The effects of game types on intensity of small-sided games among pre-adolescent youth football players.
The aim of this study was to examine the effects of variations in pitch dimensions on pre-adolescent youth soccer players' physiological responses during two different types of small-sided games (SSG). Sixteen young soccer players (age: 13.2 ± 0.6 years; body mass: 52.5 ± 7 kg; height: 163.4 ± 6 cm) participated in this study. They performed 4 vs. 4 stop-ball SSG (SB-SSG) vs. small-goals SSG (SG-SSG) with 4×4 min and 2 min of passive recovery in between, using 3 different pitch sizes (small: 10×15, medium: 15×20, and large: 20×25 m). Heart rate (HR), lactate concentration ([La-]), and rating of perceived exertion (RPE) were measured during each session. The results show that SB-SSG induced higher HR responses than SG-SSG for the 3 pitch sizes: for HR (167.2±3.0 vs. 164.5±3.0, 172.3±2.9 vs. 169.2±3.1, and 175.4±3.1 vs. 171.1±2.7 bpm; P<0.05, for small, medium, and large, respectively) and [La-] (7.1±1.0 vs. 6.5±1.04, 7.3±1.0 vs. 6.8±1.2, and 7.8±0.9 vs. 7.1±0.8 mmol·l-1; P<0.05 on small, medium, and large pitches, respectively), whereas RPE scores were significantly higher during SB-SSG compared to SG-SSG (6.2±1.0 vs. 5.8±0.9; P<0.05, respectively) on the small pitch. In the present study higher physiological responses were observed in SSG in pre-adolescent young soccer players when using the stop-ball conditions in comparison with the small-goal rule for all pitch sizes - small, medium, and large. Stop-ball conditions in comparison with the small-goal rule for all pitch sizes - small, medium, and large.


Influence of dietary fiber type and amount on energy and nutrient digestibility, fecal characteristics, and fecal fermentative end-product concentrations in captive exotic felids fed a raw beef-based diet.
Little nutritional or metabolic information has been collected from captive exotic cats fed raw diets. In particular, fiber types and concentrations for use in raw meat-based diets for captive exotic felids have not been well studied. Our objective was to evaluate the effects of fiber type and concentration on apparent total tract energy and macronutrient digestibility, fecal characteristics, and fecal fermentative end-products in captive exotic felids. Four animals of each captive exotic species (jaguar (Panthera onca), cheetah (Acinonyz jubatus), Malayan tiger (Panthera tigris corbetti), and Siberian tiger (Panthera tigris altaica) were randomized in four 4 × 4 Latin square designs (1 Latin square per species) to 1 of the 4 raw beef-based dietary treatments (94.7 to 96.7% beef trimmings): 2 or 4% cellulose or 2 or 4% beet pulp. Felid species, fiber type, and fiber concentration all impacted digestibility and fecal fermentative end-products. Inclusion of beet pulp increased (P ≤ 0.05) fecal short-chain fatty acids and fecal output in all cats. Inclusion of 2 and 4% cellulose, and 4% beet pulp increased (P ≤ 0.05) fecal bulk and diluted fecal branched-chain fatty acid concentrations compared with 2% beet pulp. Apparent total tract DM, OM, fat, and GE digestibility coefficients decreased (P ≤ 0.05) linearly with BW of cats. Additionally, fecal moisture, fecal score, and concentrations of fermentative end-products increased (P ≤ 0.05) with BW. Although the response of many outcomes was dependent on cat size, in general, beet pulp increased wet fecal weight, fecal scores, and fecal metabolites, and reduced fecal pH. Cellulose generally reduced DM and OM digestibility, but increased dry fecal weight and fecal percent DM. Although beet pulp and cellulose fibers were tested individually in this study, these data indicate that the optimum fiber type and concentration for inclusion in captive exotic felid diets is likely a combination of fermentable and nonfermentable fibers, with the optimal fiber blend being dependent on species. Smaller cats, such as cheetahs and jaguars, tolerated fermentable fibers, whereas larger cats, such as Malayan and Siberian tigers, appeared to require more insoluble fibers that limit fermentation and provide fecal bulk. Further research is required to test whether these trends hold true when fed in combination.


