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Rationale: Mycobacterial species contain high concentrations of mycolic acids in their cell wall. Mycobacteria can pose a threat to both human health and the environment. Mass spectrometry lipidomic characterization can identify bacterial species and suggest targets for microbiological interventions. Due to the complex structures of mycolic acids and the possibility of isobaric isomers, multiple levels of separation are required for complete characterization. In this study, cyclic ion mobility (cIM) mass spectrometry (MS) was used for the analysis, separation and fragmentation of mycolic acids isomers from the bacterial species Gordonia amarae and Mycobacterium bovis. Methods: Mycolic acid isomers were interrogated from cultured G. amarae biomass and commercially available M. bovis mycolic acid extracts. These were infused into a cIM‐enabled quadrupole time‐of‐flight MS. Ions of interest were non‐simultaneously selected with the quadrupole and passed around the cyclic ion mobility device multiple times. Fragment ion analysis was then performed for the resolved isomers of the quadrupole‐selected ions. Results: Repeated passes of the cIM device successfully resolved otherwise overlapping MA isomers, allowing isomer isolation and producing an ion‐specific post‐mobility fragmentation spectrum without isomeric interference. Conclusions: Mycolic acids (MA) isomers from G. amarae and M. bovis were resolved, resulting in a high mobility resolution and low interference fragmentation analysis. These revealed varying patterns of MA isomers in the two species: G. amarae's most abundant ion of each set of MA has 1–2 conformations, while the MA + 2 m/z the most abundant ion of each set has 3–6 conformations. These were resolved after 70 passes of the cyclic device. M. bovis' most abundant ion of each keto‐MA set has 2 conformations, while the keto‐MA + 2 m/z has 1–2 conformations. These were resolved after 75 passes. [ABSTRACT FROM AUTHOR]

Objectives: To describe the presentation, etiology, and outcome of dogs and cats diagnosed with gastrointestinal pneumatosis (GP). Design: Retrospective study. Setting: Three referral institutions. Animals: Twenty‐six dogs and 4 cats. Interventions: None. Measurements and Main Results: The most common sites of GP were the stomach (n = 19), followed by the colon (n = 8) and small intestine (n = 2). One case had pneumatosis of both the stomach and the colon. GP was most commonly associated with gastrointestinal disease in dogs (18/26 [69%]) and cats (3/4 [75%]), with common diagnoses including gastric dilatation and volvulus (n = 5), acute hemorrhagic diarrhea syndrome (n = 4), and gastrointestinal ulceration (n = 4). Of the 4 cases of gastrointestinal ulceration, 3 were dogs with a history of glucocorticosteroid or nonsteroidal anti‐inflammatory drug administration and vomiting and diarrhea. Six of 30 cases (20%), all of which were dogs, were determined to have a surgical indication for exploratory celiotomy, although not solely on the basis of diagnosis of GP. Five cases underwent exploratory celiotomy, of which 1 (20%) survived to hospital discharge. Of the medically managed cases, 13 of 24 (54%) survived to hospital discharge. Overall, 14 of 30 cases (47%) survived to hospital discharge. Conclusions: GP is an uncommon diagnostic imaging finding that is associated with a variety of disease processes. Its development is often related to primary gastrointestinal diseases. In the absence of other surgical disease, exploratory celiotomy based solely on the diagnosis of GP is unlikely to be indicated. [ABSTRACT FROM AUTHOR]

An 18‐month‐old male neutered Cocker Spaniel presented with a comatose mentation to its primary care veterinarian following the presumed ingestion of 216 mg/kg of phenobarbitone 2 hours prior. Following a failure to improve with intravenous fluids, lipid emulsion therapy, and supportive care, the patient was then referred to a tertiary referral centre for further treatment 30 hours following ingestion. Mechanical ventilation was initiated due to severe hypoventilation (PvCO2: 69.9 mmHg) before the patient underwent a 300‐minute prolonged veno‐venous haemodiafiltration cycle performed using a continuous haemodialysis‐based platform and a high‐flux dialyser. The patient demonstrated rapid neurological improvement being extubated after 4 hours, and becoming responsive and ambulatory 5 hours from the initiation of the treatment. The patient was discharged 60 hours following the completion of haemodialysis. This is the first report in the veterinary literature of the effective use of veno‐venous haemodiafiltration to treat phenobarbitone intoxication 30 hours after ingestion using a high‐flux dialyser. [ABSTRACT FROM AUTHOR]

