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Malnutrition in the oral and maxillofacial surgery surgical patient can have critical implications in the overall well-being and prognosis of the long-term, hospitalized, ill patient. The OMS should be capable of assessing the patient's nutritional status and nutritional requirements and developing appropriate recommendations for proper nutritional management. Knowledge of the various modalities of nutritional support should be readily available to the OMS practitioner. Show less

Harmful influence of tobacco smoke on mother and child comprises many effects, among which the best known are: impaired fecundability, increased risk of pregnancy and perinatal period complications, increased morbidity of neonates and respiratory, circulatory and neurological disorders in neonates, infants and older children. The aim of this study was to assess smoking prevalence among pregnant women and young mothers and to assess the degree of exposition of infants to environmental tobacco smoke (ETS). The study was carried out among 74 women after delivery in the F. Raszeja City Hospital in Poznan, Poland, between December 2005 and May 2006. Smoking prevalence among pregnant women was 8.1% (women with high educational level--2.6%, others--14.3%). Smoking prevalence among mothers within the first year after delivery was 4.05% (women with high educational level--0%, others--8.6%). 17.6% infants (10.3% children of mothers with high educational level and 25.7% children delivered by the other mothers) were exposed to ETS because of smokers at home (other than mother); 10 infants exposed to ETS had non-smoking mother. Show less

The cigarette smoke is a significant source of heavy metals, which after being absorbed into the human organism, may be accumulated in a calcified tissue. The accumulation process may be also a result of a passive exposure to the cigarette smoke. As the hard tissues of deciduous teeth are relatively stable in chemical composition, they are widely used as the indicators of the exposure to heavy metals in children. This project is aimed to estimate the effect of passive smoke on the levels of selected toxic and essential elements in deciduous teeth. The research material consisted of 386 deciduous teeth. Out of this, 205 teeth were from the children exposed to the cigarette smoke in the apartments. The levels of cadmium, copper, iron, manganese, lead, zinc, calcium and magnesium were determined using atomic absorption spectrometer with flame atomization. It was concluded that the exposure to the cigarette smoke in children is a factor producing the changes in the levels of selected toxic and essential elements in deciduous teeth. This particularly results in the higher levels of cadmium, copper, lead and zinc, which are the permanent constituents of the cigarette smoke, and the lower levels of manganese, calcium and magnesium. Moreover, the disturbed gradient of lead levels dependent on the tooth type is observed in the children exposed to the cigarette smoke in the apartments. Show less

Tobacco smoke contains many thousands of chemicals including a plethora of carcinogens. Most chemical carcinogens undergo metabolic activation leading to the formation of electrophilic metabolites. These highly reactive species interact with nucleophilic sites in DNA and cellular proteins. Thus as result of voluntary as well as passive tobacco smoking, the DNA and protein adducts are detected in human tissues and blood. Tobacco smoke induces also the oxidative DNA damage. This paper reviews the data indicating, that analysis of DNA and protein adducts provides mechanistic insight into association between tobacco smoke and neoplastic and degenerative diseases. Show less

The following biomarkers: cotinine, 1-hydroxypyrene (1-OHP) and carboxyhemoglobin (HbCO) were used for the estimation of the tobacco smoke exposure. In our study urine and blood sampled from 98 volunteers were subjected to assessment of the biomarkers concentrations under investigation. According to the self-reports of the volunteers, all of the volunteers were divided into three groups: non-smokers, passive and active smokers. The mean values of the biomarker concentrations were calculated for these groups. In order to verify the tobacco smoke exposure specified by volunteers, we tested the real urinary cotinine amount. Based on obtained values of this biomarker, the another volunteer's partition into the groups of nonsmokers, passive and active smokers were done. The new mean values for all of these groups were calculated. The mean values obtained for both group partitions differed. The latter partition gave opportunity to use 1-OHP for the estimation of the tobacco smoke exposure not only for the active smokers but also for the passive smokers. When HbCO was taken into consideration as the biomarker, the estimation of the tobacco smoke exposure of the passive smokers was not significant different, as compared with the group of nonsmokers. The analysis of the correlation between the concentrations of 1-OHP and cotinine gave opportunity for the estimation to tobacco smoke exposure of the passive and active smokers. The estimation of the correlation, between the HbCO and cotinine concentrations was restricted for this purpose. Show less

