Effect of supplementation with vitamins D3 and K2 on undercarboxylated osteocalcin and insulin serum levels in patients with type 2 diabetes mellitus: a randomized, double-blind, clinical trial.

BACKGROUND: Patients with type 2 diabetes mellitus (T2DM) are characterized by chronic hyperglycemia as a consequence of decreased insulin sensitivity, which contributes to bone demineralization and could also be related to changes in serum levels of osteocalcin and insulin, particularly when coupled with a deficiency in the daily consumption of vitamins D3 and K2. The objective of this study was to evaluate the effect of vitamin D3 and vitamin K2 supplements alone or in combination on osteocalcin levels and metabolic parameters in patients with T2DM. METHODS: A double-blind, randomized clinical trial was carried out in 40 patients aged between 30 and 70 years old for 3 months. Clinical and laboratory assessment was carried out at the beginning and at the end of the treatment. The patients were divided into three groups: (a) 1000 IU vitamin D3 + a calcinated magnesium placebo (n = 16), (b) 100 µg of Vitamin K2 + a calcinated magnesium placebo (n = 12), and (c) 1000 IU vitamin D3 + 100 µg vitamin K2 (n = 12). RESULTS: After treatment in the total studied population, a significant decrease in glycemia (p = 0.001), HOMA-IR (Homeostatic model assessment-insulin resistance) (p = 0.040), percentage of pancreatic beta cells (p < 0.001), uOC/cOC index and diastolic blood pressure (p = 0.030) were observed; in vitamin D3 group, differences in serum undercarboxylated osteocalcin (p = 0.026), undercarboxylated to carboxylated osteocalcin index (uOC/cOC) (p = 0.039) glucose (p < 0.001) and  % of functional pancreatic beta cells (p < 0.001) were demonstrated. In vitamin K2 group a significant decrease in glycemia (p = 0.002), HOMA-IR (p = 0.041), percentage of pancreatic beta cells (p = 0.002), and in cOC (p = 0.041) were observed, conversely cOC concentration was found high. Finally, in the vitamins D3 + K2 a significant decrease in glycemia (p = 0.002), percentage of pancreatic beta cells (p = 0.004), and in the uOC/cOC index (p = 0.023) were observed. CONCLUSION: Individual or combined supplementation with vitamins D3 and K2 significantly decreases the glucose levels and  % of functional pancreatic beta cells, while D3 and D3 + K2 treatments also induced a reduction in the uOC/cOC index. Only in the group with vitamin D3 supplementation, it was observed a reduction in undercarboxylated osteocalcin while vitamin K2 increased the carboxylated osteocalcin levels.Trial registration NCT04041492.

Enzyme-linked immunosorbent assay as complement of intradermal skin test for the detection of mycobacterium bovis infection in cattle.

Diagnostic tests based on cell-mediated immunity are used in programs for the control and eradication of bovine tuberculosis (bTB), which is mainly caused by Mycobacterium bovis. Additional serological assays could be performed as an ancillary method to detect an infected animal that fails to produce an immune response against the intradermal reaction (IDR), the official bTB test. In this study, we evaluated the effectiveness of an enzyme-linked immunosorbent assay (ELISA) that uses bovine PPD as a capture antigen as a complement to the IDR in herds with confirmed cases of bTB. The study was conducted in two stages. First, a panel of 200 serum samples was analyzed by ELISA. The sensitivity and specificity obtained were 60% and 99%, respectively. The subsequent stage consisted of evaluating 7,494 bovines from 14 selected dairy farms. The number of animals yielding a IDR negative/ELISA positive result were 200. A necropsy analysis of 33 of these IDR negative/ELISA positive animals revealed that 30 (91%) presented granulomatous lesions and positive M. bovis isolation. This finding confirmed bTB in most cases. Altogether, the results obtained in the present study suggest that the combined use of IDR and ELISA is an effective strategy to improve the control of bTB in endemic herds.

CARDIOVASCULAR RISK FACTORS IN ADOLESCENTS: ROLE OF INSULIN RESISTANCE AND OBESITY.

