D2 D3 Research Articles

Cow-level association between serum 25-hydroxyvitamin D concentration and Mycobacterium avium subspecies paratuberculosis antibody seropositivity: A pilot study

Vitamin D deficiency has been associated with various human diseases. Therefore, the objective of this study was to evaluate the cow-level association between serum 25-hydroxyvitamin D [25(OH)D] concentration and Mycobacterium avium ssp. paratuberculosis (MAP) seropositivity of dairy cows, adjusting for diet, breed, hair coat color, stage of lactation, reproductive status, and cow age. The sera of 80 MAP antibody ELISA-positive and 80 test-negative herd mates from 5 Minnesota dairy herds were analyzed for 25(OH)D and 1,25-dihydroxyvitamin D [1,25(OH)2D]. The cows’ age, production records, and hair coat color were recorded. Additionally, feed samples were obtained and analyzed for vitamin D2 and vitamin D3 content. A linear mixed model was used to identify potential predictors for serum 25(OH)D concentration, accounting for herd of origin. The majority of rations analyzed had over 22,000IU of vitamin D/day (maximum: 52,000IU/d) and the study cows’ average serum 25(OH)D concentration was 62.5±13.8ng/mL. Serum ELISA-positive cows had, on average, 5.3ng/mL lower 25(OH)D serum levels than test-negative herd mates. The reproductive status of cows was also associated with the 25(OH)D levels, with fresh cows having the lowest serum concentration. In this cross-sectional study, a temporal or causal association between MAP antibody ELISA status and serum 25(OH)D concentration could not be evaluated. In addition, the high levels of vitamin D in the rations of participating farms and the average 25(OH)D serum concentration suggest that additional supplementation with vitamin D in the ration is likely to be ineffective.

Analysis and quantification of vitamin D metabolites in serum by ultra‐performance liquid chromatography coupled to tandem mass spectrometry and high‐resolution mass spectrometry – a method comparison and validation

The aim of the work was to develop and validate a method for the quantification of vitamin D metabolites in serum using ultra-high-pressure liquid chromatography coupled to mass spectrometry (LC/MS), and to validate a high-resolution mass spectrometry (LC/HRMS) approach against a tandem mass spectrometry (LC/MS/MS) approach using a large clinical sample set. A fast, accurate and reliable method for the quantification of the vitamin D metabolites, 25-hydroxyvitamin D2 (25OH-D2) and 25-hydroxyvitamin D3 (25OH-D3), in human serum was developed and validated. The C3 epimer of 25OH-D3 (3-epi-25OH-D3) was also separated from 25OH-D3. The samples were rapidly prepared via a protein precipitation step followed by solid-phase extraction (SPE) using an HLB μelution plate. Quantification was performed using both LC/MS/MS and LC/HRMS systems. Recovery, matrix effect, inter- and intra-day reproducibility were assessed. Lower limits of quantification (LLOQs) were determined for both 25OH-D2 and 25OH-D3 for the LC/MS/MS approach (6.2 and 3.4 µg/L, respectively) and the LC/HRMS approach (2.1 and 1.7 µg/L, respectively). A Passing & Bablok fit was determined between both approaches for 25OH-D3 on 662 clinical samples (1.11 + 1.06x). It was also shown that results can be affected by the inclusion of the isomer 3-epi-25OH-D3. Quantification of the relevant vitamin D metabolites was successfully developed and validated here. It was shown that LC/HRMS is an accurate, powerful and easy to use approach for quantification within clinical laboratories. Finally, the results here suggest that it is important to separate 3-epi-25OH-D3 from 25OH-D3.

Quantum and classical correlations in quantum brachistochrone evolution

Quantum and classical correlations are investigated during quantum brachistochrone evolution (QBE) in this paper. We found some typical properties of the pair of quantum states sampled randomly by use of the Harr measure in this special kind of quantum evolution. This kind of evolution of a three-qubit system between two distinct states cannot be implemented without classical correlations (including bipartite J2 and tripartite J3) and quantum correlations (including bipartite D2 and tripartite D3). We also found that some QBEs between two distinct GHZ states do not follow the typical behaviour, and that this kind of evolution can be implemented without bipartite quantum correlations. Although the probability density function of the time-averaged bipartite classical correlation, time-averaged bipartite quantum correlations and time-averaged genuine tripartite correlations become more and more uniform with the decrease of angles of separation between an initial state and a final state, the features of their most probable values exhibit a different trend.