In Poland, tobacco smoking by women in a procreational age as well as the pregnant women is a common phenomenon. The aim of the conducted research was to assess the usefulness of cotinine markers in different biological materials--mother and newborn's urine, cord blood se. rum, placenta--as a biomarker of tobacco smoking by delivering women, and dependence between these biomarkers and the newborn's health state. 218 pregnant women (117 smokers and 101 non-smokers), who were checked in at the Perinatology and Gynecology Clinic of the University of Medical Sciences in Poznan, took part in the first stage of the research (period between the twelfth and sixteenth week of pregnancy) carried out between years 2004-2006. In the second stage, 201 pairs of women (89 smokers and 112 non-smokers) and their newborns were checked after the women came to hospital to deliver. The research that was conducted showed that both cotinine in the urine of delivering women and in the urine of newborns as well as in the cord blood serum may be used as a biomarker of exposure of a foetus to tobacco smoke. For practical reasons, it must be assumed that the delivering women urine should be the material from choice. The research did not indicate the usefulness of the determination of cotinine in the placenta, in order to assess the exposure of the foetus to the components of tobacco smoke. On the other hand, again it confirmed the influence of tobacco smoking on the newborn's birth parameters, a correlation between the birth weight, body length and cotinine concentration in the urine of a mother makes it possible to predict the lowering of the antropometric parameters of the newborn as a result of tobacco smoking by pregnant women. Show less

The aim of the paper was to evaluate the association between preconceptional and prenatal exposure to tobacco smoke and the risk of all cancers combined and separately, for brain tumors, leukemias, lymphomas and other childhood cancers. We conducted systematic search of the literature using PUBMED, MEDLINE and EBSCO database to identify all publications dealing with environmental tobacco smoke exposure and childhood cancer. Show less

Negative feedback is among the key mechanisms for regulating receptor tyrosine kinase (RTK) signaling. Human Sef, a recently identified inhibitor of RTK signaling, encodes different isoforms, including a membrane spanning (hSef-a) and a cytosolic (hSef-b) isoform. Previously, we reported that hSef-b inhibited fibroblast proliferation and prevented the activation of mitogen-activated protein kinase (MAPK), without affecting protein kinase B/Akt or p38 MAPK. Conflicting results were reported concerning hSef-a inhibition of MAPK activation, and the effect of hSef-a on other RTK-induced signaling pathways is unknown. Here we show that, in fibroblasts, similar to hSef-b, ectopic expression of hSef-a inhibited fibroblast growth factor-induced cell proliferation. Unlike hSef-b, however, the growth arrest was mediated via a MAPK-independent mechanism, and was accompanied by elevated p38 MAPK phosphorylation and inhibition of protein kinase B/Akt. In addition, hSef-a, but not hSef-b, mediated apoptosis in fibroblast growth factor-stimulated cells. Chemical inhibitor of p38 MAPK abrogated the effect of hSef-a on apoptosis. In epithelial cells, ectopic expression of hSef-a inhibited the activation of MAPK, whereas down-regulation of endogenous hSef-a significantly increased MAPK activation and accelerated growth factor-dependent cell proliferation. These results indicate that hSef-a is a multifunctional negative modulator of RTK signaling and clearly demonstrate that hSef-a can inhibit the activation of MAPK, although in a cell type-specific manner. Moreover, the differences between the activities of hSef-a and hSef-b suggest that hSef isoforms can control signal specificity and subsequent cell fate by utilizing different mechanisms to modulate RTK signaling. Show less

The Structural Proteomics In Europe (SPINE) programme is aimed at the development and implementation of high-throughput technologies for the efficient structure determination of proteins of biomedical importance, such as those of bacterial and viral pathogens linked to human health. Despite the challenging nature of some of these targets, 175 novel pathogen protein structures (approximately 220 including complexes) have been determined to date. Here the impact of several technologies on the structural determination of proteins from human pathogens is illustrated with selected examples, including the parallel expression of multiple constructs, the use of standardized refolding protocols and optimized crystallization screens. Show less