Genetic Analysis of HIV-1 Integrase Sequences from Treatment Naive Individuals in Northeastern South Africa.
Raltegravir, an integrase inhibitor, is not a component of the current South African antiretroviral treatment guidelines, but it could be introduced in the near future as cases of virological failures from current treatment regimens begin to occur. The aim of this study was to analyze the complete HIV integrase gene obtained from individuals at two treatment sites in northeastern South Africa for the presence of Raltegravir associated drug resistant mutations and viral subtypes based on the integrase gene. Examination for mutations against other integrase inhibitors, such as Elvitegravir and Dolutegravir, was also done. Viruses from 127 treatment naive individuals were analyzed. Genetic drug resistance mutations were determined using the Stanford HIV Drug Resistance Interpretation program and the International AIDS society-USA guidelines. Viral subtyping was done by phylogenetic analysis, and recombinants were determined using the REGA, jpHMM and RIP tools. No major resistance mutations were detected. However, 7% of the sequences had minor mutations and polymorphisms. The majority (99%) of the viruses were HIV-1 C. Recombination analysis showed that the polymerase gene of one virus was likely composed of HIV-1 subtype A1 and C sequences. The present study indicates that Raltegravir, Elvitegravir and Dolutegravir resistant mutations may be absent in the study communities and further indicates the presence of recombinant viruses in northeastern South Africa.


[Medical negligence].
Medical negligence is a matter of growing public interest. This review outlines various aspects of medical negligence: epidemiology, taxonomy, and the risks, causes, psychology, management and prevention of errors.


Leteinizing hormone and luteinizing hormone releasing hormone-like immunoreactivity in the jugular venous blood of sheep at various stages of the oestrous cycle.
Luteinizing hormone and LH-RH-like immunoreactivity were measured in the jugular venous plasma of Clun Forest ewes at various stages of the oestrous cycle. Blood samples were collected through jugular venous cannulae every 2 h for at least 20 days from three ewes during the breeding season. The ewes were checked twice daily for oestrus using a vasectomized ram. Plasma LH peaks of apparent height 112-192 ng NIH-LH-S17 equivalents/ml were detected at oestrus with basal levels of 2-15 ng/ml during most of the remainder of the 17-day oestrous cycle. Peaks of LH-RH-like immunoreactivity occurred at various times of the cycle. The apparent maximal level of these peaks was 220 pg/ml compared with basal levels of less than 10 pg/ml. Further ewes (two for each group) were sampled at 4 min intervals for 12 h, (1) from onset of oestrus, (2) 36-48 h after onset of oestrus or (3) on day 10 of the oestrous cycle. In the ewes sampled at oestrus, peaks of LH-RH-like immunoreactivity were detected before, during and after the preovulatory LH peak. Those detected after the LH peak were unassociated with any further increases in the plasma LH level. In the ewes sampled 36-48 h after onset of oestrus and on day 10 of the cycle, several peaks of LH-RH-like immunoreactivity unassociated with any increases in the LH level were detected. These peaks, and those detected at oestrus, had durations of only one or two samples, and in some cases reached levels of several ng/ml compared with basal levels of less than 10 pg/ml. The significance of these results is discussed.


Peruvian thanatoscape and posthumous biographies of prisoners: dispersion and re-appropriation of undesirable corpses.
This article examines the necropolitical significance of the posthumous biographies of the inmates who were executed in the repression of the Santa Barbara, Lurigancho, and El Frontón prison riots of 1986 in Lima. Of the estimated 250 victims, only 22 have been handed over to their families. The path taken by the bodies is traced, along with the means employed by state agents to conceal them. We then analyze the modalities of their recovery, identification, and return (or otherwise), beginning in the 2000s. We coin the concept of "thanatoscape" to evoke the places and dynamics of a landscape in flux, encompassing the physical circulation of the deceased and the associated imaginary. The material dimension of the body is considered, along with how it is created symbolically by society and power relations. We discuss the means used to justify and legitimize whether these mortal remains deserved to be properly mourned or would remain missing and deprived of the opportunity to receive funeral rites. The management of these undesirable bodies can be better understood by studying the process of "de-citizenization" to which prisoners held for terrorism-related offenses were subjected, and which also extends to their families.


