Exploring associations between the teat apex metagenome and Staphylococcus aureus intramammary infections in primiparous cows under organic directives.

The primary objective of this study was to identify associations between the prepartum teat apex microbiome and the presence of Staphylococcus aureus intramammary infections (IMI) in primiparous cows during the first 5 weeks after calving. We performed a case-control study using shotgun metagenomics of the teat apex and culture-based milk data collected longitudinally from 710 primiparous cows on five organic dairy farms. Cases had higher odds of having S. aureus metagenomic DNA on the teat apex prior to parturition compared to controls (OR = 38.9, 95% CI: 14.84-102.21). Differential abundance analysis confirmed this association, with cases having a 23.8 higher log fold change (LFC) in the abundance of S. aureus in their samples compared to controls. Of the most prevalent microorganisms in controls, those associated with a lower risk of post-calving S. aureus IMI included Microbacterium phage Min 1 (OR = 0.37, 95% CI: 0.25-0.53), Corynebacterium efficiens (OR = 0.53, 95% CI: 0.30-0.94), Kocuria polaris (OR = 0.54, 95% CI: 0.35-0.82), Micrococcus terreus (OR = 0.64, 95% CI: 0.44-0.93), and Dietzia alimentaria (OR = 0.45, 95% CI: 0.26-0.75). Genes encoding for Microcin B17 AMPs were the most prevalent on the teat apex of cases and controls (99.7% in both groups). The predicted abundance of genes encoding for Microcin B17 was also higher in cases compared to controls (LFC 0.26). IMPORTANCE: Intramammary infections (IMI) caused by Staphylococcus aureus remain an important problem for the dairy industry. The microbiome on the external skin of the teat apex may play a role in mitigating S. aureus IMI risk, in particular the production of antimicrobial peptides (AMPs) by commensal microbes. However, current studies of the teat apex microbiome utilize a 16S approach, which precludes the detection of genomic features such as genes that encode for AMPs. Therefore, further research using a shotgun metagenomic approach is needed to understand what role prepartum teat apex microbiome dynamics play in IMI risk.

A randomized controlled trial evaluating the efficacy of systemic ceftiofur administration for metritis therapy in dairy cows and the effect of metritis cure on economically important outcomes.

The main objective of this study was to evaluate the effect of ceftiofur on metritis cure, milk yield, reproductive performance, and culling up to 300 DIM. The secondary objective was to evaluate the effect of metritis cure at 5 (ECURE) and 14 (LCURE) d after diagnosis on milk production, reproduction, and culling. A total of 422 Holstein cows diagnosed with metritis from 4 herds located in TX, CA, and FL were enrolled in a randomized clinical trial. Cows diagnosed with metritis (fetid, watery, reddish/brownish uterine discharge) were blocked by herd and parity and were randomly allocated to receive systemic administration of ceftiofur (CEF) or to remain untreated (CON). In addition, 399 non-metritic cows (NMET) were included for comparison purposes. Metritis cure was evaluated at 5 and 14 d after diagnosis and was defined as the absence of metritis clinical signs. Logistic regression models were fitted to the data to assess the effect of treatment on metritis cure. Milk yield was analyzed using a mixed linear model, while logistic regression, Cox proportional hazard and Kaplan-Meier survival analysis models were fitted to culling and reproduction data. Cows treated with CEF had 1.86 (95% CI: 1.22 - 2.81) and 1.68 (95% CI: 1.02 - 2.75) greater odds of being cured than CON cows at 5 and 14 d after diagnosis, respectively. No effect of CEF was observed for milk yield; however, NMET cows had greater milk yield compared with metritic cows (CEF = 36.0, 95% CI = 33.8 - 38.1; CON = 36.1, 95% CI = 33.9 - 38.2; NMET = 36.9 kg/d, 95% CI = 34.8 - 39.4). Likewise, no effect of CEF was observed on reproductive performance and culling. Nonetheless, the likelihood of conceiving for NMET cows was 1.72 (95% CI = 1.41 - 2.12) and 1.64 (95% CI = 1.33 - 2.00) times greater than for CEF and CON cows, respectively. Ceftiofur-treated and CON cows had 2.93 (95% CI = 1.90 - 4.51) and 2.37 (95% CI = 1.51 - 3.71) greater hazard of culling compared with NMET, respectively. Regardless of treatment, no differences between ECURE and LCURE were observed on milk yield, reproduction, and culling throughout the entire lactation, but cows that cured at 5 or 14 d after diagnosis had greater milk production in the first 60 DIM compared with cows that did not cure (NCURE). Cows in ECURE and LCURE also had a 1.59 (95% CI = 1.16 - 2.16) and 1.49 (95% CI = 1.08 - 2.05) greater hazard of pregnancy and 0.43 (95% CI = 0.26-0.71) and 0.56 (95% CI = 0.34-0.92) hazard of culling compared with NCURE. Ceftiofur therapy increased metritis cure, but benefits to productivity and longevity were not observed. Also, cows that fail to cure have impaired lactation performance, but no differences regarding timing of cure were observed.