Origin of Lower Ordovician dolomites in eastern Laurentia: Controls on porosity and implications from geochemistry

The Catoche Formation of the St. George Group in eastern Laurentia (western Newfoundland) consists of early Ordovician (Arenigian) shallow marine platform carbonates (∼160 m thick), which were extensively dolomitized during the course of their diagenetic history. The dolomites occur as both replacement and pore-filling cements and are a major control on porosity distribution in the formation. The origin and diagenetic history of the Catoche dolomites at Daniel's Harbour (western Newfoundland) were analysed in comparison to equivalent successions at Port au Choix (PaC) and Port au Port Peninsula (PaP) to assay the reservoir potential of these dolomites in eastern Laurentia. Petrographic examination identified at least three generations of dolomites in the Catoche Formation, which are: (1) an early replacement sub- to eu-hedral micritic dolomite (<4 μm–30 μm, D1), (2) eu- to sub-hedral dolomite (70 μm–1 mm) often with cloudy cores and clear rims (D2), and (3) subhedral to anhedral saddle dolomite cement (200 μm–3 mm, D3). The micritic dolomite (D1) exhibits a dull cathodoluminescence (CL) under cathodoluminoscope, whereas dolomite D2 exhibits consistent concentric CL zonation. Some subhedral crystals of D3 appear zoned both in plane polarized light and cathodoluminoscope, otherwise D3 exhibits a dull CL. Stoichiometric dolomite occurs in all three generations with D2 as the dominant dolomite by abundance.The low strontium (47 ± 25 ppm) content coupled with depleted δ18O value of dolomitizing fluids (−10 to −11.2‰ VSMOW) and near-micritic grain size, suggests an early precipitation of dolomite D1 at low temperatures of near-surface conditions from solutions likely formed by mixing of early Ordovician sea and meteoric waters. In contrast, microthermometric measurements of primary two-phase fluid inclusions in dolomite D2 (homogenization temperatures of 102–168 °C with a salinity range of 19.8–25 eq wt% NaCl) and dolomite D3 (homogenization temperatures of 158–190 °C with a salinity range of 20.2–22.2 eq wt% NaCl), suggest that both dolomite generations were generated in mid to deep burial settings from high salinity, low temperature (<200 °C) hydrothermal fluids likely under suboxic conditions. This is consistent with the low Sr concentrations for D2 (36.4 ± 8 ppm) and D3 (38.7 ± 9 ppm), δ18O values of dolomitizing fluids for D2 (+2.1 to +8.1‰, VSMOW) and D3 (+6 to +8.1‰ VSMOW), coupled with Fe contents of D2 (1684 ± 1096 ppm) and D3 (1783.7 ± 618 ppm) as well as the respective Mn (D2 = 131.2 ± 50 and D3 = 197.5 ± 55 ppm) concentrations.ΣREE and shale normalized (REESN) values of Catoche carbonates indicate enrichment in rare earth element (REE) composition of the earliest calcite (C1) relative to those of Arenig seawater, whereas the REESN profiles of the dolomite generations mimic that of calcite C1. The Ce (Ce/Ce*)SN and La (La/La* = Pr/Pr*)SN anomalies of the Catoche dolomites are consistent with precipitation in equilibrium with source fluids in slightly oxic to suboxic conditions whereas Eu (Eu/Eu*)CN anomalies suggest similar source fluids for D2 and D3. Results of fluid inclusion gas analysis are consistent with petrographic features and geochemical compositions and support the exclusion of magmatic fluids during dolomitization. Visual estimates of porosity (ϕ) from thin sections indicate four porous (ϕ = 4–12) horizons. Vugs and intercrystalline pores are two types of porosity associated with the dolomites with the latter being the dominant type and associated mainly with dolomite D2.