[Spondylodiscitis caused by a strain of Staphylococcus aureus, in vitro sensitive to penicillin].
The case history of a man referred to our department with the diagnosis of purulent meningitis; actually, he presented spondylodiscitis of the lumbar spine with the infection spreading into the spinal canal and the left psoas muscle. The disease was caused by a strain of Staphylococcus aureus, which in vitro was sensitive to penicillin. However, treatment with crystalline penicillin in combination with ciprofloxacin failed. The paper suggests a differential diagnosis permitting to distinguish a typical purulent meningitis and an abscess in the spinal canal. The authors also discuss the suitability of administering penicillin in staphylococcal sepsis caused by a sensitive strain. The authors do not recommend crystalline penicillin in the management of severe staphylococcal infections even in cases, where the isolated S. aureus strain is really sensitive to penicillin.


Comparison of quality of life between men and women who underwent Transforaminal Percutaneous Endoscopic Discectomy for lumbar disc herniation.
Studies describing the efficacy of TPED on shortness of recovery and improvement of postoperative quality of life are limited, especially regarding gender something that has never been reported before in the literature. The purpose of this study is to evaluate possible differences of the health-related quality of life in patients who underwent TPED for LDH in accordance with sex. Seventy-six patients diagnosed and treated with TPED for LDH with 1 year follow-up were selected and divided into two groups of equal number depending on sex. Their quality of life was evaluated by using the SF-36 before the operation, six weeks, three, six and twelve months postoperatively. A statistical analysis was conducted, in order to compare the 8 scaled scores of the SF-36 combining each time two chronological phases in the total of patients, in each group and between groups. Fifty-two (68.4%) patients were ≤63 years old, while the rest 24 (31,4%) were >63 years old (mean ±SD = 56,5 ±12,1 years). Apart from the PF domain, the scores were higher in every visit for the two groups, but the change between groups was not significant. Women had a significantly higher increase of PF score in 3 months after TPED and in the interval 6 weeks-3 months comparing with men. However, in the intervals 3 months-6 months and 3 months-12 months men presented significantly higher increase compared to women. Statistically significant improvement of the quality of life for both men and women was observed. Generally, there was no significant difference between the two groups. As regards to the physical functioning, it appears to be a significant difference which is counterpoised over time. 2. TPED for LDH does not present major differences in the improvement of quality of life regarding gender.


Cardio-vascular effects of vecuronium & pancuronium in patients undergoing elective closed mitral valvotomy.
Vecuronium, a monoquaternary analogue of pancuronium, the neuromuscular blocker, was compared with pancuronium in 50 patients undergoing elective closed mitral valvotomy. The patients were randomly divided into two groups of 25 each, and the muscle relaxants were administered in a dose of 0.1 mg/kg body weight. Both the agents produced identical intubating conditions at 3 min. Vecuronium showed a significantly shorter onset of action, as compared to pancuronium. The latter significantly increased the heart rate throughout the period of study whereas vecuronium significantly decreased the heart rate, 25 min after administration. There was significant increase in the mean arterial pressure (MAP) at tracheal intubation in both the groups, which persisted throughout the period of study in pancuronium group. There was a significant fall in MAP at 30 min after relaxant in vecuronium group. The incidence of arrythmias was similar and significant in both the groups. Vecuronium, thus showed a quicker onset of action with minimal haemodynamic effects, as compared to pancuronium in patients undergoing closed mitral valvotomy.


