Gene expression of hepatic gluconeogenic and fatty acid metabolism in early-lactation dairy cows as affected by dietary starch and monensin supplementation.

Our previously published companion papers demonstrated improved production performance and energetic metabolism in cows fed diets with increased propiogenic potential in early lactation. Study objectives were to further explore effects of dietary starch content and monensin on hepatic gene expression of key enzymes related to gluconeogenesis and fatty acid metabolism in early lactation. From d 1 to 21 postpartum, primiparous (n = 16) and multiparous (n = 33) Holstein cows were fed a high (HS; 26.2% starch, 34.3% neutral detergent fiber, 22.7% acid detergent fiber, 15.5% crude protein) or low (LS; 21.5% starch, 36.9% neutral detergent fiber, 25.2% acid detergent fiber, 15.4% crude protein) starch diet with a daily topdress containing either 0 (Con) or 450 mg/d monensin (Mon). Cows were randomly assigned to treatment. Liver biopsies were obtained from cows on d 7 postpartum for DNA and RNA quantification and mRNA expression analysis. In primiparous cows, Mon supplementation decreased CPT1A expression relative to controls, whereas in multiparous cows Mon increased its expression. Cows fed HS and Mon tended to have decreased HMGCS2 expression relative to cows fed HS and Con. In multiparous cows, Mon supplementation tended to increase PC and PCK1 expression relative to controls. Correlation analysis was performed for all gene expression variables. Overall, relationships were similar in directionality and magnitude between cows fed HS and LS and Con and Mon. However, for cows fed Con there was a positive relationship between HMGCS2 and PC and HMGCS2 and PCK1, whereas for cows fed Mon there was no relationship. There was a similar lack of relationship between HMGCS2 and PC for cows fed HS. Overall, results support changes in performance and energetic metabolism reported in our companion papers, indicating that cows fed diets of different starch content in early lactation with Mon supplementation throughout the transition period had alterations in hepatic gene expression consistent with increased hepatic propionate supply.

Effect of monensin on milk production efficiency and milk composition in lactating dairy cows fed modern diets.

Since the US Food and Drug Administration's approval of monensin in 2004, significant nutritional advances have been made to increase feed efficiency and milk fat production. Recent evidence suggests monensin's adverse effect on milk fat percentage may be absent when diets are formulated to address known diet-induced milk fat depression risk factors. Thus, study objectives were to evaluate effects of monensin level on dry matter intake (DMI), milk production and composition, and efficiency of high-producing cows fed diets formulated to optimize milk fat. Ninety-six lactating Holstein cows (36 primiparous, 60 multiparous; 106 ± 17 d in milk [DIM]) were balanced by parity, DIM, and milk production and were randomly assigned to 1 of 12 pens with 8 cows per pen. All cows received 11 g/t monensin for 5 wk after which pens received 1 of 4 dietary treatments (n = 3) formulated to provide 0 (CON), 11 (R11), 14.5 (R14.5), or 18 (R18) g/t monensin for 9 wk. The basal diet was 54% forage, 27% NDF, 29% starch, and 2.3% rumen unsaturated fatty acid load. Pen was the experimental unit and data were analyzed using the Fit Model Procedure of JMP. Effects of treatment, time, and treatment × time interaction were included as fixed effects and pen as a random effect. Least squares means were determined and linear and quadratic contrasts were tested. Dry matter intake tended to decrease linearly with increasing monensin dose. Milk yield, fat percentage, and protein percentage and yield were unaffected by treatment while fat yield was quadratically increased. Milk de novo and mixed fatty acid (FA) yields (g/d) increased quadratically with monensin whereas preformed FA linearly decreased during the experimental period. Energy-corrected milk (ECM) was quadratically increased by monensin. Milk urea nitrogen concentrations increased linearly with increasing monensin dose. Monensin linearly increased feed efficiency (ECM/DMI, 3.5% fat-corrected milk/DMI, and solids-corrected milk/DMI). Body weight gain did not differ between treatments. Estimated dietary energy tended to increase linearly with increasing monensin level. These data suggest monensin improves component-corrected milk production efficiency, estimated dietary energy, and does not negatively affect milk fat percentage or FA profile.

Multiomics analysis of adaptation to repeated DNA damage in prostate cancer cells.

