner membrane of Escherichia coli that is responsible for this organism's resistance to a variety of lipophilic cations such as quaternary ammonium compounds (QACs) and interchelating dyes. EmrE is a 12-kDa protein of four transmembrane helices considered to be functional as a multimer. It is an efflux transporter that can bind and transport cytoplasmic QACs into the periplasm using the energy of the proton gradient across the inner membrane. Isothermal titration calorimetry provides information about the stoichiometry and thermodynamic properties of protein-ligand interactions, and can be used to monitor the binding of QACs to EmrE in different membrane mimetic environments. In this study the ligand binding to EmrE solubilized in dodecyl maltoside, sodium dodecyl sulfate and reconstituted into small unilamellar vesicles is examined by isothermal titration calorimetry. The binding stoichiometry of EmrE to drug was found to be 1:1, demonstrating that oligomerization of EmrE is not necessary for binding to drug. The binding of EmrE to drug was observed with the dissociation constant (K(D)) in the micromolar range for each of the drugs in any of the membrane mimetic environments. Thermodynamic properties demonstrated this interaction to be enthalpy-driven with similar enthalpies of 8-12 kcal/mol for each of the drugs in any of the membrane mimetics.


Public-private partnership (3Ps) in ensuring safe use of medicines: An Indian experience.
Adverse drug reactions (ADRs) are major concerns to the public health. To monitor ADRs and ensure patients' safety, the Pharmacovigilance Programme of India (PvPI) has been established by the Government of India in 2010. The programme is intact with the Public-Private Partnership (3Ps) in pharmacovigilance for quality services, better management of human resources and risk minimization. The present work is aimed at assessing the 3Ps engagement, performance and tangible outcomes in PvPI and also mapping of resources. The study was carried out for the period of 2011 to 2021 by assessing the various benchmarking tools such as 3Ps categorization, utilization of ADRs reporting tools, trainings, and the Individual Case Safety Reports' (ICSRs) quantity, quality and transmission for regulatory intervention (RI). Under PvPI, Central or State Government medical institutions/hospitals and public health programmes constitute public partners while private medical institutions/hospitals, pharmaceutical companies, corporate hospitals and professional bodies account for private partners. We observed that public partners extensively used ADR reporting form and toll-free helpline number while private partners used mobile based app and emails/post as preferred tools for reporting ADRs. Contribution of public sector in training programmes organized, stakeholders trained and sharing of resource materials was way higher than the private sector. The study revealed that 55.1 and 44.9% ICSRs were received from public and private partners, respectively during the study period. The quality completeness of data received from public partners was found to be 0.92/1 as compared to 0.46/1 from the private partners. The ICSRs data transmitted for RI process from the public and private partners (till 2018) was found to be 79 and 21%, respectively. In terms of sharing of resources for training and capacity building, the public sector played a major role. The 3Ps in India are enabled to establish a robust system for medicines' safety surveillance; however a more focused approach is required in mapping the resources.


Preparation of Sulfonated Poly(arylene ether nitrile)-Based Adsorbent as a Highly Selective and Efficient Adsorbent for Cationic Dyes.
In this work, a highly selective and efficient polymer adsorbent inspired by a water-soluble sulfonated poly(arylene ether nitrile) (SPEN) was successfully synthesized. Due to the distinct structure of functional carboxyl, sulfonic acid and rigid benzene rings, a facile aluminium (III) ions crosslinking method was employed to fabricate the SPEN-based adsorbents (SPEN-Al). Among the three adsorbents, SPEN-Al-2 exhibited superior adsorption capacities with uniform morphology. Subsequently, the SPEN-Al-2 was selected as the adsorbent for three cationic dyes (rhodamine B (Rh B), neutral red (NR), methylene blue (MB)) and three anionic dyes (orange G (OG), methyl orange (MO), acid fuchsin (AF)), respectively, demonstrating that the adsorbent possessing excellent selectivity toward cationic dyes. Moreover, the dye's adsorption selectivity of SPEN-Al-2 was further certificated in a binary cationic-anionic dyes mixtures (MB/OG and MB/MO) system. Taking MB as a dye model, a series of factors (contact time, concentration, temperature and pH) and adsorption models were systematically investigated in dye adsorption experiments. Results indicated that the adsorption was endothermic and the maximum adsorption capacity of SPEN-Al-2 could reach up to 877.5 mg/g; pseudo-second-model and Langmuir model were fitted to the adsorption kinetics and equilibrium isotherm, respectively, manifesting that SPEN-Al adsorbent was promising in the dyes removing field.