Genotype/subgenotype distribution of hepatitis B virus among hemodialysis patients with chronical hepatitis B

Introduction. Hepatitis B virus (HBV) infection in hemodialysis (HD) patients is a major concern, but limited information exists on the HBV genotyping in patients on maintenance HD in Turkey.Aim. To investigate the genotype and subgenotype distribution of HBV in Turkish HD patients with chronic hepatitis B.Material and methods. A total of 248 HBsAg positive patients undergoing long-term HD from all regions of Turkey were included in this study. HBV genotypes were determined by phylogenetic analysis and by genotyping tools.Results. HBV DNA was detected in 94/248 (38%) of the patients. Among the study patients, genotype D of HBV was predominant (99%) and one patient (1%) was infected with genotype G. The majority (82%) of HBV genotype D branched into subgenotype D1, and also in ayw2 HBsAg subtype clusters in the phylogenetic tree. However, 10% and 8% of the strains branched into subgenotype D2 (also in ayw3 HBsAg subtype cluster) and subgenotype D3 clusters, respectively.Conclusion. In conclusion, HBV genotyping should be routinely applied to HD patients to establish a baseline. Determination of genotypes/subgenotypes of HBV may provide robust epidemiological data related to their circulation as well as their transmissibility.

Comparison of vitamin D2 and vitamin D3 supplementation in increasing serum 25-hydroxyvitamin D status: a systematic review and meta-analysis

Comparison of vitamin D2 and vitamin D3 supplementation in increasing serum 25-hydroxyvitamin D status: a systematic review and meta-analysis

Subnuclear cyclin D3 compartments and the coordinated regulation of proliferation and immunoglobulin variable gene repression

Ubiquitously expressed D-type cyclins are required for hematopoiesis but are dispensable in other cell lineages. Furthermore, within different hematopoietic progenitor populations the D-type cyclins play nonredundant roles. The basis of this lineage and developmental specificity is unknown. In pro-B cells we demonstrate four distinct nuclear D-type cyclin compartments, including one cyclin D3 fraction associated with CDK4 and another phosphoinositide 3-kinase-regulated fraction not required for proliferation. A third fraction of cyclin D3 was associated with the nuclear matrix and repression of >200 genes including the variable (V) gene segments Igkv1-117, Iglv1, and Igh-VJ558. Consistent with different subnuclear compartments and functions, distinct domains of cyclin D3 mediated proliferation and Igk V gene segment repression. None of the cyclin D3 nuclear compartments overlapped with cyclin D2, which was distributed, unbound to CDK4, throughout the nucleus. Furthermore, compartmentalization of the cyclins appeared to be lineage restricted because in fibroblasts, cyclin D2 and cyclin D3 occupied a single nuclear compartment and neither bound CDK4 efficiently. These data suggest that subnuclear compartmentalization enables cyclin D3 to drive cell cycle progression and repress V gene accessibility, thereby ensuring coordination of proliferation with immunoglobulin recombination.

Should We Be Recommending Vitamin D Supplementation for Hypertension and Cardiovascular Disease Prevention?

Should We Be Recommending Vitamin D Supplementation for Hypertension and Cardiovascular Disease Prevention?

Guidelines for the investigation and management of mantle cell lymphoma.

The guideline group was selected to be representative of UK-based medical experts and patients representatives. Ovid MEDLINE, EMBASE and NCBI Pubmed were searched systematically for publications in English from 1980 to 2011 using the MeSH subheading 'lymphoma, mantle cell' and 'lymphoma, mantle cell' as a keyword, as well as all subheadings. In addition, all references to mantle cell lymphoma in the World Health Organization Classification of Tumours of Haematopoietic and Lymphoid Tissue (Swerdlow et al, 2008) and the British Committee for Standards in Haematology (BCSH) Guideline: Best Practice in Lymphoma Diagnosis and Reporting (Parker et al, 2010) have been included. The writing group produced the draft guideline, which was subsequently revised by consensus by members of the Haemato-oncology Task Force of the BCSH. The guideline was then reviewed by a sounding board of approximately 50 UK haematologists, the BCSH and the British Society for Haematology Committee and comments incorporated where appropriate. The 'GRADE' system was used to quote levels and grades of evidence, details of which can be found in Appendix I. The objective of this guideline is to provide healthcare professionals with clear guidance on the investigation and management of patients with mantle cell lymphoma. The guidance may not be appropriate to patients with other lymphoma sub-types and in all cases individual patient circumstances may dictate an alternative approach.