DNA damage is frequently utilized as the basis for cancer therapies; however, resistance to DNA damage remains one of the biggest challenges for successful treatment outcomes. Critically, the molecular drivers behind resistance are poorly understood. To address this question, we created an isogenic model of prostate cancer exhibiting more aggressive characteristics to better understand the molecular signatures associated with resistance and metastasis. 22Rv1 cells were repeatedly exposed to DNA damage daily for 6 weeks, similar to patient treatment regimes. Using Illumina Methylation EPIC arrays and RNA-seq, we compared DNA methylation and transcriptional profiles between the parental 22Rv1 cell line and the lineage exposed to prolonged DNA damage. Here we show that repeated DNA damage drives the molecular evolution of cancer cells to a more aggressive phenotype and identify molecular candidates behind this process. Total DNA methylation was increased while RNA-seq demonstrated these cells had dysregulated expression of genes involved in metabolism and the unfolded protein response (UPR) with Asparagine synthetase (ASNS) identified as central to this process. Despite the limited overlap between RNA-seq and DNA methylation, oxoglutarate dehydrogenase-like (OGDHL) was identified as altered in both data sets. Utilising a second approach we profiled the proteome in 22Rv1 cells following a single dose of radiotherapy. This analysis also highlighted the UPR in response to DNA damage. Together, these analyses identified dysregulation of metabolism and the UPR and identified ASNS and OGDHL as candidates for resistance to DNA damage. This work provides critical insight into molecular changes which underpin treatment resistance and metastasis.

A novel gun law strength index and influential state gun laws for firearm homicide in the United States, 1999-2018.

OBJECTIVES: Although some existing indices quantified the mixture strength of state gun laws, methodological supports are still lacking. The study aimed to build a new state gun law strength (GLS) index, evaluate the contribution of all state gun laws to the new index, and assess the association between the new index and firearm homicide mortality. STUDY DESIGN: This is a population-based ecological study. METHODS: We condensed 134 state gun laws into a mixture GLS index at the state level to quantify each state's overall GLS by using the weighted quantile sum regression in each year from 1999 to 2018. The weights of state gun laws in the GLS index revealed their influence on GLS. The model also estimated the association between the GLS index and state-level firearm homicide mortality. RESULTS: From 1999 to 2018, 15 of 134 (11.19%) state gun laws significantly contributed to the GLS index for firearm homicide every year, five of which related to the minimum age of possessing firearms. The most influential gun law was "no possession of handguns until age 21." The GLS index was significantly and negatively associated with firearm homicide mortality; however, the association gradually diminished over time. CONCLUSIONS: The GLS index has methodological support and can take different gun violence outcomes into account. Future research can adopt the GLS index to conduct additional gun violence research or apply the modeling approach to build new GLS indexes for other gun violence outcomes.

Involvement of alcohol in injury cases in rural Sri Lanka: prevalence and associated factors among in-patients in three primary care hospitals.

BACKGROUND: Injuries account for a major proportion of global morbidity and mortality related to alcohol use. Information on the prevalence of alcohol-related injury in rural Sri Lanka is limited. The aims of this study were to determine the burden of alcohol-related injury in a hospital-based sample in rural Sri Lanka and explore factors associated with an increased risk of alcohol-related injury. METHODS: Involvement of alcohol in injury amongst in-patients was assessed in three hospitals in the North Central Province of Sri Lanka over 6 months. Adult (≥ 18 years) patients were eligible. Patients were assessed for: injury characteristics, current alcohol use (in the past year) using the Alcohol Use Disorder Identification Test (AUDIT), and acute intoxication. Patients with a blood alcohol concentration (BAC) reading equivalent of 10 mg/dL (2.17 mmol/L) were considered as having an alcohol-related injury. Binary logistic regression was used to explore association between alcohol-related injury and demographic and injury characteristics. RESULTS: A total of 883 injured patients were eligible and consented to the study. No alcohol use was reported by 487 (55.2%) of patients (35.6% of men, 95.2% of women). Prevalence of alcohol-related injuries was 14.8% overall and 32.8% among current alcohol users. Almost all patients with an alcohol-related injury were male (122/123; 99.2%); 24 (18.8%) of these patients scored positive for possible alcohol dependence. Patients with an alcohol-related injury had significantly higher AUDIT scores (median = 15 vs 6, p < 0.001), were significantly more likely to be aged 26-40 (OR 2.29, 95% CI:1.11, 4.72) or 41-55 years (OR 2.76, 95% CI: 1.29, 5.90) (compared to 18-25 years), to have a transport-related injury (OR 5.14, 95% CI: 2.30, 11.49) (compared to animal/plant sting/bite), and have intentional injuries (OR 3.47, 95% CI: 1.01, 11.87). CONCLUSIONS: One in three injuries among people who drank alcohol in this sample were alcohol-related. In addition, problematic alcohol use was higher among those with alcohol-related injury. Further work is needed to explore whether this prevalence of alcohol-related injury is reflected in other rural settings in Sri Lanka.

