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Solution of cancer was obviously an important national interest for President Nixon to declare ‘War on Cancer” in 1971. Health professionals failed the challenge to put cancer away during the 5 years of intensive presidential support and in the following 46 years of exclusive medical support allocated to cancer. Cancer stem cells (CSCs) stood in the way to deny the success of cytotoxic agents, the choice of cancer establishments in the past, to put cancer away. Cancer arises due to wound not healing properly. Consequently, the process of wound healing is the most appropriate modality of cancer therapy. Cytotoxic agents creating wounds are apparently contra-indication on cancer therapy. Cytotoxic agents are, however, the most effective agents to eliminate the most outstanding feature of cancer, namely the perpetual replication of cancer cells (CCs). A critical mechanism of wound healing is the induction of terminal differentiation of progenitor stem cells (PSCs) which are the precursors of CSCs. Wound healing metabolites active as differentiation inducers (DIs) and differentiation helper inducers (DHIs) are required to direct the terminal differentiation of PSCs to heal the wound. Cell differentiation agent (CDA) formulations are preparations of wound healing metabolites most fit to take out PSCs and CSCs by directing these cells to undergo terminal differentiation, which are protected by drug resistance and anti-apoptosis mechanisms, thus unresponsive to cytotoxic agents. CDAs alone or in combination with cytotoxic agents become perfect drugs to take out both CCs and CSCs, and to restore the functionality of chemo-surveillance necessary to win the war on cancer.

Modern agri-food systems generate large amounts of crop-based biomass that are unfit for direct human consumption but potentially eligible for upcycling via livestock feeding for production of meats, milk, and eggs. This study aims to develop novel feeds for cattle from some of those biomass materials through the natural microbial-driven processes of ensiling. Fruit and vegetables resembling supermarket discards were ensiled alone or co-ensiled with corn crop residues, mushroom wastes, etc. via laboratory experiments. Longitudinal sample analyses showed that (co-)ensiling was successful, with pH and fermentation acids falling within desirable ranges (pH in vitro incubation results indicated that the ensiled products could substitute certain conventional feeds while maintaining diet digestibility. Our findings provide a proof of principle for generating quality novel feeds for cattle from food discards and under-utilized crop residues. Future scale-up studies and animal feeding trials to demonstrate the utility of this approach can help societies begin re-purposing and more effectively utilizing otherwise-untapped feedstock resources, strengthening the regenerative capacity of agri-food systems towards a more sustainable food future.

Modern agri-food systems generate large amounts of crop-based biomass that are unfit for direct human consumption but potentially suitable for livestock feeding in production of meats, milk, and eggs. This study aims to develop novel feeds for cattle from some of those biomass materials through the natural microbial-driven processes of ensiling. Fruit and vegetables resembling supermarket discards were ensiled alone or co-ensiled with corn crop residues, mushroom wastes, etc. via laboratory experiments. Longitudinal sample analyses showed that (co-)ensiling was successful, with pH and fermentation acids changing rapidly into desirable ranges (pH

This review highlights the impact of wound onto the evolution and therapy of cancer. The objective of this review is to explore the mechanism of wound healing and its relationship to the evolution and therapy of cancer. Wound healing requires the proliferation and the terminal differentiation (TD) of progenitor stem cells (PSCs), which are the precursors of cancer stem cells (CSCs).Healing wound is not a big deal. If the functionality of chemo-surveillance is intact such as healthy people who can maintain a steady level of wound healing metabolites functioning as differentiation inducers (DIs) and differentiation helper inducers (DHIs).Wounds are always successfully healed without having to put up any effort, just to let the nature to do the healing. Medications such as suture and antibiotics are subsidiary to speed up the healing or to prevent infection. Acute wound affects the functionality of chemo-surveillance only temporarily, which is quickly recovered to return to the normal state. It is the chronic wound such as persistent infectious diseases or exposure to toxic chemicals including carcinogens for a long time that produces damaging effect on the functionality of chemo-surveillance.Chronic wound prompts the production of inflammatory cytokines to cause excessive urinary excretion of wound healing metabolites to affect wound healing. Without sufficient wound healing metabolites to terminate the proliferation of PCSs, it is very easy for PSCs to evolve into CSCs. It takes only a single hit to silence TET-1 enzyme to complete the transition, which is well within the reach of PSCs equipped with abnormally active methylation enzymes (MEs).CSCs can then progress to faster growing cancer cells by the activation of oncogenes or the inactivation of (AML). Cancer due to wound not healing properly is not unique to MDS and AML. It is rather a common phenomenon. War on cancer can be easily won if the battle is conducted following the nature’s course to heal the wound, just like the success of wound healing without having to put up any effort in healthy people. Therefore, the best strategy to win the war on cancer is to restore the functionality of chemo-surveillance by the employment of DIs and DHIs and to prevent the loss of wound healing metabolites through anti-cachexia chemicals such as phenylacetylglutamine. Then the nature will take its course to stop the proliferation of cells with abnormal MEs that include CSCs, PSCs, and all cancer cells. Destruction strategy to kill cancer cells is definitely counter indication. It creates more damages to the functionality of chemo-surveillance to stop the growth of cells with abnormal MEs. Inability of destruction strategy to put away CSCs is a deciding factor to deny the success of destruction strategy to win the war on cancer.

