25-Hydroxyvitamin D status, light exposure and sleep quality in UK dwelling South Asian and Caucasian postmenopausal women.

There is a lack of research into 25-hydroxyvitamin D (25(OH)D) status, light exposure and sleep patterns in South Asian populations. In addition, results of research studies are conflicting as to whether there is an association between 25(OH)D status and sleep quality. We investigated 25(OH)D status, self-reported and actigraphic sleep quality in n = 35 UK dwelling postmenopausal women (n = 13 South Asians, n = 22 Caucasians), who kept daily sleep diaries and wore wrist-worn actiwatch (AWL-L) devices for 14 days. A subset of n = 27 women (n = 11 South Asian and n = 16 Caucasian) also wore a neck-worn AWL-L device to measure their light exposure. For 25(OH)D concentration, South Asians had a median ± IQR of 43.8 ± 28.2 nmol/L, which was significantly lower than Caucasians (68.7 ± 37.4 nmol/L)(P = 0.001). Similarly, there was a higher sleep fragmentation in the South Asians (mean ± SD 36.9 ± 8.9) compared with the Caucasians (24.7 ± 7.1)(P = 0.002). Non-parametric circadian rhythm analysis of rest/activity patterns showed a higher night-time activity (L5) (22.6 ± 14.0 vs. 10.5 ± 4.4; P = 0.0008) and lower relative amplitude (0.85 ± 0.07 vs. 0.94 ± 0.02; P < 0.0001) in the South Asian compared with the Caucasian women. More South Asians (50%) met the criteria for sleep disorders (PSQI score >5) than did Caucasians (27%) (P = 0.001, Fishers Exact Test). However, there was no association between 25(OH)D concentration and any sleep parameter measured (P > 0.05) in either ethnic group. South Asians spent significantly less time in illuminance levels over 200 lx (P = 0.009) than did Caucasians. Overall, our results show that postmenopausal South Asian women have lower 25(OH)D concentration than Caucasian women. They also have higher sleep fragmentation, as well as a lower light exposure across the day. This may have detrimental implications for their general health and further research into sleep quality and light exposure in the South Asian ethnic group is warranted.

A case-control study to identify risk factors for acute salmonellosis in New Zealand dairy herds, 2011-2012.

In late 2011 the New Zealand Ministry for Primary Industries reported an increase in confirmed laboratory diagnoses of salmonellosis in dairy herds. To identify risk factors for herd-level outbreaks of salmonellosis we conducted a case-control study of New Zealand dairy herds in 2011-2012. In a multivariable analysis, use of continuous feed troughs [adjusted odds ratio (aOR) 6·2, 95% confidence interval (CI) 2·0-20], use of pelletized magnesium supplements (aOR 10, 95% CI 3·3-33) and use of palm kernel meal as a supplementary feed (aOR 8·7, 95% CI 2·5-30) were positively associated with a herd-level outbreak of salmonellosis between 1 July 2011 and 31 January 2012. We conclude that supplementary feeds used on dairy farms (regardless of type) need to be stored and handled appropriately to reduce the likelihood of bacterial contamination, particularly from birds and rodents. Magnesium supplementation in the pelletized form played a role in triggering outbreaks of acute salmonellosis in New Zealand dairy herds in 2011-2012.

Partial deletion of both the spermine synthase gene and the Pex gene in the X-linked hypophosphatemic, gyro (Gy) mouse.

Gy, along with Hyp, is a dominant mutation of the normal gene Pex causing X-linked hypophosphatemia in the mouse. Hemizygous Gy male mice, however, have greater defects in survival, bodily growth, skeletal mineralization, and neurological function than those found in heterozygous Gy females or in Hyp mice. Since the gene for spermine synthase is immediately upstream of the homologous human gene PEX, we compared the effects of the Gy and Hyp mutations on both the spermine synthase gene and the Pex gene. Barely detectable levels of spermine (< 5% of normal) with elevated levels of its precursor, spermidine, were found in organs of Gy male mice compared to normal male littermates. Neither Gy females nor Hyp male mice were significantly affected. Four missing introns of the spermine synthase gene were identified in Gy male mice, suggesting extensive gene disruption. A pseudogene for spermine synthase was also identified in the mouse genome. Pex mRNA was found in several but not all tissues studied in adult normal mice. Pex mRNA was altered in both Gy and Hyp mice. All male Hyp mice were lacking the 3' end of the Pex message, whereas all male Gy mice were deficient at the 5' end. In summary, the Gy mutation is associated with a recessively expressed mutation of the spermine synthase gene, leading to spermine deficiency, and a dominantly expressed mutation of the Pex gene, leading to hypophosphatemia. Alterations in two contiguous genes in Gy may explain the additional phenotypic abnormalities present in the Gy male mouse.