Recombinant expression of TLR5 proteins by ligand supplementation and a leucine-rich repeat hybrid technique

Vertebrate TLR5 directly binds bacterial flagellin proteins and activates innate immune responses against pathogenic flagellated bacteria. Structural and biochemical studies on the TLR5/flagellin interaction have been challenging due to the technical difficulty in obtaining active recombinant proteins of TLR5 ectodomain (TLR5-ECD). We recently succeeded in production of the N-terminal leucine rich repeats (LRRs) of Danio rerio (dr) TLR5-ECD in a hybrid with another LRR protein, hagfish variable lymphocyte receptor (VLR), and determined the crystal structure of its complex with flagellin D1–D2–D3 domains. Although the structure provides valuable information about the interaction, it remains to be revealed how the C-terminal region of TLR5-ECD contributes to the interaction. Here, we present two methods to obtain recombinant TLR5 proteins that contain the C-terminal region in a baculovirus expression system. First, production of biologically active full-length drTLR5-ECD was substantially enhanced by supplementation of expression culture with purified flagellin proteins. Second, we designed TLR5-VLR hybrids using an LRR hybrid technology by single and double LRR fusions and were able to express diverse regions of drTLR5-ECD, allowing us to detect a previously unidentified TLR5/flagellin interaction. The drTLR5-VLR hybrid technique was also successfully applied to human TLR5-ECD whose expression has been highly problematic. These alternative TLR5 expression strategies provide an opportunity to obtain a complete view of the TLR5/flagellin interaction and can be applied to other LRR proteins.

신보광산 주변지역에서 변성광물의 성장과 변형작용 사이의 상대적인 시간관계: 우라늄 광화대의 상대적인 광화시기

옥천변성대의 남서부에 위치하는 전라북도 진안군 신보광산과 그 동부지역에서 고품위 우라늄 지화학 이상대가 보고된 바가 있다. 본 논문은 선캠브리아기 변성퇴적암류(규암, 변성이질암, 변성사질암)와 이를 관입하는 시대미상의 페그마타이트와 백악기 반암류 등이 분포하는 신보광산 동부지역에서 이들 구성암류가 경험한 변형작용과 변성광물의 성장 사이의 상대적인 시간관계에 대한 미구조의 연구결과를 보고하고, 기존 연구결과를 종합하여 광화작용의 상대적인 시기를 고찰하였다. D1 변형작용은 홍주석 반상변정 내에서 주로 신장된 석영, 흑운모, 불투명광물 등으로 정의되는 직선 형태의 내부엽리 Si 를 형성시켰다. Si 엽리를 미습곡시키고 S2 파랑엽리를 형성시키는 D2 변형작용은 전기 파랑습곡작용과 후기 파랑습곡작용으로 구분된다. 전자는 홍주석의 맨틀부에서 곡선 형태의 Si 를 형성시켰다. 후자는 홍주석을 포획하며 그 외부에서 S1-2 복합엽리를 형성시켰다. 홍주석 반상변정은 직선 Si 엽리에 해당하는 S1 엽리가 형성된 이후의 비변형작용 조건하에서 출현하여 후기 파랑습곡작용 이전까지 성장하였다. 시대미상의 페그마타이트는 D2 변형 이후에 관입하여 S1-2 복합엽리를 무질서하게 덮는 섬유상 규선석을 성장시켰다. D3 변형작용은 S1-2 복합 엽리와 D2 파랑습곡 그리고 섬유상 규선석을 미습곡시키는 F3 습곡을 형성시켰다. 이는 섬유상 규선석의 성장과 관련된 페그마타이트의 관입은 D2 변형과 D3 변형 사이의 비변형조건하에서 발생하였음을 지시한다. 흑운모의 녹니석화작용과 홍주석 반상변정 내부에서 S1-2 복합엽리와 F3 습곡축면에 평행한 두 방향의 벽개 라멜라에 의해 인지되는 후퇴변성작용은 각각 D2 후기변형과 D3 변형과 함께 발생하였다. 따라서 우라늄의 초생적 근원암을 페그마타이트로 간주하는 기존 연구결과를 고려해 볼 때, 시대미상의 페그마타이트는 지각이 상승하는 후퇴변성작용 동안에 해당하는 D2 변형과 D3 변형 사이의 비변형조건하에서 관입하여 신보광산 주변지역에 우라늄 광화대를 형성시킨 것으로 고찰된다. The geochemical high-grade uranium anormal zone has been reported in the Shinbo mine and its eastern areas, Jinan-gun, Jeollabuk-do located in the southwestern part of Ogcheon metamorphic zone, Korea. In this paper is reported the time-relationship between deformation and growth of metamorphic minerals in the eastern area of Shinbo mine, which consists of the Precambrian metasedimentary rocks (quartzite, metapelite, metapsammite) and the age-unknown pegmatite and Cretaceous porphyry which intrude them, and is considered the relative mineralization time on the basis of the previous research's result. The D1 deformation formed the straight-type Si internal foliation which is defined mainly as the arrangement of elongate quartz, biotite, opaque mineral in andalusite porphyroblast. The D2 deformation, which is defined by the microfolding of Si foliation, formed S2 crenulation cleavage. It can be divided into two sub-phases, early crenulation and late crenulation. The former occurs as the curvetype Si foliation in the mantle part of andalusite. The latter occurs as S1-2 composite foliation which warps around the andalusite. The andalusite porphyroblast began to grow under non-deformation condition after the formation of S1 foliation which corresponds to the straight-type Si foliation. It continued to grow before the late crenulation phase. The age-unknown pegmatite intruded after the D2 deformation and grew the fibrous sillimanite which random masks the S1-2 composite foliation. The D3 deformation formed F3 fold which folded the S1-2 composite foliation, D2 crenulation, fibrous sillimanite. It means that the intrusion of pegmatite related to the growth of the fibrous sillimanite took place during the inter-tectonic phase of D2 and D3 deformations. The retrograde metamorphism is recognized by the chloritization of biotite and two-way cleavage lamellae which is parallel to the S1-2 composite foliation and the F3 fold axial surface in the andalusite porphyroblast. It occurred during the D2 late crenulation phase and D3 deformation. In considering of the previous research's result inferring the most likely candidate for the uranium source rock as pegamatite, it indicates that the age-unknown pegmatite intruded during the inter-tectonic phase of D2 and D3 deformations, i.e. during the retrograde metamorphism related to the uplifting of crust, and formed the uranium ore zone around the Shinbo mine.