Working memory capacity and task goals modulate error-related ERPs.
The present study investigated individual differences in information processing following errant behavior. Participants were initially classified as high or as low working memory capacity using the Operation Span Task. In a subsequent session, they then performed a high congruency version of the flanker task under both speed and accuracy stress. We recorded ERPs and behavioral measures of accuracy and response time in the flanker task with a primary focus on processing following an error. The error-related negativity was larger for the high working memory capacity group than for the low working memory capacity group. The positivity following an error (Pe) was modulated to a greater extent by speed-accuracy instruction for the high working memory capacity group than for the low working memory capacity group. These data help to explicate the neural bases of individual differences in working memory capacity and cognitive control.


A conserved dimorphism-regulating histidine kinase controls the dimorphic switching in Paracoccidioides brasiliensis.
Paracoccidioides brasiliensis and P. lutzii, thermally dimorphic fungi, are the causative agents of paracoccidioidomycosis (PCM). Paracoccidioides infection occurs when conidia or mycelium fragments are inhaled by the host, which causes the Paracoccidioides cells to transition to the yeast form. The development of disease requires conidia inside the host alveoli to differentiate into yeast cells in a temperature-dependent manner. We describe the presence of a two-component signal transduction system in P. brasiliensis, which we investigated by expression analysis of a hypothetical protein gene (PADG_07579) that showed high similarity with the dimorphism-regulating histidine kinase (DRK1) gene of Blastomyces dermatitidis and Histoplasma capsulatum This gene was sensitive to environmental redox changes, which was demonstrated by a dose-dependent decrease in transcript levels after peroxide stimulation and a subtler decrease in transcript levels after NO stimulation. Furthermore, the higher PbDRK1 levels after treatment with increasing NaCl concentrations suggest that this histidine kinase can play a role as osmosensing. In the mycelium-yeast (M→Y) transition, PbDRK1 mRNA expression increased 14-fold after 24 h incubation at 37°C, consistent with similar observations in other virulent fungi. These results demonstrate that the PbDRK1 gene is differentially expressed during the dimorphic M→Y transition. Finally, when P. brasiliensis mycelium cells were exposed to a histidine kinase inhibitor and incubated at 37°C, there was a delay in the dimorphic M→Y transition, suggesting that histidine kinases could be targets of interest for PCM therapy.


Bacterial catabolism of acetovanillone, a lignin-derived compound.
Bacterial catabolic pathways have considerable potential as industrial biocatalysts for the valorization of lignin, a major component of plant-derived biomass. Here, we describe a pathway responsible for the catabolism of acetovanillone, a major component of several industrial lignin streams. Rhodococcus rhodochrous GD02 was previously isolated for growth on acetovanillone. A high-quality genome sequence of GD02 was generated. Transcriptomic analyses revealed a cluster of eight genes up-regulated during growth on acetovanillone and 4-hydroxyacetophenone, as well as a two-gene cluster up-regulated during growth on acetophenone. Bioinformatic analyses predicted that the hydroxyphenylethanone (Hpe) pathway proceeds via phosphorylation and carboxylation, before β-elimination yields vanillate from acetovanillone or 4-hydroxybenzoate from 4-hydroxyacetophenone. Consistent with this prediction, the kinase, HpeHI, phosphorylated acetovanillone and 4-hydroxyacetophenone. Furthermore, HpeCBA, a biotin-dependent enzyme, catalyzed the ATP-dependent carboxylation of 4-phospho-acetovanillone but not acetovanillone. The carboxylase's specificity for 4-phospho-acetophenone (kcat/KM = 34 ± 2 mM-1 s-1) was approximately an order of magnitude higher than for 4-phospho-acetovanillone. HpeD catalyzed the efficient dephosphorylation of the carboxylated products. GD02 grew on a preparation of pine lignin produced by oxidative catalytic fractionation, depleting all of the acetovanillone, vanillin, and vanillate. Genomic and metagenomic searches indicated that the Hpe pathway occurs in a relatively small number of bacteria. This study facilitates the design of bacterial strains for biocatalytic applications by identifying a pathway for the degradation of acetovanillone.