INTRODUCTION: Childhood obesity is a public health problem characterized by early insulin resistance (IR), inflammation, and oxidative stress. The presence of an uninterrupted low-grade inflammatory state impairs metabolic and cardiovascular health. The population is particularly susceptible to develop metabolic disorders related to increased body fat. METHODS: Eighty-three adolescents were recruited and grouped according to HOMA-IR and BMI in either with or without IR and obese or normal-weight respectively. Anthropometric, biochemical, immunological and hormonal variables were determined. Transverse Analytical Study. RESULTS: Obesity, dyslipidemia, IL-6, and C-reactive protein were significantly higher in the IR group than in the non-IR group. Obese adolescents showed increased insulin levels, HOMA-IR, inflammatory markers, and triglycerides; while having lower HDL-C, and adiponectin when compared to normal-weight adolescents. As expected, obesity-related anthropometric markers positively correlated with IR and inflammatory markers while negatively correlated with adiponectin levels. CONCLUSIONS: Early IR, subclinical inflammation, dyslipidemia, and hypoadiponectinemia characterize obesity in adolescents. These factors may increase the risk of future coronary heart disease (CHD) and diabetes mellitus development (DM) in early adulthood.

Lipid droplet analysis using in vitro bovine oocytes and embryos.

The aim of this study was to quantify the content of lipid droplets in bovine oocytes and embryos from Bos indicus (Bi), Bos taurus (Bt) and Bos indicus × Bos taurus (Bi × Bt). Oocytes were aspirated post-mortem and subjected to in vitro maturation, in vitro fertilization and in vitro development; the medium employed at each stage (TCM-199, TALP, SOF) was supplemented with (i) serum replacement (SR), (ii) foetal calf serum (FCS) or (iii) oestrous cow serum (ECS). The structure and distribution of the lipid droplets were established using electron microscopy, but were quantified using an optical microscope on semi-fine toluidine blue-stained sections. The highest percentage of embryos corresponded to those produced with FCS and ECS, which differed from embryos generated with SR (p < 0.05). The highest percentage of morulae and the lowest percentage of blastocysts were obtained with the SR supplement (p < 0.05). The oocytes cultured in FCS demonstrated a higher number of lipid droplets compared to those cultured in SR and ECS (p < 0.05). Less accumulation of lipids was observed in embryos supplemented with SR. The lowest and highest numbers of lipid droplets in oocytes corresponded to the Bi and Bt strain, respectively. The lowest amount of lipid droplets in embryos was observed in Bi (p < 0.05). In conclusion, supplementation of the in vitro development culture medium (synthetic oviduct fluid) with a synthetic substitute serum produced similar results in terms of embryo development compared to those obtained with FCS, but a decreased degree of lipid droplet accumulation was observed in the in vitro-cultured embryos.

Effectiveness of systematic foot and mouth disease mass vaccination campaigns in Argentina.

The objective of this paper is to evaluate the effectiveness of systematic mass vaccination campaigns against foot and mouth disease in Argentina. The analysis was based on an estimation of the proportion of protected animals and protected farms in vaccinated populations, as reflected by levels of antibodies measured in liquid-phase enzyme-linked immunosorbent assay. The analysis was carried out in 49 animal health districts in Buenos Aires province, using data collected from four cross-sectional studies, in 2004, 2007, 2008 and 2011. Cattle were assigned to one of two categories on the basis of correlation between serological titres and expected percentage protection: non-adequately protected (expected protection < 75%) and adequately protected (expected protection ≥ 75%). The proportions of adequately protected cattle and significantly non-adequately protected farms were estimated and compared among sampled locations. Protection was variable among the districts; cattle aged one to two years showed higher levels of protection than cattle six to 12 months old, and the proportion of protected cattle was higher in the more recent studies. The results of the analysis will allow the national animal health service to investigate in depth those districts where protection was lower than the regional background protection. The authors propose that this methodology could be used to evaluate the effectiveness of vaccination campaigns in other countries or zones where systematic foot and mouth disease mass vaccination campaigns are undertaken.

Fleas associated with non-flying small mammal communities from northern and central Chile: with new host and locality records.

Fleas associated with small mammals from seven localities from northern and central Chile were assessed. We captured 352 small mammals belonging to 12 species from which we obtained 675 fleas belonging to 15 different species. The most frequently captured flea species were Neotyphloceras crassispina crassispina (n = 198) and N. chilensis (n = 175). High values of flea species richness and diversity were found in Fray Jorge National Park (NP), a north-central Chilean site, whereas the highest values of mean abundance (MA) and prevalence were found in three diverse sites that include Los Molles River, a high altitude site located in north-central Chile, Fray Jorge NP and Dichato, in south-central Chile. On the other hand, high values of flea richness and diversity were found on two rodent species, Abrothrix olivacea and A. longipilis, whereas the highest values of MA and prevalence were found on Oligoryzomys longicaudatus, A. longipilis and Phyllotis xanthopygus. A total of three new host recordings, nine new localities and nine new host species and locality recordings are reported. Also, this study represents the first known record of Tetrapsyllus (Tetrapsyllus) comis in Chile and the first ecological analysis of Neotyphloceras chilensis.