Approximal caries increment in adolescents after a visual aid in combination with a comprehensive open discussion

Objective. To achieve greater motivation for behavioural changes; educating, motivating and supporting the patient's ability to change lifestyle factors related to the caries disease are important tasks in the prevention of the disease. The aim of this study was to evaluate whether a visual aid (Visual Caries Dialogue, VCD) in combination with a comprehensive open discussion has a beneficial effect on approximal caries development among a population of young adolescents. Materials and methods. The study subjects were randomized to either an intervention group (n = 118), where VCD was conducted, or a control group (n = 112) receiving traditional oral healthcare information, at the annual dental health examination. The number of caries lesions reaching through the entire enamel (D2) and dentin caries reaching through the enamel into the dentin (D3) were recorded from bitewing radiographs each year, from 2001–2004. Differences between the study groups regarding at least two new approximal surfaces with caries (D2–D3) were tested using logistic regression. Results. The caries increment was lower in the intervention group compared to the control group. During the 3-year follow-up, 18 (15.3%) patients in the intervention group and 40 (35.7%) patients in the control group demonstrated a DS-approximal increment of at least two surfaces with a risk ratio of 2.34 (95% CI = 1.43–3.83). Conclusion. Visual Caries Dialogue in combination with a comprehensive open discussion reduced approximal caries increment among young individuals. The method provides an innovative simple and low-cost way of delivering information to patients and guides busy dental healthcare personnel in the approach.

