Case report: multiple systemic disseminated tuberculosis mimicking lymphoma on 18F-FDG PET/CT.
F-fluorodeoxyglucose positron emission tomography/computed tomography (F-18 FDG PET/CT) has an important role in the diagnosis of various malignancies. However, F-18 FDG can also exhibit intense accumulation in tissues in inflammatory conditions such as active tuberculosis (TB) and sarcoidosis. We report a case of a 52-year-old female with irritable cough. CT showed a lung mass with multiple bilateral lung nodules, and sarcoidosis was suspected. F-18 FDG PET/CT was undertaken for the diagnosis and showed intense uptake of FDG in the mass in the lower lobe of the right lung, multiple lymph nodes, liver, and spleen. The maximum standardized uptake value of F-18 FDG was 43.58. This pattern of involvement most likely represents lymphomatous involvement. Histopathology suggested tubercular involvement. The patient received anti-TB treatment and recovered. Abovementioned extent and distribution of F-18 FDG in tubercular lesion is relatively rare, thus, one must be observant and aware with regards to TB being a strong mimic of lymphoma in endemic regions.


Chemical and Biochemical Basis of Cell-Bone Matrix Interaction in Health and Disease.
Bone, a calcified tissue composed of 60% inorganic component (hydroxyapatite), 10% water and 30% organic component (proteins), has three functions: providing mechanical support for locomotion, protecting vital organs, and regulating mineral homeostasis. A lifelong execution of these functions depends on a healthy skeleton, which is maintained by constant bone remodeling in which old bone is removed by the bone-resorbing cell, osteoclasts, and then replaced by new bone formed by the bone-forming cell, osteoblasts. This remodeling process requires a physical interaction of bone with these bone cells. Moreover, numerous cancers including breast and prostate have a high tendency to metastasize to bone, which is in part attributable to the capacity of the tumor cells to attach to bone. The intensive investigation in the past two decades has led to the notion that the cell-bone interaction involves integrins on cell surface and bone matrix proteins. However, the biochemical composition of bone and emerging evidence are inconsistent with this belief. In this review, I will discuss the current understanding of the molecular mechanism underlying the cell-bone interaction. I will also highlight the facts and new findings supporting that the inorganic, rather than the organic, component of bone is likely responsible for cellular attachment.


Rapid increase in salivary IgA and broad recognition of spike protein following SARS-CoV-2 vaccination.
Saliva is a key component of mucosal immunity, which protects the oral cavity from viral infections. However, salivary immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in terms of immunoglobulin dynamics and recognition, have not been investigated sufficiently. In this study, saliva samples were collected from individuals that received SARS-CoV-2 vaccine, and immunoglobulin G (IgG), IgM, and IgA against whole spike protein and S1 protein were measured. IgA against whole spike protein increased significantly following vaccination, while IgA against S1 protein did not. Of note, the IgA response was evident two weeks after the first vaccine dose and continued to rise thereafter. On the contrary, IgG antibodies against S1 increased significantly at four weeks after vaccination. These results reveal the dynamics and recognition antigens of immunoglobulins in saliva, indicating the function of IgA in the mucosal immune system. These findings may pave the way for further studies on mucosal immune response induced by vaccination.


Topical application of antisense oligonucleotide-loaded chitosan nanoparticles to rats.
Skin delivery of antisense oligonucleotides (AsODNs) has exciting potential in the treatment of skin diseases. However, the therapeutic applications of oligonucleotide-based therapies are limited by the instability of these molecules toward nucleases, short half-life in vivo, and insufficient cellular uptake. The purpose of this study was to investigate in vivo antisense effect of AsODN-loaded chitosan nanoparticles after topical application. AsODN-loaded chitosan nanoparticles were topically applied to Sprague Dawley rats (adult and baby). At 1, 3, 6, 9, and 12 days posttransfection, animals' skin samples were taken for measurement of beta-galactosidase (beta-Gal) expression and histological control. After topical application of AsODN-loaded chitosan nanoparticles in different doses, beta-Gal expression reduced significantly. Highest inhibition was observed after 6 days of transfection of nanoparticles. Free AsODNs exhibited 35% of beta-Gal inhibition on the first day. beta-Gal expression was inhibited in approximately 82-85% with transfection of nanoparticles containing 30 microg AsODNs at 6 days. The antisense effect of AsODN-loaded chitosan nanoparticle in baby skin was evaluated at 6 days: 77-86% of beta-Gal suppression was measured and differences between the doses were not significant. Thus, chitosan nanoparticles are useful carrier for delivery of AsODNs into skin cells of rats and may be used for topical application on human skin.


Identification of Sarcocystis spp. in synanthropic (Muridae) and wild (Cricetidae) rodents from Argentina.
The occurrence of Sarcocystis species was investigated in synanthropic (Muridae) and wild (Cricetidae) rodents from Argentina. Nine species were captured (n = 356). Sarcocysts were detected in muscles of 8.7% (31/356) and 3.7% (4/106) of the rodents by histopathology and direct microscopic observation, respectively. PCR-sequencing targeting the 18S rRNA, cox1, and ITS1 regions was performed on samples with positive histopathology. Four different 18S rRNA sequences or sequence groups with high intra-group identities (99.6-100%) were detected in Mus musculus, Oxymycterus rufus, Akodon azarae, and Necromys lasiurus. Eight sequences showed 99.5-99.7% identity with S. dispersa. Thirteen sequences showed low identity (95.3-96.4%) with other Sarcocystis spp. The obtained coxI sequences (n = 9) were almost identical to each other and showed a high similarity with S. strixi (99.2-99.5%) and S. lutrae (99.1%), despite the 18S rRNA sequences from the same samples suggested the occurrence of at least two species. This suggests that coxI may not show high variability in Sarcocystis spp. that use rodents as intermediate hosts. Six ITS1 sequences were obtained, showing high identity but low coverage with several Sarcocystis spp. Multilocus sequence typing and BLAST analysis did not lead to an accurate species identification. Possible reasons are the detection of new species or the limited molecular information available from previously described Sarcocystis spp. Phylogeny suggests that the detected Sarcocystis spp. may use raptor birds or snakes as definitive hosts. This study represents the first molecular identification of Sarcocystis spp. in naturally infected rodents of the Cricetidae and Muridae families in South America.


Revision total knee arthroplasty using the total condylar III prosthesis.
Implant selection for the severely damaged knee being treated by revision is difficult. Fixed or rotating hinges have provided mixed results with a high frequency of complication. An alternative to a hinge implant is the total condylar III prosthesis, which can substitute for a deficient collateral ligament. Between August 1980 and April 1987 total condylar III prostheses were used for revision of failed total knee arthroplasties (TKA) of 21 knees in 19 patients. In the same time interval, 649 TKA revisions were performed for a frequency of 3.2% using this prosthesis. The indications for using this prosthesis were bone loss in 10 knees, instability in 9 knees, supracondylar femur fracture in 1 knee, and implant malposition in one knee. At a 4-year follow-up evaluation the knee scores were excellent in 25%, good in 25%, fair in 25%, and poor in 25%. Complications occurred in 33% of the knees. The total condylar III prosthesis provided results similar to other constrained implants used for revision in patients with severe bone loss and ligamentous instability.


Continuously Cumulating Meta-Analysis and Replicability.
The current crisis in scientific psychology about whether our findings are irreproducible was presaged years ago by Tversky and Kahneman (1971), who noted that even sophisticated researchers believe in the fallacious Law of Small Numbers-erroneous intuitions about how imprecisely sample data reflect population phenomena. Combined with the low power of most current work, this often leads to the use of misleading criteria about whether an effect has replicated. Rosenthal (1990) suggested more appropriate criteria, here labeled the continuously cumulating meta-analytic (CCMA) approach. For example, a CCMA analysis on a replication attempt that does not reach significance might nonetheless provide more, not less, evidence that the effect is real. Alternatively, measures of heterogeneity might show that two studies that differ in whether they are significant might have only trivially different effect sizes. We present a nontechnical introduction to the CCMA framework (referencing relevant software), and then explain how it can be used to address aspects of replicability or more generally to assess quantitative evidence from numerous studies. We then present some examples and simulation results using the CCMA approach that show how the combination of evidence can yield improved results over the consideration of single studies.


[DNA methylation and cancer].
Tumor suppressor genes can be silenced by DNA methylation during cancer development. Aberrant DNA methylation is closely associated with histone deacetylases and histone methyltransferases that can modify histone amino-terminal lysines and develop specific histone codes, resulting in inactive chromatin formation. These processes change epigenetic information that builds up abnormal chromatin structure, and creates the unique features of cancer cells. It is well known that thousands of genes are deregulated in cancer cells. Epigenetic alterations involving aberrant DNA methylation is a possible mechanism that can explain the genome-wide abnormality of gene expression. The mechanism responsible for the aberrant DNA methylation is unclear now, however it seems that de novo DNA methyltransferases (DNMTs) play an important role in the process. DNMT3 A and DNMT3B are thought to be de novo DNMTs in human. A nucleoside analogue of cytidine induces demethylation of DNA in cancer cells by inhibiting the function of DNMTs. It is significant to elucidate precise mechanisms of aberrant DNA methylation and develop small molecules that can inhibit methylation.


Osteoporosis and osteoarthritis: a bi-directional Mendelian randomization study.
To investigate the causal relationship between low bone mineral density (BMD) and osteoarthritis (OA) using Mendelian randomization (MR) design. Two-sample bi-directional MR analyses were performed using summary-level information on OA traits from UK Biobank and arcOGEN. Sensitivity analyses including MR-Egger, simple median, weighted median, MR pleiotropy residual sum, and outlier approaches were utilized in conjunction with inverse variance weighting (IVW). Gene ontology (GO) enrichment analyses and expression quantitative trait locus (eQTL) colocalization analyses were used to investigate the potential mechanism and shared genes between osteoporosis (OP) and OA. The IVW method revealed that genetically predicted low femoral neck BMD was significantly linked with hip (β = 0.105, 95% CI: 0.023-0.188) and knee OA (β = 0.117, 95% CI: 0.049-0.184), but not with other site-specific OA. Genetically predicted low lumber spine BMD was significantly associated with OA at any sites (β = 0.048, 95% CI: 0.011-0.085), knee OA (β = 0.101, 95% CI: 0.045-0.156), and hip OA (β = 0.150, 95% CI: 0.077-0.224). Only hip OA was significantly linked with genetically predicted reduced total bone BMD (β = 0.092, 95% CI: 0.010-0.174). In the reverse MR analyses, no evidence for a causal effect of OA on BMD was found. GO enrichment analysis and eQTL analysis illustrated that DDN and SMAD-3 were the most prominent co-located genes. These findings suggested that OP may be causally linked to an increased risk of OA, indicating that measures to raise BMD may be effective in preventing OA. More research is required to determine the underlying processes via which OP causes OA.