The 100 Most Cited vs. Most Relevant Articles in the Journal of Neurosurgery: A Bibliometric Analysis.
Introduction The Journal of Neurosurgery (JNS) published its first volume in 1944 and has evolved into the top cited journal in the field of neurosurgery. The aim of this study was to determine and characterize the 100 most cited (based on the total number of citations) vs. most relevant (based on the number of citations per year) articles originating in JNS. Methods The top 100 most cited articles in JNS were determined by searching the Web of Science database. Citations per year were additionally calculated for the top 1000 articles by total citations to rank the 100 most relevant articles. Results The median number of total citations for the 100 most cited articles in JNS was 505 (range 383-2200), and the median number of citations per year for the 100 most relevant articles was 21.88 (range 17.31-82.61). The median year of publication for the 100 most cited and most relevant articles was 1990 and 1999, respectively (P < 0.0001). Most articles originated in the United States in both categories (72% and 71%, respectively). The most common topic of study was cerebrovascular on both lists, followed by trauma on the most cited list vs. tumor on the most relevant list. The most relevant list also contained considerably more articles with a higher level of evidence such as systemic reviews/meta-analyses and prospective studies. Conclusions This study highlights the key contributing factors to the growth and flourishing of JNS. It also reveals several discrepancies between the most cited and most relevant articles, with the latter including more recently published articles, more studies addressing tumor, and more level I/1 (NHMRC/CEBM) evidence. Bibliometric analysis serves as a useful tool for clinicians and researchers to appraise published literature and understand the scientific foundation of modern neurosurgery.


Factors influencing sperm-zona pellucida binding in vitro using the intact zona model.
The zona pellucida binding assay assesses the ability of spermatozoa to bind to the zona pellucida. The present study investigated the influence of: (i) prior oocyte exposure to spermatozoa on subsequent sperm-zona pellucida binding in vitro; and (ii) cryopreservation of oocytes. Only oocytes obtained from fertile donors were used and the binding capacity of non-inseminated, cryopreserved oocytes was compared with both inseminated/unfertilized, cryopreserved oocytes and inseminated/unfertilized, non-cryopreserved oocytes recovered from in-vitro fertilization cultures on sperm-zona pellucida binding using an intact zona model. There was no statistically significant difference in sperm-zona binding between non-inseminated, cryopreserved oocytes (range 9.6-23.2), inseminated/unfertilized, cryopreserved oocytes (range 15.0-16.0) and inseminated/ unfertilized, non-cryopreserved oocytes (range 3.3-23.0). The coefficient of variation for sperm binding to all oocyte groups was very large (range 37-121%). We conclude that neither prior exposure of human oocytes to human spermatozoa nor cryopreservation of human oocytes influences the subsequent binding of a different population of spermatozoa to the zona pellucida. However, large oocyte-to-oocyte variation of sperm-zona binding may diminish the usefulness of this assay in clinical practice and as a research tool.


En Route to Zero Emissions for Power and Industry with Amine-Based Post-combustion Capture.
As more countries commit to a net-zero GHG emission target, we need a whole energy and industrial system approach to decarbonization rather than focus on individual emitters. This paper presents a techno-economic analysis of monoethanolamine-based post-combustion capture to explore opportunities over a diverse range of power and industrial applications. The following ranges were investigated: feed gas flow rate between 1-1000 kg ·s-1, gas CO2 concentrations of 2-42%mol, capture rates of 70-99%, and interest rates of 2-20%. The economies of scale are evident when the flue gas flow rate is <20 kg ·s-1 and gas concentration is below 20%mol CO2. In most cases, increasing the capture rate from 90 to 95% has a negligible impact on capture cost, thereby reducing CO2 emissions at virtually no additional cost. The majority of the investigated space has an operating cost fraction above 50%. In these instances, reducing the cost of capital (i.e., interest rate) has a minor impact on the capture cost. Instead, it would be more beneficial to reduce steam requirements. We also provide a surrogate model which can evaluate capture cost from inputs of the gas flow rate, CO2 composition, capture rate, interest rate, steam cost, and electricity cost.