Contextualization of automatic alerts during electronic prescription: researchers' and users' opinions on useful context factors.
Computerized Physician Order Entry (CPOE) Systems can reduce the number of medication errors and Adverse Drug Events (ADEs). However, studies have shown that users often override alerts, as they feel these are too unspecific for the given patient context. It is unclear, however, how alerts could be contextualized, that is adapted to the clinical context. Based on a literature search, we developed a list of 20 possible context factors. We asked 69 international CPOE researchers and 120 physicians from four hospitals in two countries to judge the usefulness of each factor. Researchers judged the following factors as most important: 1.) Severity of the effect, 2.) Clinical status of the patient, 3.) Probability of occurrence, 4.) Risk factors of the patient, 5.) Strength of evidence. Physicians judged the following factors as most important: Severity of the effect, clinical status of the patients, complexity of the case, and class of drug. These top-ranked context factors could be used to re-design the way alerts are presented in CPOE systems, to increase sensitivity of alerts, to reduce overriding rates, and to improve medication safety.


(+)-cis-3,5-dimethyl-2-(3-pyridyl) thiazolidin-4-one hydrochloride (SM-12502) as a novel substrate for cytochrome P450 2A6 in human liver microsomes.
(+)-cis-3,5-dimethyl-2-(3-pyridyl)thiazolidin-4-one hydrochloride (SM-12502) was oxidized by human liver microsomes to produce the S-oxide as a sole metabolite. Indirect evidence suggested that the S-oxidation was catalyzed by cytochrome P450 (CYP). Eadie-Hofstee plots showed biphasic pattern, suggesting that at least two enzymes were involved in the S-oxidation in human liver microsomes. Kinetic parameters of the S-oxidase with high-affinity showed Km and Vmax values of 20.9 +/- 4.4 microM and 0.111 +/- 0.051 nmol/min/mg microsomal protein, respectively. The S-oxidase activity was inhibited by coumarin and anti-CYP2A antibody. Among the contents of forms of CYP 20 samples of human liver microsomes, the content of CYP2A6 correlated with S-oxidase activity measured with 50 microM SM-12502 (r = .808, P < .0005). A close correlation (r = .908, P < .0001) was observed between activities of SM-12502 S-oxidase and coumarin 7-hydroxylase. Microsomes from genetically engineered human B-lymphoblastoid cells expressing CYP2A6 metabolized SM-12502 to the S-oxide efficiently. The results indicate that CYP2A6 isozyme is a major form of CYP responsible for the S-oxidation of SM-12502 in human liver microsomes. Thus, SM-12502 will be a useful tool in further research to analyze a human genetic polymorphism of CYP2A6.


The effect of local anesthetic on quality of recovery characteristics following dental rehabilitation under general anesthesia in children.
This study is a randomized, prospective, double-blind study to evaluate the effects of the combination of local anesthetics and an intravenous nonsteroidal anti-inflammatory drug (NSAID) vs NSAID alone on quality of recovery following dental rehabilitation under general anesthesia (GA). Twenty-seven healthy children aged 3-5.5 years underwent dental rehabilitation under GA. Fifteen children in the experimental group received oral infiltration of local anesthetic in addition to intravenous ketorolac tromethamine, while 12 children in the control group received intravenous ketorolac tromethamine alone for postoperative pain management. Pain behaviors were evaluated immediately postoperatively using a FLACC scale and 4 hours postoperatively by self-report using various scales. Parents reported perception of child pain and comfort and any occurrences of postoperative cheek biting. The use of intraoral infiltration local anesthesia for complete dental rehabilitation under general anesthesia for children aged 3-5.5 years did not result in improved pain behaviors in the postanesthesia care unit (PACU), nor did it result in improved pain behaviors 4-6 hours postoperatively as measured by the FLACC scale, FACES scale, and subjective reports of parents or a PACU nurse. Those children receiving local anesthesia had a higher incidence of negative symptoms related to local anesthetic administration, including a higher incidence of lip and cheek biting, which was of clinical importance, but not statistically significant. Infiltration of local anesthetic for dental rehabilitation under general anesthesia did not improve quality of recovery in children aged 3-5.5 years.