Complex chronic conditions among children born to women with schizophrenia.

PURPOSE: Maternal schizophrenia is linked to complications in offspring near the time of birth. Whether there is also a higher future risk of the child having a complex chronic condition (CCC) - a pediatric condition affecting any bodily system expected to last at least 12 months that is severe enough to require specialty care and/or a period of hospitalization - is not known. METHODS: In this population-based health administrative data cohort study (Ontario, Canada, 1995-2018), the risk for CCC was compared in 5066 children of women with schizophrenia (the exposed) vs. 2,939,320 unexposed children. Adjusted hazard ratios (aHR) were generated for occurrence of any CCC, by CCC category, and stratified by child sex, and child prematurity. RESULTS: CCC was more frequent in the exposed (7.7 per 1000 person-years [268 children]) than unexposed (4.2 per 100 person-years [124,452 children]) - an aHR of 1.25 (95% CI 1.10-1.41). aHRs were notably higher in 5 of 9 CCC categories: neuromuscular (1.73, 1.28-2.33), cardiovascular (1.94, 1.64-2.29), respiratory (1.83, 1.32-2.54), hematology/immunodeficiency (2.24, 1.24-4.05) and other congenital or genetic defect (1.59, 1.16-2.17). The aHR for CCC was more pronounced among boys (1.32, 1.13-1.55) than girls (1.16, 0.96-1.40), and of similar magnitude in term (1.22, 1.05-1.42) and preterm infants (1.18, 0.95-1.46). CONCLUSIONS: The risk for a CCC appears to be higher in children born to women with schizophrenia. This finding introduces opportunities for targeted preconception counselling, optimization of maternal risk factors, and intervention to support a vulnerable parent population who will experience unique challenges caring for a child with CCCs.

Low dose antibiotic ingestion potentiates systemic and microbiome changes induced by silver nanoparticles.

Changes in the mammalian gut microbiome are linked to the impairment of immunological function and numerous other pathologies. Antimicrobial silver nanoparticles (AgNPs) are incorporated into numerous consumer products (e.g., clothing, cosmetics, food packaging), which may directly impact the gut microbiome through ingestion. The human health impact of chronic AgNP ingestion is still uncertain, but evidence from exposure to other antimicrobials provides a strong rationale to assess AgNP effects on organ function, immunity, metabolism, and gut-associated microbiota. To investigate this, mice were gavaged daily for 5 weeks with saline, AgNPs, antibiotics (ciprofloxacin and metronidazole), or AgNPs combined with antibiotics. Animals were weighed daily, assessed for glucose tolerance, organ function, tissue and blood cytokine and leukocyte levels. At the end of the study, we used 16S rDNA amplicon and whole-metagenome shotgun sequencing to assess changes in the gut microbiome. In mice exposed to both AgNPs and antibiotics, silver was found in the stomach, and small and large intestines, but negligible amounts were present in other organs examined. Mice exposed to AgNPs alone showed minimal tissue silver levels. Antibiotics, but not AgNPs, altered glucose metabolism. Mice given AgNPs and antibiotics together demonstrated slower weight gain, reduced peripheral lymphocytes, and elevated splenic, but not circulatory markers of inflammation. 16S rDNA profiling of cecum and feces and metagenomic sequencing of fecal DNA demonstrated that combined AgNP-antibiotic treatment also significantly altered the structure and function of the gut microbiota, including depletion of the indicator species Akkermansia muciniphila. This study provides evidence for possible biological effects from repeated ingestion of AgNP-containing consumer products when antibiotics are also being used and raises concern that an impaired gut microbiome (e.g., through antibiotic use) can potentiate the harm from chemical exposures such as AgNPs.

