Feeding nanoparticles of copper oxide coated with lysine with or without added antagonists affects the copper status but not performance of Holstein dairy cows.

The apparent absorption of copper (Cu) in ruminants is low, with between 0.01 and 0.07 g/g absorbed from sources such as copper oxide (CuO) under typical feeding conditions, resulting in high levels of excretion. Improving the bioavailability of Cu could reduce the supplemental amount required to maintain Cu status and reduce excretion, particularly in the presence of dietary antagonists such as sulfur (S) and molybdenum (Mo). The objective of our study was to determine the Cu status of cows when fed nanoparticle CuO coated with Lys compared with conventional CuO when fed without or in combination with antagonists to Cu absorption (S and Mo) in the diet of dairy cows. Fifty-six multiparous Holstein-Friesian cows that were 48 d ± 17.4 (mean ± SD) post calving and yielding 40.6 ± 6.9 kg milk/d were used in a 2 × 2 factorial design. The 4 treatment groups were; CuO (O-), CuO with added antagonists (O+), nano CuO with a lysine coating (N-), and nano CuO with a Lys coating with added antagonists (N+), fed for 16 wks. We formulated the diets to contain approximately 17 mg Cu/kg dry matter (DM) and diets with antagonists contained an additional 1 g S/kg DM and 6 mg Mo/kg DM, with Lys added to O- and O+ to provide the same daily supply as N- and N+. Blood samples were collected at wk 0, 2, 4, 6, 10 and 16, and liver biopsy samples at wk 0 and 16. We found no effect of dietary treatment on DM intake, milk yield, live weight or body condition score, with mean values of 23.3 kg/d, 40.1 kg/d, 646 kg and 2.68, but milk SCC was higher in cows fed conventional compared with non CuO, or with added antagonist. We also found no effect of treatment on blood activity of gamma glutamyl transferase, superoxide dismutase or ceruloplasmin, hematology profile, or plasma Cu and iron concentration. We found that plasma Mo concentration was increased from 0.36 µmol/L in cows fed O- or N- to 0.80 µmol/L in those receiving O+ or N+. Additional dietary antagonists also decreased the concentration of Cu in the liver of cows fed conventional CuO (C+) over the study period by 1.3 mg/kg DM/d, but in cows fed dietary antagonists and nano CuO coated with Lys (N+), liver Cu concentration was increased by 1.1 mg/kg DM/d. Our study is the first to demonstrate that reducing the particle size of CuO into the nano scale with a lysine coating improves the bioavailability of CuO in the presence of dietary antagonists in dairy cattle, and we did not observe any negative effects on performance or health.

Overfeeding copper during rearing affects the liver function and fertility of replacement dairy heifers.

BACKGROUND: Oversupply of dietary copper (Cu) is common among UK dairy herds, but studies on the long-term outcomes of this oversupply are scarce. METHODS: A longitudinal study was undertaken to determine the long-term implications when 80 Holstein‒Friesian heifers with a mean (±standard error) age of 4.1 ± 0.1 months and a mean liveweight of 137 ± 2.4 kg were fed a recommended (R; 16 mg/kg dry matter [DM]) or high (H; 32 mg/kg DM) dietary Cu concentration until 6 weeks prior to calving. RESULTS: Hepatic Cu concentrations in both treatment groups were elevated into the ranges used to diagnose chronic Cu toxicity in cattle at 6.9 months of age (798 ± 46.4 mg/kg DM for H vs. 643 ± 35.4 mg/kg DM for R), with associated evidence of liver damage. Hepatic Cu concentrations then returned to normality but remained higher (p < 0.001) for heifers fed H than for those fed R and were associated with a reduced (p = 0.044) conception rate to first and second services (73.7 ± 8.05% for H vs. 91.2 ± 7.68% for R). LIMITATION: This retrospective analysis identified pre-study liver damage, which may have affected results. CONCLUSIONS: Supplying Cu in excess of requirements resulted in liver damage and reduced conception rates.

Suppression of food intake by Glp1r/Lepr-coexpressing neurons prevents obesity in mouse models.