Electrolyte Imbalance in Children With Severe Acute Malnutrition at a Tertiary Care Hospital in Pakistan: A Cross-Sectional Study.
Background Malnutrition is a significant public health concern and a leading contributor to the global burden of children's diseases, affecting 50 to 150 million children under the age of five years worldwide. Globally, undernutrition accounts for approximately 33% of the deaths among under-fives. South Asia alone contributes to 50% and 38.8% of the world's population of wasted and stunted children, respectively. In Pakistan, malnutrition is the leading cause of childhood mortality, accounting for nearly 35% of all deaths under five years of age. Severe acute malnutrition (SAM), the most severe form of malnutrition, is often associated with electrolyte imbalances. This study aimed to determine the frequency of electrolyte imbalance in children with SAM admitted at a tertiary care hospital. Methods This cross-sectional study includes 184 patients with SAM aged between 6 and 60 months, who were admitted at the inpatient Department of Pediatrics, Dr. Ruth K. M. Pfau, Civil Hospital, Karachi, Pakistan, from January 17, 2017 to July 16, 2017. Weight and length/height were measured, and weight-for-height was calculated. Children were labeled to have SAM when weight-for-height was below -3 standard deviation (SD). Blood samples for serum electrolytes were drawn and sent to the lab. Descriptive statistics were calculated, and stratification was performed using the chi-square test. A p-value of ≤0.05 was considered statistically significant. Results The mean age of participants was 22.63 ± 12.71 months. Of the 184 patients with SAM, 172 (93.5%) patients had electrolyte imbalance. Hypokalemia was present in 79.9%, whereas hypocalcemia, hyponatremia, and hypomagnesemia were present in 71.7%, 48.9%, and 13.6%, respectively. Post-stratification results showed a significant association of electrolyte imbalance with gender (p = 0.005) and educational status of parents (p = 0.001). Conclusions Electrolyte disturbances are common in SAM. Serum electrolytes of every malnourished child admitted should be assessed and corrected to avoid fatal outcomes. We suggest that more research with better study designs should be conducted to develop policies and strategies for successfully combating malnutrition in Pakistan. In the meantime, we recommend adopting national guidelines for the management of acute malnutrition to reduce morbidity and mortality.


Early intervention with psychostimulants or antidepressants to increase methyl-CpG-binding protein 2 (MeCP2) expressions: a potential therapy for Rett syndrome.
Rett syndrome (RTT) is a severe X-linked postnatal neurodevelopmental disorder. The syndrome is caused primarily by mutations in the methyl CpG binding protein 2 (MeCP2) gene on Xq28. Most individuals with RTT are female, and female RTT is normally heterozygous for mutations in MeCP2. Patients with RTT display a normal period of development prior to the onset of symptoms, at which point they undergo a period of regression. Currently, no effective medication is available for this disorder, although animal studies have suggested that RTT symptoms are potentially reversible. For females with RTT, the severity of symptoms and progression of the disease varies a great deal, despite its homogenous genetic origin. These differences could be attributed to differences in the mutation points of MeCP2 and the skew caused by X-chromosome inactivation. Thus, the increased expression in the normal MeCP2 gene could decrease the severity of the disease. Based on findings from studies on animals indicating that fluoxetine (an antidepressant) and cocaine (a psychostimulant) can increase MeCP2 expression in the brain, it is suggested that early intervention with antidepressants or psychostimulants could increase the normal MeCP2 expression in females with RTT, who are normally heterozygous. This therapeutic hypothesis could be tested in an RTT animal model. Following the identification of the antidepressants or psychostimulants with the greatest influence on MeCP2 expression, a combination of early detection of the disorder with early intervention may result in improved therapeutic outcomes. Furthermore, a trial investigating the effects of antidepressants or psychostimulants on MeCP2 expression in lymphocyte culture from patients with RTT is suggested for clinical therapeutic prediction.