Conjugated linoleic acid trans-10, cis-12 increases the expression of genes from cell cycle progression and cis-9, trans-11 stimulates apoptotic genes in different mammary tumor explants of female dogs.

BACKGROUND: The conjugated linoleic acid (CLA) isomer cis-9, trans-11 is an anticarcinogen that inhibits cell proliferation and/or induces apoptosis of tumor cells. The objective of this study was to evaluate the expression of genes responsible for cell cycle regulation and apoptosis in tumor explants of mammary anaplastic carcinoma (AC) and mammary tubulopapillary carcinoma (TC) cultured in vitro with the CLA isomers cis-9, trans-11 and trans-10, cis-12. METHODS: In this study we used mammary explants from two adult female dogs that revealed two types of malignant tumors: (a) anaplastic mammary carcinoma (AC) and (b) mammary tubulopapillary carcinoma (TC). The explants (n = 6 per treatment) had an average weight of 80.0 ± 2.0 mg and were cultured for 24 h in 35 mm culture plates under the following treatments: (a) Control: Culture medium + fatty acid free bovine serum albumin (BSA); (b) Culture medium + cis-9, trans-11 CLA (75 µM) diluted with fatty acid free bovine serum albumin (BSA), and; (c) Culture medium + trans-10, cis-12 CLA (75 µM) diluted with fatty acid free bovine serum albumin (BSA). After that, total RNA was extracted, complementary DNA was synthesized (cDNA), and quantitative analysis by real-time polymerase chain reaction (RT-qPCR) was conducted. Data were analyzed using the MIXED procedure of SAS. RESULTS: Compared with the Control, the CLA trans-10, cis-12 treatment decreased expression of the gene encoding the p53 by 20% (P = 0.02), Caspase-3 by 25% (P = 0.06) and Bax by 51% (P = 0.001) in AC. The CLA cis-9, trans-11 increased the gene expression of proapoptotic protein Bax in TC by 68% (P = 0.01), but increased the expression of the antiapoptotic Bcl2 gene in AC by 72% (P = 0.006). CONCLUSION: The CLA cis-9, trans-11 stimulates apoptotic genes in mammary tubulopapillary carcinoma, but has a contrary effect on the anaplastic carcinoma, and the CLA trans-10, cis-12 stimulates cell cycle progression genes and may have an antiapoptotic effect, mainly in mammary anaplastic carcinoma.

Influence of Deep Sedation in Intensive Care Medicine Memories of Critical COVID-19 Survivors.

Introduction: Critical care survivors sustain a variety of sequelae after intensive care medicine (ICM) admission, and the Coronavirus Disease 2019 (COVID-19) pandemic has added further challenges. Specifically, ICM memories play a significant role, and delusional memories are associated with poor outcomes post-discharge including a delayed return to work and sleep problems. Deep sedation has been associated with a greater risk of perceiving delusional memories, bringing a move toward lighter sedation. However, there are limited reports on post-ICM memories in COVID-19, and influence of deep sedation has not been fully defined. Therefore, we aimed to evaluate ICM-memory recall in COVID-19 survivors and their relation with deep sedation. Materials/Methods: Adult COVID-19 ICM survivors admitted to a Portuguese University Hospital between October 2020 and April 2021 (second/third "waves") were evaluated 1 to 2 months post-discharge using "ICU Memory Tool," to assess real, emotional, and delusional memories. Results: The study included 132 patients (67% male; median age = 62 years, Acute Physiology and Chronic Health Evaluation [APACHE]-II = 15, Simplified Acute Physiology Score [SAPS]-II = 35, ICM stay = 9 days). Approximately 42% received deep sedation (median duration = 19 days). Most participants reported real (87%) and emotional (77%) recalls, with lesser delusional memories (36.4%). Deeply sedated patients reported significantly fewer real memories (78.6% vs 93.4%, P = .012) and increased delusional memories (60.7% vs 18.4%, P < .001), with no difference in emotional memories (75% vs 80.4%, P = .468). In multivariate analysis, deep sedation had a significant, independent association with delusional memories, increasing their likelihood by a factor of approximately 6 (OR = 6.274; 95% confidence interval = 1.165-33.773, P = .032), without influencing real (P = .545) or emotional (P = .133) memories. Conclusions: This study contributes to a better understanding of the potential adverse effects of deep sedation on ICM memories in critical COVID-19 survivors, indicating a significant, independent association with the incidence of delusional recalls. Although further studies are needed to support these findings, they suggest that strategies targeted to minimize sedation should be favored, aiming to improve long-term recovery.