Objectives: The Covid‐19 pandemic has taken a heavy toll on many people living with dementia and carers. Caring for a person living with dementia at home with limited avenues for support and a break challenged many carers. Care homes in England closed to visitors, with very few offering opportunities for a short‐stay. We investigated impact of Covid‐19 on views and expectations of carers of people living with dementia about residential respite. Methods/Design: Qualitative interviews with 35 carers were conducted March–December 2020: 30 women and 5 men, with ages ranging 30–83 years. Interviews explored experiences, views of residential respite, and expectations post‐Covid. Data were thematically analysed and salient concepts were drawn out and discussed within the research team and study advisers. Results: Three themes were identified in transcripts, relating to impact of Covid‐19 on views and expectations of respite: (1) Carers described regularly negotiating risks and stresses of Covid, weighing up how to prevent infection and changing family arrangements to facilitate caring; (2) Carers were balancing different needs, prioritising needs of their relatives while bearing the impact of cumulative caregiving responsibilities. (3) Uncertainty about future residential respite continued, in terms of availability, ongoing restrictions and trustworthy information sources. Conclusions: Residential respite is a positive, acceptable option for some carers to get a break from caring. Covid‐19 may have heighted some of caregiving stressors and there may be an increased need for a break. Views of care homes developed during the pandemic suggest that individual confidence to use respite may need to be rebuilt. Key points: The Covid‐19 pandemic increased demands on carers of people living with dementia with fewer opportunities for social contact and breaksCarers described myriad ways to negotiate risks and stresses of Covid by developing ways to prevent infection and changing family arrangements. Carers also talked of balancing different needs, ways of prioritising the needs of their relatives while facing the impact of cumulative caregiving responsibilitiesThe uncertainty about future residential respite in terms of its availability, ongoing restrictions in care homes, and which sources of information to trust was noteworthyThe Covid‐19 pandemic may have heighted some demands of caregiving, and needs for a break may be greater. Individual confidence to use residential respite may need to be rebuilt [ABSTRACT FROM AUTHOR]

Objective: To describe a case of Waterhouse–Friderichsen syndrome of adrenocortical failure in a cat with Klebsiella spp. infection. Case Summary: A 12‐year‐old male neutered domestic short‐haired cat was referred for respiratory failure requiring mechanical ventilation. The cat remained comatose despite successful weaning from the ventilator and developed a Klebsiella pneumoniae pneumonia. On day 4 of hospitalization, the cat acutely deteriorated with profound hypotension, azotemia, and hyperkalemia, which rapidly progressed to cardiac arrest. Necropsy findings revealed massive adrenal hemorrhage and intralesional bacteria, termed Waterhouse–Friderichsen syndrome. Waterhouse–Friderichsen syndrome was suspected to have been the cause of acquired adrenocortical insufficiency and sudden death of the cat. New or Unique Information: To the authors' knowledge, this is the first report of sepsis causing Waterhouse–Friderichsen syndrome in a veterinary species. [ABSTRACT FROM AUTHOR]

Objective: The decision about the best time for a person living with dementia to move to a care home involves the individual and others, particularly family. However, little is known about care professionals' views on the best time to move, particularly those with decision‐making authority. This study investigated social workers' and care home managers' views on whether there is an "optimal time" for a move. Methods: A qualitative, phenomenological approach was employed, using semi‐structured interviews with 20 social workers and 20 care home managers in England; all with experience of advising people living with dementia about a care home move and making decisions about funding or acceptance. Interviews were audio‐recorded, transcribed, and analyzed thematically. Results: Four overarching themes emerged from the data: (1) staying at home for as long as possible but avoiding crisis, (2) balancing risks proactively and anticipating triggers, (3) desires for the person living with dementia to be involved in the decision, and (4) the significance of funding in enabling choices about a care home move. Conclusions: Deciding on the timing of a care home move is context and person specific. Two professional groups with substantial experience of this among their client group both recommended proactive deliberation but funding was overall the deciding factor in the extent to which they considered choice was possible. Future research should avoid seeing all care home moves as negative and explore how practitioners can best encourage discussions prior to crisis point about care home options. [ABSTRACT FROM AUTHOR]

