The role of passive calcium influx through the cell membrane in galvanotaxis.
Passive calcium influx is one of the theories to explain the cathodal galvanotaxis of cells that utilize the electric field to guide their motion. When exposed to an electric field, the intracellular fluid becomes polarized, leading to positive charge accumulation on the cathodal side and negative charge accumulation on the anodal side. The negative charge on the anodal side attracts extracellular calcium ions, increasing the anodal calcium concentration, which is supposed to decrease the mobile properties of this side. Unfortunately, this model does not capture the Ca(2+) dynamics after its presentation to the intracellular fluid. The ions cannot permanently accumulate on the anodal side because that would build a potential drop across the cytoplasm leading to an ionic current, which would carry positive ions (not only Ca(2+)) from the anodal to the cathodal part through the cytoplasm. If the cytoplasmic conductance for Ca(2+) is low enough compared to the membrane conductance, the theory could correctly predict the actual behavior. If the ions move through the cytoplasm at a faster rate, compensating for the passive influx, this theory may fail. This paper contains a discussion of the regimes of validity for this theory.


Phototaxis of active colloids by self-thermophoresis.
Phototaxis usually refers to the ability of a motile micoorganism to move directionally within a light gradient, which is important for microorganisms to gain energy or avoid damage. Here, we show that an active Janus particle driven by self-thermophoresis from light heating is able to exhibit significant phototactic motion by means of mesoscale dynamics simulation. Depending on the particle-fluid interactions, the active particle can move along or against the light gradient, corresponding to positive or negative phototaxis, respectively. We find that the phototaxis of the active colloid is determined by various mechanisms, including alignment (polarization) of the particle to the light gradient, orientation-dependent motility and spatially inhomogeneous motility. Our results shed light on the phototactic behavior of artificial active colloids and open up a new possibility to design photo-responsive micromachines.


Production and functional evaluation of nonfat dry milk with whey proteins as fibrils.
Commercial manufacturing of dairy products involves the addition of dairy ingredients (such as nonfat dry milk and milk protein concentrates) as well as non-dairy additives (such as gums, stabilizers, emulsifiers, and texture modifiers) to get the best product appearance, maintain the product quality, and extend shelf-life. Though these non-dairy additives are not harmful, consumers do not prefer them in dairy food formulations. Therefore, the dairy industry is working on improving the inherent functionality of dairy ingredients using different processes. Recently, fibrillation emerged as a new technique to convert globular proteins such as whey proteins into fibrils, which provide enhanced viscosity, foaming, and emulsification capacity. Therefore, skim milk was subjected to microfiltration followed by ultrafiltration of microfiltration permeate to fractionate whey proteins. Then, whey proteins were selectively fibrillated and mixed back with other streams of microfiltration and ultrafiltration to get fibrillated skim milk. Fibrillated skim milk was spray-dried to get fibrillated nonfat dry milk (NDM). Visible whey protein fibrils were observed in reconstituted fibrillated NDM which showed survival of fibrils in fibrillated NDM. Fibrillated NDM showed significantly (P <0.05) higher viscosity than control NDM. Fibrillated NDM also showed higher emulsification capacity, foaming capacity, and stability than the control NDM but lower gel strength. Considering the improved functionality of fibrillated NDM, they can be used in product formulations such as ice cream mix where the thickening of a solution, good emulsification, and foaming properties are required.


Antimicrobial-resistant and ESBL-producing Escherichia coli in different ecological niches in Bangladesh.
The rapid and wide-scale environmental spread of multidrug-resistant bacteria in different ecosystems has become a serious issue in recent years. To investigate the epidemiology of antimicrobial resistance and extended spectrum beta-lactamase (ESBL) in Bangladeshi wild birds and aquatic environments, samples were taken from Open Bill Stork (Anastomus oscitans) (OBS) and the nearby water sources. Water and fresh fecal samples were collected from several locations. All samples were processed and cultured for Escherichia coli and tested for antibiotic susceptibility against commonly used antibiotics. ESBL producers were characterized at genotypic level using polymerase chain reaction (PCR), sequencing, multilocus sequence typing, and rep-PCR. A total of 76 E. coli isolates from the 170 OBS and 8 E. coli isolates from three river sources were isolated. In total, 29% of E. coli isolated from OBS and all of the E. coli isolated from water sources were resistant to at least one of the tested antimicrobials. Resistant phenotypes were observed with all antimicrobials except tigecycline, gentamicin, imipenem, and chloramphenicol. Multidrug resistance was observed in 2.6% of OBS and 37.5% of the water isolates. Also, 1.2% of the ESBL-producing E. coli were isolated from OBS, whereas 50% of the E. coli isolated from water sources were ESBL producers possessing the CTX-M-15 gene. The most concerning aspect of our findings was the presence of human-associated E. coli sequence types in the water samples, for example, ST156-complex156, ST10-complex10 and ST46. This study reports the presence of multidrug-resistant ESBL-producing E. coli in OBSs and nearby aquatic sources in Bangladesh.


Tumor-induced Stromal STAT1 Accelerates Breast Cancer via Deregulating Tissue Homeostasis.
The tumor microenvironment (TME), the dynamic tissue space in which the tumor exists, plays a significant role in tumor initiation, and is a key contributor in cancer progression; however, little is known about tumor-induced changes in the adjacent tissue stroma. Herein, tumor-induced changes in the TME were explored at the morphologic and molecular level to further understand cancer progression. Tumor-adjacent mammary glands (TAG) displayed altered branching morphology, expansion of myofibroblasts, and increased mammosphere formation, broadly suggesting a tumor-induced field effect. FACS analysis of TAGs demonstrated an increased number of Lin-CD24+/CD49+ enriched mammary gland stem cells (MaSC), suggesting deregulated tissue homeostasis in TAGs. Comparative transcriptome analysis of TAGs and contralateral control glands coupled with meta-analysis on differentially expressed genes with two breast cancer stromal patient microarray datasets identified shared upregulation of STAT1. Knockdown of STAT1 in cancer-associated fibroblast (CAF) cocultured with human breast cancer cells altered cancer cell proliferation, indicating a role for STAT1 as a stromal contributor of tumorigenesis. Furthermore, depletion of STAT1 in CAFs significantly reduced periductal reactive fibrosis and delayed early breast cancer progression in vivo Finally, cotreatment with fludarabine, a FDA-approved STAT1 activation inhibitor and DNA synthesis inhibitor, in combination with doxorubicin, showed enhanced therapeutic efficacy in treating mouse mammary gland tumors. Taken together, these results demonstrate that stromal STAT1 expression promotes tumor progression and is a potential therapeutic target for breast cancer.Implications: Tumors induce stromal STAT1-dependent cytokine secretion that promotes tumor cell proliferation and can be targeted using clinically-approved inhibitors of STAT1. Mol Cancer Res; 15(5); 585-97. ©2017 AACR.


Clinical and anatomopathological aspects of patients with hantavirus cardiopulmonary syndrome in Uberaba, Minas Gerais, Brazil.
The hantavirus cardiopulmonary syndrome is considered an emerging disease in the Americas. Since 1993, thousands of cases have been reported from different countries, but mainly from Brazil. This study aims to describe some epidemiological, clinical and anatomopathological aspects of patients with hantavirus who presented poor outcome and were autopsied in a teaching hospital in Brazil, from 2000 to 2014. Of the 10 patients included, nine were male (mean age 43.5 years) and seven reported previous contact with rodents. Fever was present in eight of ten patients, dyspnea in nine of ten and myalgia in seven of ten patients; hemoconcentration, leukocytosis, thrombocytopenia and renal involvement were evidenced in all the 10 cases. At autopsy, the main alterations were seen in the lungs: pleural effusion (8/10 cases), increased weight 2.5 to 3 times, congestion/edema (10/10), interstitial mononuclear inflammation (10/10), alveolar hemorrhage (7/10), pulmonary collapse (7/10), hyaline membranes (7/10) and alveolar neutrophilic infiltrate (2/10). Pericardial effusion (2/10), mild myocardium inflammation (4/10), right ventricle dilation (1/10), polyploidy nuclei (3/10) and pericardial diffuse petechial (1/10) were also observed. The other organs exhibited discrete and non-specific alterations. Currently, this syndrome continues to be associated with high mortality directly linked to a late diagnosis and/or a misdiagnosis in the medical centers where these patients were seen for the first time. The anatomopathological findings at autopsy revealed the final phase of the process with pulmonary alterations, allowing a direct correlation with the severity of respiratory distress observed in these patients at admission.


Killing of intracellular Leishmania donovani by human mononuclear phagocytes. Evidence for oxygen-dependent and -independent leishmanicidal activity.
Human peripheral blood monocytes were cultivated for 1-30 d before assay for H2O2 release or challenge with Leishmania donovani promastigotes (LDP) or amastigotes (LDA). 1-d cells readily generated H2O2 in response to both phorbol myristate acetate triggering (1,013 +/- 58 nmol/mg protein . 90 min) and LDP ingestion, and killed 50% of LDP within 6 h, and 90% by 24 h. In contrast, the same cells released little H2O2 during LDA ingestion, killed no LDA at 6 h and less than 30% by 24 h, and supported intracellular LDA replication. Monocyte-derived macrophages (cells first cultivated for greater than or equal to 7 d) generated less than 125 nmol H2O2/mg . 90 min after phorbol myristate acetate triggering, killed neither LDP nor LDA, and permitted both forms to replicate. The addition of mitogen- or antigen-stimulated lymphokines, however, prevented the decline in monocyte oxidative capacity, enhanced macrophage H2O2 release by more than sixfold, and, in parallel, induced 1-d monocytes to kill LDA and cultivated macrophages to display both promastigocidal and amastigocidal activity. In comparison to 1-d monocytes and lymphokine-activated macrophages from normal donors, the same cells from patients with chronic granulomatous disease (CGD) or normal cells whose oxidative activity had been impaired by catalase pretreatment or glucose deprivation exerted considerably less or no antileishmanial activity during the early (6-24 h) postphagocytic period. By 48 h after infection, however, 1-d CGD monocytes and oxidatively impaired normal cells killed 40 and greater than 80% of LDP, respectively. Although a longer period of lymphokine stimulation was required and the resulting antileishmanial effects were not as rapid as with normal cells, activated CGD monocytes and macrophages also eventually achieved promastigocidal and amastigostatic activity. These results indicate that human mononuclear phagocytes utilize both oxygen-dependent and -independent mechanisms to achieve activity against ingested Leishmania, and also demonstrate (a) the differential susceptibilities of the two forms of L. donovani to intracellular killing, (b) the key role of oxygen intermediates in effective mononuclear phagocyte antimicrobial activity, (c) the capacity of lymphocyte products to enhance oxygen-dependent as well as -independent pathways, and (d) the vulnerability of the monocyte-derived macrophage to Leishmania infection in the absence of lymphokine stimulation.