The adipose-derived hormone leptin acts via its receptor (LepRb) in the brain to control energy balance. A potentially unidentified population of GABAergic hypothalamic LepRb neurons plays key roles in the restraint of food intake and body weight by leptin. To identify markers for candidate populations of LepRb neurons in an unbiased manner, we performed single-nucleus RNA-Seq of enriched mouse hypothalamic LepRb cells, identifying several previously unrecognized populations of hypothalamic LepRb neurons. Many of these populations displayed strong conservation across species, including GABAergic Glp1r-expressing LepRb (LepRbGlp1r) neurons, which expressed more Lepr than other LepRb cell populations. Ablating Lepr from LepRbGlp1r cells provoked hyperphagic obesity without impairing energy expenditure. Similarly, improvements in energy balance caused by Lepr reactivation in GABA neurons of otherwise Lepr-null mice required Lepr expression in GABAergic Glp1r-expressing neurons. Furthermore, restoration of Glp1r expression in LepRbGlp1r neurons in otherwise Glp1r-null mice enabled food intake suppression by the GLP1R agonist, liraglutide. Thus, the conserved GABAergic LepRbGlp1r neuron population plays crucial roles in the suppression of food intake by leptin and GLP1R agonists.

Macromineral and trace element concentrations in milk from Finnish Ayrshire cows fed microalgae (Spirulina platensis) and rapeseed (Brassica napus).

Given the lack of research regarding the effect of microalgal supplementation in dairy cows on milk mineral concentrations, this study investigated the effect of feeding different protein supplements in dairy cow diets on milk, feces, and blood plasma mineral concentrations, associated milk and blood plasma transfer efficiencies, and apparent digestibility. Lactating Finnish Ayrshire cows (n = 8) were allocated at the start of the trial to 4 diets used in a replicated 4 × 4 Latin square design experiment: (1) control diet (CON), (2) a pelleted rapeseed supplement (RSS; 2,550 g/d), (3) a mixture of rapeseed and Spirulina platensis (RSAL; 1,280 g of RSS + 570 g of S. platensis per day), and (4) S. platensis (ALG; 1,130 g of S. platensis per day). In each of the 4 experimental periods, a 2-wk adaptation to the experimental diets was followed by a 7-d sampling and measurement period. Feed samples were composited per measurement period, milk, and feed samples (4 consecutive days; d 17-20), and blood plasma samples (d 21) were composited for each cow period (n = 32). Data were statistically analyzed using a linear mixed effects model with diet, period within square, square and their interaction as fixed factors, and cow within square as a random factor. Cows fed ALG were not significantly different in their milk or blood plasma mineral concentrations compared with CON, although feeding ALG increased fecal concentrations of macrominerals (Ca and Mg) and trace elements (Co, Cu, Fe, I, Mn, and Zn), and reduced their apparent digestibility, compared with CON. When compared with CON and ALG, milk from cows fed RSAL and RSS had lower milk I concentrations (-69.6 and -102.7 µg/kg of milk, respectively), but total plasma I concentrations were not affected significantly. Feeding S. platensis to dairy cows did not affect mineral concentrations in cows' blood or milk, but care should be taken when rapeseed is fed to avoid reducing milk I concentrations which may in turn reduce consumers' I intake from milk and dairy products.

Mushroom body input connections form independently of sensory activity in Drosophila melanogaster.

Associative brain centers, such as the insect mushroom body, need to represent sensory information in an efficient manner. In Drosophila melanogaster, the Kenyon cells of the mushroom body integrate inputs from a random set of olfactory projection neurons, but some projection neurons-namely those activated by a few ethologically meaningful odors-connect to Kenyon cells more frequently than others. This biased and random connectivity pattern is conceivably advantageous, as it enables the mushroom body to represent a large number of odors as unique activity patterns while prioritizing the representation of a few specific odors. How this connectivity pattern is established remains largely unknown. Here, we test whether the mechanisms patterning the connections between Kenyon cells and projection neurons depend on sensory activity or whether they are hardwired. We mapped a large number of mushroom body input connections in partially anosmic flies-flies lacking the obligate odorant co-receptor Orco-and in wild-type flies. Statistical analyses of these datasets reveal that the random and biased connectivity pattern observed between Kenyon cells and projection neurons forms normally in the absence of most olfactory sensory activity. This finding supports the idea that even comparatively subtle, population-level patterns of neuronal connectivity can be encoded by fixed genetic programs and are likely to be the result of evolved prioritization of ecologically and ethologically salient stimuli.

