HU210-Induced Downregulation in Cannabinoid CB1 Receptor Binding Strongly Correlates with Body Weight Loss in the Adult Rat

In vitro autoradiography was used to examine changes in cannabinoid CB1 receptors (targeted with [³H] CP55,940) in rats treated with the potent cannabinoid agonist HU210. Animals were administered with HU210 (25, 50, 100 μg/kg) for 4 or 14 days or received a single 100 μg/kg injection of HU210 and sacrificed 24 h later. The acute dose resulted in a decrease in binding in the caudate putamen and hippocampus. A dose dependent, region-specific reduction (P < 0.0001) in [³H] CP55,940 binding was seen in all brain regions examined after 4 and 14 days treatment. A decrease in body weight was recorded during the first 4 days of treatment but after this animals began to gain weight. Correlations (0.865 < r < 0.659, P < 0.0001) between body weight on day four and CB1 receptor binding were found in all brain regions examined suggesting that downregulation of CB1 receptors may contribute to the induction of tolerance to body weight loss induced by HU210.     

Effect of clinical mastitis and other diseases on reproductive performance of Holstein cows

The objective of this study was to evaluate the effect of clinical mastitis and (or) other diseases on reproductive performance in lactating Holstein cows. Cows (n=967) from a commercial dairy farm were divided into four groups retrospectively: cows with clinical mastitis and other diseases (MD, n=54), clinical mastitis only (M, n=154), other diseases only (D, n=187), and cows with no record of clinical mastitis or other diseases (H, n=572). Days in milk at first service (DIMFS), services per conception (S/C), days not pregnant (DNP), the rate at which animals became pregnant over time and the proportion of cows that remained non-pregnant during 224 days of lactation were evaluated. Groups MD and M had greater (P<0.05) DNP compared with H (155+/-15 and 140+/-5 vs. 88+/-2, respectively). Moreover, MD and M had greater (P<0.05) S/C compared with H (3.0+/-0.4 and 2.1+/-0.1 vs. 1.6+/-0.1, respectively). The rate at which animals became pregnant over time was less (P<0.05) for MD and M and tended (P=0.1) to be less for D when compared with H. In addition, proportion of cows that remained non-pregnant by 224 days of lactation was greater (P<0.05) in MD, M, and D compared with H. Cows with mastitis were also divided into three groups according to the day of occurrence of the first case of clinical mastitis: (1) clinical mastitis occurred before 56 days postpartum (MP1); (2) clinical mastitis occurred between 56 and 105 days after parturition (MP2); and (3) clinical mastitis occurred after 105 days postpartum (MP3) Regardless of the time of occurrence, DNP was greater (P<0.05) for cows with mastitis compared with H. Time of mastitis occurrence affected S/C in that cows in MP2 and MP3 had a greater S/C compared with H cows (P<0.05). Reproductive efficiency was decreased by the presence of clinical mastitis alone because a greater proportion of cows with mastitis remained non-pregnant over time. Moreover, a greater proportion of cows with mastitis or diseases remained non-pregnant by 224 postpartum. Furthermore, the negative effects on reproduction were exacerbated when cows experienced both clinical mastitis and other diseases.     

Structure determination of chiral sulfoxide in diastereomeric bicalutamide derivatives

We report on the synthesis and investigation of two diastereomers (5a and 5b) of a new bicalutamide analog with an asymmetric carbon atom and a chiral sulfoxide group. These bicalutamide analogs are novel androgen receptor antagonists with biological activities that depend significantly on the configuration of their stereogenic centers. We determined the absolute configuration at the SO center by combining X-ray and NMR measurements with quantum chemical calculations. Since 5a and 5b failed to yield satisfactory crystals for X-ray crystal structure determination, analogs 6a and 6b differing in only one remote functional group relative to the chiral sulfoxide were synthesized, which yielded satisfactory crystals. X-ray structure determination of 6a and 6b provided the absolute configuration at the chiral sulfoxide. Since the structural difference between 5 and 6 is remote from the chiral sulfoxide, the structural assignment was extended from the diastereomers of 6 to those of 5 provisionally. These assignments were verified with the help of measured and DFT-calculated proton and carbon NMR chemical shifts.     

