Effect of HIV infection on pre- and post-liver transplant mortality in patients with organ failure.

OBJECTIVES: Organ failure (OF), a leading cause of death in HIV-positive individuals, is common in patients undergoing liver transplantation (LT). We examined the impact of HIV infection on pre- and post-LT mortalities in cirrhotic patients stratified by the number and type of OFs. METHODS: We performed a cross-sectional study and a retrospective cohort study using the US National Inpatient Sample (NIS) and the United Network for Organ Sharing (UNOS) registry data, respectively. Patients who had not yet undergone LT from the NIS database (2010-2014) and patients undergoing LT from the UNOS database (2003-2016) were included in the study. RESULTS: Analysis of patients (201 348) from the NIS database showed that one [adjusted odds ratio (aOR) 1.531; 95% confidence interval (CI) 1.160-2.023], two (aOR 1.624; 95% CI 1.266-2.083) or three or more OFs (aOR 1.349; 95% CI 1.165-1.562) were associated with higher pre-LT mortality in HIV-infected patients compared with HIV-negative patients with the corresponding number of OFs. In patients without OF, HIV infection was not associated with increased pre-LT mortality. UNOS data for patients undergoing LT (38 942) showed that the presence of two or more OFs was associated with increased post-LT 1-year mortality in HIV-infected patients compared with non-HIV-infected patients with the corresponding number of OFs (aOR 2.342; 95% CI 1.576-3.480). However, in patients with no OF or only one OF, HIV infection was not associated with increased post-LT 1-year mortality (aOR 1.372; 95% CI 0.911-2.068). CONCLUSIONS: The results of this study emphasize the importance of preventing OF development, and justify LT for HIV-infected patients with no or only one OF.

Apparent prevalence of Mycoplasma wenyonii, Candidatus Mycoplasma haemobos, and bovine leukemia virus in Wisconsin and Michigan dairy cattle herds.

Mycoplasma wenyonii (formerly Eperythrozoon wenyonii) is a hemotrophic, epicellular bacterial parasite of cattle that has been associated with clinical disorders, including hemolytic anemia, decreased milk yield, and peripheral edema. Mycoplasma wenyonii and a related organism, Candidatus Mycoplasma haemobos, have been detected in both ill and apparently healthy cattle, but little is known about their prevalence in US dairy cattle. The objective of this prospective, cross-sectional study was to determine herd-level apparent prevalence of M. wenyonii and C. M. haemobos in dairy cattle located in Wisconsin and Michigan compared with seroprevalence of bovine leukemia virus (BLV) in the same herds. In summer 2018, researchers collected blood samples from 30 lactating cows per herd from randomly recruited farms in selected dairy-intensive counties in each state. During the farm visit, a brief survey was used to collect herd management information. Detection of M. wenyonii and C. M. haemobos were based on PCR testing, and ELISA was used to test for antibodies to BLV. Blood samples were collected from lactating cows located in 64 Wisconsin herds (n = 1,930 samples) and 18 Michigan herds (n = 591 samples). Herd-level apparent prevalence was 100% for both M. wenyonii and C. M. haemobos. Herd-level seroprevalence for BLV was 83 and 100% for Wisconsin and Michigan herds, respectively. Estimated within-herd apparent prevalence of M. wenyonii was 71.7% ± 1.0% (ranging from 23.3 to 93.5%) and for C. M. haemobos was 77.3% ± 1.0% (ranging from 16.7 to 100%). Within-herd prevalence of BLV positive samples was 39.8% ± 1.0% and ranged from 0 to 86.7%. About 22% of cows were concurrently positive for all 3 organisms. Parity and stage of lactation were recorded for 2,317 cows. Prevalence of positive cows for parity groups 1, 2, and ≥3 were 72.0, 73.8, and 67.7% for M. wenyonii; 80.9, 76.8, and 74.9% for C. M. haemobos; and 25.3, 39.7, and 55.5% for BLV, respectively. None or only minor differences in apparent prevalence were observed based on stage of lactation. This is the first report of the prevalence of hemotrophic mycoplasmas in Wisconsin and Michigan dairy herds and indicates that infection with these organisms is endemic. The impact of infection on cattle health and productivity remains unknown, and risk factors associated with infection warrant further study.

