Emergence and whole-genome sequence of Senecavirus A in Colombia.

In 2015 and 2016, Senecavirus A (SVA) emerged as an infectious disease in Brazil, China and the United States (US). In a Colombian commercial swine farm, vesicles on the snout and coronary bands were reported and tested negative for foot-and-mouth disease virus (FMDv), but positive for SVA. The whole-genome phylogenetic analysis indicates the Colombian strain clusters with the strains from the United States, not with the recent SVA strains from Brazil.

Progesterone receptor-B enhances estrogen responsiveness of breast cancer cells via scaffolding PELP1- and estrogen receptor-containing transcription complexes.

Progesterone and estrogen are important drivers of breast cancer proliferation. Herein, we probed estrogen receptor-α (ER) and progesterone receptor (PR) cross-talk in breast cancer models. Stable expression of PR-B in PR-low/ER+ MCF7 cells increased cellular sensitivity to estradiol and insulin-like growth factor 1 (IGF1), as measured in growth assays performed in the absence of exogenous progestin; similar results were obtained in PR-null/ER+ T47D cells stably expressing PR-B. Genome-wide microarray analyses revealed that unliganded PR-B induced robust expression of a subset of estradiol-responsive ER target genes, including cathepsin-D (CTSD). Estradiol-treated MCF7 cells stably expressing PR-B exhibited enhanced ER Ser167 phosphorylation and recruitment of ER, PR and the proline-, glutamate- and leucine-rich protein 1 (PELP1) to an estrogen response element in the CTSD distal promoter; this complex co-immunoprecipitated with IGF1 receptor (IGFR1) in whole-cell lysates. Importantly, ER/PR/PELP1 complexes were also detected in human breast cancer samples. Inhibition of IGF1R or phosphoinositide 3-kinase blocked PR-B-dependent CTSD mRNA upregulation in response to estradiol. Similarly, inhibition of IGF1R or PR significantly reduced ER recruitment to the CTSD promoter. Stable knockdown of endogenous PR or onapristone treatment of multiple unmodified breast cancer cell lines blocked estradiol-mediated CTSD induction, inhibited growth in soft agar and partially restored tamoxifen sensitivity of resistant cells. Further, combination treatment of breast cancer cells with both onapristone and IGF1R tyrosine kinase inhibitor AEW541 was more effective than either agent alone. In summary, unliganded PR-B enhanced proliferative responses to estradiol and IGF1 via scaffolding of ER-α/PELP1/IGF1R-containing complexes. Our data provide a strong rationale for targeting PR in combination with ER and IGF1R in patients with luminal breast cancer.

Swimming-induced pulmonary oedema--a hazard in intensive military training?

Swimming-Induced Pulmonary Oedema (SIPE) has been described in military combat swimmers in both the US and Israeli Navies. The pathophysiology is explained by the immersion in cold water, and its effects on central vascular volume. SIPE has been hypothesized to be caused by pulmonary capillary stress failure (PCSF) due to elevations in pulmonary capillary transmural pressure. This leads to mechanical impairment and leakage of blood cells and proteins from capillaries. Patients with SIPE can present with pronounced dyspnoea, cough, hypoxemia and profuse frothy haemoptysis. Physical examination and chest X-rays usually show evidence of pulmonary oedema. The treatment of choice is to recognize the symptoms, get the patient out of the water and follow with close observation for emergent problems. Soldiers prone to acquire SIPE should be identified as this medical condition has a high degree of recurrence. The awareness of the symptoms of SIPE will increase appropriate diagnosis and therefore inform treatment.

Increased understanding of intestinal drug permeability determined by the LOC-I-GUT approach using multislice computed tomography.