Artifact reduction for set theoretic super resolution image reconstruction with edge adaptive constraints and higher-order interpolants.
In this paper, we propose to improve the POCS-based super-resolution reconstruction (SRR) methods in two ways. First, the discretization of the continuous image formation model is improved to explicitly allow for higher order interpolation methods to be used. Second, the constraint sets are modified to reduce the amount of edge ringing present in the high resolution image estimate. This effectively regularizes the inversion process.


Photoperiod regulates dietary preferences and energy metabolism in young developing Fischer 344 rats but not in same-age Wistar rats.
The effects of photoperiod on dietary preference were examined using young growing Fischer 344 and Wistar rats, which are seasonal and nonseasonal breeders, respectively. Rats were provided a low-fat, high-carbohydrate diet (LFD: 66/10/24% energy as carbohydrate/fat/protein) and high-fat, low-carbohydrate diet (HFD: 21/55/24% energy as carbohydrate/fat/protein) simultaneously under long- (LD: 16 h light/day) and short-day (SD: 8 h light/day) conditions for 3 wk. Fischer 344 rats preferred the LFD to the HFD under the LD condition, whereas preference for both diets was equivalent under the SD condition. Consequently, their body weight and total energy intake exhibited 11-15 and 10-13% increases, respectively, under the LD condition. Calculation of energy intake from macronutrients revealed that rats under the LD condition consumed 20-24 and 9-13% higher energy of carbohydrates and proteins, respectively, than those under the SD condition. In contrast, Wistar rats preferred the LFD to the HFD irrespective of photoperiod and exhibited no photoperiodic changes in any parameters examined. Next, Fischer 344 rats were provided either the LFD or HFD for 3 wk under LD or SD conditions. Calorie intake was 10% higher in the rats fed the LFD than those fed the HFD under SD condition. However, rats under LD condition exhibited 5-10, 14, and 64% increases in body weight, epididymal fat mass, and plasma leptin levels, respectively, compared with those under the SD condition irrespective of dietary composition. In conclusion, photoperiod regulates feeding and energy metabolism in young growing Fischer 344 rats via the interactions with dietary macronutrient composition.


Effects of ketone bodies on amino acid metabolism in isolated rat diaphragm.
1. Diaphragms from 48h-starved rats were incubated in Krebs-Ringer bicarbonate medium at 37degreesC for 30min and then transferred into new medium and incubated for 1, 2 and 3 h. 2. The amount of free amino acids found at the end of each time of incubation was larger than the amount at the beginning of incubation, indicating that in this system proteolysis is prevailing. 3. The diaphragms was releasing mainly alanine and glutamine into the incubation medium. 4. Within the periods of incubation the release and metabolism of free amino acids was proceeding at a constant rate. 5. Addition of sodium DL-3-hydroxybutyrate decreased the tissue content of several amino acids, among which were tyrosine and phenylalanine, suggesting that proteolysis was decreased by ketone bodies. 6. In the presence of glucose (10mM) and branched-chain amino acids (0.5mM), sodium DL-3-hydroxybutyrate at concentrations of 4 or 6 mM resulted in 30% decrease in tissue alanine content and a 20% decline in alanine release. Release of taurine and glutamine was decreased by 19 and 16% respectively with 6 mM-sodium DL-3-hydroxybutyrate. Addition of sodium acetoacetate (1-3mM) also resulted in a 20-35% decrease in tissue content of alanine, glutamine and taurine and in a 15-24% decrease of alanine and glutamine release. Smaller decreases (less than 15%) in the release of glycine, threonine, proline, serine and aspartate were also observed in the presence of sodium DL-3-hydroxybutyrate or sodium acetoacetate. 7. Substitution of pyruvate (1.0mM) for glucose in the presence of acetoacetate restored alanine and glutamine production to control values. In the presence of acetoacetate, pyruvate also increased the tissue content of aspartate by 77% and decreased the tissue content of glutamate by 30%. 8. It is suggested that in diaphragms from starved rats, ketone bodies (a) in the absence of other substrates inhibit protein catabolism and (b) in the presence of glucose and branched-chain amino acids decrease alanine and glutamine production, by inhibiting glycolysis.


Attitudes of Flemish secondary school students towards euthanasia and other end-of-life decisions in minors.
The aim of the study was to investigate attitudes of secondary school students towards acceptability of requests by minors for end-of-life decisions (ELDs) with a possible life-shortening effect: non-treatment decisions, potentially life-shortening alleviation of pain and symptoms (APS) and euthanasia. A cross-sectional survey was conducted among second and fourth grades students in 20 secondary schools in Flanders, Belgium. An anonymous structured questionnaire was administered to measure attitudes towards acceptability of requests for euthanasia and other ELDs, towards the right to be informed about terminal prognosis and their own desire to be informed. In total, 1769 students participated. In case of a terminal patient, 61% found a request for euthanasia acceptable, 60% a request for APS and 69% a request for non-treatment decision, compared with 18% (euthanasia) and 50% (APS) in case of a non-terminal patient. Acceptance was highest among: boys, participants older than 14 years and participants from general as opposed to technical and vocational education. Sixty-six per cent said the parents' opinion not being asked was a circumstance that should hold back a physician from administering a lethal drug. Ninety per cent of participants thought a minor has the right to be informed about terminal prognosis of a disease while 78% would like to be informed themselves. Attitudes towards ELD requests varied with case and participant characteristics and type of ELD. The studied adolescents have a clear wish to be informed about terminal prognosis. Physicians and caregivers should adequately involve adolescents in decision making and tailor prognosis-related information to their needs and level of competency.


[Role of additional substrates in DDT degradation by cultures of Pseudomonas aeruginosa].
The activity of enzymes was compared during Pseudomonas aeruginosa 640x growth on compounds activating DDT to a different degree and on acetate which did not stimulate this process. The activity of dehydrogenases generating reduced cofactors necessary for DDT dechlorination was found to be much higher on effective additional substrates than on compounds which either had no effect on DDT degradation or stimulated this process only slightly. The activity of enzymes generating the reduced cofactors was shown to correlate with the extent of DDT degradation by this culture.


Screening for retinopathy of prematurity in South of Iran.
The purpose of this study is to report the screening data for retinopathy of prematurity (ROP) at a screening center in Southern Iran. A chart review was performed of all screened neonates who were referred to Poostchi screening center affiliated to Shiraz University of Medical Sciences from February 2006 to January 2010. Statistical analysis of data was performed with the Chi-square and independent t-test where appropriate. A P < 0.05 was considered statistically significant. Of 787 infants referred, 293 (37.2%) had some form of ROP and 77 cases (9.8%) had plus disease, only 6 (2%) patients progressed to advanced ROP stages 4 and 5. The mean gestational age (GA) of patients with ROP (ROP group) was statistically significantly lower at 29.46 ± 2.31 weeks compared to patients without ROP (non-ROP group) (31.56 ± 2.03 weeks) (P < 0.05). The mean GA of patients with plus disease was statistically significantly lower at 28.92 ± 2.18 weeks compared to patients without plus disease (30.98 ± 2.30 weeks) (P < 0.05). The mean birth weight in the ROP group was statistically significantly lower at 1248.46 ± 301.75 g compared to the non-ROP group (1485.79 ± 268.66 g) (P < 0.05). The mean birth weight of patients with plus disease was statistically significantly lower at 1207.92 ± 334.79 g compared to patients without plus disease (417.99 ± 293.19 g) (P < 0.05). There was no difference in the occurrence of ROP or plus disease between single or multiple births, normal vaginal delivery, and caesarian section and between clomiphene users and nonusers (P > 0.05, all cases). Greater cooperation between ophthalmologists, neonatologists, gynecologists, and health policy makers is necessary to optimize ROP screening programs. Data from this study can be used by health policy makers for implementation of health programs. These programs must include screening guidelines and effective coordination among the screening centers, antenatal, obstetric, and neonatal care services.


Monocyte Subset Recruitment Marker Profile Is Inversely Associated With Blood ApoA1 Levels.
Dyslipidemia promotes development of the atherosclerotic plaques that characterise cardiovascular disease. Plaque progression requires the influx of monocytes into the vessel wall, but whether dyslipidemia is associated with an increased potential of monocytes to extravasate is largely unknown. Here (using flow cytometry) we examined recruitment marker expression on monocytes from generally healthy individuals who differed in lipid profile. Comparisons were made between monocyte subsets, participants and relative to participants' lipid levels. Monocyte subsets differed significantly in their expression of recruitment markers, with highest expression being on either the classical or non-classical subsets. However, these inter-subset differences were largely overshadowed by considerable inter-participant differences with some participants having higher levels of recruitment markers on all three monocyte subsets. Furthermore, when the expression of one recruitment marker was high, so too was that of most of the other markers, with substantial correlations evident between the markers. The inter-participant differences were explained by lipid levels. Most notably, there was a significant inverse correlation for most markers with ApoA1 levels. Our results indicate that dyslipidemia, in particular low levels of ApoA1, is associated with an increased potential of all monocyte subsets to extravasate, and to do so using a wider repertoire of recruitment markers than currently appreciated.


Enhancement of the thermostability and hydrolytic activity of xylanase by random gene shuffling.
The thermostability of Streptomyces lividans xylanase B (SlxB-cat) was significantly increased by the replacement of its N-terminal region with the corresponding region from Thermomonospora fusca xylanase A (TfxA-cat) without observing a decrease in enzyme activity. In spite of the significant similarity between the amino acid sequences of the two xylanases, their thermostabilities are quite different. To facilitate an understanding of the contribution of structure to the thermostability observed, chimaeric enzymes were constructed by random gene shuffling and the thermostable chimaeric enzymes were selected for further study. A comparative study of the chimaeric and parental enzymes indicated that the N-terminus of TfxA-cat contributed to the observed thermostability. However, too many substitutions decreased both the thermostability and the activity of the enzyme. The mutants with the most desirable characteristics, Stx15 and Stx18, exhibited significant thermostabilities at 70 degrees C with optimum temperatures which were 20 degrees C higher than that of SlxB-cat and equal to that of TfxA-cat. The ability of these two chimaeric enzymes to produce reducing sugar from xylan was enhanced in comparison with the parental enzymes. These results suggest that these chimaeric enzymes inherit both their thermostability from TfxA-cat and their increased reactivity from SlxB-cat. Our study also demonstrates that random shuffling between a mesophilic enzyme and its thermophilic counterpart represents a facile approach for the improvement of the thermostability of a mesophilic enzyme.