Computed tomography of tympanic bullae in pre-weaned dairy calves diagnosed with clinical respiratory disease.

Clinical disease from otitis media in calves is a significant problem in the dairy industry and evaluation of disease severity, chronicity, and imaging remains a challenge. Our objectives were to compare imaging findings in calves with an early diagnosis of respiratory disease to calves with treatment failure. This was a prospective study of 30 Jersey heifer calves, 26-95 days of age, with elevated clinical respiratory scores. Ten clinically healthy calves served as controls for clinical scoring. Three groups of calves were selected based on elevated scores using the McGuirk respiratory scoring system and treatment history. Group A included new cases, group B included primary treatment failures, and group C included multiple treatment failures. Calves underwent a skull CT, four view radiography, post-mortem photography of the tympanic bulla and bacteriological diagnostics. Imaging and post-mortem results were evaluated using normalized scoring schemes. Computed tomography imaging of the tympanic bulla differentiated calves early in the course of disease (group A) from calves that had not responded to treatment (groups B and C). Radiographs differentiated only group C from groups A and B. Use of a 35 degree angle dorsal-right or dorsal-left ventral oblique projection for radiography allowed effective evaluation of the tympanic bulla. Clinical respiratory scores were similar among all three groups. Computed tomography imaging can differentiate early from advanced otitis media. Radiographs, which can be performed in the field, also have utility to identify advanced otitis media to aid management decisions.

Prevalence and temporal trends in antimicrobial resistance of bovine respiratory disease pathogen isolates submitted to the Wisconsin Veterinary Diagnostic Laboratory: 2008-2017.

The objective of this study was to describe the prevalence and trends in antimicrobial resistance for bacterial pathogens associated with bovine respiratory disease (BRD) isolated from samples submitted to the Wisconsin Veterinary Diagnostic Laboratory (WVDL). Data were retrospectively collected from bovine respiratory isolates including Pasteurella multocida, Mannheimia haemolytica, Histophilus somni, and Bibersteinia trehalosi identified at the WVDL between January 2008 and December 2017. Antimicrobial susceptibility testing data were queried from antimicrobial resistance databases at the WVDL. A total of 4,261 isolates were identified. Pasteurella multocida was most frequently identified, accounting for 2,094 isolates (49% of total) over the study period. Mannheimia haemolytica was the second most frequently isolated bacterial respiratory pathogen (n = 1,267, 30%) followed by H. somni (n = 749, 18%) and B. trehalosi (n = 151, 4%). Over the 10-yr period, B. trehalosi had the highest median percentage of isolates that were resistant to at least one antibiotic at 33% (interquartile range: 24, 47) followed by M. haemolytica (13%; 8, 23). For P. multocida, 10% (4, 26) of isolates were classified as resistant to at least one antibiotic, whereas H. somni had the fewest resistant isolates (9%; 3, 15). When comparing 2013-2017 to 2008-2012, the overall percentage of resistant isolates for P. multocida and B. trehalosi decreased, whereas the percentage of resistant isolates for M. haemolytica and H. somni increased. Increased resistance against florfenicol, fluoroquinolones, gentamicin, tilmicosin, and tulathromycin was observed for M. haemolytica. These data show that antimicrobial susceptibility for BRD bacterial pathogens has changed in the population served by the WVDL over this 10-yr period. For P. multocida, resistance is relatively low and has either improved or at least remained constant for the majority of drugs labeled for treatment of respiratory disease in dairy cattle. Veterinarians and producers should be aware of the bacterial pathogens most commonly associated with BRD and work toward early disease detection, proper antibiotic administration, and monitoring lung lesions to ensure that their treatment protocols improve lung health.

Influence of Nodal signalling on pluripotency factor expression, tumour cell proliferation and cisplatin-sensitivity in testicular germ cell tumours.