Vitamin D in pregnancy at high latitude in Scotland

The aims of the present study were to determine compliance with current advice on vitamin D and to assess the influence of season, dietary intake, supplement use and deprivation on vitamin D status in pregnant mothers and newborns in the north of Scotland where sunlight exposure is low. Pregnant women (n 1205) and their singleton newborns were studied in the Aberdeen Maternity Hospital (latitude 57°N) between 2000 and 2006. Plasma 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 were measured at 19 weeks of gestation in mothers and at delivery in newborns. During pregnancy, 21·0 (95 % CI 18·5, 23·5) % of women took vitamin D supplements. The median intake was 5 μg/d and only 0·6 (95 % CI 0·1, 1·0) % took the recommended 10 μg/d. Supplement use, adjusted for season, dietary intake and deprivation, significantly increased maternal 25-hydroxyvitamin D (25(OH)D) by 10·5 (95 % CI 5·7, 15·2) nmol/l (P< 0·001); however, there was no significant effect on cord 25(OH)D (1·4 (95 % CI - 1·8, 4·5) nmol/l). The biggest influence on both maternal and cord 25(OH)D was season of birth (P< 0·001). Compared with the least deprived women (top three deciles), the most deprived pregnancies (bottom three deciles) were characterised by a significantly lower seasonally adjusted 25(OH)D ( - 11·6 (95 % CI - 7·5, - 15·7) nmol/l in the mother and - 5·8 (95 % CI - 2·3, - 9·4) nmol/l in the cord), and a lower level of supplement use (10 (95 % CI 4, 17) v. 23 (95 % CI 20, 26) %). More should be done to promote vitamin D supplement use in pregnancy but the critical importance of endogenous vitamin D synthesis, and known adaptations of fat metabolism specific to pregnancy, suggest that safe sun advice may be a useful additional strategy, even at high latitude.

Long-term vitamin D3 supplementation is more effective than vitamin D2 in maintaining serum 25-hydroxyvitamin D status over the winter months

Public health recommendations do not distinguish between vitamin D2 and vitamin D3, yet disagreement exists on whether these two forms should be considered equivalent. The objective of the present study was to evaluate the effect of a daily physiological dose of vitamin D2 or vitamin D3 on 25-hydroxyvitamin D (25(OH)D) status over the winter months in healthy adults living in Dunedin, New Zealand (latitude 46°S). Participants aged 18-50 years were randomly assigned to 25 μg (1000 IU) vitamin D3 (n 32), 25 μg (1000 IU) vitamin D2 (n 31) or placebo (n 32) daily for 25 weeks beginning at the end of summer. A per-protocol approach, which included ≥ 90 % supplement compliance, was used for all analyses. Serum 25-hydroxyvitamin D3 (25(OH)D3), 25-hydroxyvitamin D2 (25(OH)D2) and parathyroid hormone (PTH) were measured at baseline and at 4, 8, 13 and 25 weeks. Geometric mean total serum 25(OH)D concentrations (sum of 25(OH)D2 and 25(OH)D3) at baseline was 80 nmol/l. After 25 weeks, participants randomised to D2 and placebo had a significant reduction in serum 25(OH)D3 concentrations over the winter months compared with vitamin D3-supplemented participants (both P< 0.001). Supplementation with vitamin D2 increased serum 25(OH)D2 but produced a 9 (95 % CI 1, 17) nmol/l greater decline in the 25(OH)D3 metabolite compared with placebo (P< 0.036). Overall, total serum 25(OH)D concentrations were 21 (95 % CI 14, 30) nmol/l lower in participants receiving vitamin D2 compared with those receiving D3 (P< 0.001), among whom total serum 25(OH)D concentrations remained unchanged. No intervention-related changes in PTH were observed. Daily supplementation of vitamin D3 was more effective than D2; however, the functional consequence of the differing metabolic response warrants further investigation.

Comparison of vitamin D2 and vitamin D3 supplementation in raising serum 25-hydroxyvitamin D status: a systematic review and meta-analysis

Background: Currently, there is a lack of clarity in the literature as to whether there is a definitive difference between the effects of vitamins D2 and D3 in the raising of serum 25-hydroxyvitamin D [25(OH)D].Objective: The objective of this article was to report a systematic review and meta-analysis of randomized controlled trials (RCTs) that have directly compared the effects of vitamin D2 and vitamin D3 on serum 25(OH)D concentrations in humans.Design: The ISI Web of Knowledge (January 1966 to July 2011) database was searched electronically for all relevant studies in adults that directly compared vitamin D3 with vitamin D2. The Cochrane Clinical Trials Registry, International Standard Randomized Controlled Trials Number register, and clinicaltrials.gov were also searched for any unpublished trials.Results: A meta-analysis of RCTs indicated that supplementation with vitamin D3 had a significant and positive effect in the raising of serum 25(OH)D concentrations compared with the effect of vitamin D2 (P = 0.001). When the frequency of dosage administration was compared, there was a significant response for vitamin D3 when given as a bolus dose (P = 0.0002) compared with administration of vitamin D2, but the effect was lost with daily supplementation.Conclusions: This meta-analysis indicates that vitamin D3 is more efficacious at raising serum 25(OH)D concentrations than is vitamin D2, and thus vitamin D3 could potentially become the preferred choice for supplementation. However, additional research is required to examine the metabolic pathways involved in oral and intramuscular administration of vitamin D and the effects across age, sex, and ethnicity, which this review was unable to verify.