Investigations into the chemical composition and nutritional value of single-cultivar rapeseed meals for broiler chickens.

A study was conducted to 1) investigate the influence of rapeseed cultivar on the range of variation in the chemical composition and feed value of rapeseed meals (RSM) for broilers and 2) to compare between single-cultivar RSM and a batch of UK-sourced proprietary rapeseed meal. Chemical composition was determined for a total of 10 single-cultivar RSM produced via prepressed solvent-extraction and a proprietary RSM. To investigate RSM digestibility a total of 360 birds were housed, six birds per pen, and between day 13 to 21 of age they received a diet containing per kg 250 g RSM and 750 g basal feed based on wheat and soybean meal. Titaniumdioxide (7 g/kg) was added as an inert marker. For the determination nitrogen-corrected metabolisable energy (AMEn) total excreta and feed intake were quantified over the last 4 d of the experiment. On the final day of the experiment ileal digesta were collected for determination of preceacal crude protein digestibility (pcCPD). With the exception of neutral detergent fibre (NDF), glucosinolates and soluble non-starch polysaccharides the coefficients of variation (CV%) for all major components were similar across all RSM (0.5 < CV% < 0.6). Glucosinolate contents were highest in the 10 single-cultivar RSM whereas NDF levels were greater in the proprietary RSM. Across the 10 single-cultivar RSM there was no significant effect of cultivar on AMEn or pcCPD. Pre-planned orthogonal contrasts indicated that the mean AMEn content and pcCPD of the proprietary RSM was significantly lower than the 10 single-cultivar RSM (p < 0.001). In view of the above, processing rather than cultivar appears to be the main driver of nutritional value in RSM fed to broilers.

Reductions in Parent Interest in Receiving Antibiotics following a 90-Second Video Intervention in Outpatient Pediatric Clinics.

OBJECTIVES: To assess the impact of a 90-second animated video on parents' interest in receiving an antibiotic for their child. STUDY DESIGN: This pre-post test study enrolled English and Spanish speaking parents (n = 1051) of children ages 1-5 years presenting with acute respiratory tract infection symptoms. Before meeting with their provider, parents rated their interest in receiving an antibiotic for their child, answered 6 true/false antibiotic knowledge questions, viewed the video, and then rated their antibiotic interest again. Parents rated their interest in receiving an antibiotic using a visual analogue scale ranging from 0 to 100, with 0 being "I definitely do not want an antibiotic," 50 "Neutral," and 100 "I absolutely want an antibiotic." RESULTS: Parents were 84% female, with a mean age of 32 ± 6.0, 26.0% had a high school education or less, 15% were black, and 19% were Hispanic. After watching the video, parents' average antibiotic interest ratings decreased by 10 points (mean, 57.0 ± 20 to M ± 21; P < .0001). Among parents with the highest initial antibiotic interest ratings (≥60), even greater decreases were observed (83.0 ± 12.0 to 63.4 ± 22; P < .0001) with more than one-half (52%) rating their interest in the low or neutral ranges after watching the video. CONCLUSIONS: A 90-second video can decrease parents' interest in receiving antibiotics, especially among those with higher baseline interest. This scalable intervention could be used in a variety of settings to reduce parents' interest in receiving antibiotics. TRIAL REGISTRATION: ClinicalTrials.gov: NCT03037112.

"It was my obligation as mother": 18-Month completion of Early Infant Diagnosis as identity control for mothers living with HIV in Kenya.