Evidence into action: a policy brief exemplar supporting attainment of nursing now.

AIM: To provide a framework for the production of policy briefs, and offer a practical example of how evidence can be turned into a succinct document to inform policy and bring about change targeted at delivering universal health coverage. INTRODUCTION: Policymakers are too busy, or do not have the necessary expertise, to read and comprehend complex scientific papers. As a result, policy briefs that capture and present the essential points are needed if evidence-informed policy is to be developed and implemented. METHOD: A two-page example of how evidence from meta-analytical and systematic reviews can be presented to identify options and recommendations to address a major global disease burden. RESULTS: The example uses a simple, seven-section template for developing a policy brief. The essential characteristics of each section are provided. The briefing, targeted at the global level, provides information on the major challenges associated with the treatment of individuals with diabetes. DISCUSSION AND CONCLUSIONS: This paper demonstrates how to use existing research evidence to address the pursuit of UHC relevant to a wide range of geographies, settings or disadvantaged groups. IMPLICATIONS FOR POLICY: Gaps in universal health coverage and major disease burdens such as diabetes can be pursued through entities such as country-based Nursing Now groups. In addition, ongoing opportunities exist through the International Council of Nurses annual International Nurses Day and WHO's regular regional meetings to inform and influence policy discussions at national and subnational levels. By focusing on a small number of global topics each year, measurable changes in addressing the burden of disease can be achieved while simultaneously keeping the nursing profession's contribution centre stage.

Evidence-based policy: nursing now and the importance of research synthesis.

AIM: This study explores how scholarship relating to meta-analytical studies and systematic and integrative reviews can inform nursing's contribution to universal health coverage. INTRODUCTION: As nursing globally embraces the 200th anniversary of the birth of Florence Nightingale, the Nursing Now social movement has called for the profession to improve universal health coverage through increasing nursing's policy voice. METHODS: In determining how the Nursing Now social movement could pursue the aim of this study, researchers undertook a comparative bibliometric analysis of scholarship relating to the systematic curation of evidence. This study uses a mixed-method analysis of the bibliometric data available through extracting and synthesizing information from one of the commercially produced indexing and citation databases. RESULTS: Generally, medicine has contributed far more synthesized contributions than nursing, except in the case of integrative reviews. Co-occurrence analysis of nursing literature through examination of key terms yielded a complex visualization of 11 specific clusters of scholarship (Care of the Older Person, Nurse Education, Emergency and Critical Care, Occupational Health and Safety, Rural Services, Anxiety and Depression, Measurement, Newborn and Post-natal Health, Cardiovascular Disease, Preventative Health and Cancer Care). DISCUSSION AND CONCLUSIONS: Bibliometric analysis of curated evidence demonstrates that there is ample nursing-relevant material to inform evidence-based policy change directed towards the attainment of universal health coverage and several of the Sustainable Development Goals. IMPLICATIONS FOR POLICY: Nursing literature is available to support policy change directed towards the pursuit of universal health coverage and sustainable development goals. Leveraging existing networks of research collaboration to increase research capacity through communities of scholarship or by twinning experienced and neophyte contributors is possible. Further work is needed to equip nurses with the competencies to navigate the policy environment and develop and deliver impactful policy messaging.

Association between ultrasonographic appearance of urine and urinalysis in dogs and cats.