Ent-hardwickiic acid from C. pubiflora and its microbial metabolites are more potent than fluconazole in vitro against Candida glabrata.

The incidence of Candida glabrata infections has rapidly grown and this species is among those responsible for causing invasive candidiasis with a high mortality rate. The diterpene ent-hardwickiic acid is a major constituent in Copaifera pubiflora oleoresin and the ethnopharmacological uses of this oleoresin by people from Brazilian Amazonian region point to a potential use of this major constituent as an antimicrobial. Therefore, the goal of this study was to evaluate the antifungal activity of ent-hardwickiic acid against Candida species and to produce derivatives of this diterpene by using microbial models for simulating the mammalian metabolism. The microbial transformations of ent-hardwickiic acid were carried out by Aspergillus brasiliensis and Cunninghamella elegans and hydroxylated metabolites were isolated and their chemical structures were determined. The antifungal activity of ent-hardwickiic acid and its metabolites was assessed by using the microdilution broth method in 96-well microplates and compared with that of fluconazole. All the diterpenes showed fungistatic effects (ranging from 19·7 to 75·2 µmol l-1 ) against C. glabrata at lower concentrations than fluconazole (163·2 µmol l-1 ) and were more potent fungicides (ranging from 39·5 to 150·4 µmol l-1 ) than fluconazole, which showed fungicidal effect at the concentration of 326·5 µmol l-1 .

Effect of injectable trace mineral supplementation on peripheral polymorphonuclear leukocyte function, antioxidant enzymes, health, and performance in dairy cows in semi-arid conditions.

The objective of this study was to evaluate the effect of subcutaneous injections of 15 mg/mL Cu, 5 mg/mL Se, 60 mg/mL Zn, and 10 mg/mL Mn on health, performance, polymorphonuclear leukocyte (PMNL) function, circulating glutathione peroxidase (GPx) and superoxide dismutase (SOD) concentrations, and inflammation of dairy cows undergoing the transition period in high temperature-humidity index. A total of 923 multiparous cows from 2 commercial dairy farms were randomly allocated into 1 of 2 treatment groups as follows: control and injectable trace mineral supplementation (ITMS). Cows in the ITMS group received 7 mL of subcutaneous injections at dry-off (208 ± 3 d of gestation), 260 ± 3 d of gestation, and at 35 ± 3 d in milk (DIM). Data regarding health traits, reproductive performance, milk yield, and survivability were extracted from farm database software, and animals were followed-up until 300 DIM. For a subset of 142 cows from one herd, blood samples were collected at enrollment, and at 3 ± 1, 7 ± 1, 10 ± 1, and 35 ± 3 DIM to evaluate hematology, PMNL function, GPx and SOD concentrations, and circulating haptoglobin. Logistic regression was used to assess health and pregnancy per artificial insemination at first service. Cox proportional hazards models were used to evaluate hazard of pregnancy and culling. Mixed linear regression models accounting for repeated measures were used to assess all continuous variables collected over time. Parity, twinning, and previous gestation length were considered as potential confounders. Farm was included as a random effect. The ITMS cows tended to have lower incidence of metritis and stillbirth compared with control group. However, ITMS treatment did not influence the incidence of other diseases (e.g., mastitis, retained placenta), milk yield, reproductive performance, culling, and leukocyte count. Neutrophil-to-lymphocyte ratio, PMNL phagocytosis, and oxidative burst as well as intensity of the oxidative burst were greater for ITMS-treated cows in comparison to control cows. The ITMS cows had decreased expression of the adhesion molecule L-selectin on PMNL surface. The serum concentration of GPx and SOD were not affected by ITMS treatment. In conclusion, ITMS tended to reduce the incidence of metritis and stillbirth parturition, improved PMNL function, and improved the inflammatory status of dairy cows undergoing the transition period in high temperature-humidity index conditions. However, these findings did not translate into improved milk yield, reproductive performance, and survivability.

The scientific literature on bats (chiroptera) in Brazil: a scientometric analysis from 1954 - 2018.