RATIONALE: Early Infant Diagnosis (EID) is critical to timely identification of HIV and rapid treatment initiation for infants found to be infected. Completing the EID cascade involves a series of age-specific tests between birth and 18 months and can be challenging for mother- infant pairs in low-resource settings. Even prior to recent increases in Kenya's testing recommendations, approximately 22% of mother-infant pairs enrolled in EID were lost to follow-up. As EID cascades become increasingly complex, identifying strategies to maximize retention becomes even more essential. Despite ongoing health system improvements, we still lack a framework for understanding the individual-level, psychosocial processes underlying EID completion-insight that could be essential for shaping strategies to support patients and close gaps in retention. OBJECTIVE: Our objective was to explain individual-level processes that lead to EID completion among mothers who successfully completed the EID cascade. METHODS: Using qualitative methods informed by grounded theory, we conducted 65 interviews with Kenyan mothers who completed EID. RESULTS: We identified three themes related to completion: (1) Negative motivation, from the consequences of infant infection, fear of postnatal transmission, and burden of maternal failure; (2) Positive motivation, from achieving a final goal, responding to provider support, and maximizing infant health; and (3) Overcoming challenges, through resolve/resiliency, planning/privatizing and rejecting stigma/leveraging support. CONCLUSION: Overall, the EID cascade served as a framing process for women to secure an identity as a good mother. Successful EID completion was the product of a strong motivational foundation channeled into strategies to surmount persistent challenges. Participant accounts of overcoming challenges highlight their resiliency as well as the outstanding need to address financial, logistical, and social barriers to care. Future EID programs may capitalize on these findings by affirming patients' quest for identity control, leveraging positive motivators, and expanding peer support structures to help mothers feel connected throughout the cascade.

The effects of supercritical carbon dioxide extraction and cold-pressed hexane extraction on the chemical composition and feeding value of rapeseed meal for broiler chickens.

The chemical characteristics of rapeseed meal (RSM) produced from two cultivars of UK-grown rapeseed, by both supercritical carbon dioxide extraction (ScCO2) and cold-pressed hexane extraction (CpHe) were examined. Their nutritional value, with and without protease, was then assessed in a broiler digestibility trial. Basal feed was substituted with one of four RSM batches (200 g/kg) following adjustments for dry matter (DM) and ether extract (EE) content. Half of each diet was supplemented with a mono-component protease derived from Bacillus subtilis (Axtra®PRO, Danisco Animal Nutrition, Malborough, UK) giving a total of eight test diets. Two control diets, with and without protease were also fed. At 13 d age male Ross 308 broilers were randomly allocated to seven replicate pens (five birds per pen) and assigned to one of 10 diets. Total excreta were collected from 17 to 21 d age and feed intake was recorded. Pre-caecal protein digestibility (pcPd) was determined using TiO2 as an indigestible marker. Colourimetrically CpHe RSM was substantially darker than ScCO2 counterparts. The influence of oil recovery method (ORM) was also evident in DM, EE, ash free neutral detergent fibre (aNDFom), neutral detergent insoluble crude protein (NDICP) and glucosinolate content (GLS). The content of DM, EE and GLS was higher in ScCO2 RSM whereas aNDFom and NDICP levels were greater in CpHe RSM. Protein solubility in KOH was greater in ScCO2 RSM whilst levels of NDICP were lower. Collectively these results suggest that less heat damage was incurred to the RSM during ScCO2 extraction. There was no significant main effect of cultivar nor were any significant interactions observed between treatment factors. Rapeseed meal ScCO2 produced greater metabolisable energy, pcPd, nitrogen retention and energy metabolisability (p < 0.05). Protease supplementation increased pcPd (p < 0.05) irrespective of ORM and cultivar. The key implications of these findings are that by adopting oil recovery methods that minimise the exposure of RSM to thermal treatments and by adding a compatible protease there is scope to increase the nutritional value of RSM for broilers and increase its utilisation in modern poultry production.

Home self-administration of intravenous antibiotics as part of an outpatient parenteral antibiotic therapy service: a qualitative study of the perspectives of patients who do not self-administer.