OBJECTIVE: To evaluate association between ultrasonographic urine echogenicity and sediment examination in dogs and cats. MATERIALS AND METHODS: Dogs and cats undergoing ultrasound-guided cystocentesis at a multidisciplinary referral hospital. Ultrasonographic images were stored and reviewed by a single, blinded, board-certified radiologist. Urine appearance was described as "echoic" or "anechoic". Urine sediment was examined for bacteriuria, pyuria, haematuria, crystalluria and urine-specific gravity and then classified as "active" or "inactive." RESULTS: Of the 194 cases included in this study, urine was echoic in 52 and anechoic in 142. Sediment was active in 52 and inactive in 142 samples. Sensitivity and specificity of echoic urine for active sediment were 40% (95% CI: 27 to 55%) and 78% (95% CI: 70 to 85%), respectively. Positive predictive value and negative predictive value of echoic urine for active sediment were 40% (CI 30 to 52%) and 78% (CI 74 to 82%), respectively. If urine-specific gravity was <1.015 urine was always described ultrasonographically as anechoic. CLINICAL SIGNIFICANCE: Association between sediment analysis and ultrasonographic appearance of urine is poor. Echoic urine had low positive predictive value for active sediment in this study, suggesting that echoic urine alone should not prompt urinary investigations in the absence of other clinical suspicion. Despite a negative predictive value of 78%, urinalysis is still indicated for anechoic urine, especially if urine specific gravity is low.

The 'Outcome Reporting in Brief Intervention Trials: Alcohol' (ORBITAL) framework: protocol to determine a core outcome set for efficacy and effectiveness trials of alcohol screening and brief intervention.

BACKGROUND: The evidence base to assess the efficacy and effectiveness of alcohol brief interventions (ABI) is weakened by variation in the outcomes measured and by inconsistent reporting. The 'Outcome Reporting in Brief Intervention Trials: Alcohol' (ORBITAL) project aims to develop a core outcome set (COS) and reporting guidance for its use in future trials of ABI in a range of settings. METHODS/DESIGN: An international Special Interest Group was convened through INEBRIA (International Network on Brief Interventions for Alcohol and Other Drugs) to inform the development of a COS for trials of ABI. ORBITAL will incorporate a systematic review to map outcomes used in efficacy and effectiveness trials of ABI and their measurement properties, using the COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) criteria. This will support a multi-round Delphi study to prioritise outcomes. Delphi panellists will be drawn from a range of settings and stakeholder groups, and the Delphi study will also be used to determine if a single COS is relevant for all settings. A consensus meeting with key stakeholder representation will determine the final COS and associated guidance for its use in trials of ABI. DISCUSSION: ORBITAL will develop a COS for alcohol screening and brief intervention trials, with outcomes stratified into domains and guidance on outcome measurement instruments. The standardisation of ABI outcomes and their measurement will support the ongoing development of ABI studies and a systematic synthesis of emerging research findings. We will track the extent to which the COS delivers on this promise through an exploration of the use of the guidance in the decade following COS publication.

The Role of Epigenetic Regulation in Transcriptional Memory in the Immune System.

The immune system is exquisitely poised to identify, respond to, and eradicate pathogens from the body, as well as to produce a more rapid and augmented response to a subsequent encounter with the pathogen. These cellular responses rely on the highly coordinated and rapid activation of gene expression programs as well as the ability of the cell to retain a memory of the initial gene response. It is clear that chromatin structure and epigenetic mechanisms play a crucial role in determining these gene responses, and in fact the immune system has proved an instructive model for investigating the multifaceted mechanisms through which the chromatin landscape contributes to gene expression programs. These mechanisms include modifications to the DNA and histone proteins, the positioning, composition, and remodeling of nucleosomes, as well as the formation of higher-order chromatin structures. Moreover, it is now apparent that epigenetic mechanisms also provide an instrument by which cells can retain memory of the initial transcriptional response, "priming" the genome so that it can respond more quickly to subsequent exposure to the signal. Here, we use the immune system as a model to demonstrate the complex interplay between transcription factors and the chromatin landscape required to orchestrate precise gene responses to external stimuli and further to demonstrate how these interactions can establish memory of past transcriptional events. We focus on what we have learnt from the immune system and how this can inform our understanding of other cellular systems.

Effect of diet fermentability and unsaturated fatty acid concentration on recovery from diet-induced milk fat depression.