BACKGROUND: Testicular germ cell tumours (TGCTs) are characterised by an overall high cisplatin-sensitivity which has been linked to their continued expression of pluripotency factors. Recently, the Nodal signalling pathway has been implicated in the regulation of pluripotency factor expression in fetal germ cells, and the pathway could therefore also be involved in regulating expression of pluripotency factors in malignant germ cells, and hence cisplatin-sensitivity in TGCTs. METHODS: We used in vitro culture of the TGCT-derived cell line NTera2, ex vivo tissue culture of primary TGCT specimens and xenografting of NTera2 cells into nude mice in order to investigate the consequences of manipulating Nodal and Activin signalling on pluripotency factor expression, apoptosis, proliferation and cisplatin-sensitivity. RESULTS: The Nodal signalling factors were markedly expressed concomitantly with the pluripotency factor OCT4 in GCNIS cells, seminomas and embryonal carcinomas. Despite this, inhibition of Nodal and Activin signalling either alone or simultaneously did not affect proliferation or apoptosis in malignant germ cells in vitro or ex vivo. Interestingly, inhibition of Nodal signalling in vitro reduced the expression of pluripotency factors and Nodal pathway genes, while stimulation of the pathway increased their expression. However, cisplatin-sensitivity was not affected following pharmacological inhibition of Nodal/Activin signalling or siRNA-mediated knockdown of the obligate co-receptor CRIPTO in NTera2 cells in vitro or in a xenograft model. CONCLUSION: Our findings suggest that the Nodal signalling pathway may be involved in regulating pluripotency factor expression in malignant germ cells, but manipulation of the pathway does not appear to affect cisplatin-sensitivity or tumour cell proliferation.

Dysregulation of FGFR signalling by a selective inhibitor reduces germ cell survival in human fetal gonads of both sexes and alters the somatic niche in fetal testes.

STUDY QUESTION: Does experimental manipulation of fibroblast growth factor 9 (FGF9)-signalling in human fetal gonads alter sex-specific gonadal differentiation? SUMMARY ANSWER: Inhibition of FGFR signalling following SU5402 treatment impaired germ cell survival in both sexes and severely altered the developing somatic niche in testes, while stimulation of FGF9 signalling promoted Sertoli cell proliferation in testes and inhibited meiotic entry of germ cells in ovaries. WHAT IS KNOWN ALREADY: Sex-specific differentiation of bipotential gonads involves a complex signalling cascade that includes a combination of factors promoting either testicular or ovarian differentiation and inhibition of the opposing pathway. In mice, FGF9/FGFR2 signalling has been shown to promote testicular differentiation and antagonize the female developmental pathway through inhibition of WNT4. STUDY DESIGN, SIZE, DURATION: FGF signalling was manipulated in human fetal gonads in an established ex vivo culture model by treatments with recombinant FGF9 (25 ng/ml) and the tyrosine kinase inhibitor SU5402 (10 µM) that was used to inhibit FGFR signalling. Human fetal testis and ovary tissues were cultured for 14 days and effects on gonadal development and expression of cell lineage markers were determined. PARTICIPANTS/MATERIALS, SETTING, METHODS: Gonadal tissues from 44 male and 33 female embryos/fetuses from first trimester were used for ex vivo culture experiments. Tissues were analyzed by evaluation of histology and immunohistochemical analysis of markers for germ cells, somatic cells, proliferation and apoptosis. Culture media were collected throughout the experimental period and production of steroid hormone metabolites was analyzed in media from fetal testis cultures by liquid chromatography-tandem mass spectrometry (LC-MS/MS). MAIN RESULTS AND THE ROLE OF CHANCE: Treatment with SU5402 resulted in near complete loss of gonocytes (224 vs. 14 OCT4+ cells per mm2, P < 0.05) and oogonia (1456 vs. 28 OCT4+ cells per mm2, P < 0.001) in human fetal testes and ovaries, respectively. This was a result of both increased apoptosis and reduced proliferation in the germ cells. Addition of exogenous FGF9 to the culture media resulted in a reduced number of germ cells entering meiosis in fetal ovaries (102 vs. 60 γH2AX+ germ cells per mm2, P < 0.05), while in fetal testes FGF9 stimulation resulted in an increased number of Sertoli cells (2503 vs. 3872 SOX9+ cells per mm2, P < 0.05). In fetal testes, inhibition of FGFR signalling by SU5402 treatment altered seminiferous cord morphology and reduced the AMH expression as well as the number of SOX9-positive Sertoli cells (2503 vs. 1561 SOX9+ cells per mm2, P < 0.05). In interstitial cells, reduced expression of COUP-TFII and increased expression of CYP11A1 and CYP17A1 in fetal Leydig cells was observed, although there were no subsequent changes in steroidogenesis. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: Ex vivo culture may not replicate all aspects of fetal gonadal development and function in vivo. Although the effects of FGF9 were studied in ex vivo culture experiments, there is no direct evidence that FGF9 acts in vivo during human fetal gonadogenesis. The FGFR inhibitor (SU5402) used in this study is not specific to FGFR2 but inhibits all FGF receptors and off-target effects on unrelated tyrosine kinases should be considered. WIDER IMPLICATIONS OF THE FINDINGS: The findings of this study suggest that dysregulation of FGFR-mediated signalling may affect both testicular and ovarian development, in particular impacting the fetal germ cell populations in both sexes. STUDY FUNDING/COMPETING INTEREST(S): This work was supported in part by an ESPE Research Fellowship, sponsored by Novo Nordisk A/S to A.JØ. Additional funding was obtained from the Erichsen Family Fund (A.JØ.), the Aase and Ejnar Danielsens Fund (A.JØ.), the Danish Government's support for the EDMaRC programme (A.JU.) and a Wellcome Trust Intermediate Clinical Fellowship (R.T.M., Grant no. 098522). The Medical Research Council (MRC) Centre for Reproductive Health (R.T.M.) is supported by an MRC Centre Grant (MR/N022556/1). The authors have no conflict of interest to disclose.