Acute inhibitory effects of 17 beta-estradiol are observed on gonadotropin secretion from perifused pituitary fragments of metestrous, but not proestrous, rats.
The in vivo suppression of LH by 17 beta-estradiol (E2) has been documented frequently. However, the demonstration of a direct inhibitory action of E2, in contrast to a stimulatory action, on the secretion of LH from the anterior pituitary has been inconsistent. The aim of this study was to determine if E2 can suppress either basal (unstimulated) or GnRH-stimulated gonadotropin secretion directly at the level of the anterior pituitary gland. Anterior pituitaries were obtained from metestrous and proestrous females rats at 0900 h, and trunk blood was collected for serum measurements of LH, FSH, E2, and progesterone (P). Each anterior pituitary was cut into eighths and placed into a microchamber for perifusion. Pituitary fragments were perifused at a rate of 10 ml/h using medium 199 (without phenol red) that contained E2 (1 nM) or ethanol as a control. Six pulses of GnRH (peak amplitude, 50 ng/ml; duration, 2 min) were administered one per h starting at 60 min. Fractions of perfusate were collected every 5 min for measurement of LH and FSH. The total amounts of LH and FSH secreted during the 1-h interval after each GnRH pulse or corresponding basal hour were calculated. Both basal and LH and FSH responses to GnRH were significantly greater from pituitaries of proestrous compared to metestrous rats. The selective suppression of LH secretion by in vitro treatment with E2 was demonstrated using pituitaries from metestrous rats receiving GnRH pulses, but not using pituitaries from proestrous rats. Thus, a negative feedback effect of E2 on LH secretion was observed only in pituitaries from donors with low serum levels of E2 and high P, but not from donors with high serum levels of E2 and low P. We believe that the in vivo steroid environment determined the subsequent responses to in vitro treatment with E2 on GnRH-stimulated gonadotropin secretion from the isolated pituitary gland.


Insulin-like growth factor binding protein-related protein 1 inhibits proliferation of MCF-7 breast cancer cells via a senescence-like mechanism.
Elevated insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) mRNA in senescent human mammary epithelial cells suggested that the IGFBP-3 gene product may inhibit cell proliferation. To test this hypothesis, we used a retroviral vector to express IGFBP-rP1 cDNA in the IGFBP-rP1-deficient MCF-7 breast cancer cell line. Compared with control vector-transduced cells, cumulative cell numbers for IGFBP-rP1-transduced polyclonal or clonal cell cultures were reduced by 39 and 74%, respectively, after 1 week. Medium conditioned by IGFBP-rP1-producing cultures reduced cumulative cell numbers in parental MCF-7 cultures by 20% compared with medium from cultures of a control vector-transduced cell line. Nuclear fragmentation analysis and cell proliferation assays completed in the presence of the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone excluded apoptosis as the responsible mechanism. The percentage of cells containing senescence-associated beta-galactosidase activity was doubled compared with control cell cultures. Flow cytometry analysis indicated that twice as many noncycling cells were present in the IGFBP-rP1-transduced MCF-7 cell cultures compared with controls. These findings indicate that IGFBP-rP1 is an inhibitor of MCF-7 breast cancer cell proliferation and may act via a cellular senescence-like mechanism.