Comparison of health status and quality of life of related versus paid unrelated living kidney donors.
The aim of this cross-sectional study was to assess the health status and quality of life (QOL) of paid unrelated versus related living kidney donors postdonation at Shiraz Transplant Center in Iran. We invited all donors (n = 580, 347 paid unrelated, 233 related) who underwent donor nephrectomy at our center from 2004 to 2010 to participate in a health survey and physical examination. Of 580 donors, 144 consented to participate; participation of paid unrelated donors was significantly lower than related (52/347 vs. 92/233; p < 0.001). Participants underwent a complete physical examination, QOL assessment (using a 36-item short form health survey [SF-36] questionnaire) and laboratory work-up. The paid unrelated donors compared with related donors were younger (34.2 ± 7.2 vs. 40.7 ± 9.7 years, p < 0.001), had shorter time since donation (2.9 ± 1.6 vs. 3.8 ± 2 years, p = 0.004), had higher estimated GFR (72.6 ± 22 vs. 63.8 ± 15.3 mL/min/1.73 m(2), p = 0.006) and had a higher percentage of patients with microalbuminuria (35% vs. 0%, p < 0.001). Additionally, general health and social functioning scores among paid unrelated donors were significantly lower (p < 0.001 and p = 0.02, respectively) than related donors. Other SF-36 scores, although lower in paid unrelated donors, did not reach statistical significance. Iranian paid unrelated donors have lower QOL and higher incidence of microalbuminuria compared with related donors.


Potential Risk Factors and Prevalence of Voice Symptoms in Students Starting Their Teacher Education.
The aim of the present study was to determine prevalence of voice problems and potential risk factors in teacher students at the start of their education. A total of 1494 students from seven teacher education schools participated in the study. The students answered a questionnaire about 11 risk factors, and one with six questions about voice symptoms, Screen6, and 30 statements in the Voice Handicap Index (VHI). Students reporting at least 2 weekly voice symptoms in Screen6 were assigned to the group with voice problems. Significance level was P < 0.05. Results: prevalence of risk factors was about the same in the seven groups of teacher students except the group with highest proportion of women that also showed the highest proportion of voice problems, 38%. Prevalence of voice problems in the total group was 17%. Comparison of students with or without voice problems showed that all factors except one were more prevalent among students with voice problems and women were overrepresented. Mean total VHI score was 22 in students with voice problems and eight in students without voice problems. Multiple regression analyses showed that frequent throat infections, hearing problems, previous speech therapy, or voice training were the potential risk factors that had the strongest association with voice symptoms as well as with total score in VHI. Conclusions: results from this study show that it is common that teacher students experience voice problems already at the start of their education and potential risk factors associated with voice problems are identified. Knowing that teaching is a high-risk profession for developing voice disorders, it is crucial that teacher students should receive compulsory preprofessional voice education including voice ergonomics and voice training.


A comparison of the effects of thymoquinone, silymarin and N-acetylcysteine in an experimental hepatotoxicity.
This study investigated the effects of thymoquinone, silymarin, and N-acetylcysteine in a rat model with carbon tetrachloride (CCl4)-induced hepatotoxicity. Although numerous similar studies are available, we aimed to compare the efficacy of these agents by considering N-acetylcysteine as a reference compound. A total of 50 male Wistar albino rats were randomly designated as 5 groups: Group I, CCl4; group II, thymoquinone and CCl4; group III, silymarin and CCl4; group IV, N-acetylcysteine and CCl4; group V, control group. CCl4 was administered intraperitoneally at a dose of 1.5 mL/kg (a mixture of CCl4: olive oil, 1:2) twice a week. Thymoquinone was administered at a dose of 10 mg/kg, silymarin was administered at a dose of 100 mg/kg, and N-acetylcysteine was administered at a dose of 100 mg/kg by daily intraperitoneal injection. At the end of four weeks, blood and liver tests were analyzed. The results were evaluated statistically via the one-way ANOVA test. A p-value <0.05 was considered statistically significant. Thymoquinone, silymarin, and N-acetylcysteine improved the levels of alanine aminotransferase, tumor necrosis factor-α, platelet-derived growth factor-BB, and interleukin-6, which were increased by CCl4. Thymoquinone and silymarin showed the positive increase in liver glutathione levels. Thymoquinone, silymarin, and N-acetylcysteine improved blood total oxidant status. In the histological examinations of liver tissue, thymoquinone decreased necrosis, and inflammation. The most positive decrease in the α-smooth muscle actin-stained hepatic stellate cell count was only observed with thymoquinone. These findings suggest that thymoquinone, silymarin, and N-acetylcysteine have potential for the treatment of diseases causing liver injury. Among these agents, thymoquinone showed the best results on most of the parameters. Since TQ appears to be at least as effective as SM and NAC in our in-vitro study, we propose that it is time for clinical studies with thymoquinone on hepatotoxicity.


Comparison of Melanoma Subtypes among Korean Patients by Morphologic Features and Ultraviolet Exposure.
Genetic alterations have been identified in melanomas according to different levels of sun exposure. Whereas the conventional morphology-based classification provides a clue for tumor growth and prognosis, the new classification by genetic alterations offers a basis for targeted therapy. The purpose of this study is to demonstrate the biological behavior of melanoma subtypes and compare the two classifications in the Korean population. A retrospective chart review was performed on patients found to have malignant melanoma in Severance Hospital from 2005 to 2012. Age, sex, location of the tumor, histologic subtype, tumor depth, ulceration, lymph node invasion, visceral organ metastasis, and overall survival were evaluated. Of the 206 cases, the most common type was acral melanoma (n=94, 45.6%), followed by nonchronic sun damage-induced melanoma (n=43, 20.9%), and mucosal melanoma (n=40, 19.4%). Twenty-one patients (10.2%) had the chronic sun-damaged type, whereas eight patients (3.9%) had tumors of unknown primary origin. Lentigo maligna melanoma was newly classified as the chronic sun-damaged type, and acral lentiginous melanoma as the acral type. More than half of the superficial spreading melanomas were newly grouped as nonchronic sun-damaged melanomas, whereas nodular melanoma was rather evenly distributed. The distribution of melanomas was largely similar in both the morphology-based and sun exposure-based classifications, and in both classifications, mucosal melanoma had the worst 5-year survival owing to its tumor thickness and advanced stage at the time of diagnosis.


Spotted fever group rickettsiae from ticks captured in Sudan.
Ticks were collected from ruminants in various areas of Sudan in 1998 and 2000. Primer pairs of rickettsial citrate synthase gene (gltA) and a spotted fever group (SFG) rickettsial 190-kDa surface antigen gene (rompA), respectively, were used for identification. Polymerase chain reaction (PCR)-positive products were used for DNA sequencing. The gltA gene was detected in 55% of the ticks examined (57/104). Among the 57 ticks studied, 19 were positive for the rompA gene. Thus, 18% of the ticks examined were found to be infected with SFG rickettsiae. The nucleotide sequences of the amplified rompA gene fragment of Hyalomma spp. and Amblyomma spp. were similar to those of Rickettsia aeschlimannii and Rickettsia africae, respectively. In this study, we succeeded in detecting the SFG rickettsiae gene in ticks, and established that there were at least two species of SFG rickettsiae in field ticks in Sudan.


Randomized trial of a depression management program in high utilizers of medical care.
High utilizers of nonpsychiatric health care services have disproportionally high rates of undiagnosed or undertreated depression. To determine the impact of offering a systematic primary care-based depression treatment program to depressed "high utilizers" not in active treatment. Randomized clinical trial. One hundred sixty-three primary care practices in 3 health maintenance organizations located in different geographic regions of the United States. A group of 1465 health maintenance organization members were identified as depressed high utilizers using a 2-stage telephone screening process. Eligibility criteria were met by 410 patients and 407 agreed to enroll: 218 in the depression management program (DMP) practices and 189 in the usual care (UC) group. The DMP included patient education materials, physician education programs, telephone-based treatment coordination, and antidepressant pharmacotherapy initiated and managed by patients' primary care physicians. Depression severity was measured using the Hamilton Depression Rating Scale (Ham-D) and functional status using the Medical Outcomes Study 20-item short form (SF-20) subscales. Outpatient visit and hospitalization rates were measured using the health plan's encounter data. Based on an intent-to-treat analysis, at least 3 antidepressant prescriptions were filled in the first 6 months by 151 (69.3%) of 218 of DMP patients vs 35 (18.5%) of 189 in UC (P < .001). Improvements in Ham-D scores were significantly greater in the intervention group at 6 weeks (P = .04), 3 months (P = .02), 6 months (P < .001), and 12 months (P < .001). At 12 months, DMP intervention patients were more improved than UC patients on the mental health, social functioning, and general health perceptions scales of the SF-20 (P < .05 for all). In depressed high utilizers not already in active treatment, a systematic primary care-based treatment program can substantially increase adequate antidepressant treatment, decrease depression severity, and improve general health status compared with usual care.


Enzyme histochemistry of rat interfascicular oligodendroglia, with special reference to 5'-nucleotidase.
The interfascicular oligodendroglia of the rat striatum nucleus is studied in control animals and under experimentally induced brain edemas. Enzyme histochemical studies were performed. Normal oligodendrocytes show two different enzymatic distributions. While Mg2+-ATPase, TPPase, TPPase, adenylate cyclase and SDH are shown to be situated at the cell bodies and some proximal processes, 5'-nucleotidase reaction is seen at the cell fine distal processes. In spite of its trong activity in oligodendroglial endings the latter enzyme is not seen at the myelin sheath. This sheath and the interfascicular cells being intimately related, 5'-nucleotidase appears to be important not only during myelination, as almost all authors emphasize, but also in the adult myelin metabolism. Experimental brain edema shows some changes in the interfasicular oligodendroglia enzymatic activites for 5'-nucleotidase, Mg2+-ATPase and adenylate cyclase. A progressive disappearance of the former enzyme is observed. These features and those of derived from local ionic alterations may be concerned with the special susceptibility of white matter for some types of brain swellings.