INTRODUCTION AND OBJECTIVE Prune Belly Syndrome (PBS) is characterized by bladder dysmyogenesis, yielding a dysfunctional compliant thick wall with excess collagen deposition. To dissect the cellular heterogeneity and gene expression networks altered in PBS, we report the cell type composition and transcriptional activity of PBS human bladder by using single cell RNA sequencing (scRNA-seq). METHODS Using IRB-approved methods, bladder dome from 2 PBS and 6 non-PBS control (CO) males underwent fresh single-cell digestion. scRNA-seq was performed and 5277 and 31828 bladder cells from PBS and CO patients was detected, respectively. Cell type clusters were graphically displayed by Uniform Manifold Approximation and Projection (UMap) plot and differentially expressed genes (DEGs) were generated to assign each cluster identity. KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis was performed for PBS affected genes. RESULTS We identified 17 distinct bladder cell clusters, including 6 fibroblast (1, 2, 3, and 4, immunofibroblast, myofibroblast), 1 smooth muscle (SM), and 2 urothelial (umbrella and basal+intermediate) clusters (Fig 1A-B). Counts of individual cell types were expressed as relative proportions, identifying significant PBS fibroblast enrichment, (67% PBS vs 40% CO). Five of 6 PBS fibroblast sub-types are proportionately fewer in number than in CO. The exception is a dominant fibroblast sub-type we label as fibroblast 4, (61% of all PBS fibroblasts vs

Antibiotic use data are critical for drawing conclusions about the epidemiological connections between antibiotic use in farms animals, antibiotic resistance, animal health, and human health. The goal of this study was to quantitatively and qualitatively characterize antibiotic use on dairy farms in Pennsylvania, the state with second largest number of dairy farms nationally. A survey was sent to 10% of the 6,580 dairy farms registered in Pennsylvania and completed by 235 producers (response rate of 36%). Data on antibiotic use in the previous month and in the previous 6 mo were collected based on farmer self-report, using either recall or treatment records. Two metrics were used to quantify antibiotic consumption: animal-defined daily doses (ADD) and days of therapy (DOT), a metric used in human medicine for purposes of antimicrobial stewardship. Across all farms, 24,444 ADD and 19,029 DOT were reported, representing treatment incidences of 4.2 ADD/1,000 animal-days and 3.3 DOT/1,000 animal-days. These rates were generally lower than those found in other states and countries. The main indication for antibiotic use was mastitis, and first-generation cephalosporins were the most commonly used class of antibiotic for all indications, followed by penicillins and third-generation cephalosporins. Trends in use were similar for ADD and DOT, but the numbers of recorded DOT and associated treatment incidences were generally lower than the number of ADD and associated treatment incidences. Rates of treatment were significantly associated with herd size. This study is the first to quantify antibiotic use on dairy farms in Pennsylvania and the first to use the DOT metric in a dairy setting.

Prune belly syndrome (PBS) is characterized by the triad of abdominal flaccidity, a variable degree of urinary tract involvement and cryptorchidism. Most cases of PBS are sporadic and have a normal karyotype, with 95% patients being male. In the last decade, mutations in known genes that regulate embryonic genitourinary myogenesis have been identified and with increasing knowledge of these critical genes involved in bladder maldevelopment, advances can be made in genetic counseling. A multidisciplinary approach is necessary and individualization of care is recommended according to phenotypic severity. Some patients require abdominal and urinary tract reconstruction while others require as little as bilateral orchiopexies. Major treatment objectives are: preservation of renal function and upper urinary tract; polyuria management; adequate bladder emptying; improvement of corporal image and quality of life; preservation of fertility and adequate sexual function. Long-term surveillance of the urinary tract is essential up to adulthood, because functional dynamics can change over time.