[Effects of hnRNP B1-specific siRNA on the expression of hnRNP B1 in lung cancer tissues in vivo].
To measure the effect of the eukaryotic expression vector of siRNA specific for hnRNP B1 on the expression of hnRNP Bx in lung cancer tissues in vivo. The eukaryotic expression vector of siRNA specific for hnRNP B1 was inoculated to nude mice with lung cancers. The mRNA and protein expressions of hnRNP B1 in the lung cancer tissues of the nude mice were detected by RT-PCR and Western blot. The optimal conditions for the expressions were identified. The eukaryotic expression vector of siRNA specific for hnRNP B1 inhibited the mRNA and protein expressions at 40 days after the inoculation. However, the inhibitive effect disappeared at 60 days after the inoculation. The expression of hnRNP B1 gene in A549 cells can be inhibited by hnRNP Br-specific siRNA expression vectors.


Deletion of the Ttf1 gene in differentiated neurons disrupts female reproduction without impairing basal ganglia function.
Thyroid transcription factor 1 (TTF1) [also known as Nkx2.1 (related to the NK-2 class of homeobox genes) and T/ebp (thyroid-specific enhancer-binding protein)], a homeodomain gene required for basal forebrain morphogenesis, remains expressed in the hypothalamus after birth, suggesting a role in neuroendocrine function. Here, we show an involvement of TTF1 in the control of mammalian puberty and adult reproductive function. Gene expression profiling of the nonhuman primate hypothalamus revealed that TTF1 expression increases at puberty. Mice in which the Ttf1 gene was ablated from differentiated neurons grew normally and had normal basal ganglia/hypothalamic morphology but exhibited delayed puberty, reduced reproductive capacity, and a short reproductive span. These defects were associated with reduced hypothalamic expression of genes required for sexual development and deregulation of a gene involved in restraining puberty. No extrapyramidal impairments associated with basal ganglia dysfunction were apparent. Thus, although TTF1 appears to fulfill only a morphogenic function in the ventral telencephalon, once this function is satisfied in the hypothalamus, TTF1 remains active as part of the transcriptional machinery controlling female sexual development.


HbA1c in Dental Practices: Enhancing Patient Health Through Blood Glucose Screenings.
Periodontal disease and diabetes are highly prevalent chronic health conditions that have far-reaching implications, impacting health, economic well-being, and overall quality of life. With half of all American adults affected by periodontal disease1 and more than 422 million individuals impacted by diabetes globally,2 the need for effective strategies to manage these conditions is significant. Incorporating hemoglobin A1c (HbA1c) testing into routine dental visits provides dental professionals an opportunity to improve patient health outcomes. This article explores the importance of HbA1c testing in dental practices and the benefits of dental-medical collaboration in managing periodontal disease and diabetes.


Renal hypouricemia caused by novel compound heterozygous mutations in the SLC22A12 gene: a case report with literature review.
Renal hypouricemia (RHUC) is a heterogeneous genetic disorder that is characterized by decreased serum uric acid concentration and increased fractional excretion of uric acid. Previous reports have revealed many functional mutations in two urate transporter genes, SLC22A12 and/or SLC2A9, to be the causative genetic factors of this disorder. However, there are still unresolved patients, suggesting the existence of other causal genes or new mutations. Here, we report an RHUC patient with novel compound heterozygous mutations in the SLC22A12 gene. A 27-year-old female presenting with recurrent hypouricemia during routine checkups was referred to our hospital. After obtaining the patient's consent, both the patient and her healthy parents were analyzed using whole-exome sequencing (WES) and Sanger sequencing to discover and validate causal mutations, respectively. The prioritization protocol of WES screened out two mutations of c.269G > A/p.R90H and c.1289_1290insGG/p.M430fsX466, which are both located in the SLC22A12 gene, in the patient. Sanger sequencing further confirmed that the patient's heterozygous c.269G > A/p.R90H mutation, which has been reported previously, derived from her mother, and the heterozygous c.1289_1290insGG/p.M430fsX466 mutation, which was found for the first time, derived from her father. p.R90H, which is highly conserved among different species, may decrease the stability of this domain and was considered to be almost damaging in silicon analysis. p.M430fsX466 lacks the last three transmembrane domains, including the tripeptide motif (S/T)XΦ (X = any amino acid and Φ = hydrophobic residue), at the C-terminal, which interact with scaffolding protein PDZK1 and thus will possibly lead to weak functioning of urate transport through the disruption of the "transporter complex" that is formed by URAT1 and PDZK1. We report a Chinese patient with RHUC, which was caused by compound heterozygous mutations of the SLC22A12 gene, using WES and Sanger sequencing for the first time. Mutation-induced structural instability or malfunction of the urate transporter complex may be the main mechanisms for this hereditary disorder.