Effect of destructive pulse duration on the detection of myocardial perfusion in myocardial contrast echocardiography: In vitro and in vivo observations.
Myocardial perfusion is detected with contrast echocardiography by comparing a contrast-enhanced image with a baseline obtained before contrast injection (true baseline) or after myocardial bubble destruction after a high-power destructive pulse (postdestructive pulse baseline). Although it is assumed that all bubbles are destroyed by a destructive pulse insuring optimal contrast detection, this assumption has not been tested. In 18 participants we compared the videointensity (VI) differences among the contrast-enhanced image, the postdestructive pulse baseline, and the true baseline using both triggered high-mechanical index imaging and real-time imaging. VI difference was significantly greater for the true baseline with both techniques at all ventricular levels. The benefit of using a true baseline was less when the duration of the destructive pulse was increased. Similarly, we quantified VI in a flow phantom using continuous Optison (commercially available perfluoropropane-filled albumin microbubbles) (Amersham, Princeton, NJ) infusion and variable durations of destructive pulses. VI decreased with the duration of the destructive pulse and reached a plateau after a duration of 8 to 15 frames. The plateau reached after a long destructive pulse was dependent on flow rate and concentration and never reached a true baseline, unless concentration (<100 microL/L) and flow rate (<0.5 cm/s) were very low. (1) in clinical studies, the difference in VI between contrast-enhanced and baseline images is greater when true baseline is used; (2) the longer the destructive pulse, the closer the postdestructive pulse baseline to true baseline; and (3) this effect exists in all regions of the left ventricle.


Integral Representation of Electrostatic Interactions inside a Lipid Membrane.
Interactions between charges and dipoles inside a lipid membrane are partially screened. The screening arises both from the polarization of water and from the structure of the electric double layer formed by the salt ions outside the membrane. Assuming that the membrane can be represented as a dielectric slab of low dielectric constant sandwiched by an aqueous solution containing mobile ions, a theoretical model is developed to quantify the strength of electrostatic interactions inside a lipid membrane that is valid in the linear limit of Poisson-Boltzmann theory. We determine the electrostatic potential produced by a single point charge that resides inside the slab and from that calculate charge-charge and dipole-dipole interactions as a function of separation. Our approach yields integral representations for these interactions that can easily be evaluated numerically for any choice of parameters and be further simplified in limiting cases.


Comparison of Different Blood Transfusion Methods in Patients Undergoing Cesarean Section.
To compare the effect of allogeneic transfusion and acute normovolemic hemodilution (ANH) autologous transfusion in patients undergoing cesarean section. Patients who underwent cesarean section and received blood transfusion therapy from February 2019 to July 2021 in our hospital were observed and divided into the allogeneic group (n = 55) who received allogeneic transfusion therapy and the autologous group (n = 55) who received ANH autologous transfusion therapy according to the mode of transfusion. Observations included vital signs [heart rate (HR), mean arterial pressure (MAP), stroke volume variation (SVV)], blood routine [red blood cells (RBC), platelets (PLT), hematocrit (HCT), hemoglobin (Hb)], T-cell subsets (CD4+, CD8+, CD4+/CD8+), immunoglobulins (IgA, IgM, IgG), inflammatory factors [C-reactive protein (CRP), tumor necrosis factor (TNF)-α, interleukin (IL)-6], and adverse effects were counted in both groups. There was no statistical significance in the intra-group and inter-group comparisons of HR, MAP, and SVV between the two groups before transfusion and transfusion for 10 min (P > 0.05). 5d after operation, the RBC, PLT, HCT, and Hb of the allogeneic group were lower than those before operation, and the autologous group was higher than that of the allogeneic group (P < 0.05). 5d after operation, the CRP, TNF-α, and IL-6 of the allogeneic group were higher than those before operation, and the autologous group was lower than that of the allogeneic group (P < 0.05). 5d after operation, the CD4+, CD4+/CD8+ of the allogeneic group were lower than before operation, and the CD8+ was higher than before operation. The CD4+ and CD4+/CD8+ of the autologous group were higher than that of the allogeneic group, and CD8+ was lower than that of the allogeneic group (P < 0.05). 5d after operation, the IgA, IgG, and IgM of the allogeneic group were lower than those before operation, and the autologous group was higher than that of the allogeneic group (P < 0.05). During blood transfusion, there was no significant difference in the adverse reaction rate between the two groups (P > 0.05). Both allogeneic transfusion and ANH autologous transfusion have little effect on the vital signs of patients undergoing cesarean section, but ANH autologous transfusion is more helpful to the stability of blood routine, T-cell subsets, immunoglobulin, and inflammation levels after surgery, which is a safe and effective way of blood transfusion.