Analysis of cattle movements in Argentina, 2005.

We describe the movement of cattle throughout Argentina in 2005. Details of farm-to-farm and farm-to-slaughter movements of cattle were obtained from the Sanitary Management System database (Sistema de Gestión Sanitaria, SGS), maintained by the National Service for Agrifood Health and Quality (SENASA). Movements were described at the regional and district level in terms of frequency, the number of stock transported, the district of origin and destination and Euclidean distance traveled. Social network analysis was used to characterize the connections made between regions and districts as a result of cattle movement transactions, and to show how these characteristics might influence disease spread. Throughout 2005 a total of 1.3 million movement events involving 32 million head of cattle (equivalent to approximately 57% of the national herd) were recorded in the SGS database. The greatest number of farm-to-farm movements occurred from April to June whereas numbers of farm-to-slaughter movement events were relatively constant throughout the year. Throughout 2005 there was a 1.1-1.6-fold increase in the number of farm-to-farm movements of cattle during April-June, compared with other times of the year. District in-degree and out-degree scores varied by season, with higher maximum scores during the autumn and winter compared with summer and spring. Districts with high in-degree scores were concentrated in the Finishing region of the country whereas districts with high out-degree scores were concentrated not only in the Finishing region but also in Mesopotamia, eastern Border and southern Central regions. Although movements of cattle from the Border region tended not to be mediated via markets, the small number of districts in this area with relatively high out-degree scores is a cause for concern as they have the potential to distribute infectious disease widely, in the event of an incursion.

A description of cattle movements in two departments of Buenos Aires province, Argentina.

We present a descriptive analysis of cattle movement information retrieved from the Argentinean animal movement database for two departments in the province of Buenos Aires during 2004. For each quarter of the year (January to March, April to June, July to September, and October to December) we report the number of on- and off-farm movement events for the purpose of finishing. Our analyses show that the distribution of the number of finishing-related movement events per farm was skewed, with the majority of farms reporting at least 1 and less than 5% of farms of reporting greater than 15 finishing related movement events throughout the year. The frequency of finishing-related movement events varied over time, with a 1.2-1.8-fold increase in reported movement events from April to September, compared with the rest of the year. These analyses indicate that cattle movement patterns in these departments are dependent on the relative mix of constituent cattle enterprise types. Departments with a mixture of breeding and finishing enterprises behave as potential recipients and distributors of infectious disease, whereas departments comprised of primarily finishing enterprises are predominantly recipients of infectious disease, rather than distributors. Data integrity audits of the Argentinean animal movement database, on a regular or intermittent basis, should allow the presence of bias in these data to be quantified in greater detail.

Seroepidemiology of Mycoplasma hyopneumoniae in pigs from farrow-to-finish farms.

A prospective study was carried out on three intensive farrow-to-finish farms. The aims were to estimate the incidence of Mycoplasma hyopneumoniae infection, to determine when pigs become infected and the pattern of transmission of infection and to verify the relationship between seroconversion and clinical signs. One batch of pigs per farm was followed from farrowing-to-slaughter. Blood samples were taken at 10, 27, 70, 94, 125 and 147 days of age, from 44, 48 and 44 pigs per farm. Colostrum and blood samples were also taken from the sows. Animals were checked clinically once a week and coughing rates were recorded. Antibodies against M. hyopneumoniae were detected by a blocking ELISA. At 27, 70 and 94 days of age most pigs on the three farms were seronegative, suggesting that no circulation of M. hyopneumoniae occurred during the growing period. Thereafter, a high proportion of pigs seroconverted, indicating that infection occurred soon after the transfer of the animals to the finishing houses. Differences were detected between farms in the incidence of seroconversion. Seropositive pigs were widely distributed among the finishing pens, suggesting that in addition to direct contact, other methods of transmission, such as indirect or airborne transmission, may have been important. Coughing started at around the same time as seroconversion. The results showed that the critical period for the transmission of M. hyopneumoniae is around the beginning of the finishing period, when pigs have low concentrations of antibodies against the agent.