Re-evaluation of Sentinel Lymph Node Biopsy for Melanoma.
The vast majority of patients newly diagnosed with melanoma present with clinically localized disease, and sentinel lymph node biopsy (SLNB) is a standard of care in the management of these patients, particularly in intermediate thickness cases, in order to provide important prognostic data. However, SLNB also has an important role in the management of patients with other subtypes of melanoma such as thick melanomas, certain thin melanomas, and specific histologic variants of melanoma such as desmoplastic melanoma. Furthermore, there have been technical advances in the SLNB technique, such as the development of newer radiotracers and use of SPECT/CT, and there is some data to suggest performing a SLNB may be therapeutic. Finally, the management of patients with a positive sentinel lymph node (SLN) has undergone dramatic changes over the past several years based on the results of recent important clinical trials. Treatment options for patients with SLN metastases now include surveillance, completion lymph node dissection, and adjuvant therapy with checkpoint inhibitors and targeted therapy. SLNB continues to play a crucial role in the management of patients with melanoma, allowing for risk stratification, potential regional disease control, and further treatment options for patients with a positive SLN.


Expression and localization of the serine proteases high-temperature requirement factor A1, serine protease 23, and serine protease 35 in the mouse ovary.
Proteolytic degradation of extracellular matrix components has been suggested to play an essential role in the occurrence of ovulation. Recent studies in our laboratory have indicated that the plasminogen activator and matrix metalloproteinase systems, which were previously believed to be crucial for ovulation, are not required in this process. In this study we have used a microarray approach to identify new proteases that are involved in ovulation. We found three serine proteases that were relatively highly expressed during ovulation: high-temperature requirement factor A1 (HtrA1), which was not regulated much during ovulation; serine protease 23 (PRSS23), which was down-regulated by gonadotropins; and serine protease 35 (PRSS35), which was up-regulated by gonadotropins. We have further investigated the expression patterns of these proteases during gonadotropin-induced ovulation in immature mice and in the corpus luteum (CL) of pseudopregnant mice. We found that HtrA1 was highly expressed in granulosa cells throughout follicular development and ovulation, as well as in the forming and regressing CL. PRSS23 was highly expressed in atretic follicles, and it was expressed in the ovarian stroma and theca tissues just before ovulation. PRSS35 was expressed in the theca layers of developing follicles. It was also highly induced in granulosa cells of preovulatory follicles. PRSS35 was also expressed in the forming and regressing CL. These data suggest that HtrA1 and PRSS35 may be involved in ovulation and CL formation and regression, and that PRSS23 may play a role in follicular atresia.


Lung ultrasonography findings in dogs with various underlying causes of cough.
To characterize lung ultrasonography (LUS) findings in dogs with a primary clinical complaint of cough. 100 client-owned coughing dogs. A standardized LUS examination was performed for all dogs to quantify the number of B lines and identify subpleural abnormalities at 4 sites on each hemithorax. The final clinical diagnosis (reference standard) was determined by medical record review, and sensitivity and specificity of LUS for the diagnosis of selected causes of cough was determined. Common underlying causes of cough included dynamic airway collapse (n = 37), cardiogenic pulmonary edema (CPE; 12), and bronchitis (10). Compared with dogs with other causes of cough, dogs with bacterial pneumonia (n = 7) were more likely to have subpleural shred signs, whereas dogs with pulmonary neoplasia (4) were more likely to have subpleural nodule signs. Dogs with CPE had higher total B-line scores and higher numbers of LUS sites strongly positive for B lines (> 3 B lines/site) than other dogs. The LUS criteria of total B-line score ≥ 10 and presence of ≥ 2 sites strongly positive for B lines were each 92% sensitive and 94% specific for CPE diagnosis. Notably, 18% (16/88) of dogs with noncardiac causes of cough had been treated previously with diuretics because of prior CPE misdiagnosis. LUS profiles in dogs with cough differed by the underlying cause. In dogs with a clinical history of cough, this imaging modality could be diagnostically useful, particularly to help exclude the possibility of underlying CPE.


Processing speed and executive attention as causes of intelligence.
Individual differences in processing speed and executive attention have both been proposed as explanations for individual differences in cognitive ability, particularly general and fluid intelligence (Engle et al., 1999; Kail & Salthouse, 1994). Both constructs have long intellectual histories in scientific psychology. This article attempts to describe the historical development of these constructs, particularly as they pertain to intelligence. It also aims to determine the degree to which speed and executive attention are theoretical competitors in explaining individual differences in intelligence. We suggest that attention is the more fundamental mechanism in explaining variation in human intelligence. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Temporal proteomic analysis and label-free quantification of viral proteins of an invertebrate iridovirus.
Invertebrate iridescent virus 6 (IIV-6) is a nucleocytoplasmic virus with a ~212 kb linear dsDNA genome that encodes 215 putative ORFs. The IIV-6 virion-associated proteins consist of at least 54 virally encoded proteins. One of our previous findings showed that most of these proteins are encoded by genes from the early transcriptional class. This indicated that these structural proteins may not only function in the formation of the virion, but also in the initial stage of viral infection. In the current study, we followed the protein expression profile of IIV-6 over time in Drosophila S2 cells by label-free quantification using a proteomic approach. A total of 95 virally encoded proteins were detected in infected cells, of which 37 were virion proteins. The expressed IIV-6 virion proteins could be categorized into three main clusters based on their expression profiles: proteins with stably low expression levels during infection, proteins with exponentially increasing expression levels during infection and proteins that were initially highly abundant, but showed slightly reduced levels after 48 h post-infection. We thus provided novel information on the kinetics of virion and infected cell-specific protein levels that assists in our understanding of gene regulation in this lesser-known DNA virus model.


Force-induced growth of adhesion domains is controlled by receptor mobility.
In living cells, adhesion structures have the astonishing ability to grow and strengthen under force. Despite the rising evidence of the importance of this phenomenon, little is known about the underlying mechanism. Here, we show that force-induced adhesion-strengthening can occur purely because of the thermodynamic response to the elastic deformation of the membrane, even in the absence of the actively regulated cytoskeleton of the cell, which was hitherto deemed necessary. We impose pN-forces on two fluid membranes, locally pre-adhered by RGD-integrin binding. One of the binding partners is always mobile whereas the mobility of the other can be switched on or off. Immediate passive strengthening of adhesion structures occurs in both cases. When both binding partners are mobile, strengthening is aided by lateral movement of intact bonds as a transient response to force-induced membrane-deformation. By extending our microinterferometric technique to the suboptical regime, we show that the adhesion, as well as the resistance to force-induced de-adhesion, is greatly enhanced when both, rather than only one, of the binding partners are mobile. We formulate a theory that explains our observations by linking the macroscopic shape deformation with the microscopic formation of bonds, which further elucidates the importance of receptor mobility. We propose this fast passive response to be the first-recognition that triggers signaling events leading to mechanosensing in living cells.


Flavan-3-ols consumption and cancer risk: A meta-analysis of epidemiologic studies.
Although numerous in vitro studies and animal model data have suggested that flavan-3-ols, the most common subclass of flavonoids in the diet, may exert protective effects against cancer, epidemiologic studies have reported inconclusive results for the association between flavan-3-ols intake and cancer risk. Therefore, we conducted this meta-analysis of epidemiologic studies to investigate the preventive effects of flavan-3-ols on various types of cancers. A total of 43 epidemiologic studies, consisting of 25 case-control and 18 prospective cohort studies, were included. A significant inverse association was shown between flavan-3-ols intake and the risk of overall cancer (relative risk (RR) 0.935, 95%CI: 0.891-0.981). When cancer types were separately analyzed, a statistically significant protective effect of flavan-3-ols consumption was observed in rectal cancer (RR 0.838, 95%CI: 0.733-0.958), oropharyngeal and laryngeal cancer (RR 0.759, 95%CI: 0.581-0.993), breast (RR 0.885, 95%CI: 0.790-0.991) in case-control studies and stomach cancer in women (RR 0.633, 95%CI: 0.468-0.858). Our analysis indicates the potential benefits of flavan-3-ols in cancer prevention.


Variation in HLA class I antigen-processing genes and susceptibility to human papillomavirus type 16-associated cervical cancer.
Persistent infection with human papillomavirus type 16 (HPV16) is a primary etiological factor for the development of cervical cancer. Genes involved in antigen processing influence both the repertoire of antigens presented by HPV16-infected cells and the nature of HPV16-specific immune responses. Genetic variation in these genes may affect protein structure and function and, consequently, the ability of an individual to clear HPV infection. Thirty-five single-nucleotide polymorphisms (SNPs) in 5 genes (LMP2, TAP1, LMP7, TAP2, and Tapasin) were investigated for association with susceptibility to HPV16-associated cervical cancer. Sequencing of these genes resulted in the discovery of 15 previously unreported SNPs. Microsphere-array flow cytometry-based genotyping was conducted on 787 samples from Hispanic and non-Hispanic white women (241 randomly selected control subjects, 205 HPV16-positive control subjects, and 341 HPV16-positive case subjects with cervical cancer). For 9 SNPs, 8 of which had not previously been reported in the context of cervical cancer, there were statistically significant differences between the genotype distribution in case subjects and that in control subjects. Haplotype analysis of 3 haplotype blocks revealed 3 haplotypes with significant differences in frequency in case-control comparisons. Both HPV16-specific and non-type-specific differences in genotype distribution were seen. Genes involved in antigen processing for HLA class I presentation may contribute to susceptibility to cervical cancer.