Background Antimicrobial resistance is a serious concern. Although the widespread use of antimicrobials in livestock has exacerbated the emergence and dissemination of antimicrobial resistance genes (ARG) in farm environments, little is known about whether antimicrobial use affects distribution of ARG in livestock systems. This study compared the distribution of microbiomes and resistomes (collections of ARG) across different farm sectors in dairy herds that differed in their use of antimicrobials. Feces from heifers, non-lactating, and lactating cows, manure storage, and soil from three conventional (antimicrobials used to treat cows) and three organic (no antimicrobials used for at least four years) farms in Pennsylvania were sampled. Samples were extracted for genomic DNA, processed, sequenced on the Illumina NextSeq platform, and analyzed for microbial community and resistome profiles using established procedures. Results Microbial communities and resistome profiles clustered by sample type across all farms. Overall, abundance and diversity of ARG in feces was significantly higher in conventional herds compared to organic herds. The ARG conferring resistance to betalactams, macrolide-lincosamide-streptogramin (MLS), and tetracyclines were significantly higher in fecal samples of dairy cows from conventional herds compared to organic herds. Regardless of farm type, all manure storage samples had greater diversity (albeit low abundance) of ARG conferring resistance to aminoglycosides, tetracyclines, MLS, multidrug resistance, and phenicol. All soil samples had lower abundance of ARG compared to feces, manure, and lagoon samples and were comprised of ARG conferring resistance to aminoglycosides, glycopeptides, and multi-drug resistance. The distribution of ARG is likely driven by the composition of microbiota in the respective sample types. Conclusions Antimicrobial use on farms significantly influenced specific groups of ARG in feces but not in manure storage or soil samples.

A term infant with prenatally noted ambiguous genitalia and nonpalpable gonads presented with life-threatening hyponatremia, hypertension, acidosis, and anuric renal failure requiring peritoneal dialysis at age 3 months.Sequencing confirmed 46, XY Denys-Drash syndrome (DDS) due to heterozygous Wilms tumor-1 exon 8 mutation encoding p.His445Arg. Renal US identified bilateral multifocal renal masses at age 8 months. Bilateral retroperitoneal nephrectomies found bilateral nephroblastomatosis without Wilms' tumor avoiding chemotherapy, followed by bilateral laparoscopic orchiopexies. We suggest monthly screening of 46, XY DSD cases for DDS by evaluating for proteinuria and electrolyte disarray starting at diagnosis of DSD to prevent acute life-threatening renal failure presentation.

Background Castration-insensitive epithelial progenitors capable of regenerating the prostate have been proposed to be concentrated in the proximal region based on facultative assays. Functional characterization of prostate epithelial populations isolated with individual cell surface markers has failed to provide a consensus on the anatomical and transcriptional identity of proximal prostate progenitors. Methods Here, we use single-cell RNA sequencing to obtain a complete transcriptomic profile of all epithelial cells in the mouse prostate and urethra to objectively identify cellular subtypes. Pan-transcriptomic comparison to human prostate cell types identified a mouse equivalent of human urethral luminal cells, which highly expressed putative prostate progenitor markers. Validation of the urethral luminal cell cluster was performed using immunostaining and flow cytometry. Results Our data reveal that previously identified facultative progenitors marked by Trop2, Sca-1, KRT4, and PSCA are actually luminal epithelial cells of the urethra that extend into the proximal region of the prostate, and are resistant to castration-induced androgen deprivation. Mouse urethral luminal cells were identified to be the equivalent of previously identified human club and hillock cells that similarly extend into proximal prostate ducts. Benign prostatic hyperplasia (BPH) has long been considered an "embryonic reawakening," but the cellular origin of the hyperplastic growth concentrated in the periurethral region is unclear. We demonstrate an increase in urethral luminal cells within glandular nodules from BPH patients. Urethral luminal cells are further increased in patients treated with a 5-α reductase inhibitor. Conclusions Our data demonstrate that cells of the proximal prostate that express putative progenitor markers, and are enriched by castration in the proximal prostate, are urethral luminal cells and that these cells may play an important role in the etiology of human BPH.

Castration-insensitive epithelial progenitors capable of regenerating the prostate are concentrated in the proximal region close to the urethra, but the identification of these cells has been limited to individual cell surface markers. Here, we use single cell RNA sequencing (scRNA-seq) to obtain a cellular anatomy of the mouse prostate and urethra and create a comparative map with the human. These data reveal that previously identified facultative progenitors marked by TROP2, Sca-1, KRT4, and PSCA are actually luminal epithelia of the urethra that extend into the proximal prostate. These mouse urethral cells are the human equivalent of previously identified club and hillock urethral cells. Castration decreases androgen-dependent prostate luminal epithelia as expected, but TROP2+ urethral luminal epithelia survive and expand into the prostate. Benign prostatic hyperplasia (BPH) has long been considered an ‘embryonic reawakening’, but the cellular origin of peri-urethral growth is unclear. We use scRNA-seq and flow cytometry to demonstrate an increase in urethral luminal epithelia within glandular nodules from patients with BPH, which are further enriched in patients treated with a 5 alpha reductase inhibitor. These data demonstrate that the putative prostate progenitors enriched by castration in the proximal prostate are an expansion of urethral luminal epithelia and that these cells may play an important role in the etiology of human BPH.Significance StatementThe prostate involutes after castration, but regrows to its original size with androgen replenishment. This observation prompted the search for a castration-insensitive prostate progenitor. Here, Joseph et al. produce a comparative cellular atlas of the prostate and urethra in the mouse vs. human, discovering an equivalent urethral luminal epithelial cell type that extends into the proximal prostatic ducts and expresses previously identified markers of facultative prostate progenitors. Urethral luminal epithelia are established before prostate budding in human and mouse development, and expand after castration in the mouse and after 5 alpha reductase inhibitor treatment in human BPH. These data suggest that luminal epithelia of the urethra are castration-insensitive cells of proximal ducts that may act as progenitors in human BPH.