Quality Control of Compounded Crystalloid Fluids for Intravenous Delivery to Horses.

BACKGROUND: Periodic lack of availability and high cost of commercially produced isotonic fluids for intravenous (IV) use in horses have increasingly led to use of home-made or commercially compound fluids by veterinarians. Data regarding the quality control and safety of compounded fluids would be of benefit to equine veterinarians. OBJECTIVES: To compare electrolyte concentrations, sterility, and endotoxin contamination of commercially available fluids to 2 forms of compounded isotonic crystalloid fluids intended for IV use in horses. METHODS: Prospective study. Two methods of preparing compounded crystalloids formulated to replicate commercial Plasma-Lyte A (Abbott, Chicago, IL) were compared. One formulation was prepared by a hand-mixed method involving chlorinated drinking water commonly employed by equine practitioners, and the other was prepared by means of ingredients obtained from a commercial compounding pharmacy. The variables for comparison were electrolyte concentrations, sterility, and presence of endotoxin contamination. RESULTS: Electrolyte concentrations were consistent within each product but different between types of fluids (P < 0.0001). Hand-mixed fluids had significantly more bacterial contamination compared to commercial Plasma-Lyte A (P = 0.0014). One of the hand-mixed fluid samples had detectable endotoxin contamination. CONCLUSIONS AND CLINICAL IMPORTANCE: Chlorinated drinking water is not an acceptable source of water to compound isotonic fluids for IV administration. Equine practitioners should be aware of this risk and obtain the informed consent of their clients.

Antimicrobial resistance patterns of bovine Salmonella enterica isolates submitted to the Wisconsin Veterinary Diagnostic Laboratory: 2006-2015.

Salmonellosis on the dairy continues to have a significant effect on animal health and productivity and in the United States. Additionally, Salmonella enterica ssp. enterica causes an estimated 1.2 million cases of human illness annually. Contributing to the morbidity and mortality in both human and domestic animal species is emergence of antimicrobial resistance by Salmonella species and increased incidence of multidrug-resistant isolates. This study describes serotype distribution and the antimicrobial resistance patterns for various Salmonella serotypes isolated from bovine samples submitted to the Wisconsin Veterinary Diagnostic Laboratory (WVDL) over the past 10 yr. Salmonella serotyping and antimicrobial susceptibility testing data were obtained from the laboratory information management system at WVDL. Data from accessions were limited to bovine samples submitted to the WVDL between January 2006 and June 2015 and those that had both a definitive serotype and complete results for antimicrobial susceptibility testing. A total of 4,976 isolates were identified. Salmonella enterica ser. Dublin was the most prevalent serotype identified among bovine samples submitted to the WVDL, accounting for a total of 1,153 isolates (23% of total isolates) over the study period. Along with Dublin, Salmonella enterica ser. Cerro (795, 16%), Newport (720, 14%), Montevideo (421, 8%), Kentucky (419, 8%), and Typhimurium (202, 4%) comprised the top 6 most commonly isolated serotypes during that time. Overall, resistance of bovine Salmonella isolates in the study population remained stable, although decreases in resistance were noted for gentamicin, neomycin, and trimethoprim sulfamethoxazole during the study period. All isolates remained susceptible to enrofloxacin. These data show that antimicrobial susceptibility for bovine Salmonella has changed in the population served by WVDL in the past 10 yr. This information is important for understanding Salmonella disease ecology in Wisconsin. Our findings are also relevant for animal and public health by improving informed antimicrobial use, new drug development, and regulation of their use in food animals.