Molecular cloning and expression of keratinocyte proline-rich protein, a novel squamous epithelial marker isolated during skin development.
We describe a novel rat cDNA named keratinocyte proline-rich protein (KPRP) isolated by RNA differential display during skin development. We determine that KPRP is expressed in stratified squamous epithelium, and its approximately 2.8-kb cDNA encodes a 699-amino acid protein with high proline content (19%). KPRP is an insoluble protein, similar to most epidermal terminal differentiation-associated proteins. Immunoblot of the protein lysate from keratinocytes, using strong reducing conditions, demonstrates two KPRP bands of approximately 76 and 55 kDa size. KPRP is expressed in stratified squamous epithelia of skin, tongue, and esophagus. The initiation of KPRP expression in fetal rat skin at E17, E18, E19, E20, and E21 was analyzed by reverse transcription-PCR. Fetal skin at E19 and later expresses KPRP. In situ hybridization of skin from E18, E19, and 4-day-old neonatal rats demonstrates that interfollicular and follicular keratinocytes express KPRP. Anti-KPRP antibody demonstrates KPRP protein localizes to all layers of stratified epithelia in skin, tongue, and esophagus. In cultured dermal keratinocytes, KPRP is diffusely distributed throughout the cytoplasm with denser staining adjacent to the nuclear and plasma membranes. Additionally, immunoreactive intracellular granules are observed during keratinocyte detachment from their plastic substrate. Rat KPRP has 89% homology to a mouse genomic DNA sequence and 56% homology to a human hypothetical protein. We conclude that KPRP may be a new epidermal terminal differentiation-related protein expressed in stratified squamous epithelia. KPRP is expressed by fetal dermal keratinocytes during late gestation and is a new marker of maturing epidermis during fetal skin development.


Effect of long-term administration of digoxin on exercise performance in chronic airflow obstruction.
We have studied the effect of long-term digoxin on exercise performance in 15 patients with pulmonary heart disease due to severe chronic airflow obstruction (FEV1/VC ratio 29 +/- 6%: mean +/- SD). Digoxin (0.25 mg/day) was given for 8 weeks in a randomized, double-blind crossover, placebo-controlled trial. All patients were ambulatory and had clinical features of right ventricular dysfunction but no clinical evidence of left ventricular dysfunction. Assessments included progressive and steady-state exercise, pulmonary function studies and evaluation of right and left ventricular function. In all patients the right ventricular ejection fraction was reduced; in 4 patients the left ventricular ejection fraction was also reduced. In patients whose left ventricular ejection fraction was initially abnormal, 8 weeks of digoxin increased left ventricular ejection fraction to normal. In spite of the improvement in resting ventricular function, exercise performance, the cardiopulmonary response to exercise, pulmonary function and general health status did not improve. We conclude that in patients with pulmonary heart disease: 1) digoxin improves ventricular function only if left ventricular function is reduced, and 2) despite the improvement in ventricular function digoxin does not improve pulmonary function, cardiopulmonary response to exercise or general feeling of well being.


[Value of the nucleolar organizers (AgNOR) in brain gliomas].
The aim of the study was to evaluate the feasibility and possible contribution of silver stained nucleolar organizer regions (AgNORs) to prognostic considerations, in a series of 55 supratentorial gliomas: eight grade II astrocytomas, twelve grade III astrocytomas, thirty grade IV astrocytomas, two glioblastomas, one anaplastic oligodendroglioma, one oligodendroglioma and one ependymoma. Silver NORs (AgNORs) were demonstrated according to the method of Crocker et al. A difference between AgNOR sizes in peritumor and tumor tissue is noted. The mean NOR numbers in the tumor areas range from 0.871 to 2.677, without overlap between peritumor gliosis and glial tumors. A comparative analysis reveals significant correlations between the mean NOR number per nucleus and histological grading. This technique can play a practical role in the diagnosis and grading of tumors sampled by stereotactic biopsies: a count higher than 0.8 is highly suggestive of malignancy. In addition, the distribution of NORs may be important: intratumoral heterogeneity expresses various degrees of transcriptional activity between different glial tumors of the same grade. This technique provides information about the biological behaviour of glial tumors supplementary to that obtained from growth fraction analysis.


AP-2 recruitment to synaptotagmin stimulated by tyrosine-based endocytic motifs.
Clathrin-mediated endocytosis is initiated by the recruitment of the clathrin adaptor protein AP-2 to the plasma membrane where the membrane protein synaptotagmin is thought to act as a docking site. AP-2 also interacts with endocytic motifs present in other cargo proteins. Peptides with a tyrosine-based endocytic motif stimulated binding of AP-2 to synaptotagmin and enhanced AP-2 recruitment to the plasma membrane of neuronal and non-neuronal cells. This suggests a mechanism by which nucleation of clathrin-coated pits is stimulated by the loading of cargo proteins.


A study on metabolic variables and its association with high sensitive C-reactive protein in obese children and adolescents.
Obesity in children and adolescents predispose to the development of obesity in adulthood and subsequent cardiovascular disease. High-sensitivity C-reactive protein (hsCRP) is a marker of low grade inflammatory state, which characterizes an atherosclerotic process. The aim of this study was to assess the metabolic abnormalities and its association with hsCRP in obese children and adolescents. A total of 62 obese children and adolescents and 24 healthy children and adolescents with a normal weight were recruited. In all subjects, anthropometric and biochemical parameters were measured. Body mass index (BMI) and blood pressure were significantly higher in the obese children and adolescents than the control. Obese children had significantly higher hsCRP levels (P < 0.001), total cholesterol, triglyceride, low-density lipoprotein-cholesterol (LDL-C) and lower high-density lipoprotein-cholesterol than the control group. Furthermore, homeostatic model assessment-insulin resistance (HOMA-IR) was significantly higher in obese children compared with the normal weight children. Furthermore, hsCRP showed a positive correlation with BMI (r = 0.357; P = 0.028), total cholesterol (r = 0.367; P = 0.008) and LDL-C (r = 0.356; P = 0.01), insulin (r = 0.311; P = 0.026) and not with HOMA-IR (r = 0.244; P = 0.084)). In conclusion, obese children and adolescents have significantly increased hsCRP compared with a normal weight group. Early intervention and prevention of obesity in children and adolescents decreases cardiovascular disease in later life.


Automated whole mount localisation techniques for plant seedlings.
Plant biology is currently experiencing a growing demand for easy and reliable mRNA and protein localisation techniques. Here, we present novel whole mount in situ hybridisation and immunolocalisation protocols, suitable to localise mRNAs and proteins in Arabidopsis seedlings. We demonstrate that these methods can be used in different organs of Arabidopsis seedlings as well as in other plant species. In order to achieve better reproducibility and higher throughput, we modified these protocols for automation to be performed by a liquid handling robot. In addition, we show that other procedures such as reporter enzyme assays and tissue clearing can be similarly automated. We present examples of application of our protocols including mRNA localisation and proteins and epitope tag (co)localisations which demonstrate that these methods provide reliable and versatile tools for expression, localisation and anatomical studies in plants.


Monitoring Piscirickettsia salmonis by denaturant gel electrophoresis and competitive PCR.
Reported strains of Piscirickettsia salmonis, a pathogen of salmonid fishes, were analyzed by amplifying part of the internal transcribed spacer (ITS) of the ribosomal RNA (rRNA) operon followed by denaturing gradient gel electrophoresis (DGGE) of the amplicons. All amplified fragments differing in sequence were distinguished by migration during DGGE. A simpler format, constant denaturant gel electrophoresis (CDGE), allowed the same diagnostic distinctions among strains. Sampling during 1997 and 1998 of salmonids from 5 different sites on and near Chiloé Island in southern Chile displaying piscirickettsiosis revealed only P. salmonis resembling LF-89, the type strain first isolated in 1989. These observations are encouraging for control strategies, which might otherwise be compromised by unpredictable shifts of P. salmonis types in salmon farms. A competitive PCR assay offered insight about the power of PCR for quantification and about specific tissue invasiveness by this intracellular pathogen. This approach revealed that the PCR could amplify approximately 1 to 10 P. salmonis genome equivalents against a background of > 99.9% salmonid DNA. It also raised the possibility that the salmonid brain is an important site for P. salmonis survival, with its bacterial load in 1 individual having been about 100 times the loads observed in liver and kidney. Pathogen detection by competitive PCR in a surface seawater sample from a netpen in use indicated a density of about 3000 to 4000 P. salmonis cells (or their DNA remnants) 1(-1). Such quantitative estimates should aid decisions about disease prevention and management as, for example, choice of netpen sites following fallow periods and certification of ova, which are known conduits of infection.


Srs2 and Sgs1 DNA helicases associate with Mre11 in different subcomplexes following checkpoint activation and CDK1-mediated Srs2 phosphorylation.
Mutations in the genes encoding the BLM and WRN RecQ DNA helicases and the MRE11-RAD50-NBS1 complex lead to genome instability and cancer predisposition syndromes. The Saccharomyces cerevisiae Sgs1 RecQ helicase and the Mre11 protein, together with the Srs2 DNA helicase, prevent chromosome rearrangements and are implicated in the DNA damage checkpoint response and in DNA recombination. By searching for Srs2 physical interactors, we have identified Sgs1 and Mre11. We show that Srs2, Sgs1, and Mre11 form a large complex, likely together with yet unidentified proteins. This complex reorganizes into Srs2-Mre11 and Sgs1-Mre11 subcomplexes following DNA damage-induced activation of the Mec1 and Tel1 checkpoint kinases. The defects in subcomplex formation observed in mec1 and tel1 cells can be recapitulated in srs2-7AV mutants that are hypersensitive to intra-S DNA damage and are altered in the DNA damage-induced and Cdk1-dependent phosphorylation of Srs2. Altogether our observations indicate that Mec1- and Tel1-dependent checkpoint pathways modulate the functional interactions between Srs2, Sgs1, and Mre11 and that the Srs2 DNA helicase represents an important target of the Cdk1-mediated cellular response induced by DNA damage.