Serum 25-Hydroxy-Calciferol Level and Failed Back Surgery Syndrome

To assess the association of serum 25-hydroxy-calciferol levels with pain and low back function in patients with failed back surgery syndrome. Records of 6 men and 3 women aged 25 to 54 (mean, 39.2) years who had failed back surgery syndrome after pedicular screw and rod instrumentation for lower lumbar degenerative diseases were reviewed. They had moderate-to-severe pain (visual analogue scale [VAS] score of >6) and low back function disability (Japanese Orthopaedic Association [JOA] back score of <10). In all patients, the serum 25-hydroxy-calciferol level was <30 ng/ ml, indicating vitamin D deficiency. Vitamin D2 (20 000 IU per day) was given for 10 days, and vitamin D3 (600 IU per day) was given for maintenance. Patients were followed up at months 3 and 6. Three men and 4 women aged 27 to 55 (mean, 41.3) years who were age- and disease-matched but achieved good outcomes (VAS score of 0-1 and mean JOA low back score of 14.7) were used as indirect referents. All 7 matched patients except one had a normal serum 25-hydroxy-calciferol level (mean, 40.6 ng/ml). In the 9 patients with failed back surgery syndrome, the mean duration of chronic pain was 2.6 years; the mean VAS score for pain was 7.7; the mean JOA low back score was 7.6; the mean number of reoperations was 2.2; and the mean serum 25-hydroxy-calciferol level was 17.0 ng/ml. Two male patients had grade-IV motor weakness and decreased sensory function based on the pin prick test. One patient had a history of prolonged (>3 months) antibiotic use after primary surgery, but had no evidence of infection. Six months after vitamin D2 and vitamin D3 supplementation, the mean serum 25-hydroxy-calciferol level improved significantly (17.0 vs. 42.5 ng/ml), as did the mean pain score (7.7 vs. 4.2) and mean JOA back score (7.6 vs. 11.1). Seven of the patients had a pain score of <6 and a JOA back score of >10, the remaining 2 patients had neurological deficits and only slight improvement. Vitamin D supplementation may be used as an adjuvant treatment for patients with failed back surgery syndrome.

Application of Ultra-High-Performance Liquid Chromatography/ Tandem Mass Spectrometry for the Measurement of Vitamin D in Infant Formula and Adult/Pediatric Nutritional Formula: First Action 2011.11

In an effort to measure vitamin D, ultra-high-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) was applied to samples. The use of UHPLC-MS/MS decreased the run time by 50%. The UHPLC-MS/MS achieved equal or better separation efficiency with complex food matrixes compared to HPLC-MS/MS. It was also observed that under the optimized conditions of UHPLC, all previtamins of vitamin D3, D2, and isotope-labeled vitamin D3 were baseline-separated from their corresponding vitamins. The sterol isomers found in complex food matrixes that interfere in the analysis were well separated from the analytes. The accuracy of the method was evaluated by analyzing National Institute of Standards and Technology Standard Reference Material 1849 infant reference material. The average vitamin D3 concentration was 0.251 +/- 0.012 microg/g. This showed excellent agreement with the certified value of 0.251 +/- 0.027 microg/g. The spike recovery study of a commercial infant formula matrix showed a range of recovery from 100 to 108%. The LOQ values determined were 0.0022 and 0.0028 microg/g for vitamins D3 and D2, respectively; LOD values were 0.00065 and 0.00083 microg/g for vitamins D3 and D2, respectively.