Rationale: Malignant pleural mesothelioma is an extremely aggressive and incurable malignancy associated with prior exposure to asbestos fibres. Difficulties remain in relation to early diagnosis, notably due to impeded identification of asbestos in lung tissue. This study describes a novel laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) imaging approach to identify asbestos within mesothelioma models with clinical significance. Methods: Human mesothelioma cells were exposed to different types of asbestos fibres and prepared on plastic slides for LA-ICP-MS analysis. No further sample preparation was required prior to analysis, which was performed using an NWR Image 266 nm laser ablation system coupled to an Element XR sector-field ICP mass spectrometer, with a lateral resolution of 2 μm. Data was processed using LA-ICP-MS ImageTool v1.7 with the final graphic production made using DPlot software. Results: Four different mineral fibres were successfully identified within the mesothelioma samples based on some of the most abundant elements that make up these fibres (Si, Mg and Fe). Using LA-ICP-MS as an imaging tool provided information on the spatial distribution of the fibres at cellular level, which is essential in asbestos detection within tissue samples. Based on the metal counts generated by the different types of asbestos, different fibres can be identified based on shape, size, and elemental composition. Detection of Ca was attempted but requires further optimisation. Conclusions: Detection of asbestos fibres in lung tissues is very useful, if not necessary, to complete the pathological dt9iagnosis of asbestos-related malignancies in the medicolegal field. For the first time, this study demonstrates the successful application of LA-ICP-MS imaging to identify asbestos fibres and other mineral fibres within mesothelioma samples. Ultimately, high-resolution, fast-speed LA-ICP-MS analysis has the potential to be integrated into clinical workflow to aid earlier detection and stratification of mesothelioma patient samples. [ABSTRACT FROM AUTHOR]

Background: Systemic hypertension (SH) is a potential complication of acute kidney injury (AKI) in dogs. Objective: To describe the prevalence of SH and hypertensive retinopathy in dogs with AKI, to investigate the relationship between SH and severity of AKI and to assess possible factors associated with SH. Animals Fifty‐two dogs with AKI. Methods: Prospective observational study of dogs presenting to a tertiary referral center that fulfilled the International Renal Interest Society (IRIS) guidelines for the diagnosis of AKI. Systolic blood pressure measurement, urine protein/creatinine ratio (UPCR), urine output, presence of hypertensive retinopathy and fluid overload (FO), survival to discharge and duration of hospitalization were subsequently assessed. The prevalence of SH was calculated and the relationship between SH and recorded factors was examined by nonparametric statistics. Results: The prevalence of SH (≥160 mm Hg) on admission or during hospitalization was 75% (39/52) and in 56% (22/39) of cases this was severe (≥180 mm Hg). Sixteen percent (7/43) of dogs had evidence of hypertensive retinopathy and 77% (24/31) dogs had UPCR >0.5. Forty‐two percent (22/52) dogs had FO on admission or during hospitalization. There was no association between SH and IRIS AKI grade, oligo/anuria, survival to discharge, duration of hospitalization or proteinuria. Dogs with FO on presentation were more likely to be hypertensive at admission compared to dogs without FO (P =.02). Dogs that did not survive to discharge were more likely to have FO (P =.007). Conclusions and Clinical Importance: Systemic hypertension is common in dogs with AKI. Systemic hypertension might be associated with FO, which itself is associated with nonsurvival. Monitoring for SH and FO is therefore warranted in dogs with AKI. [ABSTRACT FROM AUTHOR]

Three‐dimensional (3D) cell culture is a rapidly emerging field, which mimics some of the physiological conditions of human tissues. In cancer biology, it is considered a useful tool in predicting in vivo chemotherapy responses, compared with conventional two‐dimensional (2D) cell culture. We have developed a novel 3D cell culture model of osteosarcoma composed of aggregated proliferative tumour spheroids, which shows regions of tumour heterogeneity formed by aggregated spheroids of polyclonal tumour cells. Aggregated spheroids show local necrotic and apoptotic regions and have sizes suitable for the study of spatial distribution of metabolites by mass spectrometry imaging (MSI). We have used this model to perform a proof‐of‐principle study showing a heterogeneous distribution of endogenous metabolites that colocalise with the necrotic core and apoptotic regions in this model. Cytotoxic chemotherapy (doxorubicin) responses were significantly attenuated in our 3D cell culture model compared with those of standard cell culture, as determined by resazurin assay, despite sufficient doxorubicin diffusion demonstrated by localisation throughout the 3D constructs. Finally, changes to the distribution of endogenous metabolites in response to doxorubicin were readily detected by MSI. Principal component analysis identified 50 metabolites which differed most in their abundance between treatment groups, and of these, 10 were identified by both in‐software t test and mixed‐effects analysis of variance (ANOVA). Subsequent independent MSIs of identified species were consistent with principle component analysis findings. This proof‐of‐principle study shows for the first time that chemotherapy‐induced changes in metabolite abundance and distribution may be determined in 3D cell culture by MSI, highlighting this method as a potentially useful tool in the elucidation of chemotherapy responses as an alternative to in vivo testing. [ABSTRACT FROM AUTHOR]