Effects of mitogen-activated protein kinase signaling pathway inhibition on the development of cerulein-induced acute pancreatitis in mice.
Extracellular signal-regulated kinase (ERK) influences a number of pathways in all cells. The ERK cascade has long been known to be central to the activation of cellular processes such as proliferation, differentiation, and oncogenic transformation. The mitogen-activated protein (MAP) serine/threonine family of protein kinases, of which ERK is a member, is evolutionarily conserved and is activated by a mechanism that includes protein kinase cascades. The aim of this study was to investigate the effects of PD98059 [2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one], a highly selective inhibitor of MAP/ERK kinase 1 (MEK1) activation, on the development of acute pancreatitis. Pancreatitis was induced by intraperitoneal injection of cerulein (hourly × 5, 50 μg/kg) and PD98059 (10 mg/kg, 10% dimethylsulfoxide, intraperitoneally) was administrated 1 and 3 hours after cerulein administration. Cerulein injection resulted in acute necrotizing pancreatitis. On the contrary, pancreatitis histological features, amylase, lipase, pancreas edema, and immunohistochemical staining for leukocyte adhesion molecules, transforming growth factor β, and apoptosis-related proteins were found reduced in PD98059-treated mice. We propose that this study could help to clarify the role of MAPK in the regulation of the inflammatory process as acute pancreatitis.


Three protected tetrapeptides.
The structures of three protected tetrapeptides, containing the Boc-Gly-Gly-Phe-X-OMe chain, tert-butoxycarbonyl-glycy-glycl-phenylalanine-leucine methyl ester dihydrate, Boc-Gly-Gly-L-Phe-D-Leu-OMe, C25H38N4O7.2H2O, tert-butoxycarbonyl-glycy-glycl-phenylalanine-methionine methyl ester dihydrate, Boc-Gly-Gly-L-Phe-D-Met-OMe, C24H36N4O7S.2H2O and tert-butoxycarbonyl-glycy-glycl-phenylalanine-norleucine methyl ester dihydrate, Boc-Gly-Gly-D-Phe-L-Nle-OMe, C25H38N4O7.2H2O, are described. The three molecules have the same conformation on the tetrapeptide chain and display the same packing, consisting of couples of molecules linked head-to-tail by two hydrogen (N--H...O) bonds; other hydrogen bonds, also involving two water molecules of crystallization, link these couples together and give rise to a planar structure.


Firmicutes in Gut Microbiota Correlate with Blood Testosterone Levels in Elderly Men.
In males, testosterone levels have been implicated in various diseases. Recently, the influence of gut microbial-derived compounds on host metabolism has become evident, and it has been suggested that some gut bacteria may be involved in testosterone metabolism. In the present study, we examined the relationship between testosterone levels and gut microbiota in elderly Japanese men. We collected samples from Japanese male subjects suspected of having prostate cancer and underwent prostate biopsies and excluded patients with positive biopsies to avoid the effect of prostate cancer on the gut microbiota. In total, 54 Japanese males with negative biopsy results were included in our study. The gut microbiota was analyzed by 16S rRNA gene sequencing of bacterial DNA extracted from rectal swabs. Gut microbiota compositions were compared between the two groups according to the level of serum testosterone (above or below 3.5 ng /mL). The median age of the cohort was 71 years, and the quartile range was 67 to 73 years. We observed no significant difference in alpha or beta diversity, but some bacteria belonging to the phylum Firmicutes (Clostridiales, Turicibacter, and Gemella) were increased in the high testosterone group. Serum testosterone levels positively correlated with the relative amount of Firmicutes (rS=0.3323, p=0.0141), and the amount of Firmicutes affected serum testosterone levels independent of host factors (age, body mass index, triglyceride, and total cholesterol; β=0.770, p=0.0396). Some intestinal bacteria belonging to the phylum Firmicutes were associated with testosterone levels in elderly males. Therefore, the gut microbiota could affect testosterone metabolism in elderly males.