Neosporosis in a pup.

CASE: A 13-week-old female boxer pup was found to be suffering from rigidity of the left hindleg. Antibiotic and anti-inflammatory treatment over a 3-week period failed to improve the condition and the pup was humanely killed. METHODS: Serological examination for Neospora antibodies was carried out by the indirect fluorescent antibody test and for Toxoplasma gondii antibodies with a latex agglutination test. A variety of tissues were examined histologically, and the central nervous system by immunohistochemistry and the polymerase chain reaction. RESULTS: The IFAT for anti-Neospora antibodies showed a titre of 1:51 200 in the clinically affected pup while the latex agglutination test for Toxoplasma antibodies was negative. The dam and one of two tested litter-mates had anti-Neospora IFAT titres of 1:1600, the other litter mate was negative. All three were not clinically affected. Histological, immunohistochemical and polymerase chain reaction examinations of the affected pup confirmed the diagnosis of Neospora infection. CONCLUSION: In the live animal, serological examination is thought to be the most useful specific test. Post-mortem examination by traditional histology, immunohistochemistry and the polymerase chain reaction confirmed the diagnosis. The case is discussed in the context of present knowledge about Neospora infection in New Zealand.

Effects of altered diet on serum levels of 1,25-dihydroxyvitamin D and parathyroid hormone in X-linked hypophosphatemic (Hyp and Gy) mice.

X-linked hypophosphatemia is a metabolic bone disease occurring in both humans and mice. In mice, two different mutations (Hyp and Gy), occurring at separate but closely linked loci, have been proposed as models for this disease. Varying reports of the Vitamin D status of these two mutants has led us to reexamine the influence of diet on circulating calcitrophic hormones and mineral metabolism in both mutants. Hyp and Gy mice were raised on the B6C3H background, and both normal females and heterozygous mutant females were studied at 10 weeks of age. Animals were fed one of three diets at random: high (1.5% Ca and 1.0% P); medium (0.6% Ca and 0.6% P); or low (0.0% Ca and 0.6% P). After 3 days, serum and urine samples were collected. In comparison to mutant mice fed the high diet, both Hyp and Gy mice fed the low diet had decreased serum calcium levels, and further elevations in both serum alkaline phosphatase and serum parathyroid hormone (PTH). Serum 1,25-dihydroxyvitamin D levels were elevated by both the medium and low diets in all groups of mice over values obtained with the high diet. Mutant mice were significantly higher in serum PTH on all diets compared to normal mice fed the same diet. Mutant mice were not elevated in serum 1,25-dihydroxyvitamin D over normal mice when fed the high diet. However, both Hyp and Gy mice fed the medium and low diets were elevated in serum 1,25-dihydroxyvitamin D over normal mice. Serum PTH levels were correlated to serum 1,25-dihydroxyvitamin D levels with Hyp and Gy mice lying on the same line (r = 0.86; p < 0.0001). In summary, when Hyp and Gy mice are studied on the same genetic background and fed the same diet, similar responses are seen in PTH levels and 1,25-dihydroxyvitamin D levels. Both mutants should be useful in elucidating the pathophysiology of this poorly understood human disease.

A prospective, randomized comparison of the Pipelle endometrial sampling device with the Novak curette.

This prospective, randomized study compares sample adequacy, pain associated with endometrial biopsy, and correlation of endometrial histologic sampling with hysterectomy histologic results in specimens obtained by the Pipelle (Unimar Inc., Wilton, Conn.) device (N = 149) and the Novak (Miltex, Inc., Lake Success, N.Y.) curette (N = 126). During a 2-year period, patients with abnormal uterine bleeding seen in our ambulatory care facility were enrolled in this trial, unless they did not wish to participate or had a positive pregnancy test result. Patients in both groups were similar with respect to mean gravidity, parity, and menopausal status, although patients in the Novak group were older (43.4 vs 39.1 years, p = 0.005). Patients undergoing Novak biopsy had a mean pain score of 4.36 with 22 of 126 (17%) reporting severe pain, whereas patients undergoing Pipelle biopsy had a mean pain score of 3.21 with only 10 of 149 (6.7%) reporting severe pain (p less than 0.05). These pain scores were not affected by menstrual day, gravidity, parity, or menopausal status. Insufficient tissue was reported in 12.8% of patients in the Pipelle group compared with 9.5% in the Novak group (p greater than 0.05). Fifty patients underwent subsequent hysterectomy. In 48 of 50 (96%), the pathologic results at hysterectomy were in agreement with the histologic findings at endometrial sampling. This clinical trial suggests that Pipelle biopsy appears to be as effective as the Novak curette in obtaining an adequate specimen for histologic analysis and is associated with less pain.