This study further evaluated the in vivo single-pass perfusion technique (LOC-I-GUT) in three different ways. First, the intestinal radius of the human small intestinal segment was measured on plain X-ray films; second, evaluation was performed by applying multislice computed tomography investigations; and third, furosemide was used as model drug in a transport study. In total 17 (6 + 4 +7) intubation/perfusion studies were performed in healthy volunteers. Mixobar was used as a positive radiographic contrast agent in the first six volunteers when plain film examination was made, followed by four studies using multislice computed tomography. Mantel area calculations of the perfused segment after X-ray investigations using barium as contrast were determined to be 101.0 +/- 2.9 cm2. Maximal dilatation of the closed segment with room air as contrast and using MSCT revealed a mantel area of 121.30 +/- 7.0 cm2 (P < 0.01). Thus, the mantle area increased a further 20% when the bowel was fully distended, reflecting different physiologic distention patterns for air and fluid. A jejunal single-pass perfusion study was performed in a further seven volunteers. In each experiment furosemide was perfused during 200 min, and in the treatment period (100-200 min), fexofenadine was added to the perfusion solution. The mean (+/-SD) P (eff) for furosemide was 0.17 +/- 0.07 and 0.12 +/- 0.09 x 10-4 cm/s in the control and treatment period, respectively. This study showed that the calculation of human in vivo permeability is based on physiological values, which are important for the wide application of these in vivo permeability data in physiologically based pharmacokinetic modeling.

Insulin and free oestradiol are independent risk factors for benign prostatic hyperplasia.

The aetiology of benign prostatic hyperplasia (BPH) remains unclear. The objective of the present study was to test the insulin, oestradiol and metabolic syndrome hypotheses as promoters of BPH. The design was a risk factor analysis of BPH in which the total prostate gland volume was related to endocrine and anthropometric factors. The participants studied were 184 representative men, aged 72-76 years, residing in Göteborg, Sweden. Using a multivariate analysis, BPH as measured by the total prostate gland volume correlated statistically significantly with fasting serum insulin (beta=0.200, P=0.028), free oestradiol (beta=0.233, P=0.008) and lean body mass (beta=0.257, P=0.034). Insulin and free oestradiol appear to be independent risk factors for BPH, confirming both the insulin and the oestradiol hypotheses. Our findings also seem to confirm the metabolic syndrome hypothesis. The metabolic syndrome and its major endocrine aberration, hyperinsulinaemia, are possible primary events in BPH.

Single nucleotide polymorphisms for integrative mapping in the Turkey (Meleagris gallopavo).

When multiple genetic maps exist for a species, integration of these maps requires a set of common markers be genotyped across the individual mapping populations. In the turkey, three genetic maps based on separate mapping populations are available. In this study, SNP-based markers were developed for integrating the cDNA/RFLP-based map (1) with microsatellite markers of the second-generation turkey genome map (2). Forty-eight primer sets were designed and tested and 33 (69%) correctly amplified turkey genomic DNA by PCR. Putative SNPs were detected in 20 (61%) of the amplified gene fragments, and 10 SNP markers were subsequently genotyped by PCR/RFLP for segregation analysis. Eight SNP markers were incorporated into the turkey genetic map.

Using the chicken genome sequence in the development and mapping of genetic markers in the turkey (Meleagris gallopavo).

The efficacy of employing the chicken genome sequence in developing genetic markers and in mapping the turkey genome was studied. Eighty previously uncharacterized microsatellite markers were identified for the turkey using BLAST alignment to the chicken genome. The chicken sequence was then used to develop primers for polymerase chain reaction where the turkey sequence was either unavailable or insufficient. A total of 78 primer sets were tested for amplification and polymorphism in the turkey, and informative markers were genetically mapped. Sixty-five (83%) amplified turkey genomic DNA, and 33 (42%) were polymorphic in the University of Minnesota/Nicholas Turkey Breeding Farms mapping families. All but one marker genetically mapped to the position predicted from the chicken genome sequence. These results demonstrate the usefulness of the chicken sequence for the development of genomic resources in other avian species.

Rapid modulation of Ca2+ uptake in human jejunal enterocytes.