Diet-induced milk fat depression is caused by highly fermentable and high-unsaturated fatty acid (FA) diets, and results in reduced milk fat concentration and yield, reduced de novo FA, and increased trans isomers of the alternate biohydrogenation pathways. The hypothesis of the current experiment was that a diet higher in fermentability and lower in unsaturated FA (UFA) would accelerate recovery compared with a high-UFA and lower-fermentability diet. Eight ruminally cannulated and 9 noncannulated multiparous Holstein cows were randomly assigned to treatment sequences in a replicated Latin square design. During each period milk fat depression was induced for 10 d by feeding a low-fiber, high-UFA diet [25.9% neutral detergent fiber (NDF) and 3.3% C18:2]. Following the induction phase, cows were switched to recovery treatments for 18 d designed to correct dietary fermentability, UFA, or both fermentability and UFA concentration. Treatments during recovery were (1) correction of fiber and UFA diet [control; 31.8% NDF and 1.65% C18:2], (2) a diet predominantly correcting fiber, but not UFA [high oil (HO); 31.3% NDF and 2.99% C18:2], and (3) a diet predominantly correcting UFA, but not fiber concentration [low fiber (LF); 28.4% NDF and 1.71% C18:2]. Milk and milk component yield, milk FA profile, ruminal pH, and 11 rumen microbial taxa were measured every third day during recovery. Milk yield decreased progressively in HO and control, whereas it was maintained in the LF diet. Milk fat concentration increased progressively during recovery in all treatments, but was on average 9% lower in LF than control from d 12 to 18. Milk fat yield increased progressively in all treatments and was not different between control and LF at any time point, but was lower in HO than control on d 15. Milk trans-10 C18:1 and trans-10,cis-12 conjugated linoleic acid decreased progressively in all treatments, but was higher in HO than control from d 3 to 18 [136 ± 50 and 188 ± 57% (mean ± SD)], whereas LF caused a smaller increase in these FA compared with control (67 ± 25 and 90 ± 22%). Additionally, milk trans-11 C18:1 and cis-9,trans-11 conjugated linoleic acid was decreased in control and LF and increased in HO during recovery. Selected microbial species observed changed during recovery, but major treatment differences were only observed for Streptococcus bovis. The LF diet that was similar in UFA but 3.4% units lower in NDF compared with to the control had a similar decrease in alternate trans biohydrogenation intermediates in milk. The HO diet that was similar in NDF but 2.0% units higher in UFA compared with the control had higher alternate trans biohydrogenation intermediates in milk compared with control. However, recovery of milk fat yield was similar between treatments at most time points.

Heat treatment enhances the antimicrobial activity of (+)-Catechin when combined with copper sulphate.

UNLABELLED: The aim of the study was to compare the antimicrobial activities of freshly made, heat-treated (HT) and 14 day stored (+)-Catechin solutions with (+)-catechin flavanol isomers in the presence of copper sulphate. (+)-Catechin activity was investigated when combined with different ratios of Cu(2+) ; 100°C heat treatment; autoclaving; and 14 day storage against Staphylococcus aureus. Cu(2+) -(+)-Catechin complexation, isomer structure-activity relationships, and H2 O2 generation were also investigated. Freshly made, HT, and 14 day stored flavanols showed no activity. While combined Cu(2+) -autoclaved (+)-Catechin and -HT(+)-Catechin activities were similar, HT(+)-Catechin was more active than either freshly made (+)-catechin (generating more H2 O2 ) or (-)-Epicatechin (though it generated less H2 O2 ) or 14 day-(+)-Catechin (which had similar activity to Cu(2+) controls-although it generated more H2 O2 ). When combined with Cu(2+) , in terms of rates of activity, HT(+)-Catechin was lower than (-)-Epigallocatechin gallate and greater than freshly made (+)-Catechin. Freshly made and HT(+)-Catechin formed acidic complexes with Cu(2+) as indicated by pH and UV-vis measurements although pH changes did not account for antimicrobial activity. Freshly made and HT(+)-Catechin both formed Cu(2+) complexes. The HT(+)-Catechin complex generated more H2 O2 which could explain its higher antimicrobial activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Natural products attract considerable attention in the search for novel antimicrobials, prebiotics and antioxidants. Enhanced biological activity of natural products has been demonstrated with chemical and heat treatment. This article extends the few publications on heat treatments of plant products and combinations with adjuncts, to raise antimicrobial activity against pathogens such as Staphylococcus aureus. We demonstrated that heat treatment could increase the activity of (+)-Catechin, a weak antimicrobial flavanol found commonly in plants in the presence of copper sulphate. Heat treatment of readily available resources merits consideration in the development of more potent substances for use in clinical settings and agriculture.