Temozolomide (TMZ) is a chemotherapy agent used to treat Grade IV astrocytoma, also known as glioblastoma (GBM). TMZ treatment causes DNA damage that results in tumor cell apoptosis and increases the survival rate of GBM patients. However, chemoresistance as a result of TMZ‐induced autophagy significantly reduces this anticancer effects over time. Statins are competitive inhibitors of HMG‐CoA reductase, the rate‐limiting enzyme of the mevalonate (MEV) cascade. Statins are best known for their cholesterol (CH)‐lowering effect. Long‐term consumption of statins, prior to and in parallel with other cancer therapeutic approaches, has been reported to increase the survival rate of patients with various forms of cancers. In this study, we investigated the potentiation of TMZ‐induced apoptosis by simvastatin (Simva) in human GBM cell lines and patient GBM cells, using cell monolayers and three‐dimensional cell culture systems. The incubation of cells with a combination of Simva and TMZ resulted in a significant increase in apoptotic cells compared to cells treated with TMZ alone. Incubation of cells with CH or MEV cascade intermediates failed to compensate the decrease in cell viability induced by the combined Simva and TMZ treatment. Simva treatment inhibited the autophagy flux induced by TMZ by blocking autophago‐lysosome formation. Our results suggest that Simva sensitizes GBM cells to TMZ‐induced cell death in a MEV cascade‐independent manner and identifies the inhibition of autophagosome‐lysosome fusion as a promising therapeutic strategy in the treatment of GBM. [ABSTRACT FROM AUTHOR]

HepG2 cells were incubated with a 16.5:1.7:1 ratio of cholesterol:sitosterol:campesterol (CSC), a ratio of the major sterols observed in the plasma of phytosterolemia patients, or with cholesterol alone in combination with [14C]acetate for 24 h and the radioactivity incorporated into lipids determined. Cells incubated with CSC exhibited a 40% reduction in cholesterol esterification (p < 0.05) compared to cells incubated with cholesterol alone. In addition, a 17.5‐fold reduction (p < 0.05) in total cholesterol (cholesterol plus cholesteryl ester) synthesis from [14C]acetate was observed in cells incubated with CSC compared to cholesterol alone. Low‐density lipoprotein receptor (LDLR) mRNA abundance was lower in cells incubated with CSC compared to cells incubated with cholesterol alone. Our results suggest that incubation of HepG2 cells with a ratio of sterols that mimic the plasma concentration seen in phytosterolemia patients reduces cholesterol esterification, total cholesterol synthesis, and inhibits LDLR mRNA abundance. We suggest that future cell and animal‐based work on phytostosterolemia might employ this methodology to serve as a novel paradigm of the disease. [ABSTRACT FROM AUTHOR]