Photon upconversion with directed emission.

Photon upconversion has the potential to increase the efficiency of single bandgap solar cells beyond the Shockley Queisser limit. Efficient light management is an important point in this context. Here we demonstrate that the direction of upconverted emission can be controlled in a reversible way, by embedding anthracene derivatives together with palladium porphyrin in a liquid crystalline matrix. The system is employed in a triplet-triplet annihilation photon upconversion scheme demonstrating controlled switching of directional anti Stokes emission. Using this approach an emission ratio of 0.37 between the axial and longitudinal emission directions and a directivity of 1.52 is achieved, reasonably close to the theoretical maximal value of 2 obtained from a perfectly oriented sample. The system can be switched for multiple cycles without any visible degradation and the speed of switching is only limited by the intrinsic rate of alignment of the liquid crystalline matrix.

Validation of the Danish version of the Critical Care Pain Observation Tool.

BACKGROUND: Assessing pain in critically ill patients is a challenge even in an intensive care unit (ICU) with a no sedation protocol. The aim of this study was to validate the Danish version of the pain assessment method; Critical Care Pain Observation Tool (CPOT) in an ICU with a no sedation protocol. METHODS: Seventy patients were included in this study. The patients were observed during a non-nociceptive procedure (wash of an arm) and a nociceptive procedure (turning). Patients were observed before, during, and 15 min after the two interventions (six assessments). Two observers participated in the data collection and CPOT scores were blinded to each other. Calculations of interrater reliability, criterion validity and discriminant validity were performed to validate the Danish version of CPOT. RESULTS: The results indicated a good correlation between the two raters (all scores > 0.9 and P < 0.05). About 48 (68.6%) of the included patients were able to self-report pain. We found a significantly higher mean CPOT score at the nociceptive procedure than at rest or the non-nociceptive procedure (P < 0.05). No correlation was found between CPOT scores and physiological indicators. Patients self-reported pain and CPOT showed a significant correlation (P < 0.05). A CPOT score of ≥ 3 correlated with patients' self-reported pain (ROC AUC 0.83). CONCLUSION: The Danish version of CPOT can be used to assess pain in critically ill patients, also when the ICU has a no sedation protocol. CPOT scores showed a good interrater reliability and correlates well with patient's self-reported pain.

Dental pathology in conventionally fed and pasture managed dairy cattle.

Healthy teeth are important in the first stages of digestion for dairy cattle, yet little is known about bovine dental disease. This study aimed to investigate dental pathology of dairy cattle in two parts. First dairy cattle cadaver heads (n=11) were examined at the time of culling. Second, the authors performed oral exams in cattle fed a total mixed ration (TMR) (n=200) and pasture-based (n=71) grazing cattle. Cadaver heads were imaged using radiography and computed tomography before gross dissection to study dental anatomy and pathology. The most prevalent dental abnormalities were excessive transverse ridging of the occlusal surface, the presence of diastemas and third molar dental overgrowths (M3DO) in cadaver heads. Average thickness of subocclusal dentine ranged from 3.5 mm to 5.8 mm in cheek teeth but was >10 mm in maxillary teeth with M3DO. Radiographic findings were compared with oral examinations in live cattle. Prevalence of M3DO upon oral examination was 19 per cent and 28 per cent in herds of cattle fed a TMR diet and 0 per cent in a herd of grazing cattle. Dental abnormalities are prevalent in dairy cattle but due to thin subocclusal dentine in the cheek teeth, established equine dental treatment methodology is not appropriate for bovine cheek teeth with the exception of those that have developed M3DO.