Temperature-Sensitive Mutants and Revertants in the Coronavirus Nonstructural Protein 5 Protease (3CLpro) Define Residues Involved in Long-Distance Communication and Regulation of Protease Activity

Positive-strand RNA virus genomes are translated into polyproteins that are processed by viral proteases to yield functional intermediate and mature proteins. Coronaviruses (CoVs) carry genes that encode an nsp5 protease (also known as 3CLpro or Mpro) responsible for 11 maturation cleavages. The nsp5 structure contains two chymotrypsin-like domains (D1 and D2) and a unique domain (D3), and forms functional dimers. However, little is known of interactions or communication across the structure of the protease during nsp5 activity. Using reverse genetic mutagenesis of the CoV murine hepatitis virus (MHV) nsp5, we identified a new temperature-sensitive (ts) mutation in D2 of nsp5 (Ser133Ala) and confirmed a ts residue in D3 (Phe219Leu). Both D2-tsS133A and D3-tsF219L were impaired for viral replication and nsp5-mediated polyprotein processing at the nonpermissive temperature. Passage of tsS133A and tsF219L at the nonpermissive temperature resulted in emergence of multiple second-site suppressor mutations, singly and in combinations. Among the second-site mutations, a D2 His134Tyr change suppressed the ts phenotype of D2-tsS133A and D3-tsF219L, as well as the previously reported D2-tsV148A. Analysis of multiple CoV nsp5 structures, and alignment of nonredundant nsp5 primary sequences, demonstrated that ts and suppressor residues are not conserved across CoVs and are physically distant (>10 Å) from each other, from catalytic and substrate-binding residues, and from the nsp5 dimer interface. These findings demonstrate that long-distance communication pathways between multiple residues and domains of nsp5 play a significant role in nsp5 activity and viral replication, suggesting possible novel targets for non-active site inhibitors of nsp5.

Molecular characterization and diagnostics of stubby root and virus vector nematodes of the family Trichodoridae (Nematoda: Triplonchida) using ribosomal RNA genes

Plant parasitic nematodes of the family Trichodoridae cause substantial yield losses in many agricultural crops. Rapid and accurate identification of trichodorids to the species level is critical for selection of appropriate measures for control. This study analysed 99 sequences of the D2–D3 expansion segments of the 28S rRNA gene and 131 sequences of the 18S rRNA gene from the stubby nematodes belonging to the genera Nanidorus, Paratrichodorus and Trichodorus. Species delimiting was based on the integration of morphological identification, which is not provided in the present article, and molecular‐based phylogenetic inference and sequence analysis. Twenty‐two valid species and several species complexes were identified among nematodes included in the analysis. PCR‐RFLPs of the partial 18S rDNA and the D2–D3 expansion segments of the 28S rDNA were tested and proposed for identification of these nematodes. Gel PCR‐RFLP profiles and tables with restriction fragment lengths for several diagnostic enzymes are provided for identification. Some problems of taxonomy and phylogeny of nematodes of the family Trichodoridae are also discussed.

Vitamin D: The "sunshine" vitamin.

Vitamin D insufficiency affects almost 50% of the population worldwide. An estimated 1 billion people worldwide, across all ethnicities and age groups, have a vitamin D deficiency (VDD). This pandemic of hypovitaminosis D can mainly be attributed to lifestyle (for example, reduced outdoor activities) and environmental (for example, air pollution) factors that reduce exposure to sunlight, which is required for ultraviolet-B (UVB)-induced vitamin D production in the skin. High prevalence of vitamin D insufficiency is a particularly important public health issue because hypovitaminosis D is an independent risk factor for total mortality in the general population. Current studies suggest that we may need more vitamin D than presently recommended to prevent chronic disease. As the number of people with VDD continues to increase, the importance of this hormone in overall health and the prevention of chronic diseases are at the forefront of research. VDD is very common in all age groups. As few foods contain vitamin D, guidelines recommended supplementation at suggested daily intake and tolerable upper limit levels. It is also suggested to measure the serum 25-hydroxyvitamin D level as the initial diagnostic test in patients at risk for deficiency. Treatment with either vitamin D2 or vitamin D3 is recommended for deficient patients. A meta-analysis published in 2007 showed that vitamin D supplementation was associated with significantly reduced mortality. In this review, we will summarize the mechanisms that are presumed to underlie the relationship between vitamin D and understand its biology and clinical implications.