Key points: Maternal resveratrol (RESV) administration in gestational diabetes (GDM) restored normoglycaemia and insulin secretion.GDM‐induced obesity was prevented in male GDM+RESV offspring but not in females.GDM+RESV offspring exhibited improved glucose tolerance and insulin sensitivity.GDM+RESV restored hepatic glucose homeostasis in offspring.Glucose‐stimulated insulin secretion was enhanced in GDM+RESV offspring. Gestational diabetes (GDM), the most common complication of pregnancy, is associated with adverse metabolic health outcomes in offspring. Using a rat model of diet‐induced GDM, we investigated whether maternal resveratrol (RESV) supplementation (147 mg kg–1 day–1) in the third week of pregnancy could improve maternal glycaemia and protect the offspring from developing metabolic dysfunction. Female Sprague–Dawley rats consumed a high‐fat and sucrose (HFS) diet to induce GDM. Lean controls consumed a low‐fat (LF) diet. In the third trimester, when maternal hyperglycaemia was observed, the HFS diet was supplemented with RESV. At weaning, offspring were randomly assigned a LF or HFS diet until 15 weeks of age. In pregnant dams, RESV restored glucose tolerance, normoglycaemia and improved insulin secretion. At 15 weeks of age, GDM+RESV‐HFS male offspring were less obese than the GDM‐HFS offspring. By contrast, the female GDM+RESV‐HFS offspring were similarly as obese as the GDM‐HFS group. Hepatic steatosis, insulin resistance, glucose intolerance and dysregulated gluconeogenesis were observed in the male GDM offspring and were attenuated in the offspring of GDM+RESV dams. The dysregulation of several metabolic genes (e.g. ppara, lpl, pepck and g6p) in the livers of GDM offspring was attenuated in the GDM+RESV offspring group. Glucose stimulated insulin secretion was also improved in the islets from offspring of GDM+RESV dams. Thus, maternal RESV supplementation during the third trimester of pregnancy and lactation induced several beneficial metabolic health outcomes for both mothers and offspring. Therefore, RESV could be an alternative to current GDM treatments. Key points: Maternal resveratrol (RESV) administration in gestational diabetes (GDM) restored normoglycaemia and insulin secretion.GDM‐induced obesity was prevented in male GDM+RESV offspring but not in females.GDM+RESV offspring exhibited improved glucose tolerance and insulin sensitivity.GDM+RESV restored hepatic glucose homeostasis in offspring.Glucose‐stimulated insulin secretion was enhanced in GDM+RESV offspring. [ABSTRACT FROM AUTHOR]

Objective: To evaluate differences in hospital readmission risk across all payers in South Carolina (SC).Data Sources/study Setting: South Carolina Revenue and Fiscal Affairs Office (SCRFA) statewide all payer claims database including 2,476,431 hospitalizations in SC acute care hospitals between 2008 and 2014.Study Design: We compared the odds of unplanned all-cause 30-day readmission for private insurance, Medicare, Medicaid, uninsured, and other payers and examined interaction effects between payer and index admission characteristics using generalized estimating equations.Data Collection: SCRFA receives claims and administrative health care data from all SC health care facilities in accordance with SC state law.Principal Findings: Odds of readmission were lower for females compared to males in private, Medicare, and Medicaid payers. African Americans had higher odds of readmission compared to whites across private insurance, Medicare, and Medicaid, but they had lower odds among the uninsured. Longer length of stay had the strongest association with readmission for private and other payers, whereas an increased number of comorbidities related to the highest readmission odds within Medicaid.Conclusions: Associations between index admission characteristics and readmission likelihood varied significantly with payer. Findings should guide the development of payer-specific quality improvement programs. [ABSTRACT FROM AUTHOR]

Key points Gestational diabetes mellitus is a common complication of pregnancy, but its effects on the offspring are poorly understood., We developed a rat model of diet-induced gestational diabetes mellitus that recapitulates many of the clinical features of the disease, including excessive gestational weight gain, glucose intolerance, hyperinsulinaemia and mild hyperglycaemia., Compared to the offspring of lean dams, exposure to gestational diabetes mellitus during the prenatal period resulted in obesity, hepatic steatosis and insulin resistance in young rat offspring that consumed a postnatal diet that was low in fat., The combination of maternal gestational diabetes mellitus and the postnatal consumption of a high-fat diet by the offspring caused a more severe metabolic phenotype., Metabolomic profiling of the liver tissues of the offspring of gestational diabetic dams revealed accumulation of lipotoxic lipids and reduced phosphatidylethanolamine levels compared to the offspring of lean dams., The results establish that gestational diabetes mellitus is a driver of hepatic steatosis and insulin resistance in the offspring., Abstract Maternal obesity is associated with a high risk for gestational diabetes mellitus (GDM), which is a common complication of pregnancy. The influence of maternal obesity and GDM on the metabolic health of the offspring is poorly understood. We hypothesize that GDM associated with maternal obesity will cause obesity, insulin resistance and hepatic steatosis in the offspring. Female Sprague-Dawley rats were fed a high-fat (45%) and sucrose (HFS) diet to cause maternal obesity and GDM. Lean control pregnant rats received low-fat (LF; 10%) diets. To investigate the interaction between the prenatal environment and postnatal diets, rat offspring were assigned to LF or HFS diets for 12 weeks, and insulin sensitivity and hepatic steatosis were evaluated. Pregnant GDM dams exhibited excessive gestational weight gain, hyperinsulinaemia and hyperglycaemia. Offspring of GDM dams gained more weight than the offspring of lean dams due to excess adiposity. The offspring of GDM dams also developed hepatic steatosis and insulin resistance. The postnatal consumption of a LF diet did not protect offspring of GDM dams against these metabolic disorders. Analysis of the hepatic metabolome revealed increased diacylglycerol and reduced phosphatidylethanolamine in the offspring of GDM dams compared to offspring of lean dams. Consistent with altered lipid metabolism, the expression of CTP:phosphoethanolamine cytidylyltransferase, and peroxisomal proliferator activated receptor-α mRNA was reduced in the livers of GDM offspring. GDM exposure programs gene expression and hepatic metabolite levels and drives the development of hepatic steatosis and insulin resistance in young adult rat offspring. [ABSTRACT FROM AUTHOR]