OBJECTIVES: This study aimed to use a theoretical approach to understand the determinants of behaviour in patients not home self-administering intravenous antibiotics. SETTING: Outpatient care: included patients were attending an outpatient clinic for intravenous antibiotic administration in the northeast of Scotland. PARTICIPANTS: Patients were included if they had received more than 7 days of intravenous antibiotics and were aged 16 years and over. Twenty potential participants were approached, and all agreed to be interviewed. 13 were male with a mean age of 54 years (SD +17.6). OUTCOMES: Key behavioural determinants that influenced patients' behaviours relating to self-administration of intravenous antibiotics. DESIGN: Qualitative, semistructured in-depth interviews were undertaken with a purposive sample of patients. An interview schedule, underpinned by the Theoretical Domains Framework (TDF), was developed, reviewed for credibility and piloted. Interviews were audio-recorded and transcribed verbatim. Data were analysed thematically using the TDF as the coding framework. RESULTS: The key behavioural determinants emerging as encouraging patients to self-administer intravenous antibiotics were the perceptions of being sufficiently knowledgeable, skilful and competent and that self-administration afforded the potential to work while administering treatment. The key determinants that impacted their decision not to self-administer were lack of knowledge of available options, a perception that hospital staff are better trained and anxieties of potential complications. CONCLUSION: Though patients are appreciative of the skills and knowledge of hospital staff, there is also a willingness among patients to home self-administer antibiotics. However, the main barrier emerges to be a perceived lack of knowledge of ways of doing this at home. To overcome this, a number of interventions are suggested based on evidence-based behavioural change techniques.

Defining the Transcriptional Targets of Leptin Reveals a Role for Atf3 in Leptin Action.

Leptin acts via its receptor (LepRb) to modulate gene expression in hypothalamic LepRb-expressing neurons, thereby controlling energy balance and glucose homeostasis. Despite the importance of the control of gene expression in hypothalamic LepRb neurons for leptin action, the transcriptional targets of LepRb signaling have remained undefined because LepRb cells contribute a small fraction to the aggregate transcriptome of the brain regions in which they reside. We thus employed translating ribosome affinity purification followed by RNA sequencing to isolate and analyze mRNA from the hypothalamic LepRb neurons of wild-type or leptin-deficient (Lepob/ob) mice treated with vehicle or exogenous leptin. Although the expression of most of the genes encoding the neuropeptides commonly considered to represent the main targets of leptin action were altered only following chronic leptin deprivation, our analysis revealed other transcripts that were coordinately regulated by leptin under multiple treatment conditions. Among these, acute leptin treatment increased expression of the transcription factor Atf3 in LepRb neurons. Furthermore, ablation of Atf3 from LepRb neurons (Atf3LepRbKO mice) decreased leptin efficacy and promoted positive energy balance in mice. Thus, this analysis revealed the gene targets of leptin action, including Atf3, which represents a cellular mediator of leptin action.

Primary meningococcal arthritis as a presentation of nodal marginal zone lymphoma.

A 68-year-old man presented with a 4-day history of worsening knee and arm pain. On examination, there was erythema and swelling of the left knee and both wrists. Joint aspiration grew Neisseria meningitidis Blood tests showed an unusually high total protein level (100 g/L) and an IgM kappa paraprotein band of 45 g/L on protein electrophoresis. CT showed widespread lymphadenopathy, hepatosplenomegaly and multilevel thoracic vertebral collapse. A bone marrow biopsy revealed a lymphoplasmacytic infiltrate and a lymph node biopsy showed features of nodal marginal zone lymphoma with plasmacytic differentiation.

Feeding value of field beans (Vicia faba L. var. minor) with and without enzyme containing tannase, pectinase and xylanase activities for broilers.