Serum C-reactive protein as a marker for infection and inflammation in regular dialysis patients.
Serum C-reactive protein (CRP) concentration was measured in 98 hemodialysis (HD) patients and 68 continuous ambulatory peritoneal dialysis patients (CAPD). The effect, if any, of a HD session on serum CRP level and the usefulness of CRP in diagnosing and monitoring proven inflammatory disease were studied. Seventy-five percent of CAPD patients without evidence of inflammation had CRP levels within the quoted normal range (< 10 mg/l), as compared with only 47% of HD patients also without evidence of overt inflammation (p < 0.001). No significant change in the CRP level was detected during a hemodialysis session. Twenty-four patients with overt inflammation (due mainly to infection) were studied, 22 of whom had CRP levels of > 50 mg/l. These increased CRP levels fell with treatment. A CRP level of > 50 mg/l proved highly suggestive of a significant inflammatory process and a value of < 10 mg/l always excluded it. The total period of time on the regular dialysis program was not related to the CRP level. A subclinical inflammatory response resulting in a raised CRP is very common in our hemodialysis patients and less so in CAPD patients. Possible reasons for this are discussed. However, CRP is still useful in the diagnosis and monitoring of response to treatment of inflammatory disease if the level is > 50 mg/l or serial/baseline measurements are available for comparison. Whether any relationship exists between elevation of CRP and the well-documented increased risk of cardiovascular death in dialysis patients is currently unknown.


[Ultrasonography in the diagnosis of metastases of cancer of the large intestine to the liver].
Ultrasonography using a method of sectional and contact scanning is a valuable method of diagnosis of metastatic liver involvement in colon cancer. The diagnostic accuracy of the method is 95.6%. It has been noted that metastases of the same colon tumor type have different echostructure which depends on a period of disease and size of metastases. The author has defined 6 types of changes in the echostructure of metastases. In individual cases in a certain structure of metastasis changes in the liver parenchyma at ultrasonography were undetectable.


The functional consequences of the microbiome in HIV: insights from metabolomic studies.
It is critical to gain insight into the metabolic pathways by which the microbiota might influence HIV immunopathogenesis to exploit host-microbiota interactions. The aim of this review is to sketch a very broad picture of recent advances in our knowledge of how HIV might affect the microbiota, with a focus on specific gene products, particularly, metabolites produced by the microbiota that may affect HIV immunopathogenesis. First, we describe the different approaches used to explore imbalances in effector microbial products during HIV infection. Then, we review the mechanisms by which the microbiota might affect HIV immunopathogenesis. We cover several aspects of HIV immunopathogenesis, including systemic inflammation, mucosal immunity, enterocyte barrier integrity, HIV persistence and effects on HIV-specific humoral and cytotoxic responses. The altered interplay between mucosal immunity and dysbiotic bacteria helps to explain poorly understood observations in HIV infection, including susceptibility to HIV acquisition or the risk of HPV-related cancers, lung infections and cardiovascular disease. Although there is an urgent need to standardize the methods used for assessing the functional level of the microbiota, it is recognized that functional modulation of the microbiota for therapeutic purposes should be evaluated to improve HIV care.


Prevalence of SARS-CoV-2 Antibodies in a Multistate Academic Medical Center.
To estimate the seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies in health care personnel. The Mayo Clinic Serology Screening Program was created to provide a voluntary, two-stage testing program for SARS-CoV-2 antibodies to health care personnel. The first stage used a dried blood spot screening test initiated on June 15, 2020. Those participants identified as reactive were advised to have confirmatory testing via a venipuncture. Venipuncture results through August 8, 2020, were considered. Consent and authorization for testing was required to participate in the screening program. This report, which was conducted under an institutional review board-approved protocol, only includes employees who have further authorized their records for use in research. A total of 81,113 health care personnel were eligible for the program, and of these 29,606 participated in the screening program. A total of 4284 (14.5%) of the dried blood spot test results were "reactive" and warranted confirmatory testing. Confirmatory testing was completed on 4094 (95.6%) of the screen reactive with an overall seroprevalence rate of 0.60% (95% CI, 0.52% to 0.69%). Significant variation in seroprevalence was observed by region of the country and age group. The seroprevalence for SARS-CoV-2 antibodies through August 8, 2020, was found to be lower than previously reported in other health care organizations. There was an observation that seroprevalence may be associated with community disease burden.


Urokinase-type plasminogen activator and its inhibitor plasminogen activator inhibitor-1.
Tumor associated proteolysis is an essential mechanism in invasion and metastasis of cancer. The influence of the serine protease urokinase-like plasminogen activator (u-PA) and its inhibitor plasminogen activator inhibitor-1 (PAI-1) on the clinical prognosis of squamous carcinoma of the head and neck region (HNSCC) was evaluated. U-PA and PAI-1 levels were measured in tumor biopsies of 41 HNSCC patients and 6 biopsies of healthy oral mucosa using ELISA technique. Patients were followed for an average of 24 months. U-PA concentration in tumor tissue was four times higher than in healthy mucosa (4.96 ng/mg protein versus 1.32 ng/mg). PAI-1 levels were 22 times higher (69.55 ng/mg versus 3.18 ng/mg). Univariate Cox regression analysis revealed significant correlation (p=0.022) of PAI-1 with recurrence of the disease and no significance for u-PA. PAI-1 might become a new functional risk factor reflecting clinical prognosis.


Resonance instability of axially symmetric magnetostatic equilibria.
We review the evidence for and against the possibility that a strong enough poloidal field stabilizes an axisymmetric magnetostatic field configuration. We show that there does exist a class of resonant magnetohydrodynamic (MHD) waves which produce instability for any value of the ratio of poloidal and toroidal field strength. We argue that recent investigations of the stability of mixed poloidal and toroidal field configurations based on three-dimensional numerical simulations can miss this instability because of the very large azimuthal wave numbers involved and its resonant character.


Engineering of microorganisms towards recovery of rare metal ions.
The bioadsorption of metal ions using microorganisms is an attractive technology for the recovery of rare metal ions as well as removal of toxic heavy metal ions from aqueous solution. In initial attempts, microorganisms with the ability to accumulate metal ions were isolated from nature and intracellular accumulation was enhanced by the overproduction of metal-binding proteins in the cytoplasm. As an alternative, the cell surface design of microorganisms by cell surface engineering is an emerging strategy for bioadsorption and recovery of metal ions. Cell surface engineering was firstly applied to the construction of a bioadsorbent to adsorb heavy metal ions for bioremediation. Cell surface adsorption of metal ions is rapid and reversible. Therefore, adsorbed metal ions can be easily recovered without cell breakage, and the bioadsorbent can be reused or regenerated. These advantages are suitable for the recovery of rare metal ions. Actually, the cell surface display of a molybdate-binding protein on yeast led to the enhanced adsorption of molybdate, one of the rare metal ions. An additional advantage is that the cell surface display system allows high-throughput screening of protein/peptide libraries owing to the direct evaluation of the displayed protein/peptide without purification and concentration. Therefore, the creation of novel metal-binding protein/peptide and engineering of microorganisms towards the recovery of rare metal ions could be simultaneously achieved.


Blood flow through new microvessels: factors that affect regrowth of vasa vasorum.
A segment of abdominal aorta in dogs was transplanted to its original position (high PO2) or to the femoral vein (low PO2), and a segment of thoracic aorta was transplanted to its original position. Two days after grafting to each location, there was no blood flow (measured with microspheres) through vasa vasorum. After 2 and 4 wk, blood flow through vasa was restored to normal, or above normal, levels in all grafts. Vasa in grafts did not have smooth muscle, and they were not responsive to adenosine. Medial necrosis occurred in grafts of thoracic aorta but not in grafts of abdominal aorta. There was cellular degeneration and medial atrophy in grafts of abdominal aorta to the femoral vein. We conclude that 1) revascularization of the aorta by vasa vasorum is rapid; 2) new vasa do not have smooth muscle, and they are not responsive to vasoactive stimuli; and 3) medial necrosis occurs in grafts in which delivery of oxygen to media is compromised, either by low luminal PO2 or by interruption of medial vasa (in thoracic aorta), but not when the abdominal aorta is excised and reanastomosed. Absence of medial necrosis in grafts of abdominal aorta indicates that diffusion from the lumen of the vessel is adequate to nourish the vessel. This finding provides evidence that adventitial vasa, in contrast to medial vasa, are not essential for nourishment of the vessel wall.


Metal-Containing Formate Dehydrogenases, a Personal View.
Mo/W-containing formate dehydrogenases (FDH) catalyzes the reversible oxidation of formate to carbon dioxide at their molybdenum or tungsten active sites. The metal-containing FDHs are members of the dimethylsulfoxide reductase family of mononuclear molybdenum cofactor (Moco)- or tungsten cofactor (Wco)-containing enzymes. In these enzymes, the active site in the oxidized state comprises a Mo or W atom present in the bis-Moco, which is coordinated by the two dithiolene groups from the two MGD moieties, a protein-derived SeCys or Cys, and a sixth ligand that is now accepted as being a sulfido group. SeCys-containing enzymes have a generally higher turnover number than Cys-containing enzymes. The analogous chemical properties of W and Mo, the similar active sites of W- and Mo-containing enzymes, and the fact that W can replace Mo in some enzymes have led to the conclusion that Mo- and W-containing FDHs have the same reaction mechanism. Details of the catalytic mechanism of metal-containing formate dehydrogenases are still not completely understood and have been discussed here.


Recalling routes around london: activation of the right hippocampus in taxi drivers.
Functional imaging to date has examined the neural basis of knowledge of spatial layouts of large-scale environments typically in the context of episodic memory with specific spatiotemporal references. Much human behavior, however, takes place in very familiar environments in which knowledge of spatial layouts has entered the domain of general facts often referred to as semantic memory. In this study, positron emission tomography (PET) was used to examine the neural substrates of topographical memory retrieval in licensed London taxi drivers of many years experience while they recalled complex routes around the city. Compared with baseline and other nontopographical memory tasks, this resulted in activation of a network of brain regions, including the right hippocampus. Recall of famous landmarks for which subjects had no knowledge of their location within a spatial framework activated similar regions, except for the right hippocampus. This suggests that the hippocampus is involved in the processing of spatial layouts established over long time courses. The involvement of similar brain areas in routes and landmarks memory indicates that the topographical memory system may be primed to respond to any relevant topographical stimulation; however, the right hippocampus is recruited specifically for navigation in large-scale spatial environments. In contrast, nontopographical semantic memory retrieval involved the left inferior frontal gyrus, with no change in activity in medial temporal regions.