Bone marrow stem cell gene therapy of arylsulfatase A-deficient mice, using an arylsulfatase A mutant that is hypersecreted from retrovirally transduced donor-type cells.
Arylsulfatase A (ASA)-deficient mice represent an animal model for the fatal lysosomal storage disease metachromatic leukodystrophy, which is characterized by widespread intralysosomal deposition of sulfatide. Bone marrow stem cell gene therapy in mice, using a retroviral vector mediating expression of wild-type human ASA, has the potential to ameliorate the visceral pathology, but improves the prevailing brain disease and neurologic symptoms only marginally. One factor that influences the efficacy of bone marrow transplantation therapy in lysosomal storage diseases is the secretion level of the therapeutic enzyme from donor-type cells. Here we test the potential of a hypersecreted glycosylation variant of ASA. Although this mutant lacks mannose 6-phosphate residues it is taken up by cells by a mannose 6-phosphate receptor-independent pathway and causes partial metabolic correction of ASA-deficient mouse cells. Retrovirally mediated transfer of the mutant cDNA into ASA-deficient mice results in the sustained expression of the transgene. Serum levels argue for an increased secretion of the glycosylation mutant also in vivo. Tissue levels were reduced to 2% in liver and up to 40% in kidney compared with animals treated with the wild-type enzyme, indicating reduced endocytosis. Thus, the limited uptake of the variant enzyme outweighs the putative advantageous effect of improved supply. Although the mutant enzyme is able to correct the metabolic defect partially, histological examinations did not reveal any reduction of sulfatide storage in treated animals. Surprisingly, analysis of neurologic symptoms indicated a significant improvement of the gait pattern.


Creating Conditions for Professional Development through a Trauma-Informed and Restorative Practice.
Professionals in social work and nursing meet people who, due to trauma, struggle with self-regulation of emotions and social behavior. Caring for trauma survivors requires connection and compassion. Previous research has indicated that many professionals, some with their own trauma histories, do not feel sufficiently equipped to practice self-care or cope with challenging communication. To address how insecurity and trauma are rooted in deeper individual and structural causes of social injustice, social workers and nursing students (N = 29) were invited to participate in action research and a restorative circle process. The aim was to provide professionals the opportunity to challenge and reflect upon their own practices by introducing them to a trauma-informed and restorative practice based on the importance of building emotional safety and connection. The findings suggest that given time and space for reflection in a setting characterized by equality and safety, professionals can develop increased (a) self-awareness and compassion, (b) emotional safety and tolerance of stress in challenging communication, (c) trauma understanding, and (d) personal growth. A change to a more restorative and trauma-informed practice could lead to increased interpersonal competence and well-being among professionals and clients.


N-type voltage-dependent calcium channels mediate the nicotinic enhancement of GABA release in chick brain.
The role of voltage-dependent calcium channels (VDCCs) in the nicotinic acetylcholine receptor (nAChR)-mediated enhancement of spontaneous GABAergic inhibitory postsynaptic currents (IPSCs) was investigated in chick brain slices. Whole cell recordings of neurons in the lateral spiriform (SpL) and ventral lateral geniculate (LGNv) nuclei showed that cadmium chloride (CdCl2) blocked the carbachol-induced increase of spontaneous GABAergic IPSCs, indicating that VDCCs might be involved. To conclusively show a role for VDCCs, the presynaptic effect of carbachol on SpL and LGNv neurons was examined in the presence of selective blockers of VDCC subtypes. omega-Conotoxin GVIA, a selective antagonist of N-type channels, significantly reduced the nAChR-mediated enhancement of gamma-aminobutyric acid (GABA) release in the SpL by 78% compared with control responses. Nifedipine, an L-type channel blocker, and omega-Agatoxin-TK, a P/Q-type channel blocker, did not inhibit the enhancement of GABAergic IPSCs. In the LGNv, omega-Conotoxin GVIA also significantly reduced the nAChR-mediated enhancement of GABA release by 71% from control values. Although omega-Agatoxin-TK did not block the nicotinic enhancement, L-type channel blockers showed complex effects on the nAChR-mediated enhancement. These results indicate that the nAChR-mediated enhancement of spontaneous GABAergic IPSCs requires activation of N-type channels in both the SpL and LGNv.