RATIONALE Matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) provides a methodology to map the distribution of peptides generated by in situ tryptic digestion of biological tissue. It is challenging to correlate these peptides to the proteins from which they arise because of the many potentially overlapping and hence interfering peptide signals generated. METHODS A recombinant protein has been synthesised that when cleaved with trypsin yields a range of peptide standards for use as identification and quantification markers for multiple proteins in one MALDI-IMS-MSI experiment. Mass spectrometry images of the distribution of proteins in fresh frozen and formalin-fixed paraffin-embedded tissue samples following in situ tryptic digestion were generated by isolating signals on the basis of their m/z value and ion mobility drift time, which were correlated to matching peptides in the recombinant standard. RESULTS Tryptic digestion of the IMS-TAG protein and MALDI-MS analysis yielded m/z values and ion mobility drift time for the signature peptides included in it. MALDI-IMS-MSI images for the distribution of the proteins HSP90 and vimentin, in FFPE EMT6 mouse tumours, and HSP90 and plectin in a fresh frozen mouse fibrosarcoma, were generated by extracting ion images at the corresponding m/z value and drift time from the tissue samples. CONCLUSIONS The IMS-TAG approach provides a new means to confirm the identity of peptides generated by in situ digestion of biological tissue. Copyright © 2013 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Aims To investigate the prevalence of urinary and fecal incontinence in people with cognitive impairment or dementia, living at home. Method We searched electronic databases, MEDLINE, EMBASE, CINAHL, PsycINFO, BNI, and the Cochrane Library (including DARE, NTIS), from January 1, 1990 to 2012 week 13 (April 4) for studies reporting prevalence data of urinary and fecal incontinence in the population of interest. Quality assessments of studies considered risk of bias in criteria for prevalence studies. Due to the heterogeneity of the included study populations and results, meta-analysis was not appropriate and a narrative analysis was undertaken. Results From 427 references, eight studies met the inclusion criteria. Seven studies provided prevalence rates as findings incidental to their primary question. Populations and assessment tools were varied. Reported prevalence of urinary incontinence ranged from 1.1% in a general community population to 38% in those receiving home care services. Reported rates of fecal incontinence were from 0.9% in a community population to 27% in a population attending an old age psychiatry outpatient clinic. Conclusions The prevalence of incontinence in people with dementia or cognitive impairment living at home has not been clearly established. Population level data is required to inform clinicians and to reliably underpin decision-making in service planning, resource allocation and interventions for people with dementia and incontinence. Neurourol. Urodynam. 32: 314-324, 2013. © 2012 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

