Dietary Vitamin D3 Restriction Exacerbates Disease Pathophysiology in the Spinal Cord of the G93A Mouse Model of Amyotrophic Lateral Sclerosis

BackgroundDietary vitamin D₃ (D₃) restriction reduces paw grip endurance and motor performance in G93A mice, and increases inflammation and apoptosis in the quadríceps of females. ALS, a neuromuscular disease, causes progressive degeneration of motor neurons in the brain and spinal cord.ObjectiveWe analyzed the spinal cords of G93A mice following dietary D₃ restriction at 2.5% the adequate intake (AI) for oxidative damage (4-HNE, 3-NY), antioxidant enzymes (SOD2, catalase, GPx1), inflammation (TNF-α, IL-6, IL-10), apoptosis (bax/bcl-2 ratio, cleaved/pro-caspase 3 ratio), neurotrophic factor (GDNF) and neuron count (ChAT, SMI-36/SMI-32 ratio).MethodsBeginning at age 25 d, 42 G93A mice were provided food ad libitum with either adequate (AI;1 IU D₃/g feed; 12 M, 11 F) or deficient (DEF; 0.025 IU D₃/g feed; 10 M, 9 F) D₃. At age 113 d, the spinal cords were analyzed for protein content. Differences were considered significant at P ≤ 0.10, since this was a pilot study.ResultsDEF mice had 16% higher 4-HNE (P = 0.056), 12% higher GPx1 (P = 0.057) and 23% higher Bax/Bcl2 ratio (P = 0.076) vs. AI. DEF females had 29% higher GPx1 (P = 0.001) and 22% higher IL-6 (P = 0.077) vs. AI females. DEF males had 23% higher 4-HNE (P = 0.066) and 18% lower SOD2 (P = 0.034) vs. AI males. DEF males had 27% lower SOD2 (P = 0.004), 17% lower GPx1 (P = 0.070), 29% lower IL-6 (P = 0.023) and 22% lower ChAT (P = 0.082) vs. DEF females.ConclusionD₃ deficiency exacerbates disease pathophysiology in the spinal cord of G93A mice, the exact mechanisms are sex-specific. This is in accord with our previous results in the quadriceps, as well as functional and disease outcomes.     

Arsenic disrupts cellular levels of p53 and mdm2: a potential mechanism of carcinogenesis.

The antitumor protein p53 plays a critical role in DNA repair. Inorganic arsenic exposure is associated with a wide variety of human tumors, particularly of the skin. To investigate how inorganic arsenic might interfere with DNA repair and lead to greater incidence of hyperkeratosis and skin tumors, we exposed human keratinocytes (HaCaT) to environmentally relevant concentrations of arsenite for 14 days. Arsenite reduced p53 levels while concomitantly increasing the p53 regulatory protein mdm2 levels in a dose- and time-dependent manner. We propose the disruption of the p53-mdm2 loop regulating cell cycle arrest as a model for arsenic-related skin carcinogenesis and it may be important in tumors with elevated mdm2 levels.     

A model invalidation-based approach for elucidating biological signalling pathways,applied to the chemotaxis pathway in R. sphaeroides

Background  

Developing methods for understanding the connectivity of signalling pathways is a major challenge in biological research. For this purpose, mathematical models are routinely developed based on experimental observations, which also allow the prediction of the system behaviour under different experimental conditions. Often, however, the same experimental data can be represented by several competing network models.     

High levels of genetic differentiation and selfing in the Brazilian cerrado fruit tree Dipteryx alata Vog. (Fabaceae)

Dipteryx alata is a native fruit tree species of the cerrado (Brazilian savanna) that has great economic potential because of its multiple uses. Knowledge of how the genetic variability of this species is organized within and among populations would be useful for genetic conservation and breeding programs. We used nine simple sequence repeat (SSR) primers developed for Dipteryx odorata to evaluate the genetic structure of three populations of D. alata located in central Brazil based on a leaf sample analysis from 101 adults. The outcrossing rate was evaluated using 300 open-pollinated offspring from 25 seed-trees. Pollen dispersal was measured by parentage analysis. We used spatial genetic structure (SGS) to test the minimal distance for harvesting seeds in conservation and breeding programs. Our data indicate that the populations studied had a high degree of genetic diversity and population structure, as suggested by the high level of divergence among populations . The estimated outcrossing rate suggested a mixed mating system, and the intrapopulation fixation index was influenced by SGS. We conclude that seed harvesting for genetic conservation and breeding programs requires a minimum distance between trees of 196 m to avoid collecting seeds from related seed-trees.     