SPINE (Structural Proteomics In Europe) was established in 2002 as an integrated research project to develop new methods and technologies for high-throughput structural biology. Development areas were broken down into workpackages and this article gives an overview of ongoing activity in the bioinformatics workpackage. Developments cover target selection, target registration, wet and dry laboratory data management and structure annotation as they pertain to high-throughput studies. Some individual projects and developments are discussed in detail, while those that are covered elsewhere in this issue are treated more briefly. In particular, this overview focuses on the infrastructure of the software that allows the experimentalist to move projects through different areas that are crucial to high-throughput studies, leading to the collation of large data sets which are managed and eventually archived and/or deposited. Show less

Male infertility is now a major reproductive health problem because of an increasing number of environmental pollutants and chemicals, which eventually result in gene mutations. Genetic alterations caused by environmental factors account for a significant percentage of male infertility. Microarray technology is a powerful tool capable of measuring simultaneously the expression of thousands of genes expressed in a single sample. Eventually, advances in genetic technology will allow for the diagnosis of patients with male infertility due to congenital reasons or environmental factors. Since its introduction in 1994, microarray technology has made significant advances in the identification and characterization of novel or known genes possibly correlated with male infertility in mice, as well as in humans. This provides a rational basis for the application of microarray to establishing molecular signatures for the diagnosis and gene therapy targets of male infertility. In this review, the differential gene expression patterns characterized by microarray in germ and somatic cells at different steps of development or in response to stimuli, as well as a number of novel or known genes identified to be associated with male infertility in mice and humans, are addressed. Moreover, issues pertaining to measurement reproducibility are highlighted for the application of microarray data to male infertility. Show less

Failure to treat deep vein thrombosis (DVT) is associated with significant morbidity and mortality. Anticoagulation, although effective at preventing clot progression, is not able to prevent postthrombotic syndrome. Catheter-directed thrombolysis is a more aggressive alternative, with some small studies suggesting a better long-term outcome, but the associated risks are significant, and the treatment can require 2-3 days in a monitored setting. This report describes the power pulse technique, in which mechanical thrombectomy is combined with thrombolytic agents to maximize the effectiveness of the treatment and reduce the need for prolonged infusion and its associated risks. A 24-patient retrospective study showed complete thrombus removal (>90%) in 12 patients, substantial thrombus removal (50%-90%) in seven patients, and partial thrombus removal (<50%) in five patients. All 24 patients had resolution of presenting symptoms. Only two patients required blood transfusion, and one patient experienced temporary nephropathy. Show less

The feeding response following administration of the free fatty acid oxidation inhibitor, mercaptoacetate (MA) is conceptualized as an experimental model of lipoprivation, which may contribute to the understanding of inter-individual differences in the modulation of this homeostatic response. Although variation in the intake of food, water and glucoprivation as well as intake of several nutrients is known to be associated with genetic variation, it is not known whether MA-induced feeding is similarly dependent upon genotype. The present study therefore examined MA-induced feeding in mice of 11 inbred (A/J, AKR/J, BALB/cJ, CBA/J, C3H/HeJ, C57BL6/J, C57BL10/J, DBA/2J, SJL/J, SWR/J, 129P3/J) and one outbred (CD-1) strains across a wide range of previously determined effective MA doses (5, 35, 70, 100 mg/kg) and test times (1-4 h). MA produced significant dose-dependent and strain-dependent increases in food intake with strong responses noted in DBA/2J, outbred CD-1 and AKR/J mice. More limited dose-specific increases in food intake following MA occurred in C3H/HeJ, BALB/cJ, CBA/J, SJL/J, SWR/J and C57BL/6J mice. In contrast, MA failed to significantly increase food intake in A/J, C57BL/10J and 129P/3J mice. MA-induced food intake correlated significantly across strains only following the two highest doses, and intake following only the highest MA dose correlated significantly across strains with intake following only a moderate glucoprivic dose of 2-deoxy-d-glucose. Thus, these inter-strain differences suggest that lipoprivic (e.g., MA intake) and glucoprivic (e.g., 2-deoxy-d-glucose intake) responsivity operate via only partially overlapping genetic mechanisms of action. The demonstration of genotype-dependent variability in this lipoprivic response may provide the basis for the subsequent identification of trait-relevant genes. Show less