Botulinum neurotoxin serotype D attacks neurons via two carbohydrate-binding sites in a ganglioside-dependent manner.
The extraordinarily high toxicity of botulinum neurotoxins primarily results from their specific binding and uptake into neurons. At motor neurons, the seven BoNT (botulinum neurotoxin) serotypes A-G inhibit acetylcholine release leading to flaccid paralysis. Uptake of BoNT/A, B, E, F and G requires a dual interaction with gangliosides and the synaptic vesicle proteins synaptotagmin or SV2 (synaptic vesicle glycoprotein 2), whereas little is known about the cell entry mechanisms of the serotypes C and D, which display the lowest amino acid sequence identity compared with the other five serotypes. In the present study we demonstrate that the neurotoxicity of BoNT/D depends on the presence of gangliosides by employing phrenic nerve hemidiaphragm preparations derived from mice expressing the gangliosides GM3, GM2, GM1 and GD1a, or only GM3 [a description of our use of ganglioside nomenclature is given in Svennerholm (1994) Prog. Brain Res. 101, XI-XIV]. High-resolution crystal structures of the 50 kDa cell-binding domain of BoNT/D alone and in complex with sialic acid, as well as biological analyses of single-site BoNT/D mutants identified two carbohydrate-binding sites. One site is located at a position previously identified in BoNT/A, B, E, F and G, but is lacking the conserved SXWY motif. The other site, co-ordinating one molecule of sialic acid, resembles the second ganglioside-binding pocket (the sialic-acid-binding site) of TeNT (tetanus neurotoxin).


The role of female volunteers in disaster response organisations: A qualitative research.
Despite a high demand for people to participate in disaster volunteering, the extent of contribution received from female volunteers in disaster response organisations constitutes an important debate. In the Elazığ earthquake on 24 January 2020, the role of female volunteers in rescuing a female victim from the wreckage once again demonstrated the importance of considering female volunteers when creating response capacities for disaster risks. This qualitative study aims to evaluate the roles of female volunteers who play an active role in disaster response organisations in a society with gender equality issues. This qualitative study included 15 National Medical Rescue Team (UMKE) female volunteers who took an active role in the Elazig earthquake and was performed between 15.07.2020 and 16.04.2021. Data were collected using an in-depth interview method. The data were analyzed using content analysis and the comments were given descriptively. Based on the qualitative analysis, the following concepts were explored among the participants: Volunteering, volunteering in disasters, the effects of volunteering on social life, being a woman in disasters, volunteering female in disasters, the 'motherhood myth', gender inequality. Women are key actors in tackling disaster damage and increasing available social resilience to disasters. In addition, women can use special attributes such as motherhood, which is socio-culturally defined and taught as an advantage, for the common good of society. Moreover, special elements, such as the image of the 'superwoman' in relation to the responsibilities imposed on women in disaster response organisations, which distort gender equality against women, appear as possible factors of harm for women. This study shows that female volunteers can play a life-saving role in chaotic situations such as disasters, contribute to the empowerment of women, increase sensitivity to gender and, in connection with this, the development of democracy in societies where gender inequality is high. On the other hand, special situations such as the image of 'superwoman', which aims for excellence in both private and public spheres in the responsibilities imposed on women, can destroy gender equality against women and increase women's fragility.