Improvements in patient blood management for pediatric craniosynostosis surgery using a ROTEM(®) -assisted strategy - feasibility and costs.
Moderate to severe intraoperative bleeding and the presence of acquired coagulopathy remain serious problems in the management of major pediatric craniosynostosis surgery. After implementation of a ROTEM(®) -assisted patient blood management (PBM) strategy, using primarily purified coagulation factor concentrates, feasibility and costs of this new regimen were analyzed. Retrospective analysis of all consecutive children who underwent primary elective major craniofacial surgery for craniosynostosis repair was carried out at the Children's University Hospital, Zurich, between 2007 and 2013. Laboratory workup and transfusion requirements were compared. A total of 47 children (36 in the historic group and 11 after implementation of PBM) were analyzed. Although all patients in this study needed transfusion of red blood cell concentrates, there was a total avoidance of perioperative transfusion of fresh frozen plasma and a reduction in transfused platelets (one of nine children vs nine of 36 children in the historic group) after implementation of the PBM strategy. Based on a predefined ROTEM(®) threshold in the PBM group (FibTEM MCF <8 mm), administration of fibrinogen concentrate was necessary in all of these children. The mean total costs per patient consisting of transfused allogeneic blood products and coagulation factor concentrates were reduced by 17.1% after implementation of PBM (1071.82 EUR per patient before vs 888.93 EUR after implementation). The implementation of a ROTEM(®) -assisted PBM is feasible and is associated with a considerable reduction in intraoperative transfusion requirements and thereby a decrease in transfusion-related direct costs.


Youth at the intersection of parental incarceration and foster care: Examining prevalence, disparities, and mental health.
While there are various pathways by which children experience parental incarceration or foster care, involvement in either system is associated with adverse health outcomes. Despite co-occurring risk factors for parental incarceration and foster care, little is known about the prevalence or characteristics of youth navigating both of these experiences. This study details the prevalence of youth at the intersection of parental incarceration and foster care, their demographic characteristics, and heterogeneity in their mental health. Data come from the 2019 Minnesota Student Survey with 112,157 eighth-, ninth-, and eleventh-grade students. Logistic regression with interactions between parental incarceration and foster care predict associated odds of youth's anxiety and depression; self-injurious behavior, suicidal ideation and attempt; and mental health diagnoses and treatment. Nearly 2 % of students experienced both parental incarceration and foster care, with a disproportionate number of those identifying as youth of color, experiencing poverty, and/or living in rural communities. Both parental incarceration and foster care were separately linked with poor mental health, yet experiencing both was associated with higher odds of anxiety, depression, self-injury, suicidal ideation, suicide attempt, diagnosis, and treatment. Youth with proximal multiplicative exposure (recent foster care and current parental incarceration) reported the most adverse mental health symptoms. The study emboldens what is known about the inequitable distribution of parental incarceration and foster care. These findings highlight the association between dual-systems-impacted youth and mental health indicators, with important implications for increasing access to mental health services while simultaneously calling for systems change.


Virulence genotyping of Escherichia coli isolates from diarrheic and urinary tract infections in relation to phylogeny in southeast of Iran.
The purpose of this study was to determine the prevalence of virulence genes and phylogenetic groups/subgroups of Escherichia coli (E. coli) isolates from diarrheic and urinary tract infections (UTI) cases in Rigan area, southeast of Iran. One hundred thirty five E. coli were isolated from diarrheic (90 isolates) and urinary tract infections (45 isolates) samples. The confirmed isolates were examined to detect the phylogenetic group/subgroups and a selection of virulence genes including iucD, sfa/focDE, afaIBC, papEF, hly, cnfI and cdtI by PCR. The examined isolates belonged to four phylogenetic groups A (42.2%), B1 (14.1%), B2 (10.4%), and D (33.3%). Among 135 tested bacteria, 62.22% of diarrheic and 30.37% of UTI isolates had at least one of the virulence genes. In the diarrheic isolates iucD (47.77%) was the most prevalent gene. The other genes including sfa/focDE, afaIBC, papEF and cnfI/cdtI genes were detected in 15, 13, 11 and one diarrheic isolates respectively. None of the diarrheic isolates were positive for hly gene. Out of 45 UTI isolates 28.88% were positive for iucD, 13.33% for cnfI, 11.11% for afaIBC, 11.11% for papEF, 6.66% for sfa/focDE and 4.44% for cdtI genes. Several combination patterns of the virulence genes were detected in diarrheic and UTI isolates. In conclusion, the prevalence of virulence genes in diarrheic and UTI isolates differ according to phylogenetic groups, although B2 and D phylotypes have an accumulation of virulence associated genes.


Optical forces on small particles: attractive and repulsive nature and plasmon-resonance conditions.
A detailed study of time-averaged electromagnetic forces on subwavelength-sized particles is presented. An analytical decomposition of the force into gradient and scattering-plus-absorption components is carried out, on the basis of which the attractive or repulsive behavior of the force is explained. Small metallic particles are shown to experience both kinds of forces; which kind also depends on the excitation of surface plasmons. Resonances give rise to enhancements of both the scattering and the absorption forces, but the gradient force can become negligible. Also, close to resonant wavelengths, the gradient force can be maximum, while both the scattering and the absorption forces remain large. Comparisons of analytic results with rigorous calculations allow the establishment of ranges of validity of the dipolar approximation for these forces.


Novel Monthly Quality Assurance Regimen and 5-Year Analysis Using a Proton Metrology System.
To develop a novel, monthly quality assurance (QA) regimen for a proton therapy system that uses 2 custom phantoms, each housing a commercial scintillator detector and a charge-coupled device camera. The novel metrology system assessed QA trends at a pediatric proton therapy center from 2018 to 2022. The measurement system was designed to accommodate horizontal and vertical positioning of the commercial device and to enable gantry and couch isocentricity measurements (using a star shot procedure), proton spot profile verification, and imaging and radiation congruence tests to be performed simultaneously in the dual-phantom setup. Gantry angles and proton beam energies were varied and alternated each month, using gantry angles of 0°, 30°, 60°, 90°, 120°, 150°, and 180° and discrete beam energies of 69.4, 84.5, 100, 139.1, 180.4, 200.4, and 221.3 MeV after radiographic verification. A total of 1176 individual monthly QA measurements of gantry and couch isocentricity, spot size, and congruence were analyzed. Gantry and couch star shot measurements showed beam isocentricities of 0.3 ± 0.2 mm and 0.2 ± 0.2 mm, respectively, which were within the threshold of 1.0 mm. Spot sizes for each discrete energy were within the threshold of ± 10% of the baseline values for all 3 proton rooms. The imaging and radiation coincidence test results for the 1176 individual monthly QA measurements were 0.5 mm for the 50th percentile and 1.2 mm (the clinical threshold) for the 97.6th percentile. Integrating a commercial device with custom phantoms improved the quality of proton system checks compared with previous methods using radiochromic films, loose ball bearings, and foam. The scheme of alternating beam angles with discrete energies in the monthly QA-enabled, clinically meaningful verification of beam energy and gantry angle combinations while the machine performance and accuracy were being checked.


ZNF687 Mutations in an Extended Cohort of Neoplastic Transformations in Paget's Disease of Bone: Implications for Clinical Pathology.
Neoplastic transformation is a rare but serious complication of Paget's disease of bone (PDB), occurring in fewer than 1% of individuals with polyostotic disease. Their prognosis is poor, with less than 50% surviving 5 years. In 2016, the genetic alteration of giant cell tumor (GCT) complicating PDB was identified as a founder germline mutation (P937R) in the ZNF687 gene. However, the study population was exclusively of Italian descent, and patients of different ethnic origins were not studied. To fill this gap, herein we performed mutation analysis of ZNF687 in a GCT in the pelvis of a 45-year-old black American woman with polyostotic PDB. The P937R mutation in ZNF687 was found in her tumor but, as expected, the ancestral haplotype that characterizes the Italian GCT/PDB patients was not found. Furthermore, we identified two additional Italian GCT/PDB patients with this ZNF687 mutation, now constituting a cohort of 18 GCT/PDB cases, all harboring the identical mutation. We also searched for ZNF687 mutations in a unique collection of tumor tissues derived from Italian PDB patients, including 28 osteosarcomas (OS/PDB), 8 undifferentiated sarcomas (SRC/PDB), 1 fibrosarcoma (FS/PDB), and 1 chondrosarcoma (CS/PDB). We identified the P937R mutation in one SRC/PDB and a different ZNF687 mutation (R331W) in 1 of 28 pagetic osteosarcomas. Thus, whereas GCT/PDB pathogenesis globally seems to involve the P937R mutation in ZNF687, other neoplasms associated with PDB seem to be less related to mutations in this gene. Finally, we identified the G34W mutation in the H3F3A gene in the maxillary tumor masses of two PDB patients, defining them as conventional GCT rather than GCT/PDB. Thus, combined molecular analysis of H3F3A and ZNF687 is essential to clarify the origin and diagnosis of tumors in PDB. © 2020 American Society for Bone and Mineral Research.


A novel bone targeting delivery system carrying phytomolecule icaritin for prevention of steroid-associated osteonecrosis in rats.
One of the effective strategies for prevention of steroid-associated osteonecrosis (SAON) is to inhibit bone resorption and fat formation and promote bone formation at osteonecrotic sensitive skeletal sites. We identified a novel phytomolecule that showed positive effects on osteogenesis, anti-bone resorption and anti-adipogenesis in vitro and also developed a bone-targeting delivery system (BTDS) for in vivo experimental study. The study investigated if our innovative synthesized BTDS carrying this phytomolecule would be able to effectively prevent SAON in a rat model. SAON was induced by combined injections of lipopolysaccharide and methylprednisolone. SAON rats were divided into four groups, one SAON untreated control group and three SAON treatment groups with different types of delivery systems (Asp8-liposome-icaritin, liposome-icaritin and Asp8-liposome) for two weeks. SAON lesions were identified and osteoclasts activity, osteogenesis and adipogenesis at these sites were evaluated by immunohistochemistry. Ex vitro study was also designed to evaluate the osteogenic and adipogenic potential of the isolated bone marrow stromal cells (BMSCs) via real-time PCR and histochemical staining. Our results showed that as a bone surface-specific BTDS, Asp8-liposome-icaritin effectively prevented steroids-treated rats from SAON with significantly decreased osteocytes apoptosis, down-regulated osteoclatsogenesis and up-regulated osteogenesis. However, both liposome-icaritin and Asp8-liposome treatment did not show significant efficacy for SAON prevention. In summary, this proof-concept-study showed for the first time that the innovative Asp8-liposome-icaritin BTDS was effective for prevention of SAON in terms of bone resorption prevention, adipogenesis suppression, and bone-formation enhancement.