Globally, bats are of interest in many studies, beyond their ecological or epidemiological relevance or even the ecomorphological diversity of species. In Brazil, most of the indexed studies on chiropterans date from 1954, with a slow and heterogeneous progress in the publication increasing. The aim of this study was to analyze the literature on bats in Brazil, identifying patterns, tendencies and knowledge gaps in the Brazilian federal states. We carried out a sistematyzed search on the online databases Clarivate Analytics Web of Science (WoS), Scientific Electronic Library Online (SciELO), PubMed and Scopus. We used the descriptive terms "Chiroptera" and "Brazil". Besides these bases, we included data from the manuscripts published in Chiroptera Neotropical. We obtained a total of 1,115 articles, which were analyzed and classified in 22 thematic categories based on the articles' approach. We observed that each Brazilian region and state had particularities in their knowledge panoramas of bats, not being possible to generalize conditions for each federal region. Even though the increasing in the number of articles by categories, we encourage that every approach keep being developded, once no thematic could had been considered enough explored till the moment.

Effect of subclinical mastitis detected in the first month of lactation on somatic cell count linear scores, milk yield, fertility, and culling of dairy cows in certified organic herds.

It is well established that subclinical mastitis (SCM), characterized by somatic cell count (SCC) >200,000 cells/mL, has a negative effect on the productivity, reproductive performance, and survivability of cows from conventional dairy herds. However, in organic herds, where the use of antimicrobial drugs is restricted for the treatment and control of intramammary infections (IMI) in dairy cows, little is known about the effect of SCM on performance and survivability. The objective of this study was to evaluate whether SCM diagnosed during the first month of lactation was associated with SCC linear score dynamics, milk production, fertility, and culling of dairy cows in USDA-certified organic herds. We collected data from 2 organic herds in New Mexico and Texas. A total of 1,511 cows that calved between June 2018 and May 2019 were included in the study and were followed until month 10 of the current lactation. Cows with SCC >200,000 cells/mL in the first month of lactation were considered to have SCM. We used mixed linear regression models accounting for repeated measures to assess the effect of SCM on monthly milk production and SCC linear scores. We used Cox proportional hazards models to evaluate the effect of SCM on the risk of pregnancy and culling. We considered parity, farm, previous gestation length, stillbirth, twinning, dystocia, and 2- and 3-way interactions as potential confounders. Cows diagnosed with SCM during the first month of lactation produced less milk than cows without SCM. Cows with SCM had elevated SCC linear scores during their previous lactation and throughout the subsequent months of lactation compared to cows without SCM. The effect of SCM on SCC linear scores was more pronounced in multiparous than primiparous cows. Subclinical mastitis during the first month of lactation did not affect the likelihood of pregnancy during the first 300 d in milk. Cows with SCM in the first month were more likely to die or be culled during the 300 d of lactation than cows without SCM. We observed that elevated SCC in the first month of lactation had detrimental effects on the milk yield and survivability of dairy cows in USDA organic herds, but it did not affect reproductive performance. We demonstrated that cows with SCM diagnosed in the first month of lactation continued to have elevated SCC linear scores throughout their entire lactation, and that elevated SCC was carried over from the previous lactation.

Prolonged, low-grade inflammation in the first week of lactation: Associations with mineral, protein, and energy balance markers, and milk yield, in a clinically healthy Jersey cow cohort.

Our objectives were to perform a proof-of-concept study to assess the association of a prolonged inflammatory state (based on a continually elevated haptoglobin concentration at the end of the first week after parturition) with mineral, protein, and energy balance markers in the first 2 wk after parturition, and milk production in the first 60 d of lactation in clinically healthy cows. We conducted a cohort study in 1 herd in west Texas that was milking Jersey and Jersey-Holstein crosses. Only multiparous Jersey cows were eligible for enrollment. Cows were classified as having or not having elevated haptoglobin concentrations based on plasma concentrations evaluated on d 4 and 7 postpartum. We used median concentrations of haptoglobin in the reference population (i.e., before the exclusion of cows diagnosed with clinical diseases) as the limits for categorizing cows into 2 groups: cows with plasma haptoglobin concentrations greater than the median values on both d 4 (0.49 g/L) and 7 (0.35 g/L) had continually elevated haptoglobin (with eHp); and cows with plasma haptoglobin concentrations lower than or equal to the median values of the reference population on d 4 or 7 did not have continually elevated haptoglobin (without eHp). Next, cows with clinical diseases in the first 2 wk of the postpartum period were excluded, so that 233 cows remained for the final analyses. Evaluated outcomes were average daily milk production across the first 60 d of lactation, plasma Ca, Mg, and glucose concentrations on d 1, 2, 3, 4, 5, 7, 10, and 14 postpartum, and ß-hydroxybutyrate (BHB), nonesterified fatty acids (NEFA), fructosamine, albumin, urea, and creatinine concentrations on d 3, 5, 7, 10, and 14 postpartum. Rectal temperatures measured on d 4, 7, and 10 postpartum were also analyzed. We performed statistical analyses using linear mixed models while accounting for the repeated effect of sampling time (plasma analytes and rectal temperature models) and weekly milk test (milk production model). Cows with eHp had lower plasma Ca concentrations in the first 2 wk after calving, but no differences in Mg, BHB, NEFA, glucose, or rectal temperatures compared to cows without eHp. Cows with eHp had lower plasma fructosamine, albumin, and urea concentrations in a time-dependent manner. They also had lower milk production (2.3 kg/d less than cows without eHp in the first 60 DIM). Our study demonstrated that 25% of cows without clinical disorders in the first 2 wk after parturition had continually elevated haptoglobin concentrations at d 7 after parturition relative to d 4, suggestive of a prolonged, low-grade systemic inflammatory state. More observational studies are needed to more fully characterize the duration of prolonged postpartum subclinical inflammation in cows without clinical diseases, as well as its long-term effects, and to evaluate the use of other potential markers of systemic inflammation to describe this disorder.