In spite of recent evidence showing the importance of DBY (DEAD-box RNA helicase Y) in spermatogenesis in human, the biologic role of its homolog Dby (also known as Ddx3y) in the mouse is less clear. The present study aims at characterizing the molecular structure of Dby and comparing its expression with its X- and autosome-linked homologs in embryonic gonads and developing germ cells in mice. Molecular cloning by rapid amplification of 3'-cDNA ends showed that the Dby gene in the mouse gives rise to 2 transcripts that differ only in the length of the 3'-untranslated region as a consequence of the use of alternative polyadenylation signals. Measurement by quantitative real-time polymerase chain reaction showed that both transcripts were ubiquitously expressed and were present in male germ cells and Sertoli cells. They were more abundant in type A spermatogonia compared with pachytene spermatocytes and round spermatids. Expression of Dby in the embryonic gonad increased from day 10.5 and reached a peak at day 17.5. The expression level of Dby decreased after birth and remained low in adult male gonads. Although the level of expression of Dby was much lower than its X chromosome homolog, Ddx3 (also known as Ddx3x) in all samples examined, the pattern of expression of the 2 genes was comparable. In contrast, their autosomal homolog, D1Pas1(also known as PL10), was predominantly expressed in pachytene spermatocytes and round spermatids. This result is in accord with meiotic sex chromosome inactivation in that Dby and Ddx are replaced in pachytene spermatocytes by their autosomal retroposon. These observations indicate that unlike DBY in humans, the role of Dby in spermatogenesis is less obvious in the mouse and its biologic activity may be replaced by that of Ddx3 and D1Pas1. Show less

NAD(P)H quinone oxidoreductase 1 (NQO1) is a ubiquitous flavoenzyme that catalyzes two-electron reduction of quinones to hydroquinones utilizing NAD(P)H as an electron donor. NQO1 binds and stabilizes several short-lived proteins including the tumor suppressors p53 and p73 and the enzyme ornithine decarboxylase (ODC). Dicoumarol is a widely used potent competitive inhibitor of NQO1 enzymatic activity, which competes with NAD(P)H for binding to NQO1. Dicoumarol also disrupts the binding of NQO1 to p53, p73, and ODC and induces their ubiquitin-independent proteasomal degradation. We report here the crystal structure of human NQO1 in complex with dicoumarol at 2.75 A resolution. We have identified the interactions of dicoumarol with the different residues of NQO1 and the conformational changes imposed upon dicoumarol binding. The most prominent conformational changes that occur in the presence of dicoumarol involve Tyr 128 and Phe 232 that are present on the surface of the NQO1 catalytic pocket. On the basis of the comparison of the NQO1 structure in complex with different NQO1 inhibitors and our previous analysis of NQO1 mutants, we propose that the specific conformation of Tyr 128 and Phe 232 is important for NQO1 interaction with p53 and other client proteins. Show less

To explore the possible mechanism of apoptosis induced by photodynamic therapy (PDT) in human pancreatic cancer cells Capan-1 with 2-butylamino-2-demethoxy-hypocrellin B (BAHB) as photosensitizer. The localization of BAHB in Capan-1 cells was studied, apoptosis was determined by DNA gel electrophoresis after PDT. The mitochondria membrane potential (DYm) and cytochrome C release were observed by laser scan confocal microscopy and Western blotting. The low concentration photosensitizer was mainly localized in mitochondria and also in lysosomes when the concentration is high. DNA ladder analysis showed characteristic of apoptosis. The mitochondria membrane potential (DYm) showed a loss of 30% around, after 6 hours by PDT under laser scan confocal microscopy, which is caused by a sudden increase in the permeability of mitochondria membrane accompanied with apoptosis. In Western blotting, cytochrome C release was observed from the mitochondria into the cytoplasm during BAHB-induced apoptosis. The research suggests that BAHB-induced apoptosis is related to photosensitization of mitochondria. Show less

Cardiotoxin III (CTX III) is a basic polypeptide with 60 amino acid residues isolated from Naja naja atra venom. This is the first report on the mechanism of the anticancer effect of CTX III in human colorectal cancer Colo205 cells. 2. Cardiotoxin III-induced Colo205 cell apoptosis was confirmed by DNA fragmentation (DNA ladder and sub-G1 formation) with an IC(50) of 4 mg/mL at 48 h. 3. Further mechanistic analysis demonstrate that CTX III induced the loss of mitochondrial membrane potential (Dym), cytochrome c release from mitochondria into the cytosol and activation of capase-9, caspase 3, as well as markedly enhancing the expression of Bax, but not Bcl-2, protein in the cells. Moreover, the CTX III-induced apoptosis was significantly blocked by the broad-spectrum caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone. 4. However, CTX III did not generate the formation of reactive oxygen species and anti-oxidants, including N-acetylcysteine, and catalase could not block CTX III-induced apoptosis in the Colo205 cells. 5. Taken together, these results suggest that CTX III may induce apoptosis through a mitochondrial- and caspase-dependent mechanism and alteration of Bax/Bcl-2 ratio in human colorectal Colo205 cancer cells. Show less