The active metabolite of D vitamin, 1,25(OH)2D3, has been suggested to promote acute uptake of calcium through the intestinal lining in cell lines and murine models. In this study, the effects of D vitamin on the cytoplasmic Ca2+ of single human jejunal enterocytes, obtained with LOC-I-GUT technique, was analyzed in vivo in a fluorometric system using fura-2 as the Ca2+-sensing probe. Vitamin-promoted acute Ca2+ influx exhibited dual kinetics, indicating initial release from intracellular Ca2+ pools and fast entry from the extracellular space. Furthermore, providing a chemical clamp of membrane potential close to 0 mV did not activate voltage-sensitive calcium channels in the cellular membrane, neither was the hormone-induced Ca2+ influx affected by verapamil. This advocates that voltage-operated channels like L-type Ca2+ channels do not participate in the process of Ca2+ uptake. In fact, the existence of calcium-release-activated-calcium channels (I(CRAC)) was implied by the findings that irreversible depletion of intracellular Ca2+ stores by thapsigargin promoted Ca2+ entry. In the thapsigargin-treated enterocytes, D vitamin lost its ability to promote calcium entry indicating an important role for intracellular store-operated Ca2+ stores in the acute effects of 1,25(OH)2D3.

Integration of microsatellite-based genetic maps for the turkey (Meleagris gallopavo).

Integration of turkey genetic maps and their associated markers is essential to increase marker density in support of map-based genetic studies. The objectives of this study were to integrate 2 microsatellite-based turkey genetic maps--the Roslin map and the University of Minnesota (UMN) map--by genotyping markers from the Roslin study on the mapping families of the UMN study. A total of 279 markers was tested, and 240 were subsequently screened for polymorphisms in the UMN/Nicholas Turkey Breeding Farms (NTBF) mapping families. Of the 240 markers, 89 were genetically informative and were used for genotyping the F2 offspring. Significant genetic linkages (log of odds > 3.0) were found for 84 markers from the Roslin study. BLASTn comparison of marker sequences with the draft assembly of the chicken genome found 263 significant matches. The combination of genetic and in silico mapping allowed for the alignment of all linkage groups of the Roslin map with those of the UMN map. With the addition of the markers from the Roslin map, 438 markers are now genetically linked in the UMN/NTBF families, and more than 1700 turkey sequences have now been assigned to likely positions in the chicken-genome sequence.

Simulation of Sahel drought in the 20th and 21st centuries.

The Sahel, the transition zone between the Saharan desert and the rainforests of Central Africa and the Guinean Coast, experienced a severe drying trend from the 1950s to the 1980s, from which there has been partial recovery. Continuation of either the drying trend or the more recent ameliorating trend would have far-ranging implications for the economy and ecology of the region. Coupled atmosphere/ocean climate models being used to simulate the future climate have had difficulty simulating Sahel rainfall variations comparable to those observed, thus calling into question their ability to predict future climate change in this region. We describe simulations using a new global climate model that capture several aspects of the 20th century rainfall record in the Sahel. An ensemble mean over eight realizations shows a drying trend in the second half of the century of nearly half of the observed amplitude. Individual realizations can be found that display striking similarity to the observed time series and drying pattern, consistent with the hypothesis that the observations are a superposition of an externally forced trend and internal variability. The drying trend in the ensemble mean of the model simulations is attributable to anthropogenic forcing, partly to an increase in aerosol loading and partly to an increase in greenhouse gases. The model projects a drier Sahel in the future, due primarily to increasing greenhouse gases.

A comparative genetic map of the turkey genome.

Genetic markers (microsatellites and SNPs) were used to create and compare maps of the turkey and chicken genomes. A physical map of the chicken genome was built by comparing sequences of turkey markers with the chicken whole-genome sequence by BLAST analysis. A genetic linkage map of the turkey genome (Meleagris gallopavo) was developed by segregation analysis of genetic markers within the University of Minnesota/Nicholas Turkey Breeding Farms (UMN/NTBF) resource population. This linkage map of the turkey genome includes 314 loci arranged into 29 linkage groups. An additional 40 markers are tentatively placed within linkage groups based on two-point LOD scores and 16 markers remain unlinked. Total map distance contained within linkage groups is 2,011 cM with the longest linkage group (47 loci) measuring 413.3 cM. Average marker interval over the 29 linkage groups was 6.4 cM. All but one turkey linkage group could be aligned with the physical map of the chicken genome. The present genetic map of the turkey provides a comparative framework for future genomic studies.

Microsatellite loci for genetic mapping in the turkey (Meleagris gallopavo).