Short communication: Associations of serum biomarkers of stress and inflammation measured at arrival with health, mortality, and growth of calves transported within the first 4 days of life.

The objective of this study was to assess the association between serum concentrations of haptoglobin (Hp), cortisol, and l-lactate following transport of dairy heifer calves within their first week of life with health, mortality, and growth during the preweaning period. Blood samples were collected at arrival and included a cohort of 168 Jersey and Jersey-cross calves from 8 different sources located in Minnesota. All calves were raised at a single facility in New Mexico. Incidence of respiratory disease (BRD) and mortality during the preweaning period (60 d of life) were extracted from the farm's software database. Individual body weight was measured at birth and at weaning to estimate average daily gain. No simple linear correlations were found between the biomarkers. The incidence of BRD during the preweaning period was 7.7%. While cortisol and l-lactate serum concentrations were not associated with BRD, Hp was negatively associated with BRD. Receiving operating characteristic curve analysis was performed to determine the optimal threshold value for Hp [threshold ≤0.63 µmol/L (63 µg/mL), area under the curve = 0.65]. A Cox's proportional hazards model revealed that calves with Hp concentration ≤0.63 µmol/L were more likely to be diagnosed with BRD (hazard ratio = 5.0, 95% confidence interval = 1.3-19.0). A mixed linear model showed that calves with Hp concentration ≤0.63 µmol/L tended to have lower average daily gain (454.4 vs. 479.9 g/d) during the preweaning period than calves with Hp >0.63 µmol/L at arrival. Overall mortality of the cohort was 3.5%, and Hp was not associated with mortality. Although circulating concentrations of l-lactate and cortisol measured at arrival were not associated with BRD incidence during the preweaning period of heifer calves transported within the first 4 d of life, calves with serum Hp concentration >0.63 µmol/L were less likely to be diagnosed with BRD and tended to grow more than calves with Hp ≤0.63 µmol/L. Further research is needed to replicate these results in a larger cohort and to better understand the possible influence of greater inflammatory status at arrival on health and growth of calves transported to a calf-raising facility.

Association of blood calcium concentration in the first 3 days after parturition and energy balance metabolites at day 3 in milk with disease and production outcomes in multiparous Jersey cows.