CALU-3 lung cells three-dimensionally assembled onto CellFate® matrix present angiotensin-converting enzyme-2 activity.
Currently, there is a great need for the development of three-dimensional (3D) in vitro lung models. Particularly, the production of a suitable 3D model of pulmonary epithelium for understanding the pathophysiology of diseases such as the COVID-19 must consider the tissue architecture and presence, for example, of the angiotensin-converting enzyme-2 (ACE-2) in the cells. Different polymeric membranes are being used to support cell culturing, especially of lung cells, however, there is still no information about the culture of these cells onto bacterial nanocellulose (BNC) matrices. We have used the BNC matrix CellFate® as a support for the assembly of a 3D in vitro model of lung epithelium, composed of human lung fibroblasts (HLF) and lung adenocarcinoma cells (CALU-3). CellFate® matrices were made from bacterial fermentation resulting in a natural and biocompatible biopolymer. Cells were cultured onto CellFate® and maintained in a 5% CO2 humidified atmosphere at 37°C. Cell viability was assessed by the resazurin method The samples were, then, exposed to the air-liquid interface (ALI), and histologically analyzed. ACE-2 activity was verified on the hydrolyze of the fluorogenic substrate Mca-APK(Dnp)-OH, and its presence was evaluated by flow cytometry. The expression of the anionic transporter SLCO3A1 was evaluated by qPCR. Cell viability analysis indicates that CellFate® was not toxic to these cells. By flow cytometry, the presence of the ACE-2 was identified in the CALU-3 cells surface corroborating the results obtained from enzymatic activity analysis. The SLCO3A1 transporter expression was identified in cells cultured onto CellFate®, but not in cells cultured onto the transwell (control). CALU-3 cells cultivated onto CellFate® resulted in a pseudostratified organization, a typical morphology of the human respiratory tract epithelium. The current model opens perspectives for studies involving physiological characterization, improving its relevance for the understanding of the pathophysiology of diseases as well as the response to drugs.


The role of monoclonal antibodies in the treatment of severe asthma.
A number of therapeutic agents are available for the treatment of asthma, including inhaled corticosteroids, long- and short-acting beta-agonists, leukotriene-modifying agents, long- and short-acting anticholinergic agents, chromones, theophylline, allergen immunotherapy, and oral corticosteroid therapy. All available therapies, despite their proven efficacy, are purely symptomatic including the topical steroids. This issue has led to the development of several biologic agents to aid in asthma management and to potentially alter the course of the disease by interfering with specific aspects of inflammation which may modify remodeling in the airways. Monoclonal antibodies have offered a class of therapeutic agents that enhance treatment options for patients with moderate-to-severe persistent asthma. As such, this article provides an overview of present and future monoclonal antibody therapies for the treatment of patients with severe asthma.


Validation of a field test for the non-invasive determination of badminton specific aerobic performance.
To develop a badminton specific test to determine on court aerobic and anaerobic performance. The test was evaluated by using a lactate steady state test. Seventeen male competitive badminton players (mean (SD) age 26 (8) years, weight 74 (10) kg, height 179 (7) cm) performed an incremental field test on the badminton court to assess the heart rate turn point (HRTP) and the individual physical working capacity (PWC(i)) at 90% of measured maximal heart rate (HR(max)). All subjects performed a 20 minute steady state test at a workload just below the PWC(i). Significant correlations (p<0.05) for Pearson's product moment coefficient were found between the two methods for HR (r = 0.78) and velocity (r = 0.93). The HR at the PWC(i) (176 (5.5) beats/min) was significantly lower than the HRTP (179 (5.5) beats/min), but no significant difference was found for velocity (1.44 (0.3) m/s, 1.38 (0.4) m/s). The constant exercise test showed steady state conditions for both HR (175 (9) beats/min) and blood lactate concentration (3.1 (1.2) mmol/l). The data indicate that a valid determination of specific aerobic and anaerobic exercise performance for the sport of badminton is possible without HRTP determination.


[Double-blind comparative trial of trimethoprim-polymyxin B and trimethoprim-sulphacetamide-polymyxin B ear drops in the treatment of otorrhoea (author's transl)].
The purpose of his double-blind, randomized trial was to compare ear drops containing either trimethoprim and polymyxin B (TP) or trimethoprim, sulphacetamide and polymyxin B (TSP) from the points of view of effectiveness and safety. The 68 patients treated had otitis externa or recurrent otitis media with perforated ear-drum, or mastoid cavity with post-tympanoplasty infection. Satisfactory results were obtained in 60.6% of the cases with TP and in 88.6% with TSP. When evaluated by the Chi-square method with Yates' correction, the difference was statistically significant. There was no evidence of ototoxicity, fungal infection or local hypersensitivity with either preparation. The trial demonstrated that both ear drops were active and that TSP was much more effective than TP in the treatment of otorrhea.


Influence of biogas flow rate on biomass composition during the optimization of biogas upgrading in microalgal-bacterial processes.
The influence of biogas flow rate (0, 0.3, 0.6, and 1.2 m(3) m(-2) h(-1)) on the elemental and macromolecular composition of the algal-bacterial biomass produced from biogas upgrading in a 180 L photobioreactor interconnected to a 2.5 L external bubbled absorption column was investigated using diluted anaerobically digested vinasse as cultivation medium. The influence of the external liquid recirculation/biogas ratio (0.5 < L/G < 67) on the removal of CO2 and H2S, and on the concentrations of O2 and N2 in the upgraded biogas, was also evaluated. A L/G ratio of 10 was considered optimum to support CO2 and H2S removals of 80% and 100%, respectively, at all biogas flow rates tested. Biomass productivity increased at increasing biogas flow rate, with a maximum of 12 ± 1 g m(-2) d(-1) at 1.2 m(3) m(-2) h(-1), while the C, N, and P biomass content remained constant at 49 ± 2%, 9 ± 0%, and 1 ± 0%, respectively, over the 175 days of experimentation. The high carbohydrate contents (60-76%), inversely correlated to biogas flow rates, would allow the production of ≈100 L of ethanol per 1000 m(3) of biogas upgraded under a biorefinery process approach.


[Use of the bacterial flocculation test for assessing the mutagenic properties of carcinogenic nitrosamines and nitramines].
Fluctuation test developed by Green allows to detect the mutagenicity of trace and non-toxic chemical concentrations. Mutagenic activity of both carcinogenic nitramines--nitrodimethylamine, nitrodiethylamine and nitromorpholine (0.1--10 mM) and their nitrosamine analogues-nitrosodimethylamine, nitrosodiethylamine and nitrosomorpholine (0.01--0.1 mM) was detected in fluctuation test with Salmonella typhimurium TA 98 and TA 1530 as indicator organism in the presence of postmitochondrial supernatant from liver of rats treated with Aroclor 1254. Nearly all compounds appeared to be mutagenic even in the absence of co-factors of microsomal oxygenation.


The benefit to speech intelligibility of hearing a familiar voice.
Previous experience with a voice can help listeners understand speech when a competing talker is present. Using the coordinate-response measure task (Bolia, Nelson, Ericson, & Simpson, 2000), Johnsrude et al. (2013) demonstrated that speech is more intelligible when either the target or competing (masking) talker is a long-term spouse than when both talkers are unfamiliar (termed familiar-target and familiar-masker benefits, respectively). To better understand how familiarity improves intelligibility, we measured the familiar-target and familiar-masker benefits in older and younger spouses using a more challenging matrix task, and compared the benefits listeners gain from spouses' and friends' voices. On each trial, participants heard two sentences from the Boston University Gerald (Kidd, Best, & Mason, 2008) corpus ("<name> <verb> <number> <adjective> <noun>") and reported words from the sentence beginning with a target name word. A familiar-masker benefit was not observed, but all groups showed a robust familiar-target benefit and its magnitude did not differ between spouses and friends. The familiar-target benefit was not influenced by relationship length (in the range of 1.5-52 years). Together, these results imply that the familiar-target benefit can develop from various types of relationships and has already reached a plateau around 1.5 years after meeting a new friend. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


[Communication with the aphasic patient].
The consequences of aphasia with reference to the WHO ICIDH classification are evoked. The role of the physician and the behaviour of the relatives are underlined as well as the main parameters to be taken into consideration to communicate with an aphasic patient. As a rule, a dialogue in quiet surroundings will make communication easier with an aphasic patient. Our attitude will be modulated according to the behaviour of the patient, his past and the aphasiological picture. At the early stage of the disease, verbal means of communication will be mainly used, whereas later on, in a pragmatic approach, all means (verbal and non verbal) will be used to improve communication.


Prevalence of baseline HCV NS5A resistance associated substitutions in genotype 1a, 1b and 3 infection in Australia.
Direct-acting antivirals (DAA) have revolutionised hepatitis C virus (HCV) treatment, and most regimens include an NS5A inhibitor. Certain amino-acid substitutions confer resistance to NS5A inhibitors, termed resistance-associated substitutions (RAS). If present at baseline, they can reduce virological response rates. Population-based sequencing (PBS) is generally used for baseline sequencing, however next generation sequencing (NGS) reduces the threshold for detection of sequences encoding RAS from 20% to 5%. We determined the prevalence of NS5A RAS at baseline amongst Australian chronically infected with genotype (GT)1a, GT1b and GT3 HCV, using both PBS and NGS. Samples from DAA-naïve individuals were received at the Victorian Infectious Disease Reference Laboratory between June 2016 and December 2018. All samples were analysed for NS5A RAS using PBS. A subset of GT1 HCV samples were processed using NGS technology (Vela Diagnostics, Singapore) to determine the improvement in sensitivity. In total, 672 samples were analysed using PBS. The baseline prevalence of NS5A RAS was 7.6% for GT1a (n = 25/329), 15.7% for GT1b (n = 8/51) and 15.1% for GT3 (n = 44/292). NGS only marginally increased sensitivity for NS5A RAS at baseline in GT1a (16% vs 17%) and GT1b (29% vs 36%). The prevalence of NS5A RAS in GT1a HCV in Australia was low compared with international data, and was similar to other reported international prevalence for GT1b and GT3 infection. NGS at baseline only marginally increased sensitivity for the detection of NS5A RAS in patients with GT1 HCV and cannot be recommended for routine use at baseline in clinical practice.