Impact of 131I-SPECT/CT images obtained with an integrated system in the follow-up of patients with thyroid carcinoma.
The purpose of the study was to determine the diagnostic impact of 131I-SPECT/CT imaging compared with conventional scintigraphic evaluation in the follow-up of patients with thyroid carcinoma. Seventy-one patients with thyroid carcinoma underwent concurrent 131I-SPECT/CT, using an integrated imaging system, at various stages of their disease in order to evaluate foci of uptake detected on planar whole-body images. SPECT/CT imaging had an incremental diagnostic value in 57% (41/71) of patients. Uptake in the neck was evaluated in 61 patients, and SPECT/CT imaging in this region had an incremental diagnostic value in 27% of the whole patient population (19/71). Low-resolution integrated CT images allowed for the precise characterization of equivocal neck lesions on planar imaging in 14/17 patients and changed the assessment of the lesion location in five patients as compared with planar studies. Thirty-six patients underwent SPECT/CT for evaluation of foci of uptake distant from the neck. SPECT/CT imaging improved characterization of equivocal foci of uptake as definitely benign in 13% (9/71) of patients. Precise localization of malignant lesions to the skeleton was possible in 17% (12/71) and to the lungs versus the mediastinum in 6% (5/71) of patients. Integrated 131I-SPECT/CT was found to have an additional value over planar imaging in patients with thyroid cancer for correct characterization of equivocal tracer uptake seen on planar imaging as well as for precise localization of malignant lesions in the neck, chest, and skeleton. SPECT/CT optimized the localization of 131I uptake to lymph node metastases versus remnant thyroid tissue, to lung versus mediastinal metastases, and to the skeleton. It also had a further clinical impact on patient management by influencing referral for 131I treatment, tailoring of the administered radioiodine dose, and/or the addition of surgery or external radiation therapy when indicated.


p12(DOC-1) is a novel cyclin-dependent kinase 2-associated protein.
Regulated cyclin-dependent kinase (CDK) levels and activities are critical for the proper progression of the cell division cycle. p12(DOC-1) is a growth suppressor isolated from normal keratinocytes. We report that p12(DOC-1) associates with CDK2. More specifically, p12(DOC-1) associates with the monomeric nonphosphorylated form of CDK2 (p33CDK2). Ectopic expression of p12(DOC-1) resulted in decreased cellular CDK2 and reduced CDK2-associated kinase activities and was accompanied by a shift in the cell cycle positions of p12(DOC-1) transfectants ( upward arrow G(1) and downward arrow S). The p12(DOC-1)-mediated decrease of CDK2 was prevented if the p12(DOC-1) transfectants were grown in the presence of the proteosome inhibitor clasto-lactacystin beta-lactone, suggesting that p12(DOC-1) may target CDK2 for proteolysis. A CDK2 binding mutant was created and was found to revert p12(DOC-1)-mediated, CDK2-associated cell cycle phenotypes. These data support p12(DOC-1) as a specific CDK2-associated protein that negatively regulates CDK2 activities by sequestering the monomeric pool of CDK2 and/or targets CDK2 for proteolysis, reducing the active pool of CDK2.


A house finch (Haemorhous mexicanus) spleen transcriptome reveals intra- and interspecific patterns of gene expression, alternative splicing and genetic diversity in passerines.
With its plumage color dimorphism and unique history in North America, including a recent population expansion and an epizootic of Mycoplasma gallisepticum (MG), the house finch (Haemorhous mexicanus) is a model species for studying sexual selection, plumage coloration and host-parasite interactions. As part of our ongoing efforts to make available genomic resources for this species, here we report a transcriptome assembly derived from genes expressed in spleen. We characterize transcriptomes from two populations with different histories of demography and disease exposure: a recently founded population in the eastern US that has been exposed to MG for over a decade and a native population from the western range that has never been exposed to MG. We utilize this resource to quantify conservation in gene expression in passerine birds over approximately 50 MY by comparing splenic expression profiles for 9,646 house finch transcripts and those from zebra finch and find that less than half of all genes expressed in spleen in either species are expressed in both species. Comparative gene annotations from several vertebrate species suggest that the house finch transcriptomes contain ~15 genes not yet found in previously sequenced vertebrate genomes. The house finch transcriptomes harbour ~85,000 SNPs, ~20,000 of which are non-synonymous. Although not yet validated by biological or technical replication, we identify a set of genes exhibiting differences between populations in gene expression (n = 182; 2% of all transcripts), allele frequencies (76 FST ouliers) and alternative splicing as well as genes with several fixed non-synonymous substitutions; this set includes genes with functions related to double-strand break repair and immune response. The two house finch spleen transcriptome profiles will add to the increasing data on genome and transcriptome sequence information from natural populations. Differences in splenic expression between house finch and zebra finch imply either significant evolutionary turnover of splenic expression patterns or different physiological states of the individuals examined. The transcriptome resource will enhance the potential to annotate an eventual house finch genome, and the set of gene-based high-quality SNPs will help clarify the genetic underpinnings of host-pathogen interactions and sexual selection.


Four weeks versus six weeks of immobilization in a cast following closed reduction for displaced distal radial fractures in adult patients: a multicentre randomized controlled trial.
There is no level I evidence dealing with the optimal period of immobilization for patients with a displaced distal radial fracture following closed reduction. A shorter period might lead to a better functional outcome due to less stiffness and pain. The aim of this study was to investigate whether this period could be safely reduced from six to four weeks. This multicentre randomized controlled trial (RCT) included adult patients with a displaced distal radial fracture, who were randomized to be treated with immobilization in a cast for four or six weeks following closed reduction. The primary outcome measure was the Patient-Rated Wrist Evaluation (PRWE) score after follow-up at one year. Secondary outcomes were the abbreviated version of the Disability of Arm, Shoulder and Hand (QuickDASH) score after one year, the functional outcome at six weeks, 12 weeks, and six months, range of motion (ROM), the level of pain after removal of the cast, and complications. A total of 100 patients (15 male, 85 female) were randomized, with 49 being treated with four weeks of immobilization in a cast. A total of 93 completed follow-up. The mean PRWE score after one year was 6.9 (SD 8.3) in the four-week group compared with 11.6 (SD 14.3) in the six-week group. However, this difference of -4.7 (95% confidence interval -9.29 to 0.14) was not clinically relevant as the minimal clinically important difference of 11.5 was not reached. There was no significant difference in the ROM, radiological outcome, level of pain, or complications. In adult patients with a displaced and adequately reduced distal radial fracture, immobilization in a cast for four weeks is safe, and the results are similar to those after a period of immobilization of six weeks.


Docetaxel (Taxotere) and gemcitabine in combination therapy.
In patients with advanced cancers, gemcitabine and docetaxel (Taxotere; Rhône-Poulenc Rorer, Antony, France) can be administered together at 80% to 100% of their single-agent doses on days 1 and 8 of an every-3-week cycle without undue toxicity. In phase 1/11 studies, this combination has demonstrated activity against a range of tumors, including non-small cell lung cancer, and may represent an alternative to cisplatin-based chemotherapy. In advanced cancer, benefits in quality of life are likely to result. The fact that the combination of gemcitabine and docetaxel is generally well tolerated also suggests that the regimen should be assessed in the adjuvant and neoadjuvant settings.


Hypothalamic hamartoma, cerebellar hypoplasia, facial dysmorphism and very atypical combination of polydactyly: is it a new variant of oro-facio-digital syndrome?
We describe a newborn with multiple congenital anomalies consistent with an oro-facio-digital syndrome (OFDS). These are a group of inherited syndromes that have in common anomalies of the tongue (bifid or lobulated tongue with hamartomas), the face (median cleft lip) and the digits (brachydactyly, polydactyly, clinodactyly and/or syndactyly). OFDS has been classified into 11 types. The case described in this paper had manifestations overlapping with OFDS II (Mohr) and OFDS IV (Mohr-Majewski) and OFDS VI (Varadi). We propose that the present patient has a new variation of the OFDS due to the co-existence of the very atypical combination of polydactyly, cerebellar hypoplasia, hypothalamic hamartoma and classical facial findings of OFDS.


Isothermal Recombinase Polymerase Amplification-Lateral Flow Point-of-Care Diagnostic Test for Heartland Virus.
The detection of novel or re-emergent pathogens necessitates the development of rapid, easy-to-use diagnostic tests that can be readily adapted and utilized in both clinical laboratories and field settings. Heartland virus (HRTV) is the first pathogenic Phlebovirus responsible for serious and fatal cases in the United States. We developed a qualitative test based on recombinase-polymerase-amplification coupled with lateral flow reading (RPA-LF) for rapid detection of HRTV. The RPA-LF detected HRTV with a limit of detection of 1.19-1.54 plaque-forming unit equivalents/reaction. In addition, the RPA-LF was able to detect 0.6075 copies/μL of HRTV nucleoprotein gene-containing plasmid. We evaluated six clinical samples that were previously found to be real-time PCR positive for HRTV and found five out of six samples to be positive by RPA-LF, yielding 83.3% concordance with real-time PCR. All six samples had Ct values between 29 and 39 by real-time PCR. We also determined that the HRTV primers and probe do not cross-react with other tick-transmitted viruses such as Bourbon and Powassan, or other related viruses, including Lonestar tick virus and Sunday canyon virus (100% specificity). This is the first isothermal amplification test developed for a tick-borne virus, which will allow for rapid differentiation between HRTV and other pathogens producing similar clinical manifestations.


Proteomic Analysis of Lipid Rafts from RBL-2H3 Mast Cells.
Lipid rafts are highly ordered membrane microdomains enriched in cholesterol, glycosphingolipids, and certain proteins. They are involved in the regulation of cellular processes in diverse cell types, including mast cells (MCs). The MC lipid raft protein composition was assessed using qualitative mass spectrometric characterization of the proteome from detergent-resistant membrane fractions from RBL-2H3 MCs. Using two different post-isolation treatment methods, a total of 949 lipid raft associated proteins were identified. The majority of these MC lipid raft proteins had already been described in the RaftProtV2 database and are among highest cited/experimentally validated lipid raft proteins. Additionally, more than half of the identified proteins had lipid modifications and/or transmembrane domains. Classification of identified proteins into functional categories showed that the proteins were associated with cellular membrane compartments, and with some biological and molecular functions, such as regulation, localization, binding, catalytic activity, and response to stimulus. Furthermore, functional enrichment analysis demonstrated an intimate involvement of identified proteins with various aspects of MC biological processes, especially those related to regulated secretion, organization/stabilization of macromolecules complexes, and signal transduction. This study represents the first comprehensive proteomic profile of MC lipid rafts and provides additional information to elucidate immunoregulatory functions coordinated by raft proteins in MCs.