Congenital anomalies of the external genitalia (CAEG) are a prevalent and serious public health concern with lifelong impacts on the urinary function, sexual health, fertility, tumor development, and psychosocial wellbeing of affected individuals. Complications of treatment are frequent, and data reflecting long-term outcomes in adulthood are limited. To identify a path forward to improve treatments and realize the possibility of preventing CAEG, the National Institute of Diabetes and Digestive and Kidney Diseases and the American Urological Association convened researchers from a range of disciplines to coordinate research efforts to fully understand the different etiologies of these common conditions, subsequent variation in clinical phenotypes, and best practices for long term surgical success. Meeting participants concluded that a central data hub for clinical evaluations, including collection of DNA samples from patients and their parents, and short interviews to determine familial penetrance (small pedigrees), would accelerate research in this field. Such a centralized datahub will advance efforts to develop detailed multi-dimensional phenotyping and will enable access to genome sequence analyses and associated metadata to define the genetic bases for these conditions. Inclusion of tissue samples and integration of clinical studies with basic research using human cells and animal models will advance efforts to identify the developmental mechanisms that are disrupted during development and will add cellular and molecular granularity to phenotyping CAEG. While the discussion focuses heavily on hypospadias, this can be seen as a potential template for other conditions in the realm of CAEG, including cryptorchidism or the exstrophy-epispadias complex. Taken together with long-term clinical follow-up, these data could inform surgical choices and improve likelihood for long-term success.

Prune Belly Syndrome (PBS) is a congenital multisystem myopathy with mild to lethal severity. While of uncertain etiology, 95% male predominance and familial occurrence suggest a genetic basis. As copy number variations (CNVs) can cause unexplained genetic disorders, we tested for novel CNVs in a large PBS population. We genotyped 21 unrelated PBS patients by high-resolution array comparative genomic hybridization (aCGH) and phenotyped using a novel PBS severity scoring system. Available parents were screened for detected CNV via quantitative PCR (qPCR). We additionally screened for recurrence of identified novel candidate CNVs on 106 PBS probands by qPCR. We identified 10 CNVs in 8 of 21 PBS patients tested (38%). Testing confirmed inheritance from an unaffected biological parent in six patients; parental samples were unavailable in two probands. One candidate CNV includes duplication of the X-chromosome AGTR2 gene, known to function in urinary tract development. Subsequent screening of the larger PBS cohort did not identify any recurrent CNVs. Presence of CNV did not correlate with PBS severity scoring. CNVs were uncommon in this large PBS population, but analysis of identified variants may inform disease pathogenesis and reveal targets for therapeutic intervention for this rare, severe disorder.

Ruminants are dependent on the microbiota (bacteria, protozoa, archaea, and fungi) that inhabit the reticulo-rumen for digestion of feedstuffs. Nearly 70% of energy and 50% of protein requirements for dairy cows are met by microbial fermentation in the rumen, emphasizing the need to characterize the role of microbes in feed breakdown and nutrient utilization. Over the past 2 decades, next-generation sequencing technologies have allowed for rapid expansion of knowledge concerning microbial populations and alterations in response to forages, concentrates, supplements, and probiotics in the rumen. Advances in gene sequencing and emerging bioinformatic tools have allowed for increased throughput of data to aid in our understanding of the functional relevance of microbial genomes. In particular, metagenomics can identify specific genes involved in metabolic pathways, and metatranscriptomics can describe the transcriptional activity of microbial genes. These powerful approaches help untangle the complex interactions between microbes and dietary nutrients so that we can more fully understand the physiology of feed digestion in the rumen. Application of genomics-based approaches offers promise in unraveling microbial niches and respective gene repertoires to potentiate fiber and nonfiber carbohydrate digestion, microbial protein synthesis, and healthy biohydrogenation. New information on microbial genomics and interactions with dietary components will more clearly define pathways in the rumen to positively influence milk yield and components.