Unilateral Existence of Chondro-epitrochlearis: Its Embryological Perspectives and Clinical Implications.
Chondroepitrochlearis (CET) is an anomalous muscular slip that originates from the pectoralis major muscle and inserts into epicondyle of the humerus. The morphology of this variant form of pectoralis major can vary from slender to strong musculo-tendinous. In its course, it usually crosses the neurovascular structures of arm; their compression is a major complication that could be manifested by its persistence. In the present case, potentially anomalous CET muscle with the slender slip of origin, but strong tendinous insertion to the medial epicondyle of the humerus was found unilaterally. This musculo-tendinous structure was found to be compressing the brachial artery and median nerve in the arm. Detailed embryological and clinical perspective of such variant muscular slip helps the physiotherapists, orthopaedicians in their treatment strategy in complain of restricted shoulder movement. It may also help the neurologists, radiologists in their diagnostic approach of ulnar neuropathy.


Selectin Dependence of Allergic Skin Inflammation Is Diminished by Maternal Atopy.
Allergic skin inflammation requires the influx of inflammatory cells into the skin. Extravasation of leukocytes into the skin requires interactions between endothelial selectins and their glycan ligands on the surface of leukocytes. Selectin-ligand formation requires the activity of several glycosyltransferases, including Fut7 In this report, we tested the importance of Fut7 for the development of allergic skin inflammation in the Stat6VT transgenic mouse model. We observed that Fut7 deficiency was protective but did not eliminate disease. Segregation of the data by gender of the parent that transmitted the Stat6VT transgene, but not by gender of the pups, which were analyzed for disease, revealed that the protective effects of Fut7 deficiency were significantly greater when dams were Stat6VT negative. In contrast, in mice from litters of Stat6VT+ dams, Fut7 deficiency resulted in only modest protection. These findings indicate that pups from atopic dams exhibit a greater propensity for allergic disease, similar to observations in humans, and that the effect of maternal atopy is due to enhanced selectin-independent mechanisms of leukocyte recruitment in their offspring. Together, these results demonstrate that Fut7 deficiency can be protective in a model of atopic dermatitis but that maternal atopy diminishes these protective effects, suggesting alternative pathways for leukocyte recruitment in the absence of Fut7 enzyme activity. These observations have implications for understanding how the environment in utero predisposes for the development of allergic disease.


Giant-cell tumour of the tendon sheath.
An unusual case of giant-cell tumours of the tendon sheath (syn. Pigmented villonodular tenosynovitis or fibrous xanthoma) of a long thumb flexor is presented. The unusual features are: two distinctly different tumours of almost similar size and shape arising from the tendon sheath at its either ends but with a segment of normal tendon sheath in between and presence of early malignant changes in the distal tumour with a benign proximal one, confirmed microscopically. Lastly, radiograph showed definite bony involvement in the proximal phalanx of the thumb. On extensive review of the literature, I found reports of rare presentation of malignant changes and bony erosion, but none had these various features.


Investigating the Effects of Coenzyme Q10 on Human Corneal Endothelial Cells.
To investigate the effects of Coenzyme Q10 (CoQ10) treatment on immortalised human corneal endothelial cells (HCEC-12). HCEC-12 cells were cultured in different concentrations of CoQ10 (0.1%, 0.2%, 0.5%, and 1.0%) and analysed using live/dead staining assay to determine appropriate concentration for subsequent experiments. Cells were pretreated with CoQ10 before inducing apoptosis by ethanol (EtOH) treatment for 30 seconds which was followed by posttreatment with CoQ10. Viable, apoptotic, and dead cell proportions were analysed using Annexin V-FITC immunofluorescence staining. Mitochondrial intensity and respiratory functions were also investigated using MitoTracker staining and a Seahorse XFe24 analyser, respectively. Results were compared to a positive control for apoptosis. The experiments were carried out in triplicates. Graphpad prism software was used for statistical analysis where p < 0.05 was deemed statistically significant. CoQ10 treatment at 0.5% and 1% showed 92% and 30% viable cells compared with 0.1% and 0.2% that showed 96% and 94% viable cells, respectively (p=0.0562). 0.1% and 0.2% concentrations were, thus, used for subsequent experiments. Annexin V-FITC apoptotic analysis showed 2% at 0.1% and 3% at 0.2% of apoptotic cells (p=0.0824). Mitochondrial respiratory function and mitochondrial intensity increased in apoptotic cells following 0.1% CoQ10 treatment. 0.1% CoQ10 was found optimal for reducing apoptosis and increasing metabolic activity on human corneal endothelial cell line. These results support the need for further ex vivo studies to investigate the safety profile of CoQ10 as an antiapoptotic agent for human corneal endothelium.


Measurement of intracompartmental pressure with use of a new electronic transducer-tipped catheter system.
Laboratory and clinical tests were carried out to determine the clinical usefulness, validity, and safety of a new self-calibrating, battery-powered monitoring system for the measurement of intramuscular pressure with use of an electronic transducer-tipped catheter. The eight probes accurately recorded applied pressures ranging from zero to 160 millimeters of mercury (zero to 21.33 kilopascals). The system registered little temperature-induced drift (maximum, 1.25 millimeters of mercury [0.17 kilopascal]) between dry room temperature and 40 degrees Celsius. There were also minimum variations (range, -0.14 to 0.81 millimeter of mercury [0.02 to 0.11 kilopascal]) in the pressures recorded during a twenty-four-hour period. The resting pressure in the tibialis anterior muscle of twenty volunteers who had normal limbs was a mean (and standard deviation) of 13.1+/-8.3 millimeters of mercury (1.75+/-1.11 kilopascals). There was a good correlation between externally applied pressures (zero, twenty, forty, sixty, eighty, and 100 millimeters of mercury [zero, 2.67, 5.33, 8.00, 10.66, and 13.33 kilopascals] applied with use of antishock trousers) and the pressures measured in the tibialis anterior muscle of four volunteers (r = 0.997 to 0.999). The injection of sterile saline solution into the tibialis anterior muscle of a volunteer and the use of high-frequency recording during muscular activity showed a high degree of responsiveness and sensitivity to changes in intramuscular pressure. We also prospectively evaluated the clinical usefulness of the system and found it to be easy to assemble, calibrate, and use. Thus, this reusable, electronic transducer-tipped catheter system, which is based on a noninfusion technique, is simple, minimally traumatic, and highly precise. It is free of hydrostatic pressure artifacts and provides dynamic responses to changes in intramuscular pressure.


Maternal stress predicts postpartum weight retention.
Postpartum weight retention (PPWR) is a significant contributor to the development of overweight and obesity in women of childbearing age. Stress may be a key mechanism making it more difficult for mothers to lose weight in the year following delivery. The aim of this study was to assess whether specific aspects of parenting stress and life stress influence postpartum weight retention in new mothers. Women in late pregnancy or up to 2 months postpartum (n = 123) were enrolled in the study and followed through the first year postpartum. Linear regression models evaluated the associations of parenting stress (isolation, attachment and depressive symptoms) as well as overall life stress at 2, 6, and 12 months postpartum with PPWR at 6 and 12 months. During the first year postpartum, higher depression and life stress were significantly associated with greater PPWR. As the effect of depression diminished, the effect of life stress became significant. Contrary to hypothesized relationships, fewer problems with attachment and less social isolation were significantly associated with greater PPWR. Higher gestational weight gain and African American race were also significantly associated with greater PPWR at both 6 and 12 months. Different types of stress predict weight retention in first time mothers during the first year postpartum. Understanding the relationships between parenting stress, concurrent life stress and PPWR can enhance the development of future interventions that specifically target self-identified stressors, leading to improved weight related outcomes.


Regulatory significance of procaine residues in plasma and urine samples: preliminary communication.
Plasma and urinary concentrations of procaine and the duration of response to procaine after its administration as a local anaesthetic to horses were studied. Following injection of a clinical dose of procaine HCl (80 mg), the concentration of procaine in plasma was less than the lower limit of quantitation and unsuitable for threshold determination. Therefore, the urinary concentration of procaine was determined after injection of a dose of 5 mg procaine HCl, the highest no-effect dose (HNED) of this agent. Free unconjugated procaine in equine urine reached a peak concentration of 23.7 ng/mL, while total (unconjugated plus conjugated) procaine peaked at 37.9 ng/mL (mean urine pH of 8.5). Because a basic drug may concentrate substantially in acidic urine, a threshold concentration of 25 ng/mL of unconjugated procaine is a reasonable and conservative threshold for procaine at this time. Horses were administered abaxial sesamoid blocks containing 2% procaine HCl (40, 80, 160 and 320 mg) and 2% procaine HCl (40 and 320 mg) with epinephrine (1:100,000) in local anaesthetic experiments. There was a significant local anaesthetic (LA) effect for all doses of procaine HCl with the duration of effect ranging from 30 min (40 mg) to 60 min (320 mg). The addition of epinephrine significantly increased the duration of local anaesthesia to 180 min for a 40 mg dose and 420 min for a 320 mg dose. Because epinephrine may extend the duration of local anaesthesia beyond a reasonable period of confinement for horses before the starting time of a race, the increased LA effect following the addition of epinephrine to procaine has regulatory significance.