Variable selection in omics data: A practical evaluation of small sample sizes.
In omics experiments, variable selection involves a large number of metabolites/ genes and a small number of samples (the n < p problem). The ultimate goal is often the identification of one, or a few features that are different among conditions- a biomarker. Complicating biomarker identification, the p variables often contain a correlation structure due to the biology of the experiment making identifying causal compounds from correlated compounds difficult. Additionally, there may be elements in the experimental design (blocks, batches) that introduce structure in the data. While this problem has been discussed in the literature and various strategies proposed, the over fitting problems concomitant with such approaches are rarely acknowledged. Instead of viewing a single omics experiment as a definitive test for a biomarker, an unrealistic analytical goal, we propose to view such studies as screening studies where the goal of the study is to reduce the number of features present in the second round of testing, and to limit the Type II error. Using this perspective, the performance of LASSO, ridge regression and Elastic Net was compared with the performance of an ANOVA via a simulation study and two real data comparisons. Interestingly, a dramatic increase in the number of features had no effect on Type I error for the ANOVA approach. ANOVA, even without multiple test correction, has a low false positive rates in the scenarios tested. The Elastic Net has an inflated Type I error (from 10 to 50%) for small numbers of features which increases with sample size. The Type II error rate for the ANOVA is comparable or lower than that for the Elastic Net leading us to conclude that an ANOVA is an effective analytical tool for the initial screening of features in omics experiments.


Landscape effects on crop pollination services: are there general patterns?
Pollination by bees and other animals increases the size, quality, or stability of harvests for 70% of leading global crops. Because native species pollinate many of these crops effectively, conserving habitats for wild pollinators within agricultural landscapes can help maintain pollination services. Using hierarchical Bayesian techniques, we synthesize the results of 23 studies - representing 16 crops on five continents - to estimate the general relationship between pollination services and distance from natural or semi-natural habitats. We find strong exponential declines in both pollinator richness and native visitation rate. Visitation rate declines more steeply, dropping to half of its maximum at 0.6 km from natural habitat, compared to 1.5 km for richness. Evidence of general decline in fruit and seed set - variables that directly affect yields - is less clear. Visitation rate drops more steeply in tropical compared with temperate regions, and slightly more steeply for social compared with solitary bees. Tropical crops pollinated primarily by social bees may therefore be most susceptible to pollination failure from habitat loss. Quantifying these general relationships can help predict consequences of land use change on pollinator communities and crop productivity, and can inform landscape conservation efforts that balance the needs of native species and people.


p28GANK knockdown-derived reactive oxygen species induces apoptosis through mitochondrial dysfunction mediated by p38 in HepG2 cells.
Oncoprotein p28GANK knockdown by RNA interference (RNAi) can induce hepatoma cells apoptosis. However, the mechanisms have not been well defined yet. In the present study the p28GANK knockdown-induced apoptosis in HepG2 cells was prevented by caspase-9 inhibitor (Z-LEHD-FMK). During the knockdown of p28GANK, mitochondrial translocation of Bax, loss of mitochondrial transmembrane potential (DeltaPsim) and release of cytochrome c were observed. In this study, the activation of p38 was found to be critical for the p28GANK knockdown-induced apoptosis, as suggested by the finding that pharmacological inhibition of p38 with SB203580 suppressed the redistribution of Bax, the loss of DeltaPsim and the apoptosis. Moreover, generation of reactive oxygen species (ROS) contributed to the cell death because N-acetyl-L-cystenine (NAC), a ROS scavenger, suppressed the phosphorylation of p38 and the apoptosis. Our studies established the signaling pathway of p28GANK knockdown-induced apoptosis in HepG2 cells, namely, mitochondrial dysfunction mediated by p38 downstream of intracellular ROS generation.


Assembly of the TCR/CD3 complex: CD3 epsilon/delta and CD3 epsilon/gamma dimers associate indistinctly with both TCR alpha and TCR beta chains. Evidence for a double TCR heterodimer model.
The TCR/CD3 complex is composed of six subunits which are expressed on the cell surface in a coordinate fashion after assembly in the endoplasmic reticulum (ER). The TCR/CD3 complex is assembled after a series of pairwise interactions involving the formation of dimers of CD3 epsilon with either CD3 gamma or CD3 delta. These dimers assemble with TCR alpha and TCR beta chains, and finally, the CD3 zeta homodimer is added to allow export of the full complex from the ER. A model has been proposed suggesting that during assembly the CD3 epsilon/CD3 gamma dimer interacts exclusively with TCR beta and the CD3 epsilon/CD3 delta dimer with TCR alpha to form a complex with a single TCR alpha/beta heterodimer. We show in this study, by immunoprecipitation and two-dimensional gel electrophoresis, that in the human T cell line Jurkat as well as in total human thymocytes, this preferential interaction does not occur and instead, the CD3 epsilon/CD3 gamma and CD3 epsilon/CD3 delta dimers associate with both TCR chains simultaneously and indistinctly. These data are confirmed by the analysis of the TCR alpha-negative T cell line MOLT-4 in which TCR beta is found separately associated with CD3 epsilon/CD3 gamma and with CD3 epsilon/CD3 delta dimers. Indirectly, our results support a model of stoichiometry in which two TCR alpha/beta heterodimers are present in a TCR/CD3 complex. Furthermore, immunoprecipitation with anti-CD3 gamma and anti-CD3 delta antibodies from 1% NP40 and 1% Brij96 cell lysates showed that these subunits form independent partial complexes which are cross-linked through the CD3 zeta homodimer. This suggests that CD3 zeta mediates the interaction between both TCR alpha/beta heterodimers contained in the double TCR complex. Further proof for this hypothesis is obtained after analysis of a Jurkat cell transfectant containing a point mutation in the transmembrane domain of TCR beta that impairs the association of CD3 zeta. In this mutant cell line, unlike a control line with wild-type TCR beta, the CD3 gamma- and CD3 delta-containing complexes were found completely independent. Altogether, these results support a model of TCR/CD3 assembly and stoichiometry in which two TCR-alpha/beta heterodimers form two hemicomplexes containing either CD3 epsilon/gamma or CD3 epsilon/delta dimers which become associated via the CD3 zeta homodimer.


A novel animal-component-free medium for rabies virus production in Vero cells grown on Cytodex 1 microcarriers in a stirred bioreactor.
Vero cells growth and rabies production in IPT-AF medium, a property animal-component-free medium are described in this work. Kinetics of cell growth and rabies virus (strain LP 2061) production were first conducted in spinner flasks. Over eight independent experiments, Vero cell growth in IPT-AF medium, on 2 g/l Cytodex 1 was consistent. An average Cd (cell division number) of 3.3+/-0.4 and a specific growth rate micro of 0.017+/-0.006 h(-1) were achieved. Such performances were comparable to those obtained in serum-containing medium (MEM+10% FCS). Rabies virus production on Vero cells in IPT-AF medium was also optimised in spinner flasks. The effects of multiplicity of infection (MOI), regulation of glucose level at 1 g/l and cell washing step, were investigated. The highest virus titer was achieved when the cells were infected at an MOI of 0.1; this level was equal to 10(7) FFU/ml. The step of medium exchange before cell infection can be omitted; nevertheless in this case glucose level should be maintained at 1 g/l to avoid a decrease of specific virus productivity. Process optimisation in a 2-l stirred bioreactor pointed out that the aeration mode was the prominent parameter that affected cell growth in IPT-AF medium and on Cytodex 1 microcarriers. An acceptable level of cell density (cell density level of 1.5x10(6) cells/ml) was achieved when cells were grown in batch mode and using headspace aeration. Nevertheless, this aeration mode is not optimal for large-scale culture. The addition of Pluronic F68 at 0.1% at 24 h post inoculation as well as the switch from surface aeration mode to the sparged mode, 2 days after the start of the culture, had markedly improved cell growth performance. A cell density level of 5.5x10(6) cells/ml was reached when cells were grown in a 2-l bioreactor, on 3 g/l Cytodex 1 in IPT-AF medium and using the recirculation culture mode. Cell infection at an MOI of 0.1 and using perfused culture, resulted in a maximal virus titer of 3.5x10(7) FFU/ml. The activity of the pooled inactivated rabies virus harvests showed a protective activity that meets WHO requirements.


Mixed, melancholic, and anxious features in depression: a cross-sectional study of sociodemographic and clinical correlates.
Major depression (MD) is currently viewed as a heterogeneous condition, characterized by different psychopathological dimensions. Our sample was composed of 1,289 nonpsychotic bipolar/unipolar depressed patients. Participants were divided into mixed (MXD), melancholic (MEL), and anxious (ANX) depressed, according to a hierarchical functional model. Sociodemographic and clinical variables were compared across depressive subtypes by χ2 test and analysis of variance. The Young Mania Rating Scale (YMRS) and 2 subscales (melancholic [MEL-S] and psychic-somatic anxiety [PSOM-ANX]) from the Hamilton Depression Rating Scale also served as continuous outcome measures. MXD patients more frequently had bipolar I disorder (BD I), younger age of onset, and a higher familial load for mood disorders. MEL and ANX patients were more frequently diagnosed with major depressive disorder and reported a higher suicide risk. YMRS scores in depression was associated with BD I diagnosis (P < .0001) and manic polarity of the last episode (P < .0001), while a depressive polarity of the last episode (P < .0001) was associated with higher MEL-S score. No specific predictor was associated with PSOM-ANX score. Overall, our findings suggest that mixed depressive features are associated with significant hallmarks of bipolarity, and melancholic features may be influenced by previous depressive polarity. The symptom domain of anxiety appears to have no specific predictor.