Evolution of a comprehensive, orthogonal approach to sequence variant analysis for biotherapeutics.
Amino acid sequence variation in protein therapeutics requires close monitoring during cell line and cell culture process development. A cross-functional team of Pfizer colleagues from the Analytical and Bioprocess Development departments worked closely together for over 6 years to formulate and communicate a practical, reliable sequence variant (SV) testing strategy with state-of-the-art techniques that did not necessitate more resources or lengthen project timelines. The final Pfizer SV screening strategy relies on next-generation sequencing (NGS) and amino acid analysis (AAA) as frontline techniques to identify mammalian cell clones with genetic mutations and recognize cell culture process media/feed conditions that induce misincorporations, respectively. Mass spectrometry (MS)-based techniques had previously been used to monitor secreted therapeutic products for SVs, but we found NGS and AAA to be equally informative, faster, less cumbersome screening approaches. MS resources could then be used for other purposes, such as the in-depth characterization of product quality in the final stages of commercial-ready cell line and culture process development. Once an industry-wide challenge, sequence variation is now routinely monitored and controlled at Pfizer (and other biopharmaceutical companies) through increased awareness, dedicated cross-line efforts, smart comprehensive strategies, and advances in instrumentation/software, resulting in even higher product quality standards for biopharmaceutical products.


Dendritic cells in blood and urine samples from bladder cancer patients undergoing BCG immunotherapy.
Immunotherapy with BCG (Bacille Calmette-Guérin) after transurethral resection of the bladder tumor represents a highly effective primary treatment for intermediate and high-risk superficial bladder cancer. The effectiveness of this therapy has been documented, but its mechanism of action is not clear yet. In the present study, we investigated the changes of dendritic cells (DC) numbers in peripheral blood and urine of patients with superficial bladder cancer undergoing BCG intravescical therapy. We have enumerated plasmacytoid and myeloid DCs in the peripheral blood and in the urine of patients with bladder cancer in order to clarify the role of these cells in the evolution of the disease and the effect of therapy. DCs in blood and urine samples were assessed using the single-platform TruCOUNT assay with monoclonal antibodies. The study population included 37 healthy donors and 13 patients with diagnosis of primitive superficial bladder cancer. At the time of diagnosis a reduction of blood DCs was found in patients as opposed to healthy donors, while DCs were not found in the urine in the same way as in healthy subjects. Six of these patients were followed before and after weekly and monthly instillations of BCG. In the peripheral blood, we observed an immunological recovery of DCs from the third weekly instillation up to the sixth. In the urine of patients, we didn't find mDCs or pDCs at T0, but we found a statistically significant change from the third instillation up to the sixth. On the contrary, we didn't find mDCs in urine during monthly instillation. DC Count could be used in the monitoring of patients undergoing BCG therapy. Immunological restoration of mDC numbers in peripheral blood and the efflux in urine could be important for confirming the effectiveness of BCG instillation.