Socio-economic status and incident diabetes mellitus among employees in Denmark: a prospective analysis with 10-year follow-up.

AIMS: To contribute to the research on diabetes and social inequality by presenting national data on incident diabetes mellitus, stratified according to socio-economic status. METHODS: National registers were combined, linking socio-economic status with incident diabetes over a 10-year period (2001-2010). The study population comprised employees in Denmark aged 20-59 years at baseline. Poisson regression analysis was used to estimate socio-economic rate ratios. Excess fraction analysis was used to determine the proportion of cases that would not have occurred if morbidity rates in each socio-economic group had been as low as those in the reference group. Monte Carlo simulation was used to calculate 95% CIs for excess fraction estimates RESULTS: A total of 1 005 572 men and 951 039 women were included in the analysis. The follow-up yielded 43 439 cases in 9 533 199 person-years at risk among men and 29 266 cases in 9 163 405 person-years at risk among women. Using 'professionals' as a reference group, higher levels of relative risk were observed among every other socio-occupational group. The excess fraction was, 0.342 (95% CI 0.329-0.354) among men and 0.359 (95% CI 0.349-0.369) among women. CONCLUSIONS: Excess fraction analysis suggests that more than a third of cases of diabetes could be prevented if all employees were exposed to the same working conditions as the reference population. Acknowledging potential confounders, the observed levels of incident diabetes among the workforce highlight the potential gains to be had by better use of the workplace as an arena for prevention. Greater integration of occupational health and general healthcare is required to achieve this.

Anoctamin 6 differs from VRAC and VSOAC but is involved in apoptosis and supports volume regulation in the presence of Ca2+.

Anoctamin 6 (ANO6), also known as TMEM16F, has been shown to be a calcium-activated anion channel with delayed calcium activation. The cellular function of ANO6 is under debate, and different groups have come to different conclusions about ANO6's physiological role. Although it is now quite well established that ANO6 is distinct from the volume-regulated anion channel, it is still unclear whether ANO6 or other anoctamins can be activated by cell swelling. In this study, we suggest that ANO1, ANO6, and ANO10 do not contribute to the volume-activated current in ANO-overexpressing HEK293 cells. Furthermore, knock-down of ANO6 in Ehrlich ascites tumor cells (EATC) and Ehrlich-Lettre ascites (ELA) did not decrease but instead significantly increased swelling-activated membrane currents. Knock-down of ANO6 in EATC did not reduce regulatory volume decrease (RVD) in the absence of extracellular calcium, whereas it significantly reduced RVD in the presence of calcium. Interestingly, we found that knock-down of ANO6 in ELA cells resulted in a decrease in cisplatin-induced caspase-3 activity, confirming earlier findings that ANO6 is involved in apoptosis. Finally, knock-down of ANO1 and ANO6 did not affect the volume-sensitive release of taurine in ELA cells. Thus, our data provide evidence that ANO6 cannot be activated directly by cell swelling unless Ca(2+) is present. We also conclude that ANO6 carries a current during RVD, provided extracellular calcium is present. Thus, swelling activation of ANO6 requires the presence of free calcium.

Post-parturient shedding of Listeria monocytogenes in breast milk of infected mice. [corrected].

The objectives of this study were to develop an animal model to study Listeria monocytogenes infection during the peri-parturient period and identify sources of maternal shedding of the pathogen. Peri-parturient mice were infected intragastrically with L. monocytogenes that expressed bacterial luciferase. Mice were then imaged in vivo over time. Secreted breast milk samples from mice infected after parturition were enriched and plated for culture and imaging. Bioluminescence imaging technology was able to detect luciferase emitting L. monocytogenes in vaginal secretions and maternal and fetal organs at 72 and 96 h post infection in mice infected prior to, or just after, parturition. The results from this study clearly show that L. monocytogenes is shed in vaginal secretions and disseminates to the mammary chain, from which it can be shed in the milk of peri-parturient mice.