Cholesterol metabolism in the macrophage. II. Alteration of subcellular exchangeable cholesterol compartments and exchange in other cell types.
Macrophage membrane cholesterol is present in two subcellular cholesterol pools, a rapidly exchanging compartment comprising about two-thirds of the total cholesterol, and a slowly exchanging compartment comprising one-third of the total. The morphological identification of the kinetically distinguishable pools proceeded by alteration of each compartment. Trypsin treatment markedly decreased the rate of cholesterol exchange without removing cholesterol from the membrane. Recovery of normal exchange rates took more than 7 hr and required protein synthesis. This suggested that a plasma membrane receptor is involved in positioning of lipoproteins for exchange, and is consistent with the plasma membrane localization of the rapidly exchanging compartment. Extensive pinocytosis by nondegradable dextran, dextran sulfate, or sucrose resulted in the accumulation of many secondary lysosomes, thus increasing the relative proportion of intracellular membranes. The measurable granule membrane area, cholesterol content, phospholipid content, and the relative size of the slowly exchanging cholesterol compartment all increased. The amount of intracellular membrane altered by extensive phagocytosis of latex particles also increased the size of the slowly exchanging cholesterol compartment. This suggested that the slowly exchanging pool of cholesterol represented the intracellular membranes primarily of lysosomal origin. Rabbit alveolar macrophages and thioglycollate-stimulated peritoneal macrophages contain many secondary lysosomes as a result of multiple bouts of in vivo phagocytosis and pinocytosis. In both of these cells the fast and slow pools are equal in size. The increased cholesterol content was attributable to the increase in the relative size of the slowly exchanging compartment. L-cells and melanoma cells also exchange their cholesterol with that of serum lipoproteins. Both cells contain few cholesterol-rich intracellular membranes, and had lower cellular cholesterol contents. In these cells the slowly exchanging pool was a minor contribution to cell cholesterol. Studies with these cells provided further evidence for the lysosomal membrane and plasma membrane localization of the slowly and rapidly exchanging cholesterol compartments.


Uric acid levels in patients on antituberculosis drugs in the southwest Region of Cameroon.
Antituberculosis drugs (ATDs) efficiently combat Mycobacterium tuberculosis either through direct molecular interactions or those of its metabolites. However, a variety of adverse effects have been reported, leading to frequent interruptions of treatment. To investigate the possible metabolic disturbances resulting from antituberculosis (TB) treatment, the uric acid (UA) level of patients on ATDs was measured in the southwest region of Cameroon. This hospital-based cross-sectional study involved 96 TB patients on ATDs and 32 controls who were neither on ATDs nor any other treatment that could increase UA levels. The hospital records of consenting participants were reviewed for medical history and questionnaires were issued. About 2 ml venous blood was collected and analyzed using spectrophotometers to determine UA levels. Hyperuricemia was observed in 56/96 (58.3%) of the studied group as compared with 4/32 (12.5%) in the control group (P < 0.001). Our results indicated that treatment duration was significantly associated with hyperuricemia (P = 0.0016) while gender (P = 0.1275) was not. Hyperuricemia is associated with ATDs, with treatment duration being a significant factor. The disorder should be closely monitored, especially during the intensive phase of treatment.


Bladder exstrophy.
Bladder exstrophy is a very rare congenital malformation in which the anterior wall of the bladder is absent, and the posterior wall is exposed externally. The differential diagnosis includes omphalocele, gastroschisis, and cloacal exstrophy. Ultrasound and Doppler examinations are the main diagnostic tools. Although mortality is low, termination of pregnancy should be discussed due to serious morbidities.


Barriers and facilitators to the implementation of a national multisectoral action plan for the prevention and control of noncommunicable diseases in Nepal: perspectives of stakeholders.
Nepal adopted the Multisectoral Action Plan for the Prevention and Control of Non-Communicable Diseases (MSAP) in 2014. Implementation of the plan has been challenging, with limited participation from non-health sectors. The overall aim of the study was to gain the perspectives of key stakeholders involved in the Nepal MSAP on the barriers and facilitators to its implementation, through the participation of relevant sectors in the plan. We held face-to-face semi-structured interviews with 12 stakeholders working in sectors involved in the MSAP. These sectors included the Office of the Prime Minister and Council of Ministries; Ministry of Health and Population (MOHP); Ministry of Education, Science and Technology; Ministry of Forest and Environment; academia; and professional organizations. Thematic analysis of transcripts was used to identify themes on awareness of NCDs, awareness of the MSAP, and barriers and facilitators to participation in the MSAP. Participants recognised NCDs as a growing and major burden in Nepal. However, a number of participants were not familiar with the MSAP, identifying a lack of leadership and poor dissemination. Political and systemic transformation, since the adoption of the MSAP, was seen as a key barrier to implementation. International commitments to develop multisectoral action made by the Government of Nepal were identified as drivers. The recent establishment of a separate section for NCDs and Mental Health within the Department of Health Services of MOHP and the promotion of a Health in All Policies (HiAP) approach in recent national documents, were both considered to support implementation. The establishment of permanent multisectoral or multistakeholder mechanisms has been challenging despite strong political calls for their development. Moving beyond 2020, multisectoral action plans should engage with stakeholders from federal, provincial and local governments in order to develop costed action plans with specific roles and responsibilities for each sector.


Accidental sodium hypochlorite-induced skin injury during endodontic treatment.
A case of accidental skin injury caused by leakage of sodium hypochlorite solution from the rubber dam during root canal preparation is reported. After placement of a rubber dam and initiation of root canal treatment, the patient complained of a burning sensation with sodium hypochlorite irrigation. The complaints were ignored by the practitioner, and a skin rash developed on and around the patient's chin, followed by scab formation. The patient required medical treatment with topical Hamamelis virginiana extract for 2 weeks, with full recovery.


3D MDCT-based system for planning peripheral bronchoscopic procedures.
The diagnosis and staging of lung cancer often begins with the assessment of a suspect peripheral chest site. Such suspicious peripheral sites may be solitary pulmonary nodules or other abnormally appearing regions of interest (ROIs). The state-of-the-art process for assessing such peripheral ROIs involves off-line procedure planning using a three-dimensional (3D) multidetector computed tomography (MDCT) chest scan followed by bronchoscopy with an ultrathin bronchoscope. We present an integrated computer-based system for planning peripheral bronchoscopic procedures. The system takes a 3D MDCT chest image as input and performs nearly all operations automatically. The only interaction required by the physician is the selection of ROI locations. The system is computationally efficient and fits smoothly within the clinical work flow. Integrated into the system and described in detail in the paper is a new surface-definition method, which is vital for effective analysis and planning to peripheral sites. Results demonstrate the efficacy of the system and its usage for the live guidance of ultrathin bronchoscopy to the periphery.


Management of post-operative recurrent diverticulitis: a review of the literature.
Recurrent abdominal symptoms following resection for diverticular disease occur in 1-10% of patients. Not all of these patients have recurrent diverticulitis. Other conditions such as carcinoma, irritable bowel syndrome, inflammatory bowel disease and ischaemic colitis should be considered in the differential diagnosis. A thorough investigation including computerized tomography (CT) scanning, contrast studies and colonoscopy must be undertaken. The cause of recurrent diverticulitis may be the result of inadequate previous resection or progression of disease. Re-resection has been required in 0-3.1% of patients in a collected series. Re-resection may be technically demanding although permanent colostomy is usually not necessary. The best method of prevention is adequate initial resection.


The purpose and functions of immunization information systems within health care organizations.
Abroad coalition of public and private health care organizations advocate the development of computerized immunization information systems as a key national strategy for achieving and sustaining high immunization coverage levels. However, widespread adoption requires greater awareness of the purpose, functions, and value of an immunization information system within health care organizations. We propose that the purpose of an immunization information system is to increase the efficiency and effectiveness of immunization-related practices and identify 9 potential functions that accomplish this purpose through improving patient care and practice management. When implementing an immunization information system within a practice setting, health care providers must consider technological and organizational issues. Health care providers should also look beyond their particular practice setting and establish public-private partnerships to create a system that links immunization data from all health care providers.


Laboratory Reconstructions of Bicycle Helmet Damage: Investigation of Cyclist Head Impacts Using Oblique Impacts and Computed Tomography.
Although head injuries are common in cycling, exact conditions associated with cyclist head impacts are difficult to determine. Previous studies have attempted to reverse engineer cyclist head impacts by reconstructing bicycle helmet residual damage, but they have been limited by simplified damage assessment and testing. The present study seeks to enhance knowledge of cyclist head impact conditions by reconstructing helmet damage using advanced impact testing and damage quantification techniques. Damage to 18 helmets from cyclists treated in emergency departments was quantified using computed tomography and reconstructed using oblique impacts. Damage metrics were related to normal and tangential velocities from impact tests as well as peak linear accelerations (PLA) and peak rotational velocities (PRV) using case-specific regression models. Models then allowed original impact conditions and kinematics to be estimated for each case. Helmets were most frequently damaged at the front and sides, often near the rim. Concussion was the most common, non-superficial head injury. Normal velocity and PLA distributions were similar to previous studies, with median values of 3.4 m/s and 102.5 g. Associated tangential velocity and PRV medians were 3.8 m/s and 22.3 rad/s. Results can inform future oblique impact testing conditions, enabling improved helmet evaluation and design.


A randomised cross-over pilot study investigating the use of acupuncture to promote weight loss and mental health in overweight and obese individuals participating in a weight loss program.
Acupuncture is widely used as an alternative modality for weight loss. Despite its increasing use, few acupuncture studies have evaluated the effect of a weight loss program on the mental health of obese/overweight participants and none have looked at the effect on those with eating, weight and shape concerns. To investigate the feasibility of conducting an acupuncture study involving overweight or obese individuals undertaking a weight loss program with particular reference to those with eating concerns. Thirty-five overweight/obese males and females participated in a single-blinded randomised cross-over study. The two intervention phases were: (1) nutritional counselling plus Traditional Chinese Medicine (TCM) acupuncture and (2) nutritional counselling plus sham acupuncture. This study evaluates the feasibility and practicalities of the study including recruitment, retention, adverse events, effectiveness for defining eating and weight concerns, study design and statistics for power calculations. The outcome measures, the recruitment of those with eating and weight concerns and the acceptability of the intervention demonstrate a larger trial investigating the use of acupuncture for weight loss in those who have elevated eating and weight concerns is feasible.


Visualization of intracellular movement of vaccinia virus virions containing a green fluorescent protein-B5R membrane protein chimera.
We produced an infectious vaccinia virus that expressed the B5R envelope glycoprotein fused to the enhanced green fluorescent protein (GFP), allowing us to visualize intracellular virus movement in real time. Previous transfection studies indicated that fusion of GFP to the C-terminal cytoplasmic domain of B5R did not interfere with Golgi localization of the viral protein. To determine whether B5R-GFP was fully functional, we started with a B5R deletion mutant that made small plaques and inserted the B5R-GFP gene into the original B5R locus. The recombinant virus made normal-sized plaques and acquired the ability to form actin tails, indicating reversal of the mutant phenotype. Moreover, immunogold electron microscopy revealed that both intracellular enveloped virions (IEV) and extracellular enveloped virions contained B5R-GFP. By confocal microscopy of live infected cells, we visualized individual fluorescent particles, corresponding to IEV in size and shape, moving from a juxtanuclear location to the periphery of the cell, where they usually collected prior to association with actin tails. The fluorescent particles could be seen emanating from cells at the tips of microvilli. Using a digital camera attached to an inverted fluorescence microscope, we acquired images at 1 frame/s. At this resolution, IEV movement appeared saltatory; in some frames there was no net movement, whereas in others movement exceeded 2 microm/s. Further studies indicated that IEV movement was reversibly arrested by the microtubule-depolymerizing drug nocodazole. This result, together with the direction, speed, and saltatory motion of IEV, was consistent with a role for microtubules in intracellular transport of IEV.