To assess the effect of implementing an emergency surgery track for testicular torsion transfers. We hypothesized that transferring children from other facilities diagnosed with torsion straight to the operating room (STOR) would decrease ischemia time, lower costs, and reduce testicular loss.Demographics, arrival to incision time, hospital cost in dollars, and testicular outcome (determined by testicular ultrasound) at follow-up were retrospectively compared in all patients transferred to our tertiary care children's hospital with a diagnosis of testicular torsion from 2012 to 2016. Clinical data for STOR and non-STOR patients were compared by Wilcoxon rank-sum, 2-tailed t test, or Fisher exact test as appropriate.Sixty-eight patients met inclusion criteria: 35 STOR and 33 non-STOR. Children taken STOR had a shorter median arrival to incision time (STOR: 54 minutes vs non-STOR: 94 minutes, P .0001) and lower median total hospital costs (STOR: $3882 vs non-STOR: $4419, P .0001). However, only 46.8% of STOR patients and 48.4% of non-STOR patients achieved surgery within 6 hours of symptom onset. Testicular salvage rates in STOR and non-STOR patients were not significantly different (STOR: 68.4% vs non-STOR: 36.8%, P = .1), but follow-up was poor.STOR decreased arrival to incision time and hospital cost but did not affect testicular loss. The bulk of ischemia time in torsion transfers occurred before arrival at our tertiary care center. Further interventions addressing delays in diagnosis and transfer are needed to truly improve testicular salvage rates in these patients.

We investigated the photopolymerization kinetics of a low glass transition temperature (Tg) urethane acrylate network as a model soft coating system. We used real-time Fourier transform infrared (RT-FTir) spectroscopy to measure the acrylate conversion and fast sampling photorheology to measure the developing mechanical properties during cure. Experiments were conducted using a 395 nm LED ultraviolet (UV) source at different incident intensities ranging from 5 to 200 mW/cm2 at 25 °C. We found that the maximum rate of conversion and rate of complex shear modulus development scaled with incident UV intensity (I0) to an exponent (β) ≈ 0.53. This finding is consistent with the classical prediction of free radical photopolymerization that cure speed should increase to the ½ power of intensity when a bimolecular termination mechanism takes place during the reaction. A consequence of this is that the degree of conversion and complex shear modulus were not constant at a fixed energy dose (I0t) when intensity was varied illustrating that this system does not obey the exposure reciprocity rule. Instead we observed that the degree of cure and rheological properties were a universal function of I0βt. We also found that β increased only slightly throughout the reaction reaching a value of 0.65 at 95% conversion. This result indicates that bimolecular termination was predominant throughout nearly the entire reaction due to a high degree of molecular mobility in this flexible network.

Evaluating violence against women (VAW) training is essential to moving the field forward with proven approaches that can improve service provision for survivors of violence. Given existing resource constraints involved in VAW work, online training represents an economical and flexible option; however, existing evaluations of online programs in the VAW field are scant and face a variety of limitations. This study aimed to fill this gap by using a pre-/posttest design, comparison group, and mixed-method analysis to assess the effectiveness and value of an online training program. The program was intended to provide foundational knowledge in feminist antiviolence principles and values to a range of individuals working with survivors of intimate partner violence (IPV). Program participants ( N = 108) included volunteers, students, and professionals from various sectors, allowing for the application of the results to a broader field of VAW support services. This is important as individuals who work with IPV survivors may do so in a range of settings outside of the shelter context. Results demonstrate the potential for online VAW training to improve participants’ knowledge of and attitudes about VAW, which can positively inform their work with survivors. Qualitative responses provide further insight into course impact and highlight positive and negative aspects of the course. Although preliminary, these results provide justification for continued development and evaluation of online VAW training programs.

Inflammatory bowel disease (IBD) affects 1.5–3.0 million people in the United States. IBD is genetically determined and many common risk alleles have been identified. Yet, a large proportion of genetic predisposition remains unexplained. In this study, we report the identification of an ultra rare missense variant (NM_006998.3:c.230G > A;p.Arg77His) in the SCGN gene causing Mendelian early-onset ulcerative colitis. SCGN encodes a calcium sensor that is exclusively expressed in neuroendocrine lineages, including enteroendocrine cells and gut neurons. SCGN interacts with the SNARE complex, which is required for vesicle fusion with the plasma membrane. We show that the SCGN mutation identified impacted the localization of the SNARE complex partner, SNAP25, leading to impaired hormone release. Finally, we show that mouse models of Scgn deficiency recapitulate impaired hormone release and susceptibility to DSS-induced colitis. Altogether, these studies demonstrate that functional deficiency in SCGN can result in intestinal inflammation and implicates the neuroendocrine cellular compartment in IBD.