Low Level Laser Therapy in patients with chronic foot and ankle joint pain.
Chronic foot and ankle joint pain is one of the most frequent complaints which is regularly seen in the out-patient clinic of our medical institute. In previous studies we have reported on the benefits of low level laser therapy (LLLT) for chronic pain in the elbow, hand, finger and the lower back. The present study examined the effects of LLLT on chronic foot and ankle joint pain. Over the past 5 years, 17 subjects visited the out-patient clinic with complaints of chronic foot and ankle joint pain of a variety of aetiologies. The patients received LLLT using a 1000 mW semi-conductor laser device, delivering 20.1 J/cm2 per point at 830 nm in continuous wave. Each patient was given four shots per session per foot twice a week for 4 weeks. A visual analogue scale (VAS) was used to determine the effects of LLLT for the chronic pain and after the end of the treatment regimen a significant improvement was observed (p < 0.01). All but 2 of the patients showed improvement: excellent (2) and good (13). After treatment, no significant differences were observed in the ankle joint range of motion, however. Discussions with the patients revealed that it was important for them to learn how to avoid overuse of the ankle when walking, poor walking posture and a poor pacing technique that would caused them foot and ankle pain in everyday life. Following these postural guidelines could ensure continuous benefits from the treatment. The present study demonstrated that LLLT was an effective form of treatment for chronic foot and ankle joint pain, in conjunction with postural education during all activities of daily living.


Nanoscale Energy Transport Dynamics across Nonbonded Solid-Molecule Interfaces and in Molecular Thin Films.
Thermal conductance across a solid-solid interface requires an atomic- or molecular-level understanding, especially when a system is in a non-equilibrium state and/or consists of nanosized materials with prominent differences in structures, properties, and vibrational behaviors. Here, we report the lattice dynamics of graphite-supported molecular thin films of ethanol, whose layers exhibit in-plane hydrogen-bonded chains and out-of-plane van der Waals stacking with clear structural anisotropy. The direct structure-probing method of ultrafast electron diffraction reveals a surprising temperature difference of more than 400 K at pico- to sub-nanosecond times across the graphite-ethanol interface, yet the temporal behavior signifies a reasonably large thermal boundary conductance. This apparent conflict in a non-equilibrium condition can be resolved by considering the coupling of out-of-plane motions, instead of the commonly used temperature-based model, at transient times for energy transport across the interface separated by van der Waals interactions with mismatched unit sizes and no strong bonds. The importance of spatiotemporally resolved structural dynamics at the atomic or molecular level is emphasized.


Minocycline reduces proinflammatory cytokine expression, microglial activation, and caspase-3 activation in a rodent model of diabetic retinopathy.
Diabetes leads to vascular leakage, glial dysfunction, and neuronal apoptosis within the retina. The goal of the studies reported here was to determine the role that retinal microglial cells play in diabetic retinopathy and assess whether minocycline can decrease microglial activation and alleviate retinal complications. Immunohistochemical analyses showed that retinal microglia are activated early in diabetes. Furthermore, mRNAs for interleukin-1beta and tumor necrosis factor-alpha, proinflammatory mediators known to be released from microglia, are also increased in the retina early in the course of diabetes. Using an in vitro bioassay, we demonstrated that cytokine-activated microglia release cytotoxins that kill retinal neurons. Furthermore, we showed that neuronal apoptosis is increased in the diabetic retina, as measured by caspase-3 activity. Minocycline represses diabetes-induced inflammatory cytokine production, reduces the release of cytotoxins from activated microglia, and significantly reduces measurable caspase-3 activity within the retina. These results indicate that inhibiting microglial activity may be an important strategy in the treatment of diabetic retinopathy and that drugs such as minocycline hold promise in delaying or preventing the loss of vision associated with this disease.


Clinical assessment and management of the oral environment in the oncology patient.
The dental care of oncology patients is an important component of general dental practice. Oncology patients have additional requirements for their outpatient care in the dental office. Intense involvement of the general dental practitioner in the patient's overall plan of care is essential so that appropriate preventive and therapeutic strategies are followed prior to chemotherapy, radiation and other medical treatments. This paper provides an overview of the role of the dental practitioner in the pre-treatment workup and post-treatment maintenance of oncology patients, and discusses the complications which occur during the intensive and in-hospital phases of care, in the context of approaches that show promise for reducing or preventing these. The role of the general dental practitioner in the maintenance of oral health for the remainder of the patient's life is stressed, with evidence-based recommendations given for optimal use of home care products which support oral health and improve quality-of-life.


ISH-facilitated analysis of meiotic bivalent pairing.
Chiasmata constitute one of the cornerstones of sexual reproduction in most eukaryotes. They mediate the reciprocal genetic exchange between homologues and are essential to the proper orientation of the homologous centromeres in meiosis I. As markers of recombination, they offer a cytological means of mapping. Rather than trying to accurately count individual chiasmata, we have examined properties of the mathematical relationship between frequencies of nonadorned disomic configurations in meiosis (ring, rods, and univalents) and the probabilities at which arms of the respective chromosomes are chiasmate (one or more chiasma per arm). Numerical analyses indicated that conventionally analyzed bivalents with nonidentified arms yield statistically biased estimates of chiasma probabilities under a broad range of circumstances. We subsequently analyzed estimators derived from adorned configurations with ISH-marked arms, which were found to be statistically far superior, and with no assumptions concerning interference across the centromere. We applied this methodology in the study of chromosomes 16 and 23 of cotton (Gossypium hirsutum), and estimated their arm lengths in centimorgans. The results for chromosome 23, the only one of the two chromosomes with a documented RFLP map, were consistent with the literature. Similar molecular-meiotic configuration analyses can be used for a wide variety of eukaryotic organisms and purposes: for example, providing far more powerful meiotic comparisons of genomes of chromosomes, and a rapid means of evaluating effects on recombination. Key words : meiotic configurations, chiasma frequencies, in situ hybridization, cotton.


Allogenic Tendon-Autologous Cartilage Cells Transplantation Enhances Adhesive/Growth Ability and Promotes Chondrogenesis in a Rabbit Model of Glenoid Labrum Damage.
BACKGROUND Glenoid labrum injury of the shoulder commonly occurs in athletes, especially those who perform throwing motions. This study investigated the effects of the established allogenic tendon-autologous cartilage cells reconstruction approach in a rabbit model of glenoid labrum damage. MATERIAL AND METHODS The allogenic tendons were isolated and extracted using the chemical extraction method. Cartilage cells were isolated from New Zealand rabbits and identified by detecting type II collagenase. The allogenic tendon-autologous cartilage cells were transplanted to the damaged glenoid labrum. HE staining was used to observe inflammatory cells, Masson staining was used to observe muscle fibers, and scanning electron microscopy (SEM) was used to assess antigenicity of tendon tissues. PSA and AB staining were used to examine neutral protein mucopolysaccharide and acidic protein mucopolysaccharide, respectively. We assessed cartilage cell growth in autologous cartilage cells combined with allogenic tendon transplanted tissues. RESULTS Allogenic tendons were well prepared using chemical extraction method due to use of HE staining, Masson staining, and SEM. TGF-ß1 treatment induced cartilage cell formation and triggered expression of acidic and neutral protein mucopolysaccharides. HE staining, Masson staining, PAS staining, and AB staining methods showed that autologous cartilage cells combined with allogenic tendon transplanted tissues had better growth of cartilage cells. CONCLUSIONS This study establishes the allogenic tendon-autologous cartilage cells reconstruction and transplantation approach and illustrated higher adhesive ability and growth ability, and better chondrogenesis in a rabbit model of glenoid labrum damage.


Pulse transit time-based blood pressure estimation using hilbert-huang transform.
The pulse transit time (PTT) based method has been suggested as a continuous, cuffless and non-invasive approach to estimate blood pressure. It is of paramount importance to accurately determine the pulse transit time from the measured electrocardiogram (ECG) and photoplethysmo-gram (PPG) signals. We apply the celebrated Hilbert-Huang Transform (HHT) to process both the ECG and PPG signals, and improve the accuracy of the PTT estimation. Further, the blood pressure variation is obtained by using a well-established formula reflecting the relationship between the blood pressure and the estimated PTT. Simulation results are provided to illustrate the effectiveness of the proposed method.


Antimalarial properties of Goniothalamin in combination with chloroquine against Plasmodium yoelii and Plasmodium berghei growth in mice.
Malaria is a disease which is still endemic and has become a disastrous scourge because of the emergence of antimalarial drug resistant Plasmodium falciparum. A new approach in addressing this is in developing a combination drug. This study is to show the enhancement of antimalarial properties, when single compound, goniothalamin combine with standard drug, chloroquine. Based on 4 Day Test, percentage of parasite growth on treated infected mice were determined. Oral treatment with 1 mg/kg BW of chloroquine on experimental mice suppressed 70% and 76.7% of both Plasmodium yoelii and Plasmodium berghei, respectively. The infection of P. berghei in mice was inhibited less than 50% by goniothalamin individual treatment at all doses in this study. About 27.8% and 18.5% inhibition of infection were observed in P. yoelii infected mice treated with 30 mg/kg and 60 mg/kg of goniothalamin respectively and the suppression exceed more than 50% at higher doses (90 and 120 mg/kg). Combination of 1 mg/kg chloroquine with either 30 mg/kg or 60 mg/kg of goniothalamin decreased the parasitemia of P. yoelii infected mice more than 90% and prolong the survival up to 100% after treatment. Similar treatment to P. berghei infected mice only shows about 60% reduction of parasitemia. The study findings showed that antimalarial property of goniothalamin was enhanced by combination with chloroquine at lower dose of each drug.


Transcript profiling in the chl1-5 mutant of Arabidopsis reveals a role of the nitrate transporter NRT1.1 in the regulation of another nitrate transporter, NRT2.1.
Arabidopsis thaliana mutants deficient for the NRT1.1 NO(3)(-) transporter display complex phenotypes, including lowered NO(3)(-) uptake, altered development of nascent organs, and reduced stomatal opening. To obtain further insight at the molecular level on the multiple physiological functions of NRT1.1, we performed large-scale transcript profiling by serial analysis of gene expression in the roots of the chl1-5 deletion mutant of NRT1.1 and of the Columbia wild type. Several hundred genes were differentially expressed between the two genotypes, when plants were grown on NH(4)NO(3) as N source. Among these genes, the N satiety-repressed NRT2.1 gene, encoding a major component of the root high-affinity NO(3)(-) transport system (HATS), was found to be strongly derepressed in the chl1-5 mutant (as well as in other NRT1.1 mutants). This was associated with a marked stimulation of the NO(3)(-) HATS activity in the mutant, suggesting adaptive response to a possible N limitation resulting from NRT1.1 mutation. However, derepression of NRT2.1 in NH(4)NO(3)-fed chl1-5 plants could not be attributed to lowered production of N metabolites. Rather, the results show that normal regulation of NRT2.1 expression is strongly altered in the chl1-5 mutant, where this gene is no more repressible by high N provision to the plant. This indicates that NRT1.1 plays an unexpected but important role in the regulation of both NRT2.1 expression and NO(3)(-) HATS activity. Overexpression of NRT2.1 was also found in wild-type plants supplied with 1 mM NH(4)(+) plus 0.1 mM NO(3)(-), a situation where NRT1.1 is likely to mediate very low NO(3)(-) transport. Thus, we suggest that it is the lack of NRT1.1 activity, rather than the absence of this transporter, that derepresses NRT2.1 expression in the presence of NH(4)(+). Two hypotheses are discussed to explain these results: (1) NRT2.1 is upregulated by a NO(3)(-) demand signaling, indirectly triggered by lack of NRT1.1-mediated uptake, which overrides feedback repression by N metabolites, and (2) NRT1.1 plays a more direct signaling role, and its transport activity generates an unknown signal required for NRT2.1 repression by N metabolites. Both mechanisms would warrant that either NRT1.1 or NRT2.1 ensure significant NO(3)(-) uptake in the presence of NH(4)(+) in the external medium, which is crucial to prevent the detrimental effects of pure NH(4)(+) nutrition.