Perinatal Administration of a Selective Serotonin Reuptake Inhibitor Induces Impairments in Reproductive Function and Follicular Dynamics in Female Rat Offspring.

INTRODUCTION: Up to 10% of pregnant women take antidepressants, of which selective serotonin reuptake inhibitors (SSRIs) are the most commonly prescribed. Using a rodent model, we investigated the reproductive impacts of perinatal SSRI treatment on reproductive cyclicity and function in female offspring. METHODS: Virgin Wistar rats were given oral vehicle (n = 10) or fluoxetine hydrochloride (FLX, 10 mg/kg/d; n = 11) from 2 weeks prior to mating until weaning. Pubertal onset and reproductive cyclicity in offspring were assessed. Blood and ovarian tissues were collected for measures of reproductive function. RESULTS: Perinatal FLX tends to induce irregular reproductive cycles in adult offspring, which most commonly manifest as a prolonged estrus phase (FLX 34% vs control [CON] 10%) relative to CON offspring. The FLX offspring tended to have longer cycles (P = .052), had more secondary follicles (P = .0067), more total follicles (P = .0310), and increased apoptotic ovarian cells (P < .001). Prenatally exposed FLX offspring demonstrated elevated ovarian messenger RNA (mRNA) levels of ERß (P = .008), Cry1 (P = .043), and tryptophan hydroxylase 2 (P = .024), independent of stage of cycle. Ovarian mRNA levels of brain and muscle Arnt-like protein 1 (P = .046) and Pet-1 (P = .021) were increased in FLX offspring a manner that was reproductive cycle stage dependent. CONCLUSIONS: This is the first study to investigate the postnatal effects of maternal perinatal exposure to FLX on adult offspring reproduction. We show that genes that regulate serotonin signaling and action in the ovary are altered in prenatally FLX-exposed offspring, which when coupled with increased expression of components of the core Circadian Locomotor Output Cycles Kaput (CLOCK) gene regulatory loop may suggest an interaction between serotonergic signaling and clock gene signaling pathways leading to the altered reproductive phenotype.

Increased adult hippocampal neurogenesis is not necessary for wheel running to abolish conditioned place preference for cocaine in mice.

Recent evidence suggests that wheel running can abolish conditioned place preference (CPP) for cocaine in mice. Running significantly increases the number of new neurons in the hippocampus, and new neurons have been hypothesised to enhance plasticity and behavioral flexibility. Therefore, we tested the hypothesis that increased neurogenesis was necessary for exercise to abolish cocaine CPP. Male nestin-thymidine kinase transgenic mice were conditioned with cocaine, and then housed with or without running wheels for 32 days. Half of the mice were fed chow containing valganciclovir to induce apoptosis in newly divided neurons, and the other half were fed standard chow. For the first 10 days, mice received daily injections of bromodeoxyuridine (BrdU) to label dividing cells. On the last 4 days, mice were tested for CPP, and then euthanized for measurement of adult hippocampal neurogenesis by counting the number of BrdU-positive neurons in the dentate gyrus. Levels of running were similar in mice fed valganciclovir-containing chow and normal chow. Valganciclovir significantly reduced the numbers of neurons (BrdU-positive/NeuN-positive) in the dentate gyrus of both sedentary mice and runner mice. Valganciclovir-fed runner mice showed similar levels of neurogenesis as sedentary, normal-fed controls. However, valganciclovir-fed runner mice showed the same abolishment of CPP as runner mice with intact neurogenesis. The results demonstrate that elevated adult hippocampal neurogenesis resulting from running is not necessary for running to abolish cocaine CPP in mice.

Intranasal epidermoid cyst causing upper airway obstruction in three brachycephalic dogs.

This case report describes three brachycephalic dogs with intranasal epidermoid cysts that were causing additional upper airway obstruction. Although epidermoid cysts have been described in several locations in dogs, to the authors' knowledge intranasal epidermoid cysts have not been previously reported. All dogs had mucopurulent to haemorrhagic nasal discharge. Magnetic resonance imaging of the head revealed the presence of unilateral or bilateral intranasal cystic lesions obstructing the nasal cavities partially or completely, with atrophy of the ipsilateral nasal turbinates. The cystic lesions were surgically excised in all dogs using a modified lateral alveolar mucosal approach to the affected nasal cavity. Aerobic, anaerobic and fungal culture of the cystic contents were negative and histology of the excised tissue was consistent with a benign intranasal epidermoid cyst in each dog. Upper airway obstruction was clinically improved in two dogs.