Research exploring specific associations of markers of negative energy balance and Ca in postpartum Jersey cows with lactation performance is lacking. Our objectives were to evaluate the associations of total Ca concentration (tCa) measured at 1 through 3 d in milk (DIM) and free fatty acids (FFA), ß-hydroxybutyrate (BHB), and glucose measured at 3 DIM with (1) the risk of multiparous Jersey cows being diagnosed with early-lactation diseases and culling, (2) milk production in the first 9 wk of lactation, and (3) the risk of pregnancy in the first 150 DIM. A cohort study was performed in 1 dairy herd in Texas. Multivariable Poisson regression models were built to evaluate the association of the analytes of interest with the risks of early-lactation diseases and culling in the first 60 DIM (i.e., binary outcomes). Linear mixed models were used to evaluate the association of the analytes of interest with milk production within the first 9 wk of lactation, and a Cox proportional hazard model was built to assess the risk of pregnancy within 150 DIM. A total of 380 cows were used in the final analyses. Total Ca measured at 1 through 3 DIM was not associated with the risk of metritis. Cows with increased FFA and BHB had an increased risk of being diagnosed with metritis and clinical mastitis, respectively. Increased concentrations of glucose and FFA and decreased tCa at 3 DIM were associated with an increased risk of culling. Reduced tCa concentrations at 1 DIM (≤1.84 mmol/L) and 2 DIM (≤2.04 mmol/L) were associated with increased milk production across the first 9 wk of lactation compared with tCa concentrations above those thresholds. Total Ca was not associated with milk production when assessed at 3 DIM, whereas increased FFA (≥0.37 mmol/L) and decreased glucose (≤2.96 mmol/L) at 3 DIM were associated with increased milk production. None of the metabolites measured were associated with the risk of pregnancy in the first 150 DIM. Our results demonstrate that tCa concentration assessed in the first 3 DIM show temporary associations with milk production and culling in multiparous Jersey cows. Although increased concentration of FFA assessed at 3 DIM was associated with greater milk yield, it was a detrimental factor for the risk of metritis. This study attempted to better elucidate the relationship of tCa, FFA, BHB, and glucose assessed in early postpartum with health and performance of Jersey cows. Based on this study, assessments performed at 3 DIM using tCa concentration ≤1.99 mmol/L for increased risk of early-lactation culling and FFA ≥0.43 mmol/L for increased risk of metritis could be used as starting points. More studies evaluating the dynamics of energy balance markers and tCa in postpartum Jersey cows using a greater number of herds are needed to better inform dairy consultants on critical levels for exacerbated postpartum negative energy balance and subclinical hypocalcemia for the Jersey breed.

Frontal sinus as an identity and sex indicator.

The present study explores the variability of frontal sinuses volume in modern Greeks and their potential use in the forensic identification process. METHODS: The sample consisted of 102 CT scan images from a modern Greek population. 50 (49%) were male and 52 (51%) were female. Ages ranged from 19 to 101 (mean age 70.6 for males and 69.6 for females). 3D models of the skull were constructed, and volume calculations of the frontal sinus were done with the software Amira 5.4. 10 CT scans were used to extract 2D frontal and lateral images that were used as proxies of antemortem X-rays which were matched to the CT scans using manual craniofacial superimposition. RESULTS: The average of frontal sinus volume of male was 9,220 mm3 (±SD) and female was 5,880 mm3 (±SD). Mann-Witney test confirmed that mean values were significantly different (P<0.05) between them. Pearson's correlation coefficient showed no evident correlation between the volumes and ages of the skulls in both groups. Using the lateral and anterior oriented radiographs for matching the sinuses, the method produced 60% sensitivity / 99.32% specificity and 80% sensitivity / 99.97% specificity, respectively. CONCLUSIONS: Even though there are statistically significant differences in the shape and volume between sexes, these are not sufficient to be used as an indicator of sex in worldwide populations. The results indicate that using the anterior view of the frontal sinuses is more consistent in human identification, and that the method proves to be reliable, as long as the sinus is adequately observed on the radiograph.

Short communication: Characterization of molasses chemical composition.

Beet and cane molasses are produced worldwide as a by-product of sugar extraction and are widely used in animal nutrition. Due to their composition, they are fed to ruminants as an energy source. However, molasses has not been properly characterized in the literature; its description has been limited to the type (sugarcane or beet) or to the amount of dry matter (DM), total or water-soluble sugars, crude protein, and ash. Our objective was to better characterize the composition of cane and beet molasses, examine possible differences, and obtain a proper definition of such feeds. For this purpose, 16 cane and 16 beet molasses samples were sourced worldwide and analyzed for chemical composition. The chemical analysis used in this trial characterized 97.4 and 98.3% of the compounds in the DM of cane and beet molasses, respectively. Cane molasses contained less DM compared with beet molasses (76.8 ± 1.02 vs. 78.3 ± 1.61%) as well as crude protein content (6.7 ± 1.8 vs. 13.5 ± 1.4% of DM), with a minimum value of 2.2% of DM in cane molasses and a maximum of 15.6% of DM in beet molasses. The amount of sucrose differed between beet and cane molasses (60.9 ± 4.4 vs. 48.8 ± 6.4% of DM), but variability was high even within cane molasses (39.2-67.3% of DM) and beet molasses. Glucose and fructose were detected in cane molasses (5.3 ± 2.7 and 8.1 ± 2.8% of DM, respectively), showing high variability. Organic acid composition differed as well. Lactic acid was more concentrated in cane molasses than in beet molasses (6.1 ± 2.8 vs. 4.5 ± 1.8% of DM), varying from 1.6 to 12.8% of DM in cane molasses. Dietary cation-anion difference showed numerical differences among cane and beet molasses (7 ± 53 vs. 66 ± 45 mEq/100 g of DM, on average). It varied from -76 to +155 mEq/100 g of DM in the cane group and from +0 to +162 mEq/100 g of DM in the beet group. Data obtained in this study detailed differences in composition between sources of molasses and suggested that a more complete characterization could improve the use of molasses in ration formulation.