Dyggve-Melchior-Clausen syndrome (DMC) (MIM 223800) and Smith-McCort dysplasia (SMC) (MIM 607326) are rare allelic autosomal recessive spondylo-epi-metaphyseal dysplasias (SEMDs) characterized by similar skeletal manifestations. Both phenotypes have been mapped to chromosome 18q21.1 and mutations in the DYM (dymeclin) gene were identified in 13 families with DMC and in two families with SMC. Most mutations identified in DMC predict a loss of function, while those identified in SMC are mainly missense mutations, presumably associated with residual DYM activity and a less severe phenotype. We studied three consanguineous families from Turkey, Lebanon, and Georgia, one with SMC and two with DMC and identified different homozygous DYM mutations (IVS3 194-1G > A, 938₉₄₂delTGTCT) in the DMC families. No mutation was identified in the SMC family, possibly suggesting genetic heterogeneity of this disorder. Show less

Adult patients with idiopathic hypogonadotropic hypogonadism (IHH) typically present with absent puberty and therefore have prepubertal testes. IHH is recognized as one of the few curable causes of male infertility and is often effectively treated with either gonadotropins or pulsatile GnRH therapy. The objective of this study was to determine the structure of the testis prior to initiation of treatment. Eight adult IHH patients with prepubertal testes (<4 ml), with no previous gonadotropin therapy and with no history of cryptorchidism underwent open bilateral testicular biopsy prior to the initiation of hormonal treatment. The testes of all patients showed seminiferous cords separated by interstitium composed of blood vessels, connective tissue cells and collagen fibres but typical adult Leydig cells were absent. The cords contained only Sertoli cells and early type A spermatogonia. The spermatogonia mostly resided in the centre of the cords and were often large, typical of gonocytes. Sertoli cells appeared immature with ovoid nuclei devoid of infoldings and cytoplasm that lacked polarity. Tight junctional complexes commonly found connecting adult Sertoli cells were lacking. These results demonstrate that the immature testes from patients with the severe form of IHH possess early spermatogonia that could possibly reinitiate spermatogenesis with appropriate hormone stimulation. Therefore, the immature testis of this IHH subset resembles those of prepubertal boys and may provide important biologic and genetic insights into testicular development. Show less

Gene expression profiling was performed using the National Institute on Aging 15,000-cDNA microarray to reveal the differential expression pattern of 160 genes between meiotic pachytene spermatocytes and postmeiotic round spermatids of the mouse. Our results indicate that more genes are expressed in spermatids than in spermatocytes. Genes participating in cell cycle regulation and chromatin structure and dynamics are preferentially expressed in spermatocytes, while genes for protein turnover, signal transduction, energy metabolism, and intracellular transport are prevalent in spermatids. This suggests that a switch of functional requirement occurs when meiotic germ cells differentiate into haploid spermatids. Concordant expression patterns were obtained when quantitative real-time polymerase chain reaction was performed to verify the microarray data. Interestingly, the majority of the differentially expressed genes were underrepresented in mitotic type A spermatogonia, and they were preferentially expressed in the testis. Our results suggest that an even higher proportion of the mouse genome is devoted to male gamete development from meiosis than was previously estimated. We also provide evidence that underscores the advantage of using purified germ cells over whole testes in profiling spermatogenic gene expression to identify transcripts that demonstrate stage-specific expression patterns. Show less

Experimental handling stress (EHS) was applied to clinically asymptomatic farmed goldfish (Carassius auratus L.). EHS affected the gills and skin integrity of the fish and was accompanied by increased levels of plasma glucose, cortisol and interleukin-10 (IL-10). EHS application was followed by highly significant enhancement of the rate of infection with a virulent Aeromonas salmonicida isolate. Cumulative ulceration at the initial phase of the ensuing goldfish ulcerative disease (GUD) evidenced a facilitating role of EHS in the onset of GUD. Host susceptibility to the pathogen increased from 40% in unstressed fish to 90% in the stressed fish. A. salmonicida could be reisolated from the early-stage skin lesions only, whereas opportunistic strains, other than A. salmonicida (A. sobria and A. hydrophila), were recovered from progressive-stage ulcers. The implication of these findings in fish aquaculture is discussed. Show less