Context: Insulin-like peptide 3 (INSL3), a protein hormone produced by Leydig cells, may play a crucial role in testicular descent as male INSL3 knockout mice have bilateral cryptorchidism. Previous studies have measured human fetal INSL3 levels in amniotic fluid only. Objective: To measure INSL3 serum levels and mRNA in fetal umbilical cord blood and fetal testes, respectively. Design: INSL3 concentrations were assayed on 50 μl of serum from male human fetal umbilical cord blood by a non-commercial highly sensitive and specific radioimmunoassay. For secondary confirmation, quantitative real-time PCR was used to measure INSL3 relative mRNA expression in 7 age-matched human fetal testes. Setting: UT Southwestern Medical Center, Dallas, TX and Medical University of South Carolina, Charleston, SC. Patients or other Participants: Twelve human male umbilical cord blood samples and 7 human male testes were obtained from fetuses 14–21 weeks gestation. Male sex was verified by leukocyte genomic DNA SRY PCR. Interventions: None. Main Outcome Measures: Human male fetal INSL3 cord blood serum concentrations and testicular relative mRNA expression. Results: INSL3 serum concentrations during human male gestational weeks 15–20 were 2–4 times higher than published prepubertal male levels and were 5–100 times higher than previous reports of INSL3 concentrations obtained from amniotic fluid. Testicular fetal INSL3 mRNA relative expression was low from weeks 14–16, rose significantly weeks 17 and 18, and returned to low levels at week 21. Conclusions: These findings further support the role of INSL3 in human testicular descent and could prove relevant in uncovering the pathophysiology of cryptorchidism.

Background Prune belly syndrome (PBS) is a rare, multi-system congenital myopathy primarily affecting males that is poorly described genetically. Phenotypically, its morbidity spans from mild to lethal, however, all isolated PBS cases manifest three cardinal pathological features: 1) wrinkled flaccid ventral abdominal wall with skeletal muscle deficiency, 2) urinary tract dilation with poorly contractile smooth muscle, and 3) intra-abdominal undescended testes. Despite evidence for a genetic basis, previously reported PBS autosomal candidate genes only account for one consanguineous family and single cases. Methods We performed whole exome sequencing (WES) of two maternal adult half-brothers with syndromic PBS (PBS + Otopalatodigital spectrum disorder [OPDSD]) and two unrelated sporadic individuals with isolated PBS and further functionally validated the identified mutations. Results We identified three unreported hemizygous missense point mutations in the X-chromosome gene Filamin A (FLNA) (c.4952 C > T (p.A1448V), c.6727C > T (p.C2160R), c.5966 G > A (p.G2236E)) in two related cases and two unrelated sporadic individuals. Two of the three PBS mutations map to the highly regulatory, stretch-sensing Ig19–21 region of FLNA and enhance binding to intracellular tails of the transmembrane receptor β-integrin 1 (ITGβ1). Conclusions FLNA is a regulatory actin-crosslinking protein that functions in smooth muscle cells as a mechanosensing molecular scaffold, transmitting force signals from the actin-myosin motor units and cytoskeleton via binding partners to the extracellular matrix. This is the first evidence for an X-linked cause of PBS in multiple unrelated individuals and expands the phenotypic spectrum associated with FLNA in males surviving even into adulthood.

Clostridioides difficile causes severe colitis in people and is a significant enteric pathogen in many species of animals, including swine, horses, and potentially cattle. C. difficile is shed in feces, and transmission occurs horizontally via the fecal-oral route. Livestock has been suggested as a potential reservoir for C. difficile, and while studies have shown that swine and farm workers can be colonized with identical clones of C. difficile, the zoonotic transmission of C. difficile from livestock to people has not been definitively demonstrated. The goal of this study was to determine whether dairy calves and dairy farm workers harbored genetically similar isolates of C. difficile. First, we validated a glove juice protocol for detecting C. difficile on farm workers’ hands. We then visited 23 farms and collected 1) fecal samples from 92 dairy calves, 2) hand rinsates from 38 dairy farm workers, and 3) fecal samples from five of the dairy farm workers who were willing to submit them. All samples underwent anaerobic culture and qPCR to detect C. difficile. C. difficile was detected on 15 of the farms (65.2%, 95% confidence interval (CI) 42.7%–83.6%) and in 28 calves (30.4%, 95% CI 21.2–40.9%) but in none of the hand rinsates or human fecal samples. Thus, the zoonotic transmission of C. difficile on dairy farms could not be demonstrated, and dairy farmers did not appear to be at increased risk of acquiring C. difficile via the fecal-oral route.