The role of TMEM16A (ANO1) and TMEM16F (ANO6) in cell migration.

Members of the TMEM16 family have recently been described as Ca(2+)-activated Cl(-) channels. They have been implicated in cancer and appear to be associated with poor patient prognosis. Here, we investigate the role of TMEM16 channels in cell migration, which is largely unknown. We focused on TMEM16A and TMEM16F channels that have the highest expression of TMEM16 channels in Ehrlich Lettre ascites (ELA) cells. Due to the lack of specific pharmacological modulators, we employed a miRNA approach and stably knocked down the expression of TMEM16A and TMEM16F channels, respectively. Migration analysis shows that TMEM16A KD clones are affected in their directional migration, whereas TMEM16F KD clones show a 40 % reduced rate of cell migration. Moreover, TMEM16A KD clones have a smaller projected cell area, and they are rounder than TMEM16F KD clones. The morphological changes are linearly correlated with the directionality of cells. TMEM16A and TMEM16F, thus, have an important function in cell migration-TMEM16A in directional migration, TMEM16F in determination of the speed of migration. We conclude that TMEM16A and TMEM16F channels have a distinct impact on the steering and motor mechanisms of migrating ELA cells.

Characterization of periodontal biofilm in Down syndrome patients: a comparative study.

UNLABELLED: The aim of this study was to characterize the main periodontal bacterial species in Down syndrome (DS) patients with and without periodontitis. METHOD: This cross-sectional study involved 75 DS patients, 45 with and 30 without periodontitis. Informed consent, health and dental questionnaires and periodontitis diagnosis were performed PCR and LAMP assays were performed on subgingival dental plaque sample. RESULTS: Tannerella forsythia was the most frequent bacteria detected in the group with and without periodontitis (95.5 and 63.3%) followed by Treponema denticola (88.8 and 50%) and Porphyromonas gingivalis (53.3 and 25% respectively). There were statistical differences between groups (p < 0.05). Pg fimA type I was the most frequent Porphyromonas gingivalis genotype. Two different sets of primers (Aa-F/Aa-R and ltx3/ltx4) were used to detect Aggregatibacter actinomycetemcomitans and different frequencies were obtained, (68% and 14.6% respectively), they had a weak correlation (Cohen Kappa = 0.16). After sequencing of PCR products, ltx3/ltx4 showed more specificity. JP2 clone of A. actinomycetemcomitans was not detected in any sample. CONCLUSIONS: The composition of oral biofilm is fundamental for the development of periodontal disease independently of immunological alterations associated with DS. The frequency of detection of A. actinomycetemcomitans reported in the literature has a wide range, because the primers and probes applied

Prevalence of Aggregatibacter actinomycetemcomitans in Sudanese patients with aggressive periodontitis: a case-control study.

BACKGROUND AND OBJECTIVE: Aggregatibacter actinomycetemcomitans is considered a possible etiological agent for aggressive periodontitis. The aim of this study was to determine the prevalence of the JP2 clone and non-JP2 genotypes of A. actinomycetemcomitans in the subgingival plaque of patients with aggressive periodontitis and controls among Sudanese high-school students. MATERIAL AND METHODS: In a previous study we examined a large representative sample of students attending high schools in Khartoum, Sudan. In this population, 17 patients with aggressive periodontitis and 17 controls (14-19 years of age) consented to participate in the present study. The subjects underwent a clinical periodontal examination, and subgingival dental plaque samples were collected using paper points. The presence of the A. actinomycetemcomitans JP2 clone and non-JP2 genotypes were assessed using loop-mediated isothermal amplification (LAMP) and the PCR. RESULTS: The JP2 clone of A. actinomycetemcomitans was not detected in the subgingival plaque of either the cases or the controls. Non-JP2 types of A. actinomycetemcomitans were detected in the subgingival plaque of 12 (70.6%) patients with aggressive periodontitis and from only one (5.9%) control subject, showing a significantly higher frequency of detection in cases than in controls (p = 0.0001). The odds ratio for the detection of A. actinomycetemcomitans in the subgingival plaque of the patients with aggressive periodontitis was 38.4 (95% confidence interval: 4.0-373.0; p = 0.002). The PCR and LAMP methods showed identical results pertaining to the identification of non-JP2 types of A. actinomycetemcomitans. CONCLUSIONS: The JP2 clone of A. actinomycetemcomitans was not detected in the subgingival plaque of high school subjects in Sudan. The detection of non-JP2 types of A. actinomycetemcomitans may be a useful marker of increased risk for development of aggressive periodontitis in young subjects.