Ovarian cyclicity and serum progesterone and luteinizing hormone in fine-wool ewes supplemented with alfalfa or pinto beans

Effects of season and supplementation on the incidence and rate of ovulation and hormone profiles in multiparous, nonlactating ewes were investigated under range and drylot conditions during anestrus (February through August). Ninety ewes received one of six nutritional treatments: 1) range forage, 2) range forage plus 0.33 kg.hd(-1).d(-1) pinto beans, 3) 0.45 kg.hd(-1).d(-1) alfalfa pellets, 4) dry lot plus 1.33 kg.hd(-1).d(-1) prairie hay only, 5) pinto beans or 6) alfalfa pellets at rates used on the range and prairie hay. Supplemented ewes were heavier (P < 0.05) than ewes fed range forage or prairie hay during most of the study. Seasonality of ovulation, in terms of incidence and rate, was not affected (P > 0.40) by supplementation in range ewes. The ovulation rate tended to be highest in prairie hay plus pinto bean drylot ewes in February (P = 0.21) and in prairie hay plus alfalfa pellet and prairie hay plus pinto bean ewes in March (P = 0.13) compared with prairie hay ewes. Generally, seasonality of ovulation was not influenced by supplementation (P > 0.40). The incidence of ovulation approached zero for drylot ewes in April, May, June and July, while it was 43, 27, 35 and 21% for range ewes, in those same months. Supplementation did not affect serum progesterone during the estrous cycle. Luteinizing hormone (LH) concentrations were similar (P > 0.50) among drylot treatment groups before a 50-ug gonadotropin releasing hormone (GnRH) challenge. Pinto bean supplementation enhanced serum LH response to GnRH (P < 0.10).     

Breast cancer in Lebanon: Incidence and comparison to regional and Western countries

Background: Review and analyses of the 2004 Lebanese National Breast Cancer Registry (the most recently available complete national data). Methods: Crude, age-standardized rates (ASRs), and age-specific rates per 100,000 population were calculated and results were compared with estimates from Western, regional, and Arab countries. Results: Breast cancer constituted about 38.2% of all cancer cases among Lebanese females in the year 2004. The median age at diagnosis was 52.5 years. The age-standardized incidence rate per 100,000 was estimated at 71.0. ASRs remained lower than those observed in developed countries and in the Israeli Jewish population; however, they were greater than those estimated for Arab populations in the region. Five-year age-specific rates among Lebanese women were among the highest observed worldwide for the age groups 35–39, 40–44 and 45–49 years, with the exception of Israeli Jews for the age groups 35–39. Conclusions: Results endorse the new guidelines developed by the Lebanese Ministry of Public Health to start breast cancer screening with mammography at 40 years of age. Further efforts are needed from different stakeholders in order to realize a comprehensive and full database, and to enhance awareness for early detection at all age groups.     

Novel bicyclic lactam inhibitors of thrombin: highly potent and selective inhibitors

The potency and selectivity of a previous series of low molecular weight thrombin inhibitors were improved through modifications of the P1 and P3 residues. Introduction of diphenyl substituted sulfonamides in the P3 moiety led to highly efficacious compounds. By correctly selecting the combination of P1 and P3 residues, high levels of potency, selectivity and in vivo efficacy were obtained.     

Human sulfate kinetics

Electrospray tandem mass spectrometry was used to determine steady-state serum and urinary inorganic sulfate and sulfate ester kinetic profiles of nine normal men after intravenous injection of the stable isotope sodium [34S]sulfate. Sulfate ester appearance was traced by eliminating inorganic sulfate from samples, followed by hydrolysis of sulfate esters to inorganic sulfate for analysis. Whole body inorganic sulfate turnover in steady state was calculated using standard tracer techniques. Rate of appearance and disappearance of inorganic sulfate was 841 +/- 49 micromol/h. Average urinary inorganic sulfate excretion was 609 +/- 41 micromol/h, and the whole body sulfation rate (total rate of disappearance minus rate of urinary excretion) was 232 +/- 36 micromol/h. Tracer-labeled sulfate esters appeared in serum and urine within 1 h of tracer injection. The kinetics of inorganic sulfate and sulfate esters were linked by means of a compartmental model. The appearance and excretion of sulfate esters accounted for approximately 50% of the total sulfation rate. These results indicate that human whole body sulfation accounts for approximately 27% of inorganic sulfate turnover and that extracellular inorganic sulfate is an important pool for intracellular sulfation. A substantial fraction of newly synthesized sulfate esters promptly enters the extracellular space for excretion in the urine.