Effects of field beans with various tannin content and exogenous enzyme mixture containing tannase, pectinase and xylanase activities on N-corrected dietary apparent metabolisable energy (AMEn), coefficients of dry matter (DMR) and nitrogen retention (NR), fat digestibility, gastrointestinal tract (GIT) development, jejunal villus morphometry, ileal digesta viscosity and sialic acid were examined. Birds' growth performance and energy conversion ratio (ECR) were also measured. Birds were fed one of eight mash diets. The Control diet contained as major ingredients wheat (400 g/kg) and soybean meal (SBM) (127 g/kg and 221 g crude protein/kg and 12.83 MJ AMEn/kg. To reduce nutrient density, the Control diet also contained washed sand at 119 g/kg. Another three diets containing 300 g/kg of each of three experimental field bean cultivar samples in replacement for SBM and sand were also mixed. Each diet was fed to nine pens with two male Ross 308 broilers. Diets high in tannin had low AMEn, ECR, DMR and NR (p < 0.001). Feeding field beans increased (p < 0.001) the weights of the pancreas and the proventriculus and gizzard (PG) of the birds. Supplementing diets with the enzyme mixture improved (p < 0.001) feed conversion efficiency, AMEn and all nutrient utilisation coefficients despite the tannins in diets. The enzyme mixture reduced ileal digesta viscosity (p < 0.001) and the weight of pancreas, total GIT and PG (p < 0.05) of the birds. It can be concluded that the feeding value of field beans with different tannin contents may vary when fed to broilers. The supplementation of the enzyme mixture improved the feeding value of diets for broilers. The beneficial effect of the addition of the enzyme mixture seems to be mediated through reduced ileal digesta viscosity and improved nutrient availability.

Nutritional value of raw and micronised field beans (Vicia faba L. var. minor) with and without enzyme supplementation containing tannase for growing chickens.

An experiment examined the effects of two field bean cultivar samples with different tannin contents, the effect of heat treatment (micronising) and the effect of dietary supplementation of a proprietary enzyme preparation containing tannase, pectinase, and xylanase activities on metabolisable energy (ME), total tract dry matter digestibility (DMD) and ether extract digestibility (EED), nitrogen retention (NR), tannin degradability, gastrointestinal tract (GIT) development, and endogenous mucin losses excretion in broiler chickens. The Control diet contained per kg 221 g crude protein and 12.83 MJ ME. Four additional diets contained 300 g/kg of each of the two untreated or micronised experimental field bean cultivar samples. Each diet was then split into two batches and one of them was supplemented with 3400 units tannase per kg diet resulting in 10 diets in total. Each diet was fed to seven pens with two randomly selected male broilers each. Birds fed the high tannin bean sample had a lower weight gain (p < 0.001), and a lower determined apparent ME (p < 0.05), and DMD (p < 0.001) but a higher tannin degradability (p < 0.001). Compared to the Control diet, feeding field beans increased (p < 0.001) the weights of the proventriculus and gizzard of the birds, and also increased endogenous mucin losses (p < 0.05). Supplementing diets with the tannase-containing enzyme preparation improved dietary ME (p < 0.001), DMD (p < 0.001), NR (p < 0.001) and DEE (p < 0.05), but did not change tannin degradability. Heat treatment of the beans reduced the degradability of condensed tannins and increased endogenous mucin losses (p < 0.05). The differences in the feeding value of the different field bean samples were not improved by heat treatment, but enzyme supplementation improved the feeding value of all diets regardless of the bean samples or heat treatment. Further research is warranted to study the effectiveness of tannase supplementation in poultry diet formulations by dose response trials with purified tannase preparations.

ERα in Tac2 Neurons Regulates Puberty Onset in Female Mice.