Pathogenicity of retroviruses containing either the normal human c-Ha-ras1 gene or its mutated form derived from the bladder carcinoma EJ/T24 cell line.

Cellular ras oncogenes transduced by retroviruses carry mutations in amino acids 12, 59 and 122. Similar mutations have been observed in ras oncogenes activated during induction of neoplasia in both humans and experimental animals. The unmutated normal rat or human c-Ha-ras-1 genes have the ability to transform NIH 3T3 cells in culture when activated by a RNA synthesis promoter. These findings raise the question of whether the mutations are necessary for the ras oncogenes to induce the neoplastic phenotype in vivo. To address this question, we inserted the normal human c-Ha-ras-1 or its mutated counterpart EJ/T24 bladder carcinoma oncogene independently into a retrovirus vector derived from the M1 strain of Moloney murine sarcoma virus (MoMuSV). Both recombinant clones induced foci of transformed cells in an NIH 3T3 cell transfection assay. Infectious virus particles were rescued from cloned transformants carrying a single copy of the integrated provirus using the nonpathogenic amphotrophic wild mouse leukemia virus (WMLV) as helper. The pseudotypes rescued from the EJ/T24-containing transformants had higher titers than the normal c-Ha-ras-1 pseudotypes as determined by a focus assay and gave rise to larger and earlier detected foci upon infection of NIH 3T3 cells. The two pseudotypes were tested for in vivo pathogenicity by inoculation into newborn NFS mice and were compared to the pseudotype WMLV/Harvey murine sarcoma virus (HaMuSV) (positive control) and WMLV (negative control). While the WMLV/EJ/T24 and the WMLV/HaMuSV pseudotypes induced erythroleukemias and sarcomas with a latency period of 6-9 weeks, the WMLV/c-Ha-ras-1 pseudotype induced only mild splenomegaly. As expected the WMLV negative control induced no pathology. Tumor-bearing animals that were not euthanized at 6-9 weeks died within 2-3 months following virus inoculation.

Comparison of isoflurane and halothane safety margins in rats.

In rat experiments, the dose-effect curves for three different end points of anesthesia [loss of righting reflex (RR), prevention of movement (PM), and heart rate response (HR) to noxious stimuli] and for the lethal effect (LE) due to cardiovascular depression were determined with isoflurane and halothane. The obtained data were used to calculate LD50/ED50 ratios and standard safety margins (SSM) for assessment of each agent's safety. It was found that isoflurane provides an equal degree of separation between dose-effect curves for different end points of anesthesia as halothane does. However, isoflurane provides greater margins of safety. The margin between the highest of anesthetic doses--the loss of HR response--and the lethal dose for isoflurane was twice that for halothane (LD50/HR ED50 4.3 vs. 2.2, P less than 0.01). The standard safety margin for the loss of HR response was also greater with isoflurane (142 vs. 43, P less than 0.05). These results agreed that isoflurane may provide greater cardiovascular safety for anesthesia than halothane does.

Anesthetic potencies of isoflurane, halothane, and diethyl ether for various end points of anesthesia.

In experiments with rats, the authors compared potency ratios and slopes of dose-effect curves of isoflurane, halothane, and diethyl ether for three end points of anesthesia: loss of righting reflex (RR), abolition of purposeful movement (PM) response to painful stimuli, and abolition of heart rate (HR) response to painful stimuli. Determinations of potency were based on the direct measurement of brain concentrations of anesthetics with the use of gas chromatography. It was found that the ratio of the PM ED50 to RR ED50 was 2.41 for isoflurane, 1.74 for halothane, and 1.25 for diethyl ester. They were significantly different for all three agents. Differences between the slopes of the dose-effect curves for RR and PM were significant only with diethyl ether (7 vs. 28). The ratios of HR ED50 to PM ED50 were not significantly different for the studied agents and there were no differences found between the slopes of the dose-effect curves for PM and HR. The results suggest that heart rate response to a noxious stimuli in contrast to the righting reflex is depressed by inhalation anesthetics through a mechanism similar to that underlying the depression of purposeful movement response to a noxious stimuli. Heart rate response to a noxious stimuli might be used as an alternative index for the measurement of anesthetic potency.