Rationale & Objective Although maintaining high fluid intake is an effective low-risk intervention for the secondary prevention of urinary stone disease, many patients with stones do not increase their fluid intake. Study Design We describe the rationale and design of the Prevention of Urinary Stones With Hydration (PUSH) Study, a randomized trial of a multicomponent behavioral intervention program to increase and maintain high fluid intake. Participants are randomly assigned (1:1 ratio) to the intervention or control arm. The target sample size is 1,642 participants. Setting & Participants Adults and adolescents 12 years and older with a symptomatic stone history and low urine volume are eligible. Exclusion criteria include infectious or monogenic causes of urinary stone disease and comorbid conditions precluding increased fluid intake. Interventions All participants receive usual care and a smart water bottle with smartphone application. Participants in the intervention arm receive a fluid intake prescription and an adaptive program of behavioral interventions, including financial incentives, structured problem solving, and other automated adherence interventions. Control arm participants receive guideline-based fluid instructions. Outcomes The primary end point is recurrence of a symptomatic stone during 24 months of follow-up. Secondary end points include changes in radiographic stone burden, 24-hour urine output, and urinary symptoms. Limitations Periodic 24-hour urine volumes may not fully reflect daily behavior. Conclusions With its highly novel features, the PUSH Study will address an important health care problem. Funding National Institute of Diabetes and Digestive and Kidney Diseases. Trial Registration Registered at ClinicalTrials.gov with study number NCT03244189 .

The TWIST (Testicular Workup for Ischemia and Suspected Torsion) score uses urological history and physical examination to assess risk of testis torsion. Parameters include testis swelling (2 points), hard testis (2), absent cremasteric reflex (1), nausea/vomiting (1) and high riding testis (1). While TWIST has been validated when scored by urologists, its diagnostic accuracy among nonurological providers is unknown. We assessed the usefulness of the TWIST score when determined by nonurological nonphysician providers, mirroring emergency room evaluation of acute scrotal pain.Children with unilateral acute scrotum were prospectively enrolled in a National Institutes of Health clinical trial. After undergoing basic history and physical examination training, emergency medical technicians calculated TWIST score and determined Tanner stage per pictorial diagram. Clinical torsion was confirmed by surgical exploration. All data were captured into REDCap™ and ROC curves were used to evaluate the diagnostic usefulness of TWIST.Of 128 patients (mean age 11.3 years) 44 (13.0 years) had torsion. TWIST score cutoff values of 0 and 6 derived from ROC analysis identified 31 high, 57 intermediate and 40 low risk cases (positive predictive value 93.5%, negative predictive value 100%).TWIST score assessed by nonurologists, such as emergency medical technicians, is accurate. Low risk patients do not require ultrasound to rule out torsion. High risk patients can proceed directly to surgery, with more than 50% avoiding ultrasound. In the future emergency medical technicians and/or emergency room triage personnel may be able to calculate TWIST score to guide radiological evaluation and immediate surgical intervention at initial assessment long before urological consultation.

The microbial ecology of the rumen microbiome is influenced by the diet and the physiological status of the dairy cow and can have tremendous influence on the yield and components of milk. There are significant differences in milk yields between first and subsequent lactations of dairy cows, but information on how the rumen microbiome changes as the dairy cow gets older has received little attention. We characterized the rumen microbiome of the dairy cow for phylogeny and functional pathways by lactation group and stage of lactation using a metagenomics approach. Our findings revealed that the rumen microbiome was dominated by Bacteroidetes (70%), Firmicutes (15-20%) and Proteobacteria (7%). The abundance of Firmicutes and Proteobacteria were independently influenced by diet and lactation. Bacteroidetes contributed to a majority of the metabolic functions in first lactation dairy cows while the contribution from Firmicutes and Proteobacteria increased incrementally in second and third lactation dairy cows. We found that nearly 70% of the CAZymes were oligosaccharide breaking enzymes which reflect the higher starch and fermentable sugars in the diet. The results of this study suggest that the rumen microbiome continues to evolve as the dairy cow advances in lactations and these changes may have a significant role in milk production.

Purpose: Bladder outlet procedures without augmentation cystoplasty remain controversial. We hypothesized that bladder outlet procedures without augmentation cystoplasty may lead to unfavorable bladder dynamics, upper tract changes and/or continued incontinence. We reviewed long-term urodynamic, upper tract and continence outcomes following bladder outlet procedures without augmentation cystoplasty.Materials and Methods: We retrospectively reviewed all patients who underwent bladder neck reconstruction/closure/sling without augmentation cystoplasty between 2000 and 2014. Because of variation in length of followup, we calculated the cumulative incidence and proportion of cases of upper tract and urodynamic changes, augmentation cystoplasty and subsequent continence procedures. Preoperative factors were compared between patients with and without adverse outcomes.Results: A total of 109 patients underwent bladder outlet procedures without augmentation cystoplasty at a mean age of 8.5 years. At a mean of 4.9 ...