Surveillance of neonatal group B streptococcal infection in Sunderland.
Surveillance of neonatal group B streptococcus infection in Sunderland identified 15 confirmed cases (with positive cultures from blood or CSF) in three years from 1995 to 1997, equivalent to an incidence of 1.42 per 1000 live births, not much lower than the estimate of 1.8 used in the United States to justify the introduction of preventative policies. Confirmed early-onset cases may represent only a fraction of the true number of cases, and a modified risk factor-based policy was introduced in Sunderland in June 1998.


Neuro-ophthalmic manifestations of Lyme disease.
Lyme disease is a multisystem disorder caused by infection with the Borrelia burgdorferi spirochete. The diagnosis of Lyme disease usually is based on several clinical criteria, with supportive data from laboratory testing. The presence of the bullseye skin lesion, erythema migrans, is the single pathognomonic criterion. In the 20 years since the initial description of Lyme disease in the United States, B. burgdorferi has been implicated as an etiologic agent in numerous ophthalmic and neuro-ophthalmic syndromes, involving most structures from the cornea to the cranial nerves. Neuro-ophthalmic and ocular manifestations of Lyme disease include meningitis with papilledema, cranial neuropathies, follicular conjunctivitis, nummular keratitis, and intraocular inflammation. Although an association with Lyme disease has been purported for numerous other syndromes, a definite causal relationship has not been proved in many cases. During a period of rapidly increasing awareness of Lyme disease, a high index of suspicion and poorly defined criteria for its presence have resulted in over-diagnosis of Lyme disease. In the authors' experience, the incorrect diagnosis of Lyme disease initially has been made in patients with allergic conjunctivitis, keratoconus, morning glory syndrome, craniopharyngioma, meningioma, CNS lymphoma, paraneoplastic syndrome, multiple sclerosis, sarcoid, syphilis, and functional illness. Nevertheless, this treatable infection must be an important consideration in the differential diagnosis of certain ocular or neurologic diseases.


The effects of yoga versus stretching and resistance training exercises on psychological distress for people with mild-to-moderate Parkinson's disease: study prxotocol for a randomized controlled trial.
Psychological distress is prevalent among people with Parkinson's disease (PD) and aggravates their motor symptoms, thereby leading to increased disability, high healthcare costs, and poor health-related quality of life (HRQoL). The under-recognition and adverse effects of the pharmacological management of anxiety and depression among the PD population are considerable. Thus, adopting a Complementary and Alternative Management (CAM) approach to address this problem is important. Yoga, one of the most common "mind-body" CAM therapies, can improve the psychological wellbeing of people with chronic illnesses. However, limited research on the effects of yoga in people with PD has been conducted. This study will determine the effects of yoga on the psychological wellbeing of people with mild-to-moderate PD and will compare these effects with those of stretching and resistance training exercises. A community-based, single-blind, randomized trial will be conducted. A total of 126 subjects will be recruited and randomly divided into yoga (n = 63) or stretching and resistance exercise (n = 63) groups. For 8 weeks, the yoga group will receive a weekly 90-min session of yoga, and the control group will receive a weekly 60-min session of stretching and resistance exercises. The primary outcome will be the level of psychological distress measured using the Hospital Anxiety and Depression Scale. The secondary outcomes will include the severity of motor symptoms measured by the Movement Disorders Society - Unified Parkinson's Disease Scale - Part III Motor Examination; mobility, balance, and fall risk measured by the Timed Up and Go test; spiritual wellbeing measured by the Holistic Wellbeing Scale; and HRQoL measured by the Parkinson's Disease Questionnaire-8. Assessment will be conducted at baseline, 8th, and 20th weeks of follow-ups. This study will be the first randomized trial to compare the effect of yoga versus stretching and resistance training exercises in a PD population. Results will contribute to the value of yoga as a therapeutic option for managing psychological distress in PD patients. Multiple outcomes including psychological, physiological, and spiritual and HRQoL will also be measured to elucidate the potential mechanisms of yoga. The effect of yoga on people with chronic illnesses will further be elucidated. This information should contribute to future research, practice, and policy related to PD management. WHO Primary Registry - Chinese Clinical Trials Registry (ChiCTR): CUHK_CCRB00522 Registered on 8 October 2016; date of approval 19 August 2016.


Dyscalculia, dyslexia, and medical students' needs for learning and using statistics.
Much has been written on the learning needs of dyslexic and dyscalculic students in primary and early secondary education. However, it is not clear that the necessary disability support staff and specialist literature are available to ensure that these needs are being adequately met within the context of learning statistics and general quantitative skills in the self-directed learning environments encountered in higher education. This commentary draws attention to dyslexia and dyscalculia as two potentially unrecognized conditions among undergraduate medical students and in turn, highlights key developments from recent literature in the diagnosis of these conditions. With a view to assisting medical educators meet the needs of dyscalculic learners and the more varied needs of dyslexic learners, a comprehensive list of suggestions is provided as to how learning resources can be designed from the outset to be more inclusive. A hitherto neglected area for future research is also identified through a call for a thorough investigation of the meaning of statistical literacy within the context of the undergraduate medical curriculum.


Highly cross-linked polyethylene in cemented THA: randomized study of 61 hips.
Highly cross-linked polyethylene (PE) has been introduced as an alternative bearing material in total hip arthroplasty (THA) because of high wear resistance in laboratory tests but the clinical experience of this material is limited. We evaluated a highly cross-linked PE (warm irradiated adiabatic melting, absorbed dose, 95 kGy) in a randomized study of cemented THAs. Cups of the same design but made of conventionally gamma irradiated PE (absorbed dose, 25-40 kGy) constituted the control group. Sixty-one hips (30 women, 30 men) with a median age of 55 years (range, 35-70 years) were included. All patients received a Spectron stem with 28-mm CoCr head. Radiostereometric examinations with the patient supine or standing were done at regular intervals. Wear was measured with the patient in the supine position from the first postoperative week, whereas examinations done with the patient standing were initiated 3 months after the operation. Dual x-ray absorptiometry and conventional radiography were used to evaluate the bone mineral density and the radiolucencies around the acetabular component. Fifty-two patients (53 hips; 25 highly cross-linked, 28 control) have been followed up for 2 years. At the 2-year followup, the highly cross-linked cups showed 50% reduction of proximal wear compared with the control group, when the patients were studied standing. When evaluated supine, the difference in proximal wear did not reach significance. The migration of the socket, the relative changes of periprosthetic bone mineral density, and the progression of radiolucencies between the immediately postoperative followup and 2-year followup did not differ. Highly cross-linked PE showed increase resistance to wear. Different mechanical properties of the two types of PE studied did not alter the performance of the cup in terms of fixation, periprosthetic bone loss, and radiographic appearance. However, the followup is short and these results are preliminary.


Cerebral palsy in childhood.
Cerebral palsy is the most common cause of physical disability in childhood. While some children have only a motor disorder, others have a range of problems and associated health issues. This article describes the known causes of cerebral palsy, the classification of motor disorders and associated disabilities, health maintenance, and the consequences of the motor disorder. The importance of multidisciplinary assessment and treatment in enabling children to achieve their optimal potential and independence is highlighted. General practitioners play an important role in the management of children with cerebral palsy. Disability is a life-long problem which impacts on the child, their parents and their siblings. After transition to adult services, the GP may be the only health professional that has known the young person over an extended period, providing important continuity of care.


Hospital-directed feedback to Emergency Medical Services improves prehospital performance.
A potential way to improve prehospital stroke care and patient handoff is hospital-directed feedback for emergency medical service (EMS) providers. We evaluated whether a hospital-directed EMS stroke follow-up tool improved documentation of adherence to the Rhode Island state prehospital stroke protocol for EMS providers. A standardized, 10-item feedback tool was developed in 2012 and sent to EMS directors for every transported patient with a discharge diagnosis of ischemic stroke. We reviewed patient charts meeting these criteria between January 2008 and December 2013. Performance on the tool was compared between the preintervention (January 2008 through January 2012) and postintervention (February 2012 through December 2013) periods. We identified 1176 patients with ischemic stroke who arrived by EMS in the study period: 668 in the preintervention period and 508 in the postintervention period. The overall score for the preintervention group was 5.31 and for the postintervention group 6.42 (P<0.001). Each of the 10 items, except checking blood glucose, showed statistically significant improvement in the postintervention period compared with the preintervention period. Hospital-directed feedback to EMS was associated with improved overall compliance with state protocols and documentation of 9 out of 10 individual items. Future confirmatory studies in different locales and studies on the impact of this intervention on actual tissue-type plasminogen activator administration rates and EMS personnel knowledge and behavior are needed.


Cochlear Impairment and Autoimmune Ear Disorder in a Patient with Breast Cancer.
The purpose of this study was to consider the possible role of autoimmune diseases and paraneoplastic syndrome in the genesis of tinnitus. The incidence of autoimmune inner ear disease (AIED) is rare, accounting for <1% of all cases of hearing impairment and dizziness. In presence of auditory and vestibular deficit in oncological patients, a paraneoplastic syndrome with cochleovestibulopathy should be considered. We described a 50-year-old Caucasian woman came to our attention with complaints of severe disabling bilateral tinnitus (Tinnitus Handicap Inventory, THI: 96), ear fullness and headache. The onset of tinnitus was associated to the last breast implant and prolonged antibiotic therapy. Serological autoimmunity tests were positive and a diagnosis of mixed connective tissue disease with notes of fibromyalgia was made. Pure tone audiometry testing revealed bilateral fluctuating mild hearing loss on high frequencies. The tinnitus was successfully treated with bilateral wideband sound generators (listening 8-9 h for day) regulated at the mixing point. At 12 months follow up THI has shrunk considerably (THI: 4) and the patient has continued treatment only with the sound pillow. In conclusion significant progress is needed to better understand the role of autoantibodies in the pathogenesis and diagnosis of paraneoplastic cochleovestibulopathy. To our knowledge, our study is the first in which hearing loss and tinnitus is considered as a manifestation of a paraneoplastic syndrome.