Acute and sub-acute oral toxicity studies of standardized extract of Nasturtium officinale in Wistar rats.

Nasturtium officinale (watercress) is a perennial dicotyledonous plant, rich in vitamins, minerals and chemical compounds. The leaves of this plant, which contain glucosinolate, are used for its diuretic and hypoglycemic effects. The purpose of the study was to investigate the safety of the standardized extract of Nasturtium officinale (SENO) with phenylethyl glucosinolate 5.0 mg/ml-1, using acute and sub-acute oral dosage in Wistar rats. High-Performance Liquid Chromatography (HPLC) analyzed the chemical composition, from aerial parts of watercress. In the acute toxicity study, dose estimated was LD50 in the range of 2000-5000 mg/kg, signs of mortality and toxicity on female rats were observed for 14 days, after single doses of 2000 and 5000 mg/kg. In the sub-acute study, female and male rats, age 10 weeks, were supplemented at doses of 250, 500 and 1000 mg/kg for 28 days. On the 29th day, rats were fasted, anesthetized, euthanized, then their blood used for hematological and biochemical evaluation. No significant changes in general behavior were reported regarding the acute study, while the sub-acute study demonstrated no toxicity of the hematopoietic and biochemical systems. The results showed that SENO at dosage up to 5000 mg/kg in acute study was safe, and NOAEL (no-observed-adverse-effect levels) in the sub-acute, was up to 1000 mg/kg.

Demonstration of Single-Barium-Ion Sensitivity for Neutrinoless Double-Beta Decay Using Single-Molecule Fluorescence Imaging.

A new method to tag the barium daughter in the double-beta decay of ^{136}Xe is reported. Using the technique of single molecule fluorescent imaging (SMFI), individual barium dication (Ba^{++}) resolution at a transparent scanning surface is demonstrated. A single-step photobleach confirms the single ion interpretation. Individual ions are localized with superresolution (∼2 nm), and detected with a statistical significance of 12.9σ over backgrounds. This lays the foundation for a new and potentially background-free neutrinoless double-beta decay technology, based on SMFI coupled to high pressure xenon gas time projection chambers.

Multicentre evaluation of a direct agglutination test prototype kit (DAT-LPC) for diagnosis of visceral leishmaniasis.

In this study, we assessed the sensitivity, specificity, and diagnostic accuracy of a previously developed direct agglutination test (DAT) using a freeze-dried antigen derived from Leishmania infantum promastigotes and composed in a prototype kit for visceral leishmaniasis (VL) diagnosis, named DAT-LPC. To evaluate DAT-LPC reproducibility, the kit was used to analyse 207 serum samples from VL patients and 80 serum samples from patients with other parasitic infections or healthy subjects in four laboratories from different public health institutions in Brazil. DAT-LPC showed sensitivity between 96·2 and 99·5% (P = 0·14), specificity ranging from 96·2 to 97·5% (P = 0·95), and diagnostic accuracy ranging from 96·5 to 99% (P = 0·34). The inter-laboratory reproducibility of qualitative results was classified as excellent (κ index: 0·94-0·97). The reproducibility of the end-titre results in relation to the reference laboratory, ranged from 31 to 85%. These results demonstrate an excellent performance of the DAT-LPC, and validate it for the diagnosis of VL that could replace the immunofluorescent antibody test as the routine diagnostic test in the Brazilian public health system.

Wheat gluten intake increases weight gain and adiposity associated with reduced thermogenesis and energy expenditure in an animal model of obesity.