Next Stop a Longer Life.
Aim A request was made to the Department of Public Health in early 2019 for some interesting statistics (funtistics) for a planned health promotion campaign encouraging public transport users to increase their physical activity levels by alighting one stop earlier and walking to their destination. For a novel presentation of the benefits of increasing physical activity it was decided to calculate the potential increase in life-expectancy that a given amount of physical activity would correspond to (at a population level). Method Estimated increase in weekly walking time was calculated for the Dublin Bus commuter walking the last stop of their journey. The reduced risk of mortality was estimated for this increase in physical activity and applied to Irish life tables to calculate change in life expectancy. Results Alighting from a bus one stop earlier in Dublin would lead to an average of 4.42 minutes additional walking (44.21 minutes additional walking per week for a commuter). In the Dublin Bus commuter population, this leads to an estimated 50 day increase in life expectancy (male population). Conclusion At the lower end of the dose response curve for physical activity, health benefits include: reduced risk of cardiovascular disease, reduced risk of diabetes, psychosocial benefit, reduced risk of musculoskeletal problems. For the prevention of weight gain and some cancers, activity at the upper end of the range (1000 MET.min/week, approximately 300 minutes of walking/week) is thought to be required.


Morphologic and molecular characterization of renal cell carcinoma in children and young adults.
A new WHO classification of renal cell carcinoma has been introduced in 2004. This classification includes the recently described renal cell carcinomas with the ASPL-TFE3 gene fusion and carcinomas with a PRCC-TFE3 gene fusion. Collectively, these tumors have been termed Xp11.2 or TFE3 translocation carcinomas, which primarily occur in children and young adults. To further study the characteristics of renal cell carcinoma in young patients and to determine their genetic background, 41 renal cell carcinomas of patients younger than 22 years were morphologically and genetically characterized. Loss of heterozygosity analysis of the von Hippel-Lindau gene region and screening for VHL gene mutations by direct sequencing were performed in 20 tumors. TFE3 protein overexpression, which correlates with the presence of a TFE3 gene fusion, was assessed by immunohistochemistry. Applying the new WHO classification for renal cell carcinoma, there were 6 clear cell (15%), 9 papillary (22%), 2 chromophobe, and 2 collecting duct carcinomas. Eight carcinomas showed translocation carcinoma morphology (20%). One carcinoma occurred 4 years after a neuroblastoma. Thirteen tumors could not be assigned to types specified by the new WHO classification: 10 were grouped as unclassified (24%), including a unique renal cell carcinoma with prominently vacuolated cytoplasm and WT1 expression. Three carcinomas occurred in combination with nephroblastoma. Molecular analysis revealed deletions at 3p25-26 in one translocation carcinoma, one chromophobe renal cell carcinoma, and one papillary renal cell carcinoma. There were no VHL mutations. Nuclear TFE3 overexpression was detected in 6 renal cell carcinomas, all of which showed areas with voluminous cytoplasm and foci of papillary architecture, consistent with a translocation carcinoma phenotype. The large proportion of TFE3 "translocation" carcinomas and "unclassified" carcinomas in the first two decades of life demonstrates that renal cell carcinomas in young patients contain genetically and phenotypically distinct tumors with further potential for novel renal cell carcinoma subtypes. The far lower frequency of clear cell carcinomas and VHL alterations compared with adults suggests that renal cell carcinomas in young patients have a unique genetic background.


Resveratrol inhibits cell cycle progression by targeting Aurora kinase A and Polo-like kinase 1 in breast cancer cells.
The Aurora protein kinase (AURKA) and the Polo-like kinase-1 (PLK1) activate the cell cycle, and they are considered promising druggable targets in cancer therapy. However, resistance to chemotherapy and to specific small‑molecule inhibitors is common in cancer patients; thus alternative therapeutic approaches are needed to overcome clinical resistance. Here, we showed that the dietary compound resveratrol suppressed the cell cycle by targeting AURKA and PLK1 kinases. First, we identified genes modulated by resveratrol using a genome-wide analysis of gene expression in MDA-MB-231 breast cancer cells. Transcriptional profiling indicated that 375 genes were modulated at 24 h after resveratrol intervention, whereas 579 genes were regulated at 48 h. Of these, 290 genes were deregulated in common at 24 and 48 h. Interestingly, a significant decrease in the expression of genes involved in the cell cycle, DNA repair, cytoskeleton organization, and angiogenesis was detected. In particular, AURKA and PLK1 kinases were downregulated by resveratrol at 24 h. In addition the BRCA1 gene, an AURKA/PLK1 inhibitor, was upregulated at 24 h of treatment. Moreover, two well-known resveratrol effectors, cyclin D1 (CCND1) and cyclin B1 (CCNB1), were also repressed at both times. Congruently, we found that resveratrol impaired G1/S phase transition in both MDA-MB-231 and MCF-7 cells. By western blot assays, we confirmed that resveratrol suppressed AURKA, CCND1 and CCNB1 at 24 and 48 h. In summary, we showed for the first time that resveratrol regulates cell cycle progression by targeting AURKA and PLK1. Our findings highlight the potential use of resveratrol as an adjuvant therapy for breast cancer.


Genome editing enables reverse genetics of multicellular development in the choanoflagellate Salpingoeca rosetta.
In a previous study, we established a forward genetic screen to identify genes required for multicellular development in the choanoflagellate, Salpingoeca rosetta (Levin et al., 2014). Yet, the paucity of reverse genetic tools for choanoflagellates has hampered direct tests of gene function and impeded the establishment of choanoflagellates as a model for reconstructing the origin of their closest living relatives, the animals. Here we establish CRISPR/Cas9-mediated genome editing in S. rosetta by engineering a selectable marker to enrich for edited cells. We then use genome editing to disrupt the coding sequence of a S. rosetta C-type lectin gene, rosetteless, and thereby demonstrate its necessity for multicellular rosette development. This work advances S. rosetta as a model system in which to investigate how genes identified from genetic screens and genomic surveys function in choanoflagellates and evolved as critical regulators of animal biology.


Long term storage in liquid nitrogen leads to only minor phenotypic and gene expression changes in the mammary carcinoma model cell line BT474.
Cancer cell lines are indispensible surrogate models in cancer research, as they can be used off-the-shelf, expanded to the desired extent, easily modified and exchanged between research groups for affirmation, reproduction or follow-up experiments.As malignant cells are prone to genomic instability, phenotypical changes may occur after certain passages in culture. Thus, cell lines have to be regularly authenticated to ensure data quality. In between experiments these cell lines are often stored in liquid nitrogen for extended time periods.Although freezing of cells is a necessary evil, little research is performed on how long-term storage affects cancer cell lines. Therefore, this study investigated the effects of a 28-year long liquid nitrogen storage period on BT474 cells with regard to phenotypical changes, differences in cell-surface receptor expression as well as cytokine and gene expressional variations. Two batches of BT474 cells, one frozen in 1986, the other directly purchased from ATCC were investigated by light microscopy, cell growth analysis, flow cytometry and cytokine as well as whole-transcriptome expression profiling. The cell lines were morphologically indifferent and showed similar growth rates and similar cell-surface receptor expression. Transcriptome analysis revealed significant differences in only 26 of 40,716 investigated RefSeq transcripts with 4 of them being up-regulated and 22 down-regulated. This study demonstrates that even after very long periods of storage in liquid nitrogen, cancer cell lines display only minimal changes in their gene expression profiles. However, also such minor changes should be carefully assessed before continuation of experiments, especially if phenotypic alterations can be additionally observed.


Bezlotoxumab for Prevention of Recurrent Clostridium difficile Infection.
Clostridium difficile is the most common cause of infectious diarrhea in hospitalized patients. Recurrences are common after antibiotic therapy. Actoxumab and bezlotoxumab are human monoclonal antibodies against C. difficile toxins A and B, respectively. We conducted two double-blind, randomized, placebo-controlled, phase 3 trials, MODIFY I and MODIFY II, involving 2655 adults receiving oral standard-of-care antibiotics for primary or recurrent C. difficile infection. Participants received an infusion of bezlotoxumab (10 mg per kilogram of body weight), actoxumab plus bezlotoxumab (10 mg per kilogram each), or placebo; actoxumab alone (10 mg per kilogram) was given in MODIFY I but discontinued after a planned interim analysis. The primary end point was recurrent infection (new episode after initial clinical cure) within 12 weeks after infusion in the modified intention-to-treat population. In both trials, the rate of recurrent C. difficile infection was significantly lower with bezlotoxumab alone than with placebo (MODIFY I: 17% [67 of 386] vs. 28% [109 of 395]; adjusted difference, -10.1 percentage points; 95% confidence interval [CI], -15.9 to -4.3; P<0.001; MODIFY II: 16% [62 of 395] vs. 26% [97 of 378]; adjusted difference, -9.9 percentage points; 95% CI, -15.5 to -4.3; P<0.001) and was significantly lower with actoxumab plus bezlotoxumab than with placebo (MODIFY I: 16% [61 of 383] vs. 28% [109 of 395]; adjusted difference, -11.6 percentage points; 95% CI, -17.4 to -5.9; P<0.001; MODIFY II: 15% [58 of 390] vs. 26% [97 of 378]; adjusted difference, -10.7 percentage points; 95% CI, -16.4 to -5.1; P<0.001). In prespecified subgroup analyses (combined data set), rates of recurrent infection were lower in both groups that received bezlotoxumab than in the placebo group in subpopulations at high risk for recurrent infection or for an adverse outcome. The rates of initial clinical cure were 80% with bezlotoxumab alone, 73% with actoxumab plus bezlotoxumab, and 80% with placebo; the rates of sustained cure (initial clinical cure without recurrent infection in 12 weeks) were 64%, 58%, and 54%, respectively. The rates of adverse events were similar among these groups; the most common events were diarrhea and nausea. Among participants receiving antibiotic treatment for primary or recurrent C. difficile infection, bezlotoxumab was associated with a substantially lower rate of recurrent infection than placebo and had a safety profile similar to that of placebo. The addition of actoxumab did not improve efficacy. (Funded by Merck; MODIFY I and MODIFY II ClinicalTrials.gov numbers, NCT01241552 and NCT01513239 .).