Biochemical and bioimaging markers for risk assessment and diagnosis in major cardiovascular diseases: a road to integration of complementary diagnostic tools.
This report from the first International Course on Integrated Biomarkers, Biochemical and Bioimaging Endpoints in Cardiovascular Diagnosis, Prevention, Therapy and Drug Development provides the basis for optimizing diagnostic, prognostic and therapeutic information in four areas of cardiovascular medicine: primary prevention of cardiovascular diseases, acute coronary syndromes, heart failure and stroke. Risk stratification and treatment strategies can be refined and enhanced through integration of bioimaging and biochemical markers to characterize sub-clinical and clinical atherosclerosis. For the integrative approach to be useful, each of the biomarkers must be validated and cost-effective. Clinical decision is the primary level of integration and is based on clinical evaluation and the use of a combination of bioimaging and biochemical markers. The decision to initiate preventive or therapeutic intervention must take into account the factors affecting the levels of expression of the biomarker and the potential input the biomarker has on metabolic processes or modulation of other biomarkers. The optimal approach to intervention must take into consideration the risk-benefit and cost-effectiveness ratios.


Nanomaterial-based bioorthogonal nanozymes for biological applications.
Bioorthogonal transformations are chemical reactions that use pathways which biological processes do not access. Bioorthogonal chemistry provides new approaches for imaging and therapeutic strategies, as well as tools for fundamental biology. Bioorthogonal catalysis enables the development of bioorthogonal "factories" for on-demand and in situ generation of drugs and imaging tools. Transition metal catalysts (TMCs) are widely employed as bioorthogonal catalysts due to their high efficiency and versatility. The direct application of TMCs in living systems is challenging, however, due to their limited solubility, instability in biological media and toxicity. Incorporation of TMCs into nanomaterial scaffolds can be used to enhance aqueous solubility, improve long-term stability in biological environment and minimize cytotoxicity. These nanomaterial platforms can be engineered for biomedical applications, increasing cellular uptake, directing biodistribution, and enabling active targeting. This review summarizes strategies for incorporating TMCs into nanomaterial scaffolds, demonstrating the potential and challenges of moving bioorthogonal nanocatalysts and nanozymes toward the clinic.


Evaluation of Chios mastic gum on lipid and glucose metabolism in diabetic mice.
Chios mastic gum (MG), a resin produced from Pistacia lentiscus var. Chia, is reported to possess beneficial cardiovascular and hepatoprotective properties. This study investigated the effect of crude Chios MG on metabolic parameters in diabetic mice. Streptozotocin-induced diabetic 12-week-old male C57bl/6 mice were assigned to three groups: NC (n=9) control; LdM (n=9) animals receiving low dose mastic for 8 weeks (20 mg/kg body weight [BW]); and HdM (n=9) animals receiving high dose mastic (500 mg/kg BW) for the same period. Serum lipid and glucose levels were determined at baseline, at 4 and 8 weeks. Serum total protein, adiponectin, and resistin levels were also measured at the end of the experiment. Histopathological examination for liver, kidney, aorta, and heart lesions was performed. After 4 weeks, MG administration resulted in decreased serum glucose and triglyceride levels in both LdM and HdM, whereas BW levels were reduced in LdM group compared with controls. At the end of the experiment, LdM presented significantly lower serum glucose, cholesterol, low-density lipoprotein cholesterol, and triglyceride levels and improved high-density lipoprotein cholesterol levels compared with control group. HdM group had ameliorated serum triglyceride levels. Hepatic steatosis observed in control group was partially reversed in LdM and HdM groups. MG administered in low dosages improves glucose and lipid disturbances in diabetic mice while alleviating hepatic damage.


Comparative study of home and office blood pressure in hypertensive patients treated with enalapril/HCTZ 20/6 mg: the ESPADA study.
The introduction and generalization of 24-h ambulatory blood pressure (BP) monitoring has shown the clinical relevance of home BP. The aim of this study was to assess the validity of home-measured BP for monitoring and controlling patients with arterial hypertension while on a homogeneous treatment. An additional objective was to establish the degree of office BP control obtained. This was a prospective, longitudinal, observational and multicenter study in a cohort of 156 patients of both sexes, aged over 18 years and with essential hypertension. All of them received the fixed combination enalapril/HCTZ 20/6 mg as the only hypertensive agent for at least 4 weeks previously. Office BP was the average of three measurements. For home BP, a semi-automated device (OMRON HEM 705 CP) was used. The patients measured their BP twice a day for 2 consecutive days. The average differences between the two measuring methods were low, but significant: 3.99 mmHg for systolic BP (SBP; p < 0.05), 2.02 mmHg for diastolic BP (DBP; p < 0.05). Pearson's regression coefficient between the office and home values was highly significant (p < 0.0001) for SBP, DBP and heart rate. Home BP measurement was highly reproducible as shown by the high within-class correlation coefficient for individual measurements on the first day compared with the second: 0.88 (95% confidence interval, CI 0.82-0.92; p < 0.00001) for SBP and 0.89 for DBP (95% CI 0.83-0.93; p < 0.00001). The percentage of patients with strict office DBP and SBP control (< 140/90 mmHg) was 61.3% and with DBP control (<90 mmHg) 92%. In conclusion, in the ESPADA study, the application of home BP measurement is valid, reproducible and shows a high correlation with office BP.


Blebs and apoptotic bodies are B cell autoantigens.
Mounting evidence suggests that systemic lupus erythematosus autoantigens are derived from apoptotic cells. To characterize the potential interactions between apoptotic cells and B cells, the D56R/S76R variant of 3H9, a murine autoantibody that binds to DNA, chromatin, and anionic phospholipids, was compared with DNA4/1, a human anti-DNA autoantibody. Flow cytometry revealed that only D56R/S76R bound to Jurkat cells treated with either of three distinct proapoptotic stimuli, Ab binding was dependent on caspase activity, and immunoreactivity developed subsequent to annexin V binding. Confocal microscopy established a structural basis for the distinct kinetics of binding. D56R/S76R preferentially bound to membrane blebs of apoptotic cells, whereas annexin V binding did not require blebs. Inhibition of ROCK I kinase, an enzyme that stimulates nuclear fragmentation and fragment distribution into blebs, significantly reduced Ab binding. Because members of the collectin and pentraxin families of serum proteins bind to blebs on apoptotic cells and assist in the clearance of cellular remains, our results suggest that Abs to blebs could affect the recognition of apoptotic cells by cells of the innate immune system and thus modify tolerance to nuclear Ags.


Performance Evaluation of a Nanofluid-Based Direct Absorption Solar Collector with Parabolic Trough Concentrator.
Application of solar collectors for hot water supply, space heating, and cooling plays a significant role in reducing building energy consumption. For conventional solar collectors, solar radiation is absorbed by spectral selective coating on the collectors' tube/plate wall. The poor durability of the coating can lead to an increased manufacturing cost and unreliability for a solar collector operated at a higher temperature. Therefore, a novel nanofluid-based direct absorption solar collector (NDASC) employing uncoated collector tubes has been proposed, and its operating characteristics for medium-temperature solar collection were theoretically and experimentally studied in this paper. CuO/oil nanofluid was prepared and used as working fluid of the NDASC. The heat-transfer mechanism of the NDASC with parabolic trough concentrator was theoretically evaluated and compared with a conventional indirect absorption solar collector (IASC). The theoretical analysis results suggested that the fluid's temperature distribution in the NDASC was much more uniform than that in the IASC, and an enhanced collection efficiency could be achieved for the NDASC operated within a preferred working temperature range. To demonstrate the feasibility of the proposed NDASC, experimental performances of an NDASC and an IASC with the same parabolic trough concentrator were furthermore evaluated and comparatively discussed.


Full-mouth disinfection for the treatment of adult chronic periodontitis.
In an attempt to enhance treatment outcomes, alternative protocols for anti-infective periodontal therapy have been introduced. To evaluate the effectiveness of full-mouth disinfection or full-mouth scaling compared to conventional quadrant scaling for periodontitis. Data sources included electronic databases, handsearched journals and contact with experts. The Cochrane Oral Health Group Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE and EMBASE were searched. Reference lists from relevant articles were scanned and the authors of eligible studies were contacted to identify trials and obtain additional information. Date of most recent searches: December 2006: (CENTRAL) (The Cochrane Library 2006, Issue 4). Randomised controlled trials were selected with at least 3 months follow up comparing full-mouth scaling and root planing within 24 hours with (FMD) or without (FMS) the adjunctive use of an antiseptic (chlorhexidine) with conventional quadrant scaling and root planing (control). The methodological quality of the studies was assessed within the data extraction form, mainly focusing on: method of randomisation, allocation concealment, blindness of examiners and completeness of follow up. Data extraction and quality assessment were conducted independently by multiple review authors. The primary outcome measure was tooth loss, secondary outcomes were reduction of probing depth, bleeding on probing and gain in probing attachment. The Cochrane Collaboration statistical guidelines were followed. The search identified 216 abstracts. Review of these abstracts resulted in 12 publications for detailed review. Finally, seven randomised controlled trials (RCTs) which met the criteria for eligibility were independently selected by two review authors. None of the studies included reported on tooth loss. All treatment modalities led to significant improvements in clinical parameters after a follow up of at least 3 months. For the secondary outcome, reduction in probing depth, the mean difference between FMD and control was 0.53 mm (95% confidence interval (CI) 0.28 to 0.77) in moderately deep pockets of single rooted teeth and for gain in probing attachment 0.33 mm (95% CI 0.04 to 0.62) in moderately deep single and multirooted teeth. Comparing FMD and FMS the mean difference in one study for gain in probing attachment amounted to 0.74 mm in favour of FMS (95% CI 0.17 to 1.31) for deep pockets in multirooted teeth, while another study reported a mean difference for reduction in bleeding on probing of 18% in favour of FMD (95% CI -33.74 to -2.26) for deep pockets of single rooted teeth. No significant differences were observed for any of the outcome measures, when comparing FMS and control. In patients with chronic periodontitis in moderately deep pockets slightly more favourable outcomes for pocket reduction and gain in probing attachment were found following FMD compared to control. However, these additional improvements were only modest and there was only a very limited number of studies available for comparison, thus limiting general conclusions about the clinical benefit of full-mouth disinfection.