The prevention of experimentally induced postoperative adhesions.
For many years, various substances have been used to prevent postoperative adhesions, with suboptimal effectiveness. Saline has been postulated to have a positive effect, but it is reabsorbed from the abdominal cavity within 24 hours. To determine whether a longer-lasting intraperitoneal solution would be effective in preventing postoperative adhesions, 36 rats were divided into three groups: in 15 rats, a 2.5 per cent hyperosmolar solution of peritoneal dialysis fluid was injected intraperitoneally immediately following exploratory laparotomy, which included traumatization of the intestinal serosa at two sites; in 12 rats, normal saline was injected by the same procedure; and in 9 rats, no fluid was injected after the same operation. After 3 weeks, the group receiving the hyperosmolar peritoneal dialysis solution achieved the best results in both sites, the group that received normal saline showed marginal improvement, and the control group fared the worst.


How should Helicobacter pylori negative patients be managed?
Dyspepsia is a digestive syndrome distinct from (although frequently overlapping with) gastro-oesophageal reflux disease (GORD) and irritable bowel syndrome (IBS), which is characterised by various combinations of painful and non-painful symptoms arising from the epigastrium. Dyspepsia can be secondary to a variety of diseases, but in most instances it is idiopathic. Helicobacter pylori infection is responsible for the majority of peptic ulcers and of other diseases potentially associated with dyspepsia. Nevertheless, a causal role for H pylori infection in symptom occurrence has not been established. Experimental data indicate that H pylori eradication does not improve symptoms in the majority of dyspeptic patients. It has been proposed recently that H pylori negative patients should be managed according to their clinical presentation. Some reports suggest that taking into consideration the most relevant or "predominant" symptom may help to identify distinct subgroups among dyspeptic patients with different underlying pathophysiological abnormalities and different responses to treatment. Well designed and conducted prospective studies are needed to verify whether treatment of H pylori negative dyspeptic patients based on the predominant symptom actually is a cost effective approach.


Characterization of flow cytometer instrument sensitivity.
Fluorescence sensitivity, measured in terms of resolution, allows the researcher to more accurately answer the question of how dim a cell can be and still be resolvable from another population. This measure focuses on the width, i.e., the standard deviation, of the population distributions and not on the location, i.e., the mean intensity, of populations on a histogram scale relative to the background. To determine the fluorescence sensitivity in terms of the resolution, both the detection efficiency, Q, and optical background, B, need to be measured. Both factors affect the ability to resolve dimly fluorescent subpopulations, and both factors are required to uniquely characterize the performance of a flow cytometer. Using Q and B, it is possible to determine the minimum fluorescence intensity that is resolvable from the background or from another population. This unit describes a practical and robust approach to measure the critical factors that determine the ability of a flow cytometer to analyze dimly fluorescent particles.


Rapid intraplate strain accumulation in the new madrid seismic zone.
Remeasurement of a triangulation network in the southern part of the New Madrid seismic zone with the Global Positioning System has revealed rapid crustal strain accumulation since the 1950s. This area experienced three large (moment magnitudes >8) earthquakes in 1811 to 1812. The orientation and sense of shear is consistent with right-lateral strike slip motion along a northeast-trending fault zone (as indicated by current seismicity). Detection of crustal strain accumulation may be a useful discriminant for identifying areas where potentially damaging intraplate earthquakes may occur despite the absence of large earthquakes during historic time.


Prediction of Lower Extremity Multi-Joint Angles during Overground Walking by Using a Single IMU with a Low Frequency Based on an LSTM Recurrent Neural Network.
The joint angle during gait is an important indicator, such as injury risk index, rehabilitation status evaluation, etc. To analyze gait, inertial measurement unit (IMU) sensors have been used in studies and continuously developed; however, they are difficult to utilize in daily life because of the inconvenience of having to attach multiple sensors together and the difficulty of long-term use due to the battery consumption required for high data sampling rates. To overcome these problems, this study propose a multi-joint angle estimation method based on a long short-term memory (LSTM) recurrent neural network with a single low-frequency (23 Hz) IMU sensor. IMU sensor data attached to the lateral shank were measured during overground walking at a self-selected speed for 30 healthy young persons. The results show a comparatively good accuracy level, similar to previous studies using high-frequency IMU sensors. Compared to the reference results obtained from the motion capture system, the estimated angle coefficient of determination (R2) is greater than 0.74, and the root mean square error and normalized root mean square error (NRMSE) are less than 7° and 9.87%, respectively. The knee joint showed the best estimation performance in terms of the NRMSE and R2 among the hip, knee, and ankle joints.


Complementary DNA clones of chicken proto-oncogene c-ets: sequence divergence from the viral oncogene v-ets.
The avian acute leukemia virus E26 induces erythroblastosis and myeloblastosis in chickens. The oncogene of this virus includes sequences derived from the cellular gene designated c-ets, which is normally expressed in lymphoid cells and whose product is a protein of apparent molecular weight ca. 54,000 daltons. Complementary DNA clones representing the major transcript of the chicken c-ets proto-oncogene were isolated from a spleen cell library. Sequence analysis of the cDNA revealed that it contains an open reading frame encoding a polypeptide of 441 amino acids with a molecular weight of 49,932 daltons. This open reading frame can be transcribed and translated in vitro into a 50 kd protein that is specifically immunoprecipitated with antiserum to the v-ets oncogene product. Within the central region of homology between c-ets and v-ets, there are only 5 nucleotide substitutions resulting in 4 amino acid changes. However, coding sequences at the 5' and 3' ends of the v-ets oncogene and the chicken c-ets cDNA differ from one another. These changes may be responsible for the differential functions of c-ets and v-ets in cells of different hematopoietic lineages and may account for the pathogenic properties of the v-ets oncogene.


Unexpectedly High Beta-Diversity of Root-Associated Fungal Communities in the Bolivian Andes.
Bolivia is one of the most biologically diverse countries on the planet. Between the Andes and the Amazon drainage basin spans the Yungas, a vast forested region shown to be extremely species rich in macro-organisms. However, it remains unclear whether this high diversity is also reflected in microbial diversity. Here we assess the genetic, taxonomic and functional diversity of root-associated fungi surrounding Cinchona calisaya trees, a typical element of the intermediate altitudes of the Bolivian Yungas. We determine the relative effects of edaphic properties, climate, and geography in regulating fungal community assembly. We show that α-diversity for these fungal communities was similar to temperate and arid ecosystems, averaging 90.1 operational taxonomic units (OTUs) per sample, with reads predominantly assigned to the Ascomycota phylum and with a saprotrophic lifestyle. ß-diversity was calculated as the distance-decay rate, and in contrast to α-diversity, was exceptionally high with a rate of -0.407. Soil properties (pH and P) principally regulated fungal community assembly in an analogous manner to temperate environments, with pH and phosphorus explaining 7.8 and 7.2% of community variation respectively. Surprisingly, altitude does not influence community formation, and there is limited evidence that climate (precipitation and temperature) play a role. Our results suggest that sampling should be performed over a wide geographical and environmental range in order to capture the full root-associated fungal diversity in subtropical regions. This study sheds further light on the diversity and distribution of the world's "hidden biodiversity."


Bleeding risk of cerebrovascular malformations in hereditary hemorrhagic telangiectasia.
Hereditary hemorrhagic telangiectasia (HHT) is a rare autosomal-dominant vascular dysplasia with a high prevalence of cerebrovascular malformations (CVMs), mostly manifested as arteriovenous malformations (AVMs). The natural history and bleeding risk of these CVMs is unknown. The authors investigated the risk of bleeding in conjunction with clinical and radiological features in patients with HHT and proven CVMs. Intravenous digital subtraction (DS) angiography was used to screen 196 patients with HHT for the presence of CVMs. Patients with abnormal results on DS angiography were asked to undergo a conventional cerebral angiographic study. All patients with a proven CVM were assessed by a neurologist. The bleeding risk was retrospectively and prospectively calculated for patients with AVMs only, as well as for the whole cohort of patients with CVMs. Twenty-four patients (12.2%; 16 female and eight male), aged 14 to 66 years (mean 35.4 years) with one or more CVMs were identified. Fifteen patients (62.5%) had a CVM and a pulmonary AVM. Eleven patients (45.8%) exhibited no neurological signs of their CVM; six (25%) had headache or migraine; four (16.7%) had seizures; and three (12.5%) had an intracranial hemorrhage. Twenty-two patients had at least one AVM (with a total of 28 AVMs), whereas two patients only had telangiectases. Twenty-seven AVMs were small (96%), 36% were located in eloquent areas of the brain, and 82% had superficial venous drainage. One third of the patients had multiple CVMs. The bleeding risk for patients with at least one AVM ranged from 0.41 to 0.72% per year, and for the whole cohort the range was 0.38 to 0.69% per year. Calculation of the bleeding risk as determined by lesion-years ranged from 0.36 to 0.56% per year for patients with AVMs and from 0.27 to 0.46% per year for all patients with CVMs. Patients with HHT have a high risk of harboring a CVM, especially in the presence of a pulmonary AVM. These CVMs are mostly low-grade AVMs (Spetzler-Martin Grade I or II), are frequently multiple, and have a lower risk of bleeding than that associated with sporadic AVMs. Female patients are more often affected than male patients. The inherent low sensitivity of DS angiography screening for CVMs may yield false negative results.