Genital hygiene practices of fishermen targeted for a topical microbicide intervention against sexually transmitted infections in Kisumu, Kenya.
Research on hygiene has been relatively limited in the current era of rigorous observational studies and clinical trials. We set out to investigate the perception and practices of genital hygiene among fishermen working on the beaches along Lake Victoria, targeted for a topical male microbicide hygiene intervention. We conducted 12 focus group discussions involving fishermen (n = 130), recording the discussions in Dholuo (the local language) and transcribing them verbatim before translating into English. Transcripts were double-coded and analysed using constant comparative analysis. Despite easy access to lake water and recognition of a link that may exist between poor genital hygiene and the risk of penile infection and poor sexual relationships, few fishermen regularly washed their genitalia due to fear/embarrassment from cleaning their genitalia in public, traditional Luo beliefs such as that washing with soap would reduce the fish catch, lack of time because of their busy schedules, laziness and lack of responsibility, and excessive consumption of alcohol and illicit drugs. Hygiene practices of the fishermen were poor and could contribute to genital infections including sexually transmitted infections. Given the fishermen's poor genital hygiene practices, they may benefit from hygiene intervention, including that provided by penile microbicides, which can be applied in the privacy of their bedrooms.


Immersing lungs in hydrogen-rich saline attenuates lung ischaemia-reperfusion injury.
Anti-oxidant effects of hydrogen have been reported in studies examining ischaemia-reperfusion injury (IRI). In this study, we evaluated the therapeutic efficacy of immersing lungs in hydrogen-rich saline on lung IRI. Lewis rats were divided into three groups: (i) sham, (ii) normal saline and (iii) hydrogen-rich saline. In the first experiment, the left thoracic cavity was filled with either normal saline or hydrogen-rich saline for 1 h. Then, we measured the hydrogen concentration in the left lung using a sensor gas chromatograph ( N = 3 per group). In the second experiment, lung IRI was induced by occlusion of the left pulmonary hilum for 1 h, followed by reperfusion for 3 h. During the ischaemic period, the left thoracic cavity was filled with either normal saline or hydrogen-rich saline. After reperfusion, we assessed lung function, histological changes and cytokine production ( N = 5-7 per group). Immersing lungs in hydrogen-rich saline resulted in an elevated hydrogen concentration in the lung (6.9 ± 2.9 μmol/1 g lung). After IRI, pulmonary function (pulmonary compliance and oxygenation levels) was significantly higher in the hydrogen-rich saline group than in the normal saline group ( P < 0.05). Similarly, pro-inflammatory cytokine levels (interleukin-1β and interleukin-6) in the left lung were significantly lower in the hydrogen-rich saline group than in the normal saline group ( P < 0.05). Immersing lungs in hydrogen-rich saline delivered hydrogen into the lung and consequently attenuated lung IRI. Hydrogen-rich solution appears to be a promising approach to managing lung IRI.


Intraganglionic signaling as a novel nasal-meningeal pathway for TRPA1-dependent trigeminovascular activation by inhaled environmental irritants.
Headache is the most common symptom associated with air pollution, but little is understood about the underlying mechanism. Nasal administration of environmental irritants activates the trigeminovascular system by a TRPA1-dependent process. This report addresses questions about the anatomical pathway involved and the function of TRP channels in this pathway. TRPV1 and TRPA1 are frequently co-localized and interact to modulate function in sensory neurons. We demonstrate here that resiniferatoxin ablation of TRPV1 expressing neurons significantly reduces meningeal blood flow responses to nasal administration of both TRPV1 and TRPA1 agonists. Accordingly resiniferatoxin also significantly reduces TRPV1 and CGRP immunostaining and TRPV1 and TRPA1 message levels in trigeminal ganglia. Sensory neurons of the trigeminal ganglia innervate the nasal epithelium and the meninges, but the mechanism and anatomical route by which nasal administration evokes meningeal vasodilatation is unclear. Double retrograde labeling from the nose and meninges reveals no co-localization of fluorescent label, however nasal and meningeal labeled cells are located in close proximity to each other within the trigeminal ganglion. Our data demonstrate that TRPV1 expressing neurons are important for TRPA1 responses in the nasal-meningeal pathway. Our data also suggest that the nasal-meningeal pathway is not primarily by axon reflex, but may instead result from intraganglionic transmission.


Hemodynamic factors changing blood flow velocity waveform and profile in normal human brachial artery.
We have investigated the influence of changes of perfusion pressure and local peripheral resistance on blood flow velocity waveform and profile in normal human peripheral arteries. Blood flow velocity and profile were recorded from the distal end of the left brachial artery in ten normal subjects by means of an ultrasonic device. The records were obtained in basal conditions and after blood pressure in the brachial artery and local peripheral vascular resistance were changed, separately or together, by progressive inflation of two arm cuffs, one encircling the proximal half of the left arm and the other the middle part of the left forearm. Both blood flow velocity waveform and profile were shown to be markedly modified by changes in perfusion pressure and local peripheral vascular resistance. Reduction of perfusion pressure decreased both forward and reverse peak velocities, but had the largest effect upon reverse velocity. The upslope and the downslope of the forward velocity wave were left unchanged. Increase in local peripheral vascular resistance markedly augmented reverse peak velocity, whether perfusion pressure was normal or reduced. Increased resistance only slightly influenced peak forward velocity.


[Obesity and developing countries of the south].
An adult is considered as overweight if his body mass index is 25.0 kg/m2 or more and as obese if it is 30.0 kg/m2 or more. Since excess weight is a predisposing factor for many chronic diseases, e.g. diabetes, an increase in its incidence in the population is cause for concern. Until now, excess weight has been problem of epidemic proportions only in developed countries, but it has recently spread to the developing world. More than 30% of the population in Latin America, the Caribbean, the Middle East, and Northern Africa is overweight. Populations living on Pacific and Indian Ocean islands have the highest prevalence of obesity in the world. In Asia and Black Africa, the overall prevalence of overweight is still low but incidence is high in urban areas. In most of these countries, both underweight and overweight people can now be seen. In many countries, the increase in the number of overweight people has occurred within the last few years. Excess weight appears first among the affluent and then among low-income classes including young children and teenagers. The main causes are a nutrition transition to lipid-rich diets and, above all, reduced physical activity in city dwellers. Obesity and associated diseases could become major problems in the future since malnutrition during fetal development and early childhood are predisposing factors. Already overweight is creating an extra burden for countries where malnutrition and nutritional deficiencies are still observed in young children. Given the economic costs of management of obesity-related diseases, surveillance and prevention programs are needed to stem the growth of this problem.


Zfat-deficiency results in a loss of CD3ζ phosphorylation with dysregulation of ERK and Egr activities leading to impaired positive selection.
The human ZFAT gene was originally identified as a susceptibility gene for autoimmune thyroid disease. Mouse Zfat is a critical transcriptional regulator for primitive hematopoiesis and required for peripheral T cell homeostasis. However, its physiological roles in T cell development remain poorly understood. Here, we generated Zfat (f/f)-LckCre mice and demonstrated that T cell-specific Zfat-deletion in Zfat (f/f)-LckCre mice resulted in a reduction in the number of CD4(+)CD8(+)double-positive (DP) cells, CD4(+)single positive cells and CD8(+)single positive cells. Indeed, in Zfat (f/f)-LckCre DP cells, positive selection was severely impaired. Defects of positive selection in Zfat-deficient thymocytes were not restored in the presence of the exogenous TCR by using TCR-transgenic mice. Furthermore, Zfat-deficient DP cells showed a loss of CD3ζ phosphorylation in response to T cell antigen receptor (TCR)-stimulation concomitant with dysregulation of extracellular signal-related kinase (ERK) and early growth response protein (Egr) activities. These results demonstrate that Zfat is required for proper regulation of the TCR-proximal signalings, and is a crucial molecule for positive selection through ERK and Egr activities, thus suggesting that a full understanding of the precise molecular mechanisms of Zfat will provide deeper insight into T cell development and immune regulation.


Assessing DSM-5 nonsuicidal self-injury disorder in a clinical sample.
The entry for nonsuicidal self-injury (NSI) disorder in the 5th edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) provides a criterion-based definition of clinically relevant NSI. NSI disorder is currently classified in the DSM-5 as a condition requiring further study. The present study aimed to examine the reliability, validity, and clinical utility of a self-report measure of NSI disorder, the Alexian Brothers Assessment of Self-Injury (ABASI). The sample included 511 patients admitted to an acute care treatment program designed to treat NSI. Patients were administered the ABASI as part of a clinical assessment and routine outcome evaluation. The sample included a broad age range, as well as sufficient numbers of males and Hispanics to examine sociodemographic differences. The ABASI demonstrated adequate internal consistency and test-retest reliability, and the factor structure reflects NSI disorder criteria. Among patients being treated for NSI, 74% met criteria for NSI disorder. No differences in the rate of NSI disorder were observed by sex, ethnicity, or age. Although NSI disorder is associated with a worse presentation of self-injurious behavior, NSI disorder provides limited clinical utility as a dichotomous diagnosis, at least when compared with common NSI characteristics such as number of methods of NSI and the urge to self-injure. Instead, findings support a dimensional approach to NSI disorder. Analyses of specific symptoms of NSI disorder indicate concerns with Criterion B as currently defined by the DSM-5. Recommendations for a more parsimonious revision of NSI disorder are discussed.


Determination of loyalty among high school students to retain in the same university for higher education: An integration of Self-Determination Theory and Extended Theory of Planned Behavior.
Student loyalty generally refers to the formed bond between the student and a university. This relationship between a university and its students proves essential in a university's success in the competitive field of higher education institutions. The aim of this study was to determine the factors affecting students' loyalty among high school students to pursue their college or higher education in their current universities by utilizing Self-Determination Theory and Extended Theory of Planned Behavior. A total of 1224 high school students voluntarily participated and answered an online questionnaire that consist of 80 questions. Structural Equation Modeling (SEM) showed that competency had the highest direct significant effect on perceived behavioral control which subsequently led to student satisfaction, followed by relatedness and empathy. In addition, student satisfaction had the highest direct effect significant effect on student loyalty, followed by university image and effectiveness. Interestingly, university ranking, programs offered, and kinship patronage also had significant indirect effects on student loyalty. This new framework may be a theoretical foundation for universities to enhance student loyalty and student recruitment. Considering students as customers, the satisfaction of students would result in an increase in the application which would present an increase in population, sales, marketability, and profitability of the university.