Liposarcomas in young patients: a study of 82 cases occurring in patients younger than 22 years of age.
Liposarcomas typically occur in middle aged to older adults. Altogether, approximately 50 bona fide liposarcomas have been reported in children and adolescents, most of which have represented myxoid liposarcomas, with a good prognosis. We undertook a retrospective study of 82 liposarcomas occurring in patients below 22 years of age. Clinicopathologic and follow-up information was obtained. Fluorescence in situ hybridization for FUS, EWSR1, CHOP (DDIT3), and MDM2 was performed in 30 cases. The tumors occurred in 28 males and 54 females (5 to 22 y of age) and involved many locations. Fifty-six cases were typical myxoid liposarcomas, including 2 with round cell areas. The tumors were grade 1 (56 cases) and grade 3 (2 cases). Thirty-seven of 38 patients with follow-up are alive without disease and 1 is alive with disease (median 59 mo follow-up duration, range: 8 to 108 mo). Six cases showed myxoid liposarcoma with spindled growth ("spindle cell myxoid liposarcoma"); these arose in 5 females and 1 male (median age 14 y) and involved the thigh in 40% of cases. All were grade 1. Follow-up (4 of 6 patients) showed local recurrences in 2 cases and metastases in 1 case. Twelve tumors consisted of conventional myxoid liposarcoma and pleomorphic liposarcoma ("pleomorphic myxoid liposarcoma"); these arose in 4 males and 8 females (10 to 22 y of age) and often involved the mediastinum. Tumor grades were 2 (4 cases) and 3 (8 cases). Follow-up (10 patients) showed 7 dead of disease, 1 alive with disease, and 2 disease free. Four atypical lipomatous tumors were seen including 2 with low-grade dedifferentiation. Two local recurrences were seen; all patients are disease free. Two conventional pleomorphic liposarcomas were seen; 1 patient with follow-up is disease free. FUS-CHOP and EWSR1-CHOP rearrangements were identified by fluorescence in situ hybridization in 15/23 and 2/23 conventional myxoid liposarcomas, respectively, and in no other tumors. Amplification for MDM2 was absent in all cases. We conclude that conventional myxoid liposarcoma is by far the most common subtype of liposarcoma in young patients, with an excellent prognosis. Two apparently novel subtypes of liposarcoma, termed pleomorphic myxoid liposarcoma and spindle cell myxoid liposarcoma comprise considerable percentages of liposarcomas in this age group and should be distinguished from conventional myxoid liposarcoma and conventional pleomorphic liposarcoma. Pleomorphic myxoid liposarcoma and spindle cell myxoid liposarcoma most likely represent high-grade and low-grade variants of myxoid liposarcoma, respectively. Additional study of such cases will be necessary for definitive classification.


In vitro assays of vesicular transport.
Movement of proteins and lipids between the various compartments of eukaryotic cells is fundamental to the maintenance of cellular homeostasis, and an understanding of the molecular mechanisms that govern these processes remains a key goal of cell biological research. This aim has been greatly facilitated by the development of assays that recapitulate specific events in vitro. In the following article we provide an overview of some of the currently used assays that measure the movement of proteins within the exocytic and endocytic pathways, and provide a starting point for those wishing to establish their own systems to study other vesicular transport steps.


US outbreak investigations highlight the need for safe injection practices and basic infection control.
Current understanding of viral hepatitis transmission in United States health care settings indicates progress over the past several decades with respect to the risks from transfusions or blood products. Likewise, risks to health care providers from sharps injuries and other blood and body fluid exposures have been reduced as a consequence of widespread hepatitis B vaccination and the adoption of safer work practices. Increasing recognition of outbreaks involving patient-to-patient spread of hepatitis B and hepatitis C virus infections, however, has uncovered a disturbing trend. This article highlights the importance of basic infection control and the need for increased awareness of safe injection practices.


Electrokinetics of diffuse soft interfaces. IV. Analysis of streaming current measurements at thermoresponsive thin films.
Streaming current measurements were performed on poly(N-isopropylacrylamide)-co-N-(1-phenylethyl) acrylamide [P(NIPAAm-co-PEAAm)] thermoresponsive thin films above and below the transition temperature of the polymer (i.e., at 22 and 4 degrees C, respectively). Electrokinetic measurements (ionic strength 0.01-10 mM KCl, pH 2.5-9.5 in 1 mM KCl) revealed that the charging of the polymer/aqueous solution interface is determined by unsymmetrical adsorption of hydroxide and hydronium ions onto the Teflon AF substrate that supports the hydrogel film. The magnitude of the streaming current significantly decreased with decreasing temperature, that is, when the hydrogel was swelling. The pH- and ionic strength-dependent data for unswollen and swollen films were interpreted on the basis of the here-reported general theory for the electrokinetics of diffuse soft gel layers. The formalism based on the Debye-Brinkman equation for hydrodynamics and the nonlinear Poisson-Boltzmann equation for electrostatics extends previous theoretical studies by considering the most general situation of a charged gel layer supported by a charged rigid surface. Full analytical expression is provided for the streaming current in the limit of homogeneous distribution of segments under low potential conditions. Numerical analysis of the governing transport and electrostatic equations allows for the computation of streaming current for cases where analytical developments are not possible. The theory successfully reproduces the electrokinetic data for the P(NIPAAm-co-PEAAm) copolymer film at 22 and 4 degrees C over the whole range of pH and ionic strength examined. It is found that the 3-fold increase of the hydrogel film thickness with decreasing temperature from 22 to 4 degrees C (i.e., from 23 to 70 nm as measured by ellipsometry), is in line with homogeneous swelling and an increase of the hydrodynamic penetration length (1/lambdao) by a factor of approximately 1.6. Additionally, the hydrodynamic thicknesses (deltaH) of the swollen and unswollen hydrogels are evaluated in terms of their respective hydrodynamic penetration length and electrosurface characteristics of the supporting Teflon AF surface.


A model T-cell receptor system for studying memory T-cell development.
When T-cell clones were first grown in long-term cell culture, each clone was considered to be capable of displaying a limited range of functional activities, constrained by the clones' coreceptor, CD4 or CD8, and the specificity of its antigen-specific receptor (TCR) for one or a few peptides in association with a class I or class II MHC molecule. Subsequent studies, especially with transgenic mice, have shown, however, that T cells expressing the same receptor can be obtained in a variety of differentiated states, including naïve cells, activated effector cells, memory cells, and anergic or tolerant cells, as well as cells with or without a coreceptor. In each of these states T cells can display distinctly different responses to the peptide-MHC (pepMHC) complexes the TCR recognizes. Recently, memory T cells have received considerable attention, in part because of the likelihood that they confer long-term protective immunity against diverse infectious agents and possibly against some forms of cancer. Here we review some recent studies that our colleagues and we have carried out on memory CD8(+) T cells. These studies have made extensive use of cells that express the TCR called 2C. The diverse set of cells expressing the 2C TCR, in mice and in cultured clones and cell lines, are referred to as the 2C system. Before reviewing the studies of memory cells, we summarize the 2C system's main features, including evidence that a large and diverse array of pepMHC complexes, involving at least four class I MHC proteins, can stimulate TCR-mediated responses of 2C cells.


[Complementary therapies in the hospital:'if you can't beat them, join them'?].
Some major cancer hospitals in the United States have established a department for complementary medicine as a service to their patients. Surveys in the Netherlands have shown that notably nursing staff and consumers consider the availability of complementary therapy in health care to be very important. Nevertheless, this appears to be an undesirable development as the effectiveness of these therapies has not been demonstrated and the therapists involved might interfere with regular patient therapy.


Successful use of topical vitamin E solution in the treatment of nail changes in yellow nail syndrome.
A double-blind controlled study was performed on a patient with long-standing yellow nail syndrome to investigate the potential beneficial role of topical vitamin E solution for the nail changes seen in this disorder. Three nails received active solution containing DL-alpha-tocopherol in dimethyl sulfoxide; three nails received dimethyl sulfoxide only; and the remainder were untreated. After 6 months of therapy, the three nails treated with active solution showed marked clinical improvement and a corresponding increase in nail growth rates. Dimethyl sulfoxide alone produced a small increase in nail growth rate and slight clinical improvement. Active solution was then applied to all of the nails for a further 6 months, resulting in clinical improvement and increased nail growth rates in the remaining nails. These beneficial effects require further study.


The role of the toothbrush in the abrasion process.
To evaluate the relative abrasivity of different toothbrushes both qualitatively and quantitatively. Acrylic plates were exposed to brushing in a brushing machine with ten different toothbrushes with water alone and with a toothpaste. The results were evaluated using a profilometer after one and 6 h of brushing (corresponding to 2000 and 12,000 double strokes, respectively). A surface roughness value (Ra-value) and also a volume loss value were calculated from the profilometer measurements. These values were then compared to each other. The results showed that brushing with water alone caused less abrasion than when a toothpaste was added. Six-hour brushing with water caused less abrasion than 1 h with a toothpaste. The number of filaments or filament diameter influenced the results in various ways. When brushing with water, the harder toothbrush (Jordan Medium) caused more abrasion (higher Ra-value), but when adding the toothpaste, the softer toothbrush (Jordan soft) caused more abrasion after 12,000 double strokes. Besides supporting the fact that a toothpaste is needed to create a significant abrasion, this study also showed that a softer toothbrush can cause as much and in some cases more abrasion than harder ones. When conducting abrasivity studies, it is important to look at both the quantitative and qualitative aspect of abrasivity.