Choroidal structural changes in patients with early diabetic nephropathy.
To determine alterations of the choroidal thickness (CT) and the choroidal vascularity index (CVI) in patients with glomerular hyperfiltration, a marker of early diabetic nephropathy (DN). Twenty-two patients with type 2 diabetes (T2D) with glomerular hyperfiltration (early DN group) and 28 patients with T2D without DN (NDN group) were included in the study. Patients with diabetic retinopathy were excluded. Parameters including subfoveal CT, the subfoveal choroidal vascularity index (CVI), and total CVI were measured using spectral-domain enhanced depth imaging optical coherence tomography method. The early DN group included 22 patients and the NDN group comprised 28 patients. The groups were similar in terms of age and sex (p>0.05). The CT values were statistically significantly lower in the early DN group than in the NDN group (p < 0.001). There was no significant difference between the early DN group and the NDN group in terms of total and subfoveal CVI (p>0.05). The choroidal thickness decreased in patients with T2D with glomerular hyperfiltration, but there were no differences in CVI when they were compared with patients with T2D without DN.


Single-bead, single-molecule, single-cell fluorescence: technologies for drug screening and target validation.
According to many current reports, the pharmaceutical business will hit a wall over the next few years. The generic competition is expected to wipe out a double-digit billion-dollar amount from top companies' annual sales between 2007 and 2012 (Wall Street Journal, online, December 6, 2007). The industry's science engine has stalled, new blockbusters are lacking, and patent expirations are a big problem. Also, the U.S. Food and Drug Administration is pulling back on approvals, requesting larger safety studies. Among the different approaches taken throughout the industry to improve productivity and to reduce the attrition rate of compounds in the drug discovery process, an extended application of quantitative biology and biophysical methods is ranked very high. Fluorescence spectroscopy and imaging represented the main detection technologies for assays and screening methods in recent years. Today, label-free detection methods, such as isothermal titration calorimetry, differential scanning calorimetry, tandem mass spectrometry (MS(n)), light scattering, or interferometry, start to provide viable alternative readouts for physicochemical characterization of leads and hit list triaging. However, the multidimensional nature of fluorescence along with its high sensitivity and single-molecule resolution remains an unparalleled source of molecular parameters to extract all different kinds of information on molecules and ligand-protein complexes in solution. Although fluorescence-based methods are currently applied throughout the different stages of the industrial drug discovery process, they are usually applied in an unconnected way. We have developed a fully integrated hit and lead discovery process combining bead-based synthesis and screening methods with confocal fluorescence microspectroscopy. The primary on-bead screening process provides fluorescent ligands that after a multistep characterization process ultimately leads to fully mechanistically characterized cellularly validated binders and inhibitors of target protein interactions. The unlabeled small-molecular inhibitors represent chemical starting points in drug discovery and target validation.


Asymptomatic Cryptococcal Antigenemia in People Living with HIV (PLHIV) with Severe Immunosuppression: Is Routine CrAg Screening Indicated in India?
Cryptococcal meningitis (CM) is a common, life-threatening opportunistic infection (OI) among people living with HIV (PLHIV) in India. Serum Cryptococcal antigen (CrAg) positivity is predictive of future occurrence of CM and pre-emptive treatment reduces its mortality. Routine CrAg screening among PLHIV is not adopted by India's national programme. This study evaluated the prevalence of CrAg and assessed CrAg positivity in predicting all-cause mortality among PLHIV. This prospective study was conducted in a tertiary care, public health facility in New Delhi, India. Prevalence of CrAg was assessed in 128 ART naive adult PLHIV with CD4 < 100 cells/mm3 using a latex agglutination test. Age, gender, weight, body mass index (BMI), CD4 count, haemoglobin, serum albumin, and presence of other OI were evaluated as determinants of CrAg positivity. Subjects were followed up for occurrence of CM and mortality (all-cause) at 12 weeks and 6 months. The mean age of the subjects was 36.2±9.48 years, 73.4% were men, 21.09% women and 5.46% were transgender. The mean BMI was 18.4±2.53 kg/m2 and 64% of subjects belonged to the lower socio-economic strata. Mean CD4 counts of the subjects was 54.9±26.58 cells/mm3 and 42.97% had CD4 < 50 cells/mm3. The prevalence of CrAg in the subjects was 3.125 % (4/128). None of the factors assessed showed statistically significant difference between the 2 groups, though CD4 count <50 cells/mm3, low serum albumin and presence of oral candidiasis had a stronger association with CrAg positivity. None of the subjects developed CM during follow up. At 12 weeks, 3/4 (75%) CrAg positive patients were alive compared to 118/124 (95.16%) of CrAg negative subjects. At 6 months, 50% (2/4) CrAg positive patients had died compared to 10.48% (13/124) CrAg negative (p<0.01). Though CrAg prevalence in PLHIV with CD4<100 cells/mm3 is moderate, asymptomatic CrAg positivity among PLHIV with CD4 < 100cells/mm3 is significantly associated with higher all-cause mortality. CrAg testing is very cost effective and India's National AIDS Control Programme should seriously consider routine screening among the severely immunosuppressed PLHIV.


Partial cystectomy for urothelial carcinoma of the bladder: Practice patterns and outcomes in the general population.
Partial cystectomy (PC) for urothelial carcinoma (UC) in selected patients may avoid the morbidity of radical cystectomy (RC). We describe use and outcomes of PC for UC in routine clinical practice. All patients with urothelial carcinoma of the bladder (UCB) undergoing PC or RC in Ontario from 1994-2008 were identified using the Ontario Cancer Registry and linked electronic records. Pathology reports were reviewed. Variables associated with PC use were identified using logistic regression. Cox proportional hazards model identified factors affecting cancer-specific (CSS) and overall survival (OS). A total of 3320 patients underwent PC (n=181; 5%) or RC (n=3139; 95%) from 1994-2008. PC patients were older (36% 80+ years vs. 19%; p<0.001) and more likely to have organ-confined (<pT3) disease (54% vs. 36% RC; p<0.001). Two-thirds (67%) of PC patients did not undergo lymph node dissection (24% for RC; p<0.001). Factors associated with having PC included older age (odds ratio [OR] 1.55; 95% confidence interval [CI] 0.96-2.51 for 70+ years), moderate comorbidity (OR 1.95; 95% CI 1.13-3.37), and surgery outside of a comprehensive cancer centre (OR 1.44; 95% CI 1.03-2.01). Unadjusted five-year OS for PC and RC cases was 34% and 33%, respectively (p=0.455); CSS at five years was 43% and 37% (p=0.045). On adjusted analysis, PC was associated with comparable CSS (hazard ratio [HR] 0.87, 95% CI 0.70-1.09) and OS (HR 0.95, 95% CI 0.79-1.14) as RC. In routine clinical practice, PC is not common. A substantial proportion of patients treated with PC achieve long-term survival. PC remains a treatment option in selected patients with UCB.


Aldosterone, Renin, and Aldosterone-to-Renin Ratio Variability in Screening for Primary Aldosteronism.
The plasma aldosterone concentration (PAC), renin, and aldosterone-to-renin ratio (ARR) are used to screen for primary aldosteronism (PA). Substantial intra-individual variability of PAC and ARR using plasma renin activity in the context of usual antihypertensive therapy has been described, but there is no data on ARR variability calculated using direct renin concentration (DRC). To describe the intra-individual variability of PAC, DRC, and ARR in the absence of interfering medications in patients with and without PA. Retrospective cohort study. Hypertensive patients referred for investigation of PA, with at least 2 ARR measurements while off interfering medications. Endocrine hypertension service of a tertiary center, from May 2017 to July 2021. PAC, DRC, and ARR variability was calculated as coefficient of variation (CV) and percent difference (PD). Analysis of 223 patients (55% female, median age 52 years), including 162 with confirmed PA, demonstrated high variability with a sample CV of 22-25% in the PAC and sample CV of 41% to 42% in the DRC and ARR in both the PA and non-PA groups. The degree of variability was substantially higher than the assays' analytical CV. Sixty-two patients (38%) with PA had at least one ARR below 70 pmol/L:mU/L (2.4 ng/dL:mU/L), a cut-off for first-line screening of PA. Significant intra-individual variability in PAC, DRC, and hence ARR occurs in a large proportion of patients being investigated for PA. These findings support the need for at least 2 ARR before PA is excluded or further investigated.


Over-the-counter Acne Treatments: A Review.
Acne is a common dermatological disorder that most frequently affects adolescents; however, individuals may be affected at all ages. Many people who suffer from acne seek treatment from both prescription and over-the-counter acne medications. Due to convenience, lower cost, and difficulty getting an appointment with a dermatologist, the use of over-the-counter acne treatments is on the rise. As the plethora of over-the-counter acne treatment options can be overwhelming, it is important that dermatologists are well-versed on this subject to provide appropriate information about treatment regimens and potential drug interactions and that their patients see them as well-informed. This article reviews the efficacy of various over-the-counter acne treatments based on the current literature. A thorough literature review revealed there are many types of over-the-counter acne treatments and each are designed to target at least one of the pathogenic pathways that are reported to be involved in the development of acne lesions. Many of the key over-the-counter ingredients are incorporated in different formulations to broaden the spectrum and consumer appeal of available products. Unfortunately, many over-the-counter products are not well-supported by clinical studies, with a conspicuous absence of double-blind or investigator-blind, randomized, vehicle-controlled studies. Most studies that do exist on over-the-counter acne products are often funded by the manufacturer. Use of over-the-counter acne treatments is a mainstay in our society and it is important that dermatologists are knowledgeable about the different options, including potential benefits and limitations. Overall, over-the-counter acne therapies can be classified into the following five major groups: cleansers, leave-on products, mechanical treatments, essential oils, and vitamins.


Rif1 Functions in a Tissue-Specific Manner To Control Replication Timing Through Its PP1-Binding Motif.
Replication initiation in eukaryotic cells occurs asynchronously throughout S phase, yielding early- and late-replicating regions of the genome, a process known as replication timing (RT). RT changes during development to ensure accurate genome duplication and maintain genome stability. To understand the relative contributions that cell lineage, cell cycle, and replication initiation regulators have on RT, we utilized the powerful developmental systems available in Drosophila melanogaster We generated and compared RT profiles from mitotic cells of different tissues and from mitotic and endocycling cells of the same tissue. Our results demonstrate that cell lineage has the largest effect on RT, whereas switching from a mitotic to an endoreplicative cell cycle has little to no effect on RT. Additionally, we demonstrate that the RT differences we observed in all cases are largely independent of transcriptional differences. We also employed a genetic approach in these same cell types to understand the relative contribution the eukaryotic RT control factor, Rif1, has on RT control. Our results demonstrate that Rif1 can function in a tissue-specific manner to control RT. Importantly, the Protein Phosphatase 1 (PP1) binding motif of Rif1 is essential for Rif1 to regulate RT. Together, our data support a model in which the RT program is primarily driven by cell lineage and is further refined by Rif1/PP1 to ultimately generate tissue-specific RT programs.