A variety of data suggest that estrogen action on kisspeptin (Kiss1)-containing arcuate nucleus neurons (which coexpress Kiss1, neurokinin B (the product of Tac2) and dynorphin (KNDy) neurons restrains reproductive onset and function, but roles for estrogen action in these Kiss1 neurons relative to a distinct population of rostral hypothalamic Kiss1 neurons (which does not express Tac2 or dynorphin) have not been directly tested. To test the role for estrogen receptor (ER)α in KNDy cells, we thus generated Tac2(Cre) and Kiss1(Cre) knock-in mice and bred them onto the Esr1(flox) background to ablate ERα specifically in Tac2-expressing cells (ERα(Tac2)KO mice) or all Kiss1 cells (ERα(Kiss1)KO mice), respectively. Most ERα-expressing Tac2 neurons represent KNDy cells. Arcuate nucleus Kiss1 expression was elevated in ERα(Tac2)KO and ERα(Kiss1)KO females independent of gonadal hormones, whereas rostral hypothalamic Kiss1 expression was normal in ERα(Tac2)KO but decreased in ERα(Kiss1)KO females; this suggests that ERα in rostral Kiss1 cells is crucial for control of Kiss1 expression in these cells. Both ERα(Kiss1)KO and ERα(Tac2)KO females displayed early vaginal opening, early and persistent vaginal cornification, increased gonadotropins, uterine hypertrophy, and other evidence of estrogen excess. Thus, deletion of ERα in Tac2 neurons suffices to drive precocious gonadal hyperstimulation, demonstrating that ERα in Tac2 neurons typically restrains pubertal onset and hypothalamic reproductive drive.

Ventral tegmental area neurotensin signaling links the lateral hypothalamus to locomotor activity and striatal dopamine efflux in male mice.

Projections from the lateral hypothalamic area (LHA) innervate components of the mesolimbic dopamine (MLDA) system, including the ventral tegmental area (VTA) and nucleus accumbens (NAc), to modulate motivation appropriately for physiologic state. Neurotensin (NT)-containing LHA neurons respond to multiple homeostatic challenges and project to the VTA, suggesting that these neurons could link such signals to MLDA function. Indeed, we found that pharmacogenetic activation of LHA NT neurons promoted prolonged DA-dependent locomotor activity and NAc DA efflux, suggesting the importance of VTA neurotransmitter release by LHA NT neurons for the control of MLDA function. Using a microdialysis-mass spectrometry technique that we developed to detect endogenous NT in extracellular fluid in the mouse brain, we found that activation of LHA NT cells acutely increased the extracellular concentration of NT (a known activator of VTA DA cells) in the VTA. In contrast to the prolonged elevation of extracellular NAc DA, however, VTA NT concentrations rapidly returned to baseline. Intra-VTA infusion of NT receptor antagonist abrogated the ability of LHA NT cells to increase extracellular DA in the NAc, demonstrating that VTA NT promotes NAc DA release. Thus, transient LHA-derived NT release in the VTA couples LHA signaling to prolonged changes in DA efflux and MLDA function.

Invasive Staphylococcus Aureus Infections in Diabetes Mellitus : Review

Staphylococcus aureus; the most virulent of the many staphylococcal species; has remained a major cause of morbidity and mortality despite the availability of numerous effective anti-staphylococcal antibiotics. S. aureus causes disease through both toxin-mediated and non-toxin-mediated mechanisms. This organism is responsible for both healthcare-associated and community-based infections ranging from relatively minor skin and soft tissue infections to severe life-threatening systemic infections. Patients with diabetes mellitus are at increased risk of invasive S aureus infections. This article focuses on the spectrum of invasive S aureus infections and discusses the clinical features; investigations and management of these infections in patients with diabetes mellitus

Lactobacillus plantarum 299v Prevents Caspase-Dependent Apoptosis In Vitro.