BACKGROUND/OBJECTIVES: The association between gluten and body weight is inconsistent. Previously, we showed that a gluten-free diet reduces weight gain without changing food intake in mice fed high-fat diets. In the present study, we investigated the effects of gluten intake on fat metabolism, thermogenesis and energy expenditure in mice fed a standard or high-fat diet. METHODS: Mice were fed four different experimental diets during 8 weeks: a control-standard diet (CD), a CD added with 4.5% of wheat gluten (CD-G), a high-fat diet (HFD) and a HFD added with 4.5% of wheat gluten (HFD-G). After 8 weeks, the mice received (99m)Tc-radiolabeled gluten orally to study gluten absorption and biodistribution or they underwent indirect calorimetry. After killing, subcutaneous and brown adipose tissues (SAT and BAT) were collected to assess thermogenesis-related protein expression. Lipid metabolism was studied in adipocyte cultures from the four groups. RESULTS: Despite having had the same energy intake, CD-G and HFD-G mice exhibited increased body weight and fat deposits compared with their respective controls. (99m)Tc-GLU or its peptides were detected in the blood, liver and visceral adipose tissue, suggesting that gluten can even reach extraintestinal organs. Uncoupling protein-1 expression was reduced in the BAT of HFD-G and in the SAT of CD-G and HFD-G mice. Indirect calorimetry showed lower oxygen volume consumption in CD-G and HFD-G groups compared with their controls. In HFD mice, daily energy expenditure was reduced with gluten intake. Gluten also reduced adiponectin, peroxisome proliferator-activated receptor (PPAR)-α and PPARγ and hormone-sensitive lipase in cultures of isolated adipocytes from HFD mice, whereas in the CD-G group, gluten intake increased interleukin-6 expression and tended to increase that of tumor necrosis factor. CONCLUSIONS: Wheat gluten promotes weight gain in animals on both HFD and CD, partly by reducing the thermogenic capacity of adipose tissues.

Clinical algorithms to aid osteoarthritis guideline dissemination.

BACKGROUND: Numerous scientific organisations have developed evidence-based recommendations aiming to optimise the management of osteoarthritis (OA). Uptake, however, has been suboptimal. The purpose of this exercise was to harmonize the recent recommendations and develop a user-friendly treatment algorithm to facilitate translation of evidence into practice. METHODS: We updated a previous systematic review on clinical practice guidelines (CPGs) for OA management. The guidelines were assessed using the Appraisal of Guidelines for Research and Evaluation for quality and the standards for developing trustworthy CPGs as established by the National Academy of Medicine (NAM). Four case scenarios and algorithms were developed by consensus of a multidisciplinary panel. RESULTS: Sixteen guidelines were included in the systematic review. Most recommendations were directed toward physicians and allied health professionals, and most had multi-disciplinary input. Analysis for trustworthiness suggests that many guidelines still present a lack of transparency. A treatment algorithm was developed for each case scenario advised by recommendations from guidelines and based on panel consensus. CONCLUSION: Strategies to facilitate the implementation of guidelines in clinical practice are necessary. The algorithms proposed are examples of how to apply recommendations in the clinical context, helping the clinician to visualise the patient flow and timing of different treatment modalities.

Tuning the Stoichiometry of Ag2S Thin Films for Resistive Switching Applications.

In this work silver-rich and sulfur-rich silver sulfide (Ag2S) thin films were fabricated using a diversified set of experimental methods, namely ion beam deposition and atmosphere- and solution-based sulfurizations. The composition of the Ag2S thin films was studied using X-ray diffraction, Raman spectroscopy, scanning electron microscopy and energy dispersive X-ray spectroscopy. We found that it strongly depends on the fabrication conditions, such as sulfurization time and temperature. These conditions, in turn, affect the electrical characteristics of the thin films, namely the resistivity and resistive switching. We were able to control the Ag2S stoichiometry and infer its dependence on the fabrication parameters for all the followed methods.

Polymorphisms and alternative splicing of the luteinizing hormone receptor of dairy cattle.

The aim of this study was to screen for variability in the luteinizing hormone/choriogonadotropin receptor (LHCGR) and to determine the occurrence of LHCGR mRNA isoforms in two dairy breeds of cattle. Granulosa cells from dominant ovarian follicles were recovered from 16 Gir and 16 Holstein cows, and total RNA was extracted. Complementary DNA was synthesized and PCR was used to generate amplicons for sequencing. Chromatograms were evaluated, and multiple sequences were aligned and analyzed for the presence of polymorphisms, allele frequency, polymorphic information content (PIC), and Hardy-Weinberg equilibrium (HWE). Twenty-one single nucleotide polymorphisms (SNPs) were identified in LH receptor mRNA. Seventeen SNPs were identified in Gir cattle (seven exclusively), and 14 were found in Holstein cattle (four exclusively). Seven of the 21 polymorphisms found did not alter which amino acid was translated. Eight SNPs caused a change to an amino acid in a different chemical group. Classification of SNPs according to PIC values identified 12 as being highly informative in Gir cattle and five in Holstein. Eight SNPs deviated from HWE in Gir compared with 11 in Holstein, and eight in both breeds. Two isoforms were also identified, one in exon 1, which lacks 30 nucleotides beginning at position 118, and the other in exon 10. Taken together, these data show that LHCGR in dairy cattle breeds has a high frequency of polymorphism and exists in multiple isoforms resulting from alternative splicing.