New microsatellite loci for the turkey (Meleagris gallopavo) were developed from two small insert DNA libraries. Polymorphism at these new loci was examined in domestic birds and two resource populations designed for genetic linkage mapping. The majority of loci (152 of 168) was polymorphic in domestic turkeys and informative in two mapping resource populations and thus will be useful for genetic linkage mapping.

The effect of amiloride on the in vivo effective permeability of amoxicillin in human jejunum: experience from a regional perfusion technique.

The purpose of this human intestinal perfusion study (in vivo) was twofold. Firstly, we aimed to determine the effective in vivo jejunal permeability (P(eff)) of amoxicillin and to classify it according to the Biopharmaceutics Classification System (BCS). Secondly, we investigated the acute effect of amiloride on the jejunal P(eff) of amoxicillin. Amoxicillin, a beta-lactam antibiotic, has been reported to be absorbed across the intestinal mucosa by both passive diffusion and active transport. A regional single-pass perfusion of the jejunum was performed using a Loc-I-Gut perfusion tube in 14 healthy volunteers. Each perfusion lasted for 200 min and was divided into two periods of 100 min each. The concentration of amoxicillin entering the jejunal segment was 300 mg/l in both periods, and amiloride, an inhibitor of the Na+/H+ exchanger, was added at 25 mg/l in period 2. The concentrations of amoxicillin and amiloride in the outlet jejunal perfusate were measured with two different HPLC-methods. Antipyrine and [14C]PEG 4000 were added as internal standards to the perfusion solution. Amiloride had no significant effect on the jejunal P(eff) of amoxicillin. The human in vivo jejunal P(eff) for amoxicillin was 0.34+/-0.11 x 10(-4) and 0.46+/-0.12 x 10(-4) cm/s in periods 1 and 2, respectively. The high jejunal P(eff) for amiloride was 1.63+/-0.51 x 10(-4) cm/s which predicts an intestinal absorption of more than 90%. Following the BCS amoxicillin was classified as a low P(eff) drug, and amiloride had no acute effect on the in vivo jejunal P(eff) of amoxicillin.

Axon collaterals of Meynert cells diverge over large portions of area V1 in the macaque monkey.

Patchy intrinsic connections, originating mainly from horizontal collaterals of pyramidal neurons, have been demonstrated in area V1 and many other cortical areas. In this article, we identify a network of intrinsic connections concentrated in layer 6 of area V1. These are visualized by extracellular injections of anterograde tracers in V1, which label small clusters of large terminal boutons in layer 6, in conjunction with thick axon segments. These segments can be traced back to infragranular Meynert cells (n = 10), which are retrogradely labeled from the injections. By using serial section analysis, we identified the following features of this distinctive system of Meynert cell collaterals: (1) terminal clusters are relatively small (<100 microm); (2) each cluster has a small number of rather large boutons (up to 3.0 microm); (3) there is typically a termination-free zone in the immediate vicinity (0.5-2.0 mm) of the cell body; (4) a single neuron has multiple branches that can extend up to 8.0 mm from the soma; and (5) the collaterals are concentrated in layer 6. These features are different from those of horizontal intrinsic connections in the supragranular layers of area V1. They are consistent with fast dynamics and a possible role in wide-field motion processing, such as has been associated with Meynert cells from other studies.

BPH with coexisting overactive bladder dysfunction--an everyday urological dilemma.

The aim of this study was to use a systematic schedule, including urodynamics, to describe the rate of coexisting overactive bladder (OB) in patients with bladder outlet obstruction (BOO) caused by benign prostatic hyperplasia (BPH). We also identified differences between the patients with pure BOO compared with those with BOO combined with OB (BOO + OB). One hundred and sixty-two men referred to our clinic due to LUTS were included. Patients with a history that might affect their bladder function were excluded. After cystometry and pressure-flow studies, the patients were divided into pure BOO and BOO + OB. Of the 162 men, 55% had pure BOO. BOO + OB was found in 45%. Age, s-PSA, voided volume, and obstruction grade differed significantly between the groups. The patients with BOO + OB were older, had a higher s-PSA, voided smaller volumes, and were more obstructed. We found no differences in TRUS-volume, Q-max, IPS score, or PVR. There was a strong association between OB and BOO, the percentage of OB increasing with increased obstruction. TRUS-volume, Q-max, IPS score, and PVR did not predict whether the patients had a combined BOO + OB or not. These findings indicate that BOO is a progressive disease, which in time causes pronounced obstruction and perhaps in itself contributes to the development of OB.