BACKGROUND: Alloimmunization to red blood cell (RBC) blood group antigens is a major complication for patients with sickle cell disease (SCD), which limits the usefulness of RBC transfusion. Here, we report our experiences with extended RBC antigen matching for SCD patients. STUDY DESIGN AND METHODS: Records for 99 SCD patients transfused only with the extended matching protocol between 1993 and 2006 were reviewed. Patients and donors were phenotyped for 20 blood group antigens and RBC units that were negative for antigens not expressed by the recipient were provided. When necessary, mismatches were allowed at Lea, Leb, Fyb, and MNSs to meet requirements for antigens regarded as the most clinically significant. Matched RBC units (6946) were provided to 99 patients (mean, 70 units/patient; range, 1-519 units/patient). Eliminating mismatches, 90% of the transfusions matched all other negative antigens. RESULTS: Seven alloantibodies were detected in seven patients resulting in 7% alloimmunized at a rate of 0.1 antibodies per 100 units transfused. Three recipients who developed antibodies were D mosaic and would have been mistyped with serologic techniques. Alloimmunization was decreased compared to ABO and/or D matching at our institution and others. Twelve autoantibodies and no severe hemolytic transfusion reactions were reported. CONCLUSION: Exact matching for ABO, Rhesus, Kell, Kidd, and Fya and extending this match whenever possible is an effective strategy to reduce alloimmunization to RBC antigens. Consideration should be given to exploring this conclusion further with a controlled, multi-institutional trial to determine efficacy, cost-benefit analysis, and reproducibility of this approach. [ABSTRACT FROM AUTHOR]

Context: Rates of preterm birth (PTB) and low birth weight (LBW) vary by region, with disparities particularly evident in the Appalachian region of the South. Community conditions related to rurality likely contribute to adverse birth outcomes in this region. Purpose: This study examined associations between rurality and related community conditions, and newborn outcomes in southern Appalachia, and explored whether pregnancy smoking explained such associations. Methods: Data for all births in a southern Appalachian county over a 2-year period were extracted from hospital records. Findings: Data were available for 4,144 births, with 45 different counties of residence. Babies born to women from completely rural counties, on average, weighed 700 g less, were 1.5 inches shorter, and were born over 3 weeks earlier than less rural infants. In addition, these babies were 4.5 times more likely to be LBW, 4 times more likely to be PTB, and 5 times more likely to be admitted to the neonatal intensive care unit (NICU). Effects were also found for per capita income, poverty rate, and unemployment rate, all of which were associated with rurality. Some, but not all of the association was explained by elevated rates of pregnancy smoking. Conclusions: Babies born to women residing in rural and economically depressed counties in southern Appalachia are at substantially increased risk for poor birth outcomes. Improving these outcomes in the rural South will likely require addressing access to health services and information, health care provider retention, transportation services, employment opportunities, and availability of public health services including smoking cessation assistance. [ABSTRACT FROM AUTHOR]

Objective Case Summary New or Unique Information Provided To describe a case of concurrent septic pyothorax, peritonitis, and cholecystitis due to Staphylococcus pseudintermedius in a dog.A 10‐year‐old neutered male, medium‐sized, mixed‐breed dog presented with an acute abdomen. Initial investigations found the presence of a septic pyothorax, septic peritonitis, and bacterial cholecystitis. Exploratory celiotomy did not identify the source of infection, and no macroscopic communication between pleural and peritoneal space was detected. An abdominal Jackson‐Pratt drain and bilateral small‐bore thoracostomy tubes were placed for further management, with the dog receiving antimicrobial therapy and intensive supportive care. Subsequently, microbiological cultures confirmed scant growth of S. pseudintermedius in bile, pleural, and peritoneal fluid. The dog was discharged after 1 week of hospitalization. Antimicrobial therapy was discontinued 4 weeks after discharge due to an improvement of clinical signs and normalization of C‐reactive protein concentration. A diagnosis of hyperadrenocorticism was made 6 weeks after discharge, and appropriate therapy was initiated.Septic peritonitis and septic pyothorax are both common life‐threatening conditions in veterinary medicine. To the authors’ knowledge, this is the first report describing septic bicavitary effusion associated with a biliary staphylococcal infection in a dog. The exact mechanism by which the infection spread remains unclear, but hematogenous or lymphatic routes are considered the most probable. However, the presence of microscopic diaphragmatic defects and an anicteric gallbladder rupture cannot completely be excluded. The concurrent hyperadrenocorticism may have also contributed to an immunocompromised state, predisposing the dog to the development of infection. [ABSTRACT FROM AUTHOR]