The overwhelming number and complexity of domestic violence cases in criminal and family courts has resulted in the development of education programmes to assist judges. There is limited research on judicial education in this area. This paper reviews one such initiative entitled ‘Enhancing Judicial Skills in Domestic Violence Cases’ (EJS) that has been developed and implemented over the last 20 years by the National Judicial Institute on Domestic Violence, a partnership of the US Department of Justice Office on Violence Against Women, National Council of Juvenile and Family Court Judges and Futures Without Violence. We present findings of a preliminary evaluation of the programme based on the self-reports of 480 judges who had taken the four-day workshop between 2006 and 2010. Overall, judges reported the programme to be engaging and effective. At a six-month follow‐up, most of the judges identified specific benefits and behavior changes in the areas of access to justice, judicial leadership, victim safety, and abuser accountability as a result of participating in the programme. Critical issues in judicial education are highlighted based on the authors’ experiences in the development and implementation of this programme.

OBJECTIVE To design a novel system of scoring prune belly syndrome (PBS) phenotypic severity at any presenting age and apply it to a large pilot cohort. PATIENTS AND METHODS From 2000 to 2017, patients with PBS were recruited to our prospective PBS study and medical records were cross-sectionally analysed, generating individualised RUBACE scores. We designed the pragmatic RUBACE-scoring system based on six sub-scores (R: renal, U: ureter, B: bladder/outlet, A: abdominal wall, C: cryptorchidism, E: extra-genitourinary, generating the acronym RUBACE), yielding a potential summed score of 0-31. The 'E' score was used to segregate syndromic PBS and PBS-plus variants. The cohort was scored per classic Woodard criteria and RUBACE scores compared to Woodard category. RESULTS In all, 48 males and two females had a mean (range) RUBACE score of 13.8 (8-25) at a mean age of 7.3 years. Segregated by phenotypic categories, there were 39 isolated PBS (76%), six syndromic PBS (12%) and five PBS-plus (10%) cases. The mean RUBACE scores for Woodard categories 1, 2, and 3 were 20.5 (eight patients), 13.8 (25), and 10.6 (17), respectively (P

Bladder agenesis is an extremely rare congenital anomaly of the genitourinary tract. Two female patients with known diagnoses of bladder agenesis presented for prerenal transplant evaluation and neobladder creation. Similar unique pelvic arterial malformations were identified through preoperative imaging and intraoperative examination. With these similar findings, it could be proposed that such anatomical variants are products of the same insult or involve a causal relationship, with vascular aberrancies potentially provoking pelvic organ maldevelopment. We advocate detailed mapping of the pelvic arterial tree preoperatively in bladder agenesis cases to prevent arterial injury and to safely delineate anatomy for appropriate transplant placement.

Wildlife rehabilitation can contribute to species conservation by releasing healthy individuals back into the wild and educating the public about threatening processes. Rehabilitation has substantial financial costs, however, and thus it is important to understand the success rates of these potential conservation management actions. We quantified the success rates for 1700 sea turtles admitted to rehabilitation facilities in Florida (USA) between 1986 and 2004. Rehabilitation success was low: 61.5% of turtles died in rehabilitation and only 36.8% were released back into the wild. A further 1.6% of turtles were maintained in captivity permanently due to the severe nature of their injuries. Most mortality occurred early during the rehabilitation process (within a few weeks), and successful rehabilitation often took several months to more than 3 yr. Loggerhead turtles (Caretta caretta) were most likely to survive rehabilitation, followed by Kemp’s ridleys (Lepidochelys kempii) and green turtles (Chelonia mydas); for all three species, larger individuals had an increased chance of successful rehabilitation. At face value, the low rates of rehabilitated turtles successfully released back into the wild may contribute only modestly to conservation in terms of contributing to population viability. However, many rehabilitation facilities provide important educational experiences that increase public awareness of the threats facing animals and highlight potential conservation solutions. Media coverage highlighting the release of rehabilitated animals further extends the conservation value of these efforts. Wildlife rehabilitation provides important direct benefits that, combined with social benefits, together may justify the expense and difficulty of rehabilitating individual animals.

Background The purpose of this study was to compare the rumen bacterial composition in high and low yielding dairy cows within and between two dairy herds. Eighty five Holstein dairy cows in mid-lactation (79–179 days in milk) were selected from two farms: Farm 12 (M305 = 12,300 kg; n = 47; 24 primiparous cows, 23 multiparous cows) and Farm 9 (M305 = 9700 kg; n = 38; 19 primiparous cows, 19 multiparous cows). Each study cow was sampled once using the stomach tube method and processed for 16S rRNA gene amplicon sequencing using the Ion Torrent (PGM) platform. Results Differences in bacterial communities between farms were greater (Adonis: R2 = 0.16; p