Selective microbes used as probiotics can enhance epithelial cell protection. We have previously shown that a Lactobacillus plantarum strain 299v (Lp299v) has the ability to induce mucin genes. In the current study, we utilized a cytokine model of inflammation in cell culture to study the modulation of apoptosis by this probiotic. HT-29 cells were pre-incubated with the Lp299v or L. plantarum strain adh- (Lpadh-), a non-adherent derivative of Lp299v. Cells were challenged with a mixture of cytokines (TNF-α, IFN-γ, and IL-1a) to imitate conditions of inflammation. To assess for cell death, we evaluated TUNEL, multi-caspase, and caspase-3 and caspase-7 activity assays. There was a marked decrease in apoptosis as measured by TUNEL(+) cells in samples pre-treated with Lp299v (18.7 ± 4.1%, p < 0.01) and Lpadh- (16.6 ± 3.2%, p < 0.05) prior to cytokine exposure when compared to cells (43.6 ± 6.2%) exposed to the cytokine mixture. Lp299v pre-incubation with HT-29 cells reduced caspase(+) cells in the multi-caspase activity assay (3.6 ± 0.6%, p < 0.05) compared to cells exposed to cytokines (68.9 ± 5.1%) whereas Lpadh- did not (46.8 ± 17.5%, p > 0.05). Similarly, caspase-3, caspase-7 activity was also reduced by Lp299v. Selected probiotics may confer an exogenous protective effect at the mucosal-luminal interface for intestinal epithelial cells via alteration of caspase-dependent apoptotic pathways.

Pulse probiotic administration induces repeated small intestinal Muc3 expression in rats.

Upon ingestion, probiotics may act to protect the host through a number of protective mechanisms including modulation of genes involved in intestinal innate mucosal defense such as epithelial cell-derived mucin glycoproteins and inhibitor of apoptosis proteins. To determine the specificity of effect and sustainability of response in vivo, Lactobacillus plantarum 299v (Lp299v), Lactobacillus rhamnosus R0011 (LrR0011), and Bifidobacterium bifidum R0071 (BbR0071) were added repeatedly or intermittently to the drinking water of Sprague-Dawley rats. After killing the rats via CO2 suffocation, Muc2, Muc3, neuronal apoptosis inhibitor protein (NAIP), human inhibitor of apoptosis protein 1/cellular inhibitor of apoptosis 2 (HIAP1/cIAP2), and human inhibitor of apoptosis protein 2/cellular inhibitor of apoptosis 1 (HIAP2/cIAP1) mRNA and protein levels were analyzed via RT-PCR and immunohistochemistry. Live Lp299v, BbR0071, and LrR0011 increased Muc3 protein and mRNA expression in jejunum and ileum. Heat-killed and a nonadherent derivative of Lp299v failed to induce Muc3 expression. Lp299v did induce expression of HIAP2/cIAP1 and NAIP expression. Muc3 mucin expression was elevated for 5 d after oral administration of Lp299v; however, this effect was not sustained despite ongoing daily ingestion of a probiotic. Intermittent pulse ingestion of probiotics, however, was found to repeatedly increase Muc3 expression. We conclude that selected probiotics can induce protective genes of mucosal intestinal epithelial cells, an effect that is reproducible with pulse probiotic administration.

Mcl-1 is a key regulator of apoptosis during CNS development and after DNA damage.

Despite the importance of Mcl-1, an anti-apoptotic Bcl-2 family member, in the regulation of apoptosis, little is known regarding its role in nervous system development and injury-induced neuronal cell death. Because germline deletion of Mcl-1 results in peri-implantation lethality, we address the function of Mcl-1 in the nervous system using two different conditional Mcl-1 mouse mutants in the developing nervous system. Here, we show for the first time that Mcl-1 is required for neuronal development. Neural precursors within the ventricular zone and newly committed neurons in the cortical plate express high levels of Mcl-1 throughout cortical neurogenesis. Loss of Mcl-1 in neuronal progenitors results in widespread apoptosis. Double labeling with active caspase 3 and Tuj1 reveals that newly committed Mcl1 deficient neurons undergo apoptosis as they commence migration away from the ventricular zone. Examination of neural progenitor differentiation in vitro demonstrated that cell death in the absence of Mcl1 is cell autonomous. Although conditional deletion of Mcl-1 in cultured neurons does not trigger apoptosis, loss of Mcl-1 sensitizes neurons to an acute DNA damaging insult. Indeed, the rapid reduction of Mcl-1 mRNA and protein levels are early events after DNA damage in neurons, and maintaining high Mcl-1 levels can protect neurons against death. Together, our results are the first to demonstrate the requirement of Mcl-1, an anti-apoptotic Bcl-2 family protein, for cortical neurogenesis and the survival of neurons after DNA damage.