The overall goal of this study is to unravel the role(s) played by glial cell line-derived neurotrophic factor (GDNF) in the fate of spermatogonial stem cells. There is great interest in the biology of spermatogonial stem cells, or A(single) spermatogonia, because of their importance in the treatment of infertility, the development of contraceptives, and the understanding of the etiology of testicular cancer, particularly seminoma. In the mouse, spermatogonial stem cells express GFRalpha-1, the receptor for GDNF, and respond to this growth factor in vivo and in vitro. GDNF is produced by the adjacent Sertoli cells, which are part of the germ-line stem cell niche in vertebrates. We specifically isolated GFRalpha-1-positive spermatogonia using an immunomagnetic bead technique. We then stimulated the cells with 100 ng/mL of rGDNF for 10 hours; unstimulated cells served as negative controls. Microarray analysis, immunocytochemistry, and Western blotting revealed that Numb, a regulator of the Notch pathway, is upregulated by GDNF in spermatogonial stem cells. There are indications that in rats, mice, and humans, the Notch pathway promotes spermatogonial differentiation. We observed that an increase in Numb expression is concomitant with Notch degradation in these cells. Thus, through Numb, GDNF might inhibit differentiation and allows the maintenance of the stem cell pool in the mouse seminiferous epithelium. Show less

The objective of this study was to compare the expression of Col1a1, Col1a2, and procollagen I in the seminiferous tubules of immature and adult mice and to characterize the cellular expression pattern of procollagen I in germ cells during spermatogenesis in order to provide necessary groundwork for further functional studies in the process of spermatogenesis. Microarray analysis demonstrated that Col1a1 and Col1a2 were abundantly expressed in the seminiferous tubules of 6-day-old mice compared with 60-day-old mice, and the expression levels of Col1a1 and Col1a2 mRNA were validated using a semi-quantitative RT-PCR assay. Western blot analysis further confirmed that procollagen I was expressed at a higher level in the seminiferous tubules of 6-day-old mice compared with 60-day-old mice. Immunohistochemical analysis revealed that type A spermatogonia were positive for procollagen I in the testis of 6-day-old mice, whereas Sertoli cells were negative for this protein. The in vivo procollagen I staining in type A spermatogonia was corroborated in spermatogonia exhibiting a high potential for proliferation and the ability to form germ cell colonies in in vitro culture. Moreover, procollagen I was also detected in type A spermatogonia, intermediate spermatogonia, type B spermatogonia, and preleptotene spermatocytes in the adult mouse testes, but positive staining disappeared in more differentiated germ cell lineages detaching from the basement membrane, including leptotene spermatocytes, pachytene spermatocytes, round spermatids and elongated spermatids. These data suggest that Col1a1, Col1a2 and procollagen I are associated with type A spermatogonia and play a potential role in mediating the detachment and migration of germ cells during spermatogenesis. Show less

Dyggve Melchior Clausen syndrome (DMC) is a severe autosomal recessive skeletal dysplasia associated with mental retardation. Direct sequencing of genomic DNA has identified causative mutations in the gene Dymeclin (chromosome 18q12-21), with the majority predicting the generation of a truncated protein product. To carry out molecular genetic studies in three DMC kindreds. Two novel nonsense mutations and two complex genomic duplication events resulting in exon repetition were identified. Exon dosage assessment or mRNA analysis, in addition to direct genomic DNA sequencing, should be employed in the investigation of DMC affected individuals. Genomic duplication may be the causative mutation mechanism in other autosomal recessive disorders. Show less