Effect of monensin on recovery from diet-induced milk fat depression.

The objective of the present experiment was to investigate the effect of monensin (MN) on the time course of recovery from diet-induced milk fat depression. Milk fat depression was induced in all cows (n = 16) during the first phase of each period by feeding a low-fiber, high-unsaturated fat diet [25.3% neutral detergent fiber (NDF), 6.9% fatty acids (FA), and 3.24% C18:2] with MN (450mg/cow per day) for 10 to 14d. A recovery phase of 18d followed, where cows were switched to a higher-fiber and lower unsaturated fat diet (31.2% NDF, 4.3% FA, and 1.7% C18:2). According to a crossover design, treatments during recovery were (1) control (no MN supplementation) or (2) continued MN supplementation. Milk yield, milk composition, and milk FA profile were measured every 3d during recovery. No effect was observed of MN on dry matter intake or yield of milk, milk protein, and lactose. Milk fat concentration and yield increased progressively during recovery in both treatments. Monensin decreased milk fat yield from d 6 to 15, but it was the same as the control on d 18. A treatment by time interaction on milk fat concentration was detected, which was decreased by MN only on d 3 and 6. The yield of milk de novo synthesized FA increased progressively in both treatments and was not affected by treatment. Similarly, yield of 16-C FA increased progressively, but was decreased by MN on d 6 and 9. Preformed FA yield was lower in the MN group from d 6 to 15, but was not different from the control on d 18. Importantly, milk FA concentration of trans-10 C18:1 and trans-10,cis-12 conjugated linoleic acid rapidly decreased in both groups; however, MN slightly increased trans-10 C18:1 concentration above baseline on d 15 and 18. In conclusion, MN supplementation had minimal effect on recovery of normal rumen biohydrogenation and de novo FA synthesis during recovery from milk fat depression by correction of dietary starch, NDF, and polyunsaturated FA concentration, but moderately decreased recovery of preformed FA in milk.

Characterization of the adverse effects of nicotine on placental development: in vivo and in vitro studies.

In utero exposure to nicotine is associated with increased risk of numerous adverse fetal and neonatal outcomes, which suggests that it acts directly to affect placental development and the establishment of the fetomaternal circulation (FC). This study used both in vivo [Wistar rats treated with 1 mg/kg nicotine from 2 wk prior to mating until gestational day (GD) 15] and in vitro (RCHO-1 cell line; treated with 10(-9) to 10(-3)M nicotine) models to examine the effects of nicotine on these pathways. At GD 15, control and treated placentas were examined for the impact of nicotine on 1) trophoblast invasion, proliferation, and degree of hypoxia, 2) labyrinth vascularization, 3) expression of key genes of placental development, and 4) expression of placental angiogenic factors. The RCHO-1 cell line was used to determine the direct effects of nicotine on trophoblast differentiation. Our in vivo experiments show that nicotine inhibits trophoblast interstitial invasion, increases placental hypoxia, downregulates labyrinth vascularization as well as key transcription factors Hand1 and GCM1, and decreases local and circulating EG-VEGF, a key placental angiogenic factor. The in vitro experiments confirmed the inhibitory effects of nicotine on the trophoblast migration, invasion, and differentiation processes and demonstrated that those effects are most likely due to a dysregulation in the expression of nicotine receptors and a decrease in MMP9 activity. Taken together, these data suggest that adverse effects of maternal smoking on pregnancy outcome are due in part to direct and endocrine effects of nicotine on the main processes of placental development and establishment of FC.

Imaging diagnosis and minimally-invasive management of necrotizing fasciitis in a dog.

Necrotising fasciitis is a rapidly progressive, aggressive bacterial infection of the subcutis associated with significant morbidity and mortality in both man and domestic animals. To the best of our knowledge, this is the first veterinary report of magnetic resonance imaging findings in necrotising fasciitis, and the first reported case in a dog to be successfully treated with minimally invasive surgical intervention.