Soybean fermentation with Bacillus licheniformis increases insulin sensitizing and insulinotropic activity.
Traditionally fermented soybeans (chungkookjang; TFC) may have potent anti-diabetic activity, depending on the ambient microorganisms and conditions. We hypothesized that one of the major Bacillus species in TFC contributes to the anti-diabetic activity and could be used to standardize a highly functional TFC. We tested the hypothesis by using cell-based studies to evaluate insulin sensitizing and insulinotropic action of chungkookjangs fermented with various Bacillus spp. and fermentation periods. The 70% methanol and water extracts of chungkookjang fermented with Bacillus licheniformis (BL) for 48 h contained similar profiles of isoflavonoids and peptides to methanol and water extracts of TFC with potent anti-diabetic activity. Water extracts (mainly containing peptides) of TFC and BL fermented for 48 h and 72 h had a better insulin sensitizing action via activating peroxisome proliferator-activated receptor-γ (PPAR-γ) and increased the expression of PPAR-γ in 3T3-L1 adipocytes better than unfermented cooked soybeans (CSB). The 70% methanol extracts (predominantly isoflavone aglycones) of BL fermented for 48 h and 72 h improved glucose-stimulated insulin secretion and protected β-cell viability better than CSB in insulinoma cells, and the improvement by BL was similar to TFC. In conclusion, the BL water extract fermented for 48 h exhibited equal insulin sensitization as TFC and BL methanol extract exerted similar insulinotropic actions to those of TFC. B. licheniformis may be one of the major microorganisms responsible for anti-diabetic actions of chungkookjang. It is important to make chungkookjang that retains the anti-diabetic properties of the most efficacious traditional chungkookjang using a standardized method.


Nucleotide clusters in deoxyribonucleic acids: sequence analysis of DNA using pyrimidine oligonucleotides as primers in the DNA polymerase I repair reaction.
Pyrimidine oligonucleotides have been shown to prime the E. coli DNA polymerase I repair reaction, specifically and reproducibly. DNA molecules up to 30 nucleotides long have been obtained from the extension of oligopyrimidine primers, 9 to 11 nucleotides long isolated from the complementary (minus) strand of bacteriophage S13 RFDNA using S13 viral DNA as the template molecule. The sequences of the extended primers were determined from mobility shift following separation of partially extended primers by ionophoresis and homochromatography, and by a modification of the "plus" system of Sanger and Coulson (1975). The 3' leads to 5' exonuclease activity of E. coli DNA polymerase was utilized for the "plus" system in the presence of single dNTPs and also with two dNTPs in the reaction, to give a nearest neighbor type of analysis for sequence confirmation. The ready availability of oligopyrimidine primers from any DNA and the simplification of the "plus" method broaden the range of applicability of the primed DNA polymerase I repair reaction for DNA sequence analysis.


Creatine kinase isoforms following isometric exercise.
The present study assessed creatine kinase (CK) activity, CK MM isoforms, and muscle soreness following an exercise regimen designed to induce skeletal muscle damage. Eight college-age subjects performed 40 maximal isometric contractions of the knee extensor muscles (10-second contraction/20-second rest). Serum samples and soreness ratings were taken prior to and 2, 6, 18, and 24 hours after the exercise. The CK MM1 and CK MM3 isoforms were determined by flatbed isoelectric focusing (IEF). In serum, the MM1 isoform (the pure gene product) is considered to be evidence of newly released CK from muscle, as upon entering the plasma, the CK MM1 is converted to MM2 and then MM3. A significant increase in serum CK activity was found at 6 hours and remained elevated at 24 hours. CK MM1 increased significantly at 2 hours, peaked at 6 hours, then approached baseline. Soreness did not increase significantly until 18 hours. Analysis of CK isoforms in serum can provide an earlier indicator of skeletal muscle damage than total CK or perception of soreness and may be useful in tracking the time course of skeletal muscle damage and repair.


Studies on the nutritional physiology of zoophilic dermatophytes isolated from horses in Ibadan, Nigeria.
The effects of the various growth factors with regard to the nutritional physiology of zoophilic dermatophytes isolated from horses are reported. The optimum temperature, pH and growth period for the fungal isolates were found to be 30 degrees-35 degrees C, 5-6 and 7-12 days respectively while the carbon and nitrogen sources utilised by the organisms were sucrose, fructose, maltose, L-arginine and calcium nitrate. The use of these factors in preparation of efficacious fungicides used in the treatment of ringworm infections in man and animals is discussed.


Malaria vaccine trials in pregnant women: An imperative without precedent.
Pregnant women are highly susceptible to Plasmodium falciparum malaria, leading to substantial maternal, perinatal, and infant mortality. While malaria vaccine development has made significant progress in recent years, no trials of malaria vaccines have ever been conducted in pregnant women. In December 2016, an expert meeting was convened at NIAID, NIH, in Rockville, Maryland to deliberate on the rationale and design of malaria vaccine trials in pregnant women. The discussions highlighted the progress made over recent years in the field of maternal immunization for other infectious diseases, and the evolving regulatory and ethical environment, all of which support a new emphasis on testing malaria vaccines that offer direct benefits to pregnant women. Initial safety and immunogenicity studies of malaria vaccines will be conducted in non-pregnant adult volunteers. Subsequently, efficacy trials involving pregnant women will likely be conducted in malaria-endemic and often resource-poor environments where sufficiently high malaria incidence will allow vaccine activity to be measured. Such trials will need to meet all international standards to ensure the safety of mother and offspring, under oversight of appropriate ethical and regulatory bodies. The convened experts drafted a clinical development plan to test a malaria vaccine product during pregnancy, using as a case study PfSPZ Vaccine being developed by Sanaria Inc. that is currently in phase 2 testing. Following the expert recommendations, a pregnancy registry has been initiated in Ouelessebougou, Mali, to provide baseline information on maternal and fetal outcomes as a context for evaluating PfSPZ Vaccine safety in the future, and new regimens are being assessed that will be suitable for evaluation in pregnant women.


Longitudinal evidence for a relation between depressive symptoms and quality of life in schizophrenia using structural equation modeling.
Patients diagnosed with schizophrenia often report a low quality of life (QoL). The purpose of this study was to investigate whether we could replicate a cross-sectional model by Alessandrini et al. (2016, n = 271) and whether this model predicts QoL later in life. This model showed strong associations between schizophrenia spectrum symptoms and depressive symptoms on QoL, but lacked follow-up assessment. This model was adapted in the current study and the robustness was investigated by using a longitudinal design in which the association between baseline variables (including IQ, depression, schizophrenia spectrum symptoms as well as social functioning) and QoL during 3-years of follow-up was investigated. We included patients with a non-affective psychotic disorder (n = 744) from a prospective naturalistic cohort-study. In the cross-sectional model, with good measure of fit, both depression as well as social functioning was associated with QoL (direct path coefficient -0.28 and 0.41, respectively). Additionally, the severity of schizophrenia spectrum symptoms was highly associated with social functioning (direct path coefficient -0.70). Importantly, the longitudinal model showed good measures of fit, which strengthens the validity of the initial model and highlights that depression prospectively affect QoL while schizophrenia spectrum symptoms prospectively influence QoL via social functioning. The negative, longitudinal impact of a depression on QoL highlights the need to focus on treatment of this co-morbidity.


Effect of ischemia and reperfusion in vivo on energy metabolism of rat sciatic-tibial and caudal nerves.
Our model of severe nerve ischemia consistently results in extinction of the compound nerve and muscle action potentials (NAP; CMAP) within 30 min. Since impulse transmission may depend on nerve energy metabolism (NEM), we studied the effects of ischemia with reperfusion on sciatic-tibial nerve NEM in vivo and compared these results with NEM of this nerve in deoxygenated Ringer's solution in vitro and postmortem. Ischemia for 30 min postmortem or in deoxygenated Ringer's solution resulted in marked depletion of adenosine triphosphate (ATP) and creatine phosphate (CP) and an increase in lactate (LAC) of sciatic-tibial nerve of adult male Sprague-Dawley rats. In vivo ischemia for up to 3 h to sciatic-tibial nerve was sufficient to extinguish CMAP but not NAP and did not deplete ATP, CP, or GLU nor did it increase LAC. Ischemia sufficient to extinguish NAP resulted in reduction of energy substrates to about 50% of resting. Muscle fails to conduct impulses before nerve and in vivo reductions of energy substrates are milder than in vitro changes. These changes are explainable in terms of energy requirements and supply. These findings support an energetic basis of ischemic conduction failure.


[Progress Report on Cases of Peripherally Inserted Central Catheter (PICC) Management in Our Hospital, Including Elderly Patients].
In our hospital, the number of cases in which peripherally inserted central catheters (PICCs) are used has increased; these patients include elderly people who cannot take medications orally, patients with low levels of awareness, and patients with dysphagia. We report the situation at this time mainly with regard to the number of days on which PICCs were used. Fifteen elderly patients (male, n=7; female, n=8; average age, 89.3±5.3 years) underwent PICC insertion at our hospital from August 2016 to October 2018. Among these patients, 6 had cerebrovascular disorders, 5 had aspiration pneumonia, 2 had Parkinson's syndrome, 1 had consciousness disorder resulting from asphyxia caused by foreign body aspiration, and 1 had interstitial pneumonia. Seven patients received home management. In each patient, we measured the length of time that the PICC was in place. The average duration for which a PICC was in place was 92.9±25.4 days (in cases of home care management, 159.5±48.3 days). The longest duration was 342 days, in a patient with aspiration pneumonia. The endpoints were death and discharge from hospital. Complications/accidents occurred as a result of catheter infection in 2 cases (both patients recovered after catheter removal) and as a result of self-extraction in 1 case. These results suggest that PICC is useful and does not cause serious adverse effects, even in elderly patients who require central parenteral nutrition management.


B and T lymphocyte attenuator down-regulation by HIV-1 depends on type I interferon and contributes to T-cell hyperactivation.
Nonspecific T-cell hyperactivation is the main driving force for human immunodeficiency virus (HIV)-1 disease progression, but the reasons why the excess immune response is not properly shut off are poorly defined. Eighty-five HIV-1-infected individuals were enrolled to characterize B and T lymphocyte attenuator (BTLA) expression and function. Infection and blockade assays were used to dissect the factors that influenced BTLA signaling in vitro. BTLA expression on overall CD4(+) and CD8(+) T cells was progressively decreased in HIV-1 infection, which was directly correlated with disease progression and CD4(+) T-cell differentiation and activation. BTLA(+)CD4(+) T cells from HIV-1-infected patients also displayed an altered immune status, which was indicated by reduced expression of naive markers but increased activation and exhaustion markers. Cross-linking of BTLA can substantially decrease CD4(+) T-cell activation in vitro. This responsiveness of CD4(+) T cells to BTLA-mediated inhibitory signaling was further found to be impaired in HIV-1-infected patients. Furthermore, HIV-1 NL4-3 down-regulated BTLA expression on CD4(+) T cells dependent on plasmacytoid dendritic cell (pDC)-derived interferon (IFN)-α. Blockade of IFN-α or depletion of pDCs prevents HIV-1-induced BTLA down-regulation. HIV-1 infection potentially impairs BTLA-mediated signaling dependent on pDC-derived IFN-α, which may contribute to broad T-cell hyperactivation induced by chronic HIV-1 infection.