The principal goal of the Israel Structural Proteomics Center (ISPC) is to determine the structures of proteins related to human health in their functional context. Emphasis is on the solution of structures of proteins complexed with their natural partner proteins and/or with DNA. To date, the ISPC has solved the structures of 14 proteins, including two protein complexes. It has adopted automated high-throughput (HTP) cloning and expression techniques and is now expressing in Escherichia coli, Pichia pastoris and baculovirus, and in a cell-free E. coli system. Protein expression in E. coli is the primary system of choice in which different parameters are tested in parallel. Much effort is being devoted to development of automated refolding of proteins expressed as inclusion bodies in E. coli. The current procedure utilizes tagged proteins from which the tag can subsequently be removed by TEV protease, thus permitting streamlined purification of a large number of samples. Robotic protein crystallization screens and optimization utilize both the batch method under oil and vapour diffusion. In order to record and organize the data accumulated by the ISPC, a laboratory information-management system (LIMS) has been developed which facilitates data monitoring and analysis. This permits optimization of conditions at all stages of protein production and structure determination. A set of bioinformatics tools, which are implemented in our LIMS, is utilized to analyze each target. Show less

Mouse strain differences for intake of sucrose and saccharin have been reported across studies, and some of these differences have been related to variants of the Tas1r3 taste receptor gene. However, several methodological concerns remain, including use of relatively few strains and/or a limited number of palatable concentrations in previous analyses. The present study examined strain differences in sucrose intake among 11 inbred (A/J, AKR/J, BALB/cJ, CBA/J, C3H/HeJ, C57BL6/J, C57BL10/J, DBA/2J, SJL/J, SWR/J, 129P3/J) and one outbred (CD-1) mouse strains across nine different sucrose concentrations (0.0001-20%) using two-bottle 24-h preference tests which controlled for sucrose concentration presentation effects, sucrose and water bottle positions, and measurement of kilocalorie intake as sucrose or chow. A/J, C57BL/6J, CD-1 and SWR/J strains consumed the greatest (11.6-22 ml) amount of sucrose, whereas the A/J, C57BL/10J, SJL/J and SWR/J strains consumed the greatest (44-56%) percentages of kilocalories as sucrose. The AKR/J, CBA/J, C3H/HeJ and DBA/2J strains consumed the least (6.9-7.9 ml) amount of sucrose, and displayed lower (20-30%) percentages of kilocalories consumed as sucrose. Whereas A/J, C57BL/6J, C57BL/10J, CD-1, SWR/J and SJL/J strains all displayed the most pronounced compensatory decreases in chow intake as the percentage of kilocalories consumed as sucrose increased, the AKR/J, C3H/HeJ and DBA/2J strains failed to significantly alter chow intake even at high sucrose concentrations. There was a paucity of significant correlations in the percentage of sucrose intake between sucrose concentrations, but percentage of sucrose intake at lower concentrations did correlate with previous descriptions of saccharin intake and variants of the Tas1r3 taste receptor gene. These data demonstrate clear mouse strain differences across a range of measures in sucrose intake across a wide range of concentrations, but caution against extrapolating between extremely high and low concentrations. The identification of strains with diverging abilities to regulate kilocalorie intake when presented with high sucrose concentrations may lead to the successful QTL mapping of this trait. Show less

An unusual case is discussed in which a child presented with a significant facial deformity that was thought to be a soft tissue tumor. After surgical exploration it was found to be a fibrous encapsulated mass containing a foreign body. Show less

Spondyloepimetaphyseal dysplasias (SEMD) represent a heterogeneous group of conditions composed of at least 15 well-defined entities. The classification is based on clinical, radiological and molecular findings. Among them, several conditions also include a mental retardation (MR) syndrome, namely Wolcott-Rallison syndrome, Dyggve-Melchior-Clausen syndrome (DMC) and lysosomal storage disorders. Here, we report on a novel form of SEMD with MR in two Pakistani sisters born to first-cousin parents. SEMD, MR, microcephaly, ataxia, facial dysmorphism and hirsutism of back and legs were noted in the two children. Skeletal findings included flat vertebral bodies with irregular vertebral plates, irregular and flared metaphyses with vertical striations, small and irregular epiphyses, small carpal bones and narrow iliac wings without lacy pelvis iliac crest. Similarities with DMC prompted us to test and eventually exclude the DMC gene, dymeclin, by direct sequencing. Similarly, we excluded the PAPSS2 gene (3'-alpha phosphoadenosine 5'-phosphosulphate synthase 2) responsible for SEMD Pakistani type. The combination of features observed in the two sisters does not fit with any previously reported SEMD and represents therefore a novel form of autosomal recessive SEMD with MR. Show less