A multifunctional, modified rigid neuroendoscopic system: clinical experience with 83 procedures. Technical note.
The authors combined a monopolar electrode and a suction/irrigation channel with a 0 degrees, 4-mm Hopkins rigid telescope into a single multifunctional unit. This three-in-one instrument is inserted through a lightweight 7.5-mm outer sheath, which is fixed separately. A fourth instrument (for example, a balloon catheter or a biopsy forceps) can be introduced and manipulated independently with the other hand. All endoscopic procedures were performed with a trephine to create a 15-mm craniotomy. After opening the dura mater, ventricles were tapped with a brain needle, which was followed by the insertion of the rigid scope for visualization. The telescope was then withdrawn momentarily; the outer sheath was introduced into the ventricle and fixed over the area of interest. The definitive procedure was then performed with ease by using the multifunctional three-in-one instrument in one hand and a fourth instrument in the other hand. This novel neuroendoscopic system has been used in clinical testing at the Vidyasagar Institute of Mental Health and Neurosciences since May 1998. Thus far, 83 neuroendoscopic procedures have been successfully performed with the aid of this instrumentation system, which has proven to be safe, versatile, and cost-effective, allowing a greater degree of freedom for the neurosurgeon.


Propylthiouracil-induced hepatic damage.
To report a case of propylthiouracil-induced hepatic damage. A 64-year-old white woman with hyperthyroidism received propylthiouracil 250 mg/d for 1 year. She developed hepatitis after 1 year of therapy. Alcohol and drug abuse were ruled out and all serologic tests for hepatitis A, B, and C were negative. Cytomegalovirus and Epstein-Barr virus infection were also ruled out. Antinuclear antibody, antimitochondrial antibody, and antismooth muscle antibody were negative. The clinical picture was similar to that of viral hepatitis characterized by nausea, vomiting, and jaundice. Histologic examination of a liver biopsy specimen showed chronic active hepatitis. The patient developed cirrhosis during follow-up. Propylthiouracil is widely used in the treatment of hyperthyroidism. Despite its widespread use, there have been only a few reported cases of propylthiouracil-induced hepatotoxicity. The precise mechanism of the injury is unknown, although immunologic factors are suggested. Hepatic damage induced by propylthiouracil is a rare complication. However, the danger of permanent hepatic damage should be kept in mind. The best way of preventing propylthiouracil hepatotoxicity is careful screening of patients considered for treatment.


Domain wall conductivity in La-doped BiFeO3.
The transport physics of domain wall conductivity in La-doped bismuth ferrite (BiFeO3) has been probed using variable temperature conducting atomic force microscopy and piezoresponse force microscopy in samples with arrays of domain walls in the as-grown state. Nanoscale current measurements are investigated as a function of bias and temperature and are shown to be consistent with distinct electronic properties at the domain walls leading to changes in the observed local conductivity. Our observation is well described within a band picture of the observed electronic conduction. Finally, we demonstrate an additional degree of control of the wall conductivity through chemical doping with oxygen vacancies, thus influencing the local conductive state.


Attitudes of physicians, patients, and village health workers toward glaucoma and diabetic retinopathy in rural China: a focus group study.
OBJECTIVE To understand the knowledge and attitudes of rural Chinese physicians, patients, and village health workers (VHWs) toward diabetic eye disease and glaucoma. METHODS Focus groups for each of the 3 stakeholders were conducted in 3 counties (9 groups). The focus groups were recorded, transcribed, and coded using specialized software. Responses to questions about barriers to compliance and interventions to remove these barriers were also ranked and scored. RESULTS Among 22 physicians, 23 patients, and 25 VHWs, knowledge about diabetic eye disease was generally good, but physicians and patients understood glaucoma only as an acutely symptomatic disease of relatively low prevalence. Physicians did not favor routine pupillary dilation to detect asymptomatic disease, expressing concerns about workflow and danger and inconvenience to patients. Providers believed that cost was the main barrier to patient compliance, whereas patients ranked poorly trained physicians as more important. All 3 stakeholder groups ranked financial interventions to improve compliance (eg, direct payment, lotteries, and contracts) low and preferred patient education and telephone contact by nurses. All the groups somewhat doubted the ability of VHWs to screen for eye disease accurately, but patients were generally willing to pay for VHW screening. The VHWs were uncertain about the value of eye care training but might accept it if accompanied by equipment. They did not rank payment for screening services as important. CONCLUSIONS Misconceptions about glaucoma's asymptomatic nature and an unwillingness to routinely examine asymptomatic patients must be addressed in training programs. Home contact by nurses and patient education may be the most appropriate interventions to improve compliance.


The VP4 and VP7 of bovine rotavirus VMRI are antigenically and genetically closely related to P-type 5, G-type 6 strains.
We have previously reported the isolation of a bovine rotavirus, designated VMRI, with a super-short electropherotype. We have characterized this strain further as it has shown antigenic differences with the prototype G6 strain NCDV-Lincoln. In this communication, we report the antigenic and molecular characterization and the nucleotide sequence of the VP4 and VP7 genes of this strain. Virus neutralization tests indicated 2- to 13-fold differences in the titers between NCDV-Lincoln, B641 and VMRI strains. Northern blot hybridization results indicated a degree of heterogeneity in the VP4 gene of these strains which can be detected under conditions of high stringency. The VP4 and VP7 genes of the VMRI strain were cloned and sequenced and compared with the published sequences of other bovine rotavirus strains. The VP4 gene of VMRI had a high degree of homology with that of UK and B641 strains but differed significantly from that of both NCDV-Lincoln and B223 strains. Sequence analysis of the VP7 gene of VMRI and other strains indicated a high degree of conservation and the amino acid identity between the different strains was 96%. Sequence information regarding these strains and field isolates will assist in the generation of effective vaccination strategies for control of neonatal calf diarrhea.


Variations in Physical Activity and Screentime According to School Format During the COVID-19 Pandemic.
Early studies indicate children's physical activity (PA) decreased during the pandemic but may vary by school format and season. We longitudinally assessed changes in PA and screentime (ST) behaviors throughout 1 year of the COVID-19 pandemic and according to changes in school format. Parents of children (5-18 years) completed a survey about school format, home characteristics, PA, and ST. Initial data were collected in November 2020 with follow-up in February, May, and August 2021. Physical activity and ST were compared by season and school format using a 4 (format) × 4 (season) repeated measures analysis of variance. Children were active for at least 60 minutes/day less often in winter (2.4 ± 0.3 days; p < .001; n = 176) than in other seasons. Longitudinal analysis (n = 61) showed that PA was highest among students attending school fully in-person and lowest among students attending remotely, though school format differences were not significant. Non-school ST did not differ by season or school format. These results suggest that changes in school format influence students' lifestyle behaviors. The impact on children's lifestyle behaviors should be considered in planning for safe, in-person operation of schools in the event of future pandemics or natural disasters.


Growth of artificially fed infant rats: effect of supplementation with insulin-like growth factor I.
Insulin-like growth factor I (IGF-I), a potent mitogenic peptide, is present in considerable quantities in most mammalian milks, but its importance for the neonate is unknown. To test the hypothesis that milk-borne IGF-I is an important factor in the regulation of neonatal growth, as well as that of the gastrointestinal tract, rat pups were fed a rat milk substitute (RMS) devoid of growth factors via gastrostomy. These animals were compared with those given RMS supplemented with recombinant human IGF-I added at a concentration of 500 ng/ml. Animals given RMS + IGF-I gained mere weight than controls, although skeletal growth as represented by elongation of the tail was no different. Animals fed RMS + IGF-I had increased brain and liver wet weights as well as increased liver and small intestine protein contents. Serum IGF-I concentrations in the IGF-I-supplemented group were more than twofold above RMS controls and were similar to dam-fed rat pups. Semiquantification of serum IGF-binding proteins (IGFBP) in these animals documented that in IGF-I-supplemented pups the amount of 38- to 40-kDa molecular mass IGFBP species was also greater than in RMS controls. The rate of migration of enterocytes from crypts in duodenum and proximal jejunum was greater in IGF-I-supplemented animals than in rats fed RMS alone. These studies suggest that milk-borne IGF-I is important in modulation of somatic and gastrointestinal tract growth in the neonatal rat.


A Fibrous Hybrid Patch Couples Cell-Derived Matrix and Poly(l-lactide-co-caprolactone) for Endothelial Cells Delivery and Skin Wound Repair.
Skin wound healing is an intricate orchestration that involves different cell types, an extracellular matrix (ECM), cytokines, and growth factors. On the basis of the great benefits of cell-derived ECM in regenerative applications, here we propose an electrospun fibrous membrane that combines poly(l-lactide-co-caprolactone) (PLCL) and human fibroblast-derived ECM (hFDM). hFDM-deposited PLCL (hFDM-PLCL) was obtained via decellularization of a confluent layer of fibroblasts cultivated on PLCL. An organized assembly of fibrillar structure on hFDM-PLCL was notable via immunostaining. As human umbilical vein endothelial cells (HUVECs) were seeded on the hFDM-PLCL, they proliferated faster and exhibited more elongated, capillary-like morphology than those on PLCL or fibronectin-coated PLCL (Fn-PLCL). HUVECs have a relatively large aspect ratio, spreading area, and vinculin-positive area per cell on the fibrillary structure of hFDM-PLCL. In addition, transwell cell migration assay showed the chemoattractant effect of hFDM for HUVECs and human dermal fibroblasts. Furthermore, HUVECs-loaded hFDM-PLCL membranes showed the most promising therapeutic effects on a mouse skin wound model as proved via the wound closure rate, neovascularization effect, regenerated epidermis, and skin appendage. This study shows that biodegradable PLCL fibers not only support the weak mechanical properties of hFDM but also allow hFDM to reserve ECM macromolecules and to maintain structural integrity. Current results also demonstrate the critical role of hFDM with biochemical and biophysical cues on HUVECs adhesion, proliferation, and vascular morphogenesis in vitro and even on the wound healing process in vivo. Taken together, hFDM-functionalized PLCL patch should be a promising platform for cell delivery and regenerative applications.