Deregulation of apoptotic volume decrease and ionic movements in multidrug-resistant tumor cells: role of chloride channels.

Changes in cell volume and ion gradients across the plasma membrane play a pivotal role in the initiation of apoptosis. Here we explore the kinetics of apoptotic volume decrease (AVD) and ion content dynamics in wild-type (WT) and multidrug-resistant (MDR) Ehrlich ascites tumor cells (EATC). In WT EATC, induction of apoptosis with cisplatin (5 muM) leads to three distinctive AVD stages: an early AVD(1) (4-12 h), associated with a 30% cell water loss; a transition stage AVD(T) ( approximately 12 to 32 h), where cell volume is partly recovered; and a secondary AVD(2) (past 32 h), where cell volume was further reduced. AVD(1) and AVD(2) were coupled to net loss of Cl(-), K(+), Na(+), and amino acids (ninhydrin-positive substances), whereas during AVD(T), Na(+) and Cl(-) were accumulated. MDR EATC was resistant to cisplatin, showing increased viability and less caspase 3 activation. Compared with WT EATC, MDR EATC underwent a less pronounced AVD(1,) an augmented AVD(T), and a delay in induction of AVD(2). Changes in AVD were associated with inhibition of Cl(-) loss during AVD(1), augmented NaCl uptake during AVD(T), and a delay of Cl(-) loss during AVD(2). Application of the anion channel inhibitor NS3728 inhibited AVD and completely abolished the differences in AVD, ionic movements, and caspase 3 activation between WT and MDR EATC. Finally, the maximal capacity of volume-regulated anion channel was found to be strongly repressed in MDR EATC. Together, these data suggest that impairment of AVD, primarily via modulation of NaCl movements, contribute to protection against apoptosis in MDR EATC.

Stability of the JP2 clone of Aggregatibacter actinomycetemcomitans.

The JP2 clone of Aggregatibacter actinomycetemcomitans is strongly associated with aggressive periodontitis. To obtain information about colonization dynamics of the JP2 clone, we used PCR to examine its presence in 365 Moroccan juveniles from whom periodontal plaque samples were collected at baseline and after one and two years. Periodontal attachment loss was measured at baseline and at the two-year follow-up. At baseline, 43 (12%) carriers of the JP2 clone were found. Nearly half (44 %) of these were persistently colonized with the clone. The relative risk for the development of aggressive periodontitis, adjusted for the concomitant presence of other genotypes of A. actinomycetemcomitans, was highest for individuals continuously infected by the JP2 clone (RR = 13.9; 95% CI, 9.0 to 21.4), indicating a relationship between infectious dose and disease, which further substantiates the evidence for the JP2 clone as a causal factor in aggressive periodontitis.

Analytical assessment of the osteoinductive material COLLOSSE.

In this study, the growth factors in COLLOSSE were analyzed, using ELISA tests, mass spectrometry, western blotting, and a 24-day cell culture experiment using osteoblast-like cells. The results of the ELISA testing, mass spectrometry, and western blotting all confirmed that TGF-beta1 was the main growth factor in COLLOSSE at 55 ng/mg. The results from the culture test showed that the cell proliferation, alkaline phosphatase activity, and matrix calcification were all drastically changed by the addition of COLLOSSE, mirroring the effects of addition of TGF-beta1. We conclude that COLLOSSE is not only a rich source of TGFbeta-1, but also contains the growth factors TGFbeta-2, BMP-2, BMP-3, BMP-7, IGF-1, and possibly VEGF. Other growth factors might be present in COLLOSSE, but were not identified due to inherent detection limits of the used ELISA and mass spectrometry techniques. The number of osteoinductive factors in COLLOSSE causes a synergistic effect, explaining the new bone formation found in previously described in vivo studies, with much lower growth factor concentrations when compared with recombinant BMPs.