Invasive urodynamic studies are well tolerated by the patients and associated with a low risk of urinary tract infection.

OBJECTIVE: An essential part of investigation of the lower urinary tract is pressure/flow studies (pQS). In fact, pQS is the only way of diagnosing bladder outlet obstruction. There is controversy regarding whether or not prophylactic antibiotic treatment is necessary. This prospective study was carried out in order to determine the frequency of infections and/or distress after pQS performed without the use of antibiotic prophylaxis. MATERIAL AND METHODS: One hundred and twenty-three patients were included in the present study, all males. They were requested to answer a questionnaire I week after pQS. Questions were asked concerning symptoms of voiding disorders, dysuria, hematuria, incidence of fever and the patient's acceptance of the investigation after the pQS procedure. Urine was obtained for culture immediately before the investigation and 3 and 7 days after the pQS. RESULTS: Forty-six per cent of the patients experienced some degree of transient dysuria after pQS. and 18.5% experienced voiding problems of varying nature. Five per cent of the patients had hematuria and 2.5% reported fever. Fifty per cent of the patients experienced some degree of discomfort during the pQS investigation, and 4.1% had positive culture and symptoms of UTI requiring antibiotic treatment. CONCLUSIONS: PQS is well accepted by the patients and the regular use of propylactic antibiotics is not indicated. We recommend, though, that patients at risk for serious complications from infections (e.g. those with prosthetic heart valves) should receive prophylactic antibiotics.

Can urodynamic assessment of outflow obstruction predict outcome from watchful waiting?--A four-year follow-up study.

OBJECTIVE: One of the most common "treatment" alternatives in suspected outflow obstruction due to bladder outlet obstruction (BOO) is watchful waiting (WW). The aim of this study was to see whether there were any differences in outcome between patients with slight, moderate or severe obstruction due to BOO as classified by transrectal ultrasound (TRUS) and urodynamics. MATERIAL AND METHODS: Thirty-seven men with lower urinary tract symptoms (LUTS) and suspected BOO were included. All of the patients were investigated by a routine investigation schedule, including TRUS and urodynamics with pressure-flow measurement (pQS) at baseline. Patients with cancer in the urinary tract, prostatitis, history of detrusor hyperreflexia (peripheral or central diseases or trauma to the nervous system affecting the bladder) and serious systemic diseases were excluded. Patients were examined at baseline, then checked again after 1 year and 4 years. Patients who did not want to continue with WW were listed as treatment failures. RESULTS: At baseline, 43.2% of the patients were urodynamically severely obstructed and 32.3% were moderately obstructed. Thirty-five per cent of the patients were found to have previously unknown detrusor hyperactivity/overactivity. The prevalence of detrusor hyperactivity/overactivity increased with BOO. After 1 year, IPSS had decreased at unchanged Qmax and postresidual volume. These findings persisted at 4 years. The failure rate increased in the more obstructed patients and was significantly higher with more severe obstruction. Complications were found in 13.5%, with no significant differences between patients with minor BOO [Detrusor Adjusted Mean PURR Factor (DAMPF) scale <42], moderate BOO (DAMPF 42-65) and severe BOO (DAMPF >65). CONCLUSIONS: In patients with severe BOO, the LUTS and failure rate increase over time. The percentage of patients with detrusor hyperactivity/overactivity was higher in the severely obstructed group. By including full urodynamics when investigating patients with BOO, it seems possible to predict the failure rate according to the patients' obstruction grade. This gives an opportunity to treat the patient with minimal invasion, and to give the individual patient a more precise prognosis if WW is preferred.

Pressure-flow studies for patient selection in the treatment of symptomatic BPH--a one-year follow-up study.