Phenotypic comparison of Pseudomonas aeruginosa strains isolated from a variety of clinical sites.
Pseudomonas aeruginosa elaborates a number of extracellular products which have been shown to play a role in the pathogenesis of disease caused by this organism. In this study, we showed that the host environment markedly affects the levels of exoproducts produced. We compared the phenotypes of a number of P. aeruginosa strains obtained from a variety of clinical sources, including burn wounds, skin wounds, urine, cystic fibrosis sputum, acute pneumonia sputum, and blood. The clinical isolates were examined quantitatively for levels of total protease, elastase, phospholipase C, exotoxin A, and exoenzyme S produced in vitro under defined conditions. The exoproduct levels varied significantly, depending on the site of isolation. Elevated levels of elastase were demonstrated in strains isolated from acute lung infections, phospholipase C levels were elevated in urinary tract and blood isolates, exotoxin A levels were elevated in blood isolates, and exoenzyme S levels were increased in acute pneumonia isolates. Isolates from cystic fibrosis sputum produced low amounts of virtually all of the tested exoproducts, particularly as compared with sputum isolates from acute P. aeruginosa lung infections.


Do you see what we see? The complex effects of perceptual distance between leaders and teams.
Previous distance-related theories and concepts (e.g., social distance) have failed to address the sometimes wide disparity in perceptions between leaders and the teams they lead. Drawing from the extensive literature on teams, leadership, and cognitive models of social information processing, the authors develop the concept of leader-team perceptual distance, defined as differences between a leader and a team in perceptions of the same social stimulus. The authors investigate the effects of perceptual distance on team performance, operationalizing the construct with 3 distinct foci: goal accomplishment, constructive conflict, and decision-making autonomy. Analyzing leader, member, and customer survey responses for a large sample of teams, the authors demonstrate that perceptual distance between a leader and a team regarding goal accomplishment and constructive conflict have a nonlinear relationship with team performance. Greater perceptual differences are associated with decreases in team performance. Moreover, this effect is strongest when a team's perceptions are more positive than the leader's are (as opposed to the reverse). This pattern illustrates the pervasive effects that perceptions can have on team performance, highlighting the importance of developing awareness of perceptions in order to increase effectiveness. Implications for theory and practice are delineated. (PsycINFO Database Record (c) 2009 APA, all rights reserved).


Regulation of hepatic lipase activity by sphingomyelin in plasma lipoproteins.
Hepatic lipase (HL) is an important enzyme in the clearance of triacylglycerol (TAG) from the circulation, and has been proposed to have pro-atherogenic as well as anti-atherogenic properties. It hydrolyzes both phospholipids and TAG of lipoproteins, and its activity is negatively correlated with HDL levels. Although it is known that HL acts preferentially on HDL lipids, the basis for this specificity is not known, since it does not require any specific apoprotein for activity. In this study, we tested the hypothesis that sphingomyelin (SM), whose concentration is much higher in VLDL and LDL compared to HDL, is an inhibitor of HL, and that this could explain the lipoprotein specificity of the enzyme. The results presented show that the depletion of SM from normal lipoproteins activated the HL roughly in proportion to their SM content. SM depletion stimulated the hydrolysis of both phosphatidylcholine (PC) and TAG, although the PC hydrolysis was stimulated more. In the native lipoproteins, HL showed specificity for PC species containing polyunsaturated fatty acids at sn-2 position, and produced more unsaturated lyso PC species. The enzyme also showed preferential hydrolysis of certain TAG species over others. SM depletion affected the specificity of the enzyme towards PC and TAG species modestly. These results show that SM is a physiological inhibitor of HL activity in lipoproteins and that the specificity of the enzyme towards HDL is at least partly due to its low SM content.


The feasibility study of stress position device simulating standing weight-bearing applied in computed tomography examination.
In order to simulate weight-bearing Computed Tomography (CT) examination, this study designed a simple stress position device. By analyzing the relevant data of stress position footprints and weight-bearing position footprints, the feasibility of the stress position device to simulate standing weight-bearing was verified. This study randomly selected 25 volunteers for standing weight-bearing and stress position footprints collection, and measured the relevant indicators of stress position footprints and standing weight-bearing position footprints. Two foot and ankle surgeons conducted two measurements respectively on the footprints. Intra-observer and inter-observer reliability were calculated using intra-class correlation coefficients (ICC). Pearson correlation coefficient, ICC, scatter plot analysis, and paired t-test were used to analyze the stress and weight-bearing position data. The intra-observer and inter-observer measurement values were reliable. There was a certain degree of correlation between the stress position footprints and weight-bearing position footprints in terms of Pearson correlation coefficient, ICC, and scatter plot analysis. Paired t-tests showed significant differences in Clarke angle (t 2.636, p .012), C-S index (t 10.568, p .000), arch indx (t 2.176, p .035), and arch lower angle (t 6.246, p .000). The stress position device can generate a certain degree of stress, and after further optimization and improvement of the stress position device, it is feasible to apply it to weight-bearing CT examination in clinical settings.


Characterization of Glutamate-Mediated Hormonal Regulatory Pathway of the Drought Responses in Relation to Proline Metabolism in Brassica napus L.
Proline metabolism influences the metabolic and/or signaling pathway in regulating plant stress responses. This study aimed to characterize the physiological significance of glutamate (Glu)-mediated proline metabolism in the drought stress responses, focusing on the hormonal regulatory pathway. The responses of cytosolic Ca2+ signaling, proline metabolism, and redox components to the exogenous application of Glu in well-watered or drought-stressed plants were interpreted in relation to endogenous hormone status and their signaling genes. Drought-enhanced level of abscisic acid (ABA) was concomitant with the accumulation of ROS and proline, as well as loss of reducing potential, which was assessed by measuring NAD(P)H/NAD(P)+ and GSH/GSSG ratios. Glu application to drought-stressed plants increased both salicylic acid (SA) and cytosolic Ca2+ levels, with the highest expression of calcium-dependent protein kinase (CPK5) and salicylic acid synthesis-related ICS1. The SA-enhanced CPK5 expression was closely associated with further enhancement of proline synthesis-related genes (P5CS1, P5CS2, and P5CR) expression and a reset of reducing potential with enhanced expression of redox regulating genes (TRXh5 and GRXC9) in a SA-mediated NPR1- and/or PR1-dependent manner. These results clearly indicate that Glu-activated interplay between SA- and CPK5-signaling as well as Glu-enhanced proline synthesis are crucial in the amelioration of drought stress in Brassica napus.


Seed treatment with clothianidin induces changes in plant metabolism and alters pollinator foraging preferences.
Neonicotinoids, systemic insecticides that are distributed into all plant tissues and protect against pests, have become a common part of crop production, but can unintentionally also affect non-target organisms, including pollinators. Such effects can be direct effects from insecticide exposure, but neonicotinoids can affect plant physiology, and effects could therefore also be indirectly mediated by changes in plant phenology, attractiveness and nutritional value. Under controlled greenhouse conditions, we tested if seed treatment with the neonicotinoid clothianidin affected oilseed rape's production of flower resources for bees and the content of the secondary plant products glucosinolates that provide defense against herbivores. Additionally, we tested if seed treatment affected the attractiveness of oilseed rape to flower visiting bumblebees, using outdoor mesocosms. Flowers and leaves of clothianidin-treated plants had different profiles of glucosinolates compared with untreated plants. Bumblebees in mesocosms foraged slightly more on untreated plants. Neither flower timing, flower size nor the production of pollen and nectar differed between treatments, and therefore cannot explain any preference for untreated oilseed rape. We instead propose that this small but significant preference for untreated plants was related to the altered glucosinolate profile caused by clothianidin. Thereby, this study contributes to the understanding of the complex relationships between neonicotinoid-treated crops and pollinator foraging choices, by suggesting a potential mechanistic link by which insecticide treatment can affect insect behavior.


Integrating Biological Advances Into the Clinical Management of Breast Cancer Related Lymphedema.
Breast cancer-related lymphedema (BCRL) occurs in a significant number of breast cancer survivors as a consequence of the axillary lymphatics' impairment after therapy (mainly axillary surgery and irradiation). Despite the recent achievements in the clinical management of these patients, BCRL is often diagnosed at its occurrence. In most cases, it remains a progressive and irreversible condition, with dramatic consequences in terms of quality of life and on sanitary costs. There are still no validated pre-surgical strategies to identify individuals that harbor an increased risk of BCRL. However, clinical, therapeutic, and tumor-specific traits are recurrent in these patients. Over the past few years, many studies have unraveled the complexity of the molecular and transcriptional events leading to the lymphatic system ontogenesis. Additionally, molecular insights are coming from the study of the germline alterations involved at variable levels in BCRL models. Regrettably, there is a substantial lack of predictive biomarkers for BCRL, given that our knowledge of its molecular milieu remains extremely puzzled. The purposes of this review were (i) to outline the biology underpinning the ontogenesis of the lymphatic system; (ii) to assess the current state of knowledge of the molecular alterations that can be involved in BCRL pathogenesis and progression; (iii) to discuss the present and short-term future perspectives in biomarker-based patients' risk stratification; and (iv) to provide practical information that can be employed to improve the quality of life of these patients.


Clinical triage for colonoscopy is useful in young women.
Colonoscopy is an invasive procedure and a limited resource. It is therefore desirable to restrict its use to those in whom it yields an important diagnosis, without missing pathology in others. The aim of this study was to determine whether standard clinical criteria can be used to reliably distinguish when colonoscopy is advisable in women 30 years and younger. A retrospective audit was performed at a single centre of 100 consecutive colonoscopies performed in women 30 years old and younger. The indications for the colonoscopy were recorded, and divided into clear and relative indications. The primary outcome of whether an endoscopic diagnosis was made was compared between the two groups. Clear indications for colonoscopy included overt rectal bleeding, elevated inflammatory markers, anaemia, iron deficiency and strong family history of colorectal cancer. Relative indications included abdominal pain or discomfort, bloating and altered bowel habit/motions. The average age was 23 years. Sixty women had both relative and clear indications. Eleven had only clear indications and 28 only relative indications. Altogether, 58 colonoscopies were normal, and 17 showed inflammatory bowel disease. No subject with only relative indications had an abnormal finding (0/28). The diagnostic yield was significantly different between those with only relative indications (0%) versus those with at least one clear indication (59%; P < 0.0001). Standard clinical criteria can be used to restrict safely the use of colonoscopy in young women. This will avoid performing procedures in people without clear indications, saving costs, resources and complications.