L5240 --> Introduction: Tafazzin is a transacylase enzyme that remodels the phospholipid cardiolipin with fatty acids. Mutations in tafazzin result in the X‐linked genetic disease Barth Syndrome. Some Barth syndrome patients exhibit a lean phenotype and tafazzin knockdown mice, an animal model of Barth Syndrome, are protected from diet‐induced obesity. We hypothesized that tafazzin deficiency in both the liver and beta cells of the pancreas contribute to this lean phenotype. Methods: Embyros from mice containing loxP‐flanked Taz (flox) were previously generated by using sperm from a Taz flox male (Taz‐fl/Y) for in vitro fertilization of wild type C57BL/6J oocytes (Dr. Douglas Strathdee, Beaton Institute, Glasgow, UK). The resulting heterozygous female (Taz‐fl/+) and hemizygous males (Taz‐fl/Y) were bred >9 times to a C57BL/6 background. To specifically delete Taz in the liver, mice homozygous/hemizygous for the floxed Taz allele are mated with B6.Cg‐Speer6‐ps1Tg(Alb‐cre)21Mgn/J strain of mice transgenic for cre recombinase under the albumin (Albumin) promoter (AlbuminCre). The B6.Cg‐Speer6‐ps1Tg(Alb‐cre)21Mgn/J strain of mice are obtained from Jackson Laboratories. Subsequent breeding took place between female mice heterozygous for the floxed Taz allele (Taz‐fl/+, without AlbuminCre) with male mice which are wildtype (+/+) as well as male mice which are hemizygous for the floxed Taz allele (Taz‐fl/Y) and heterozygous for the AlbuminCre transgene. This generated male and female mice which are hemizygous for the floxed Taz allele (Taz‐fl/Y) and homozygous for the floxed allele (Taz‐fl/Taz‐fl) with AlbuminCre expression (liver specific knockout of Taz). Littermate controls included both male and female floxed Taz (Taz‐fl/Y, Taz‐fl/Taz‐fl) without AlbuminCre expression (flox controls) and wildtype Taz with AlbuminCre (cre recombinase controls). The same protocol was used to generate pancreatic beta cell‐specific TazKO mice driven under the Ins1 promoter using B6.Cg‐Tg(Ins1‐cre/ERT)1Lphi/J strain of mice. In addition, liver‐ and pancreatic beta cell‐specific double knockout mice were generated. 4 Month old male mice were fed a high‐fat (23%, w/w) (Envigo Teklad, Rodent diet cat# TD.130261) for up to 8 weeks. Body weights were measured weekly and fat accumulating tissues weighed after 8 weeks. Results: Liver‐specific or pancreatic beta cell‐specific male tafazzin knockout mice accumulated weight gain (≍40% increase in body weight) at the same rate as control animals (Figure 1). However, the liver‐ and beta cell‐specific double tafazzin knockout mice exhibited a reduced rate of weight gain by 8 weeks (≍26% increase in body weight) compared to control or the single tafazzin knockout animals. In addition, at 8 weeks the double tafazzin knockout mice exhibited reduced weight gain in tissues known to accumulate fat including the liver, the gonadal, inguinal and perirenal white adipose tissues and the brown adipose tissue. Conclusion: The data suggest that liver‐ and pancreatic beta cell‐specific double tafazzin knockout male mice are protected from high fat diet induced weight gain and fat accumulation. [ABSTRACT FROM AUTHOR]

Phosphatidylcholine (PC) is the predominant phospholipid in mammalian plasma lipoproteins and cellular membranes. A liver-specific pathway for PC biosynthesis is catalyzed by phosphatidylethanolamine N-methyltransferase (PEMT). We have found using quantitative polymerase chain reaction that PEMT mRNA increases in the liver by 96-fold during the perinatal period in mice. To determine regions within the Pemt promoter which are important for gene activation, we prepared sections of the Pemt promoter upstream of a luciferase reporter gene. Our preliminary data suggests that activation elements may exist between -471 and -371bp (relative to the transcriptional start site). This section of the Pemt promoter (-471 to -371bp) also binds nuclear factors from perinatal mouse liver nuclear extract as determined by electromobility shift assays (EMSA). We have identified, within this region, a conserved putative binding site (GC box) for a member of the Sp family of transcription factors. An oligonucleotide containing a GC box competed out the formation of a nuclear protein-Pemt promoter complex by EMSA. Furthermore, mutation of the GC box but not another region within the oligonucleotide eliminated this effect. These experiments suggest that a GC-rich element may regulate the perinatal expression of the mouse Pemt gene. [ABSTRACT FROM AUTHOR]