OBJECTIVE: Many different treatments for lower urinary tract symptoms (LUTS) due to bladder outlet obstruction (BOO) are available today. To select the most suitable method for each patient is therefore a delicate task. The aim of this study has been to use a standardised systematic investigation schedule including pressure flow studies (pQS) in order to try to use graded treatment according to obstruction. METHODS: Ninety-nine patients were systematically examined with routine investigations and pQS to select between 3 treatment options, TURP, TUMT 2.0 (low energy) and watchful waiting (WW). Patients with severe BOO were recommended TURP, patients with moderate BOO were treated with TUMT and patients with no or minor BOO were recommended WW. RESULTS: TURP produced the best improvement in maximum free flow (Q-max), IPS-score and PVR, but only TURP had serious complications. TUMT treatment produced a more moderate improvement in flow rate, IPS-score and PVR, and all of the complications were minor. WW did not improve PVR or Q-max but the IPS-score decreased significantly. One UTI was the only complication in the WW group. CONCLUSIONS: pQS can be used to allocate patients with LUTS due to suspected BOO into different treatment arms; TURP, TUMT, WW, all with known different effects of BOO and with different severity of complications. Good symptomatic effect in Q-max, PVR and IPS-score with less serious complications and at low failure rate can thereby be obtained.

Feedback connections from area MT of the squirrel monkey to areas V1 and V2.

Area MT/V5 is reciprocally connected with both V1 and V2; but, despite extensive anatomical and physiological investigations, detailed information on the feedback component of these connections is still not available. The present report uses serial section reconstruction of single axons, labeled by anterograde tracers injected in area MT of squirrel monkeys, to characterize these connections further. As with other feedback systems, MT axons terminating in both areas V1 (n = 9) and V2 (n = 6) are widely divergent. In area V1, MT fields are larger than those from V2 and are about comparable to those from V4 or TEO. Terminations in V1, unlike other feedback connections described so far, terminate in several laminar combinations: only layer 1 (n = 2); only layer 4B (n = 3); layers 1 and 4B (n = 1); and layers 1, 4B, and 6 (n = 3). In V2, they occur mainly in layers 1 and 5 or 6. Terminations have two patterns even within a single axon: strung along collateral segments and grouped within small clusters. There are no apparent differences in the size, shape, or density of terminal specializations in V1 or V2, and, consistently with previous double-labeling experiments (Kennedy and Bullier [1985] J Neurosci 5:2815-2830), some axons can branch to both areas. This result, along with the laminar evidence for subtypes of feedback connections, argues against an exclusively hierarchical organization based on "pairwise" connectivity. For V1 and MT, there may be directly reciprocal loops between feedforward and feedback projecting neurons, but this is less likely to be so for V2 and MT.

The effect of ketoconazole on the jejunal permeability and CYP3A metabolism of (R/S)-verapamil in humans.

AIMS: The purpose of this human intestinal perfusion study was to investigate the effect of ketoconazole on the jejunal permeability and first-pass metabolism of (R)- and (S)-verapamil in humans. METHODS: A regional single-pass perfusion of the jejunum was performed using a Loc-I-Gut(R) perfusion tube in six healthy volunteers. Each perfusion lasted for 200 min and was divided into two periods of 100 min each. The inlet concentration of (R/S)-verapamil was 120 mg l-1 in both periods, and ketoconazole was added at 40 mg l-1 in period 2. (R/S)-verapamil was also administered as a short intravenous infusion of 5 mg, over a period of 10 min. The appearance ratios of the CYP3A formed metabolites (R)- and (S)-norverapamil were also estimated in the outlet jejunal perfusate. RESULTS: The effective jejunal permeability (Peff) of both (R)- and (S)-verapamil was unaffected by the addition of ketoconazole in period 2 suggesting that ketoconazole had no effect on the P-glycoprotein mediated efflux. However, the appearance ratio of both (R)- and (S)-norverapamil in the outlet jejunal perfusate decreased in the presence of ketoconazole. The rate of absorption into plasma of (R)- and (S)-verapamil increased despite the low dose of ketoconazole added, indicating an inhibition of the gut wall metabolism of (R/S)-verapamil by ketoconazole. CONCLUSIONS: Ketoconazole did not affect the jejunal Peff of (R/S)-verapamil, but it did increase the overall transport into the systemic circulation (bioavailability), probably by inhibition of the gut wall metabolism of verapamil. This might be due to ketoconazole being less potent as an inhibitor of P-glycoprotein than of CYP3A4 in vivo in humans.