PPIB Mutations Cause Severe Osteogenesis Imperfecta

Deficiency of cartilage-associated protein (CRTAP) or prolyl 3-hydroxylase 1(P3H1) has been reported in autosomal-recessive lethal or severe osteogenesis imperfecta (OI). CRTAP, P3H1, and cyclophilin B (CyPB) form an intracellular collagen-modifying complex that 3-hydroxylates proline at position 986 (P986) in the α1 chains of collagen type I. This 3-prolyl hydroxylation is decreased in patients with CRTAP and P3H1 deficiency. It was suspected that mutations in the PPIB gene encoding CyPB would also cause OI with decreased collagen 3-prolyl hydroxylation. To our knowledge we present the first two families with recessive OI caused by PPIB gene mutations. The clinical phenotype is compatible with OI Sillence type II-B/III as seen with COL1A1/2, CRTAP, and LEPRE1 mutations. The percentage of 3-hydroxylated P986 residues in patients with PPIB mutations is decreased in comparison to normal, but it is higher than in patients with CRTAP and LEPRE1 mutations. This result and the fact that CyPB is demonstrable independent of CRTAP and P3H1, along with reported decreased 3-prolyl hydroxylation due to deficiency of CRTAP lacking the catalytic hydroxylation domain and the known function of CyPB as a cis-trans isomerase, suggest that recessive OI is caused by a dysfunctional P3H1/CRTAP/CyPB complex rather than by the lack of 3-prolyl hydroxylation of a single proline residue in the α1 chains of collagen type I.     

Immunological Interactions between 2 Common Pathogens,Th1-Inducing Protozoan Toxoplasma gondii and the Th2-Inducing Helminth Fasciola hepatica

Background

The nature of the immune response to infection is dependent on the type of infecting organism. Intracellular organisms such as Toxoplasma gondii stimulate a Th1-driven response associated with production of IL-12, IFN-γ, nitric oxide and IgG2a antibodies and classical activation of macrophages. In contrast, extracellular helminths such as Fasciola hepatica induce Th2 responses characterised by the production of IL-4, IL-5, IL-10 and IgG1 antibodies and alternative activation of macrophages. As co-infections with these types of parasites commonly exist in the field it is relevant to examine how the various facets of the immune responses induced by each may influence or counter-regulate that of the other.

Principal Findings

Regardless, of whether F. hepatica infection preceded or succeeded T. gondii infection, there was little impact on the production of the Th1 cytokines IL-12, IFN-γ or on the development of classically-activated macrophages induced by T. gondii. By contrast, the production of helminth-specific Th2 cytokines, such as IL-4 and IL-5, was suppressed by infection with T. gondii. Additionally, the recruitment and alternative activation of macrophages by F. hepatica was blocked or reversed by subsequent infection with T. gondii. The clinical symptoms of toxoplasmosis and the survival rate of infected mice were not significantly altered by the helminth.

Conclusions

Despite previous studies showing that F. hepatica suppressed the classical activation of macrophages and the Th1-driven responses of mice to bystander microbial infection, as well as reduced their ability to reject these, here we found that the potent immune responses to T. gondii were capable of suppressing the responses to helminth infection. Clearly, the outcome of particular infections in polyparasitoses depends on the means and potency by which each pathogen controls the immune response.     

Effects of a Low‐intensity Intervention That Prescribed a Low‐carbohydrate vs. a Low‐fat Diet in Obese,Diabetic Participants

Low‐carbohydrate diets have been associated with significant reductions in weight and HbA_1c in obese, diabetic participants who received high‐intensity lifestyle modification for 6 or 12 months. This investigation sought to determine whether comparable results to those of short‐term, intensive interventions could be achieved over a 24‐month study period using a low‐intensity intervention that approximates what is feasible in outpatient practice. A total of 144 obese, diabetic participants were randomly assigned to a low‐carbohydrate diet (<30 g/day) or to a low fat diet (≤30% of calories from fat with a deficit of 500 kcal/day). Participants were provided weekly group nutrition education sessions for the first month, and monthly sessions thereafter through the end of 24 months. Weight, HbA_1c, glucose, and lipids were measured at baseline and 6, 12, and 24 months. Of the 144 enrolled participants, 68 returned for the month 24 assessment visit. Weights were retrieved from electronic medical records for an additional 57 participants (total, 125 participants) at month 24. All participants with a baseline measurement and at least one of the three other measurements were included in the mixed‐model analyses (n = 138). The low‐intensity intervention resulted in modest weight loss in both groups at month 24. At this time, participants in the low‐carbohydrate group lost 1.5 kg, compared to 0.2 kg in the low‐fat group (P = 0.147). Lipids, glycemic indexes, and dietary intake did not differ between groups at month 24 (or at months 6 or 12) (ClinicalTrials.gov number, NCT00108459).