Three-dimensional computer-guided implant placement in oligodontia.
The aim of computer-designed surgical templates is to attain higher precision and accuracy of implant placement, particularly for compromised cases. The purpose of this study is to show the benefit of a full three-dimensional virtual workflow to guide implant placement in oligodontia cases where treatment is challenging due compromised bone quantity and limited interdental spaces. A full, digitalized workflow was performed for implant placement in two oligodontia patients. Accuracy was assessed by calculating the coordinates of the entry point (shoulder) and apex (tip) as well as the angular deviation of the planned and actual implants. Implant placement could be well performed with the developed computer-designed templates in oligodontia. Mean shoulder deviation was 1.41 mm (SD 0.55), mean apical deviation was 1.20 mm (SD 0.54) and mean angular deviation was 5.27° (SD 2.51). Application of computer-designed surgical templates, as described in this technical advanced article, aid in predictable implant placement in oligodontia where bone quantity is scarce and interdental spaces are limited.


Loss-of-function variants in ZEB1 cause dominant anomalies of the corpus callosum with favourable cognitive prognosis.
The neurodevelopmental prognosis of anomalies of the corpus callosum (ACC), one of the most frequent brain malformations, varies extremely, ranging from normal development to profound intellectual disability (ID). Numerous genes are known to cause syndromic ACC with ID, whereas the genetics of ACC without ID remains poorly deciphered. Through a collaborative work, we describe here ZEB1, a gene previously involved in an ophthalmological condition called type 3 posterior polymorphous corneal dystrophy, as a new dominant gene of ACC. We report a series of nine individuals with ACC (including three fetuses terminated due to ACC) carrying a ZEB1 heterozygous loss-of-function (LoF) variant, identified by exome sequencing. In five cases, the variant was inherited from a parent with a normal corpus callosum, which illustrates the incomplete penetrance of ACC in individuals with an LoF in ZEB1. All patients reported normal schooling and none of them had ID. Neuropsychological assessment in six patients showed either normal functioning or heterogeneous cognition. Moreover, two patients had a bicornuate uterus, three had a cardiovascular anomaly and four had macrocephaly at birth, which suggests a larger spectrum of malformations related to ZEB1. This study shows ZEB1 LoF variants cause dominantly inherited ACC without ID and extends the extraocular phenotype related to this gene.


Neurobiochemical mechanisms of a ketogenic diet in refractory epilepsy.
A ketogenic diet is an important therapy used in the control of drug-refractory seizures. Many studies have shown that children and adolescents following ketogenic diets exhibit an over 50% reduction in seizure frequency, which is considered to be clinically relevant. These benefits are based on a diet containing high fat (approximately 90% fat) for 24 months. This dietary model was proposed in the 1920s and has produced variable clinical responses. Previous studies have shown that the mechanisms underlying seizure control involve ketone bodies, which are produced by fatty acid oxidation. Although the pathways involved in the ketogenic diet are not entirely clear, the main effects of the production of ketone bodies appear to be neurotransmitter modulation and antioxidant effects on the brain. This review highlights the impacts of the ketogenic diet on the modulation of neurotransmitters, levels of biogenic monoamines and protective antioxidant mechanisms of neurons. In addition, future perspectives are proposed.


The haematology of hyperthyroidism: abnormalities of erythrocytes, leucocytes, thrombocytes and haemostasis.
The abnormalities of erythrocytes, leucocytes, thrombocytes and coagulation that have been reported, particularly in more recent years, to be associated with hyperthyroidism are surveyed. Several areas are highlighted where further investigations could lead to clinically useful insights, improved information about the haematological processes involved or to a better understanding of thyroid hormone action.


[Anesthetic management by psoas compartment block for adult inguinal herniorrhaphy].
Spinal anesthesia has been often used for inguinal herniorrhaphy. With the advancement of surgical technique, tension free inguinal herniorrhaphy is now performed under local anesthesia. Local anesthesia, however, does not always offer sufficient anesthesia. We performed psoas compartment block with 1% lidocaine in a 78-year-old man undergoing tension free inguinal herniorrhaphy. During the operation, the patient felt pain and needed additional local anesthesia once. But the pain was not severe, and he did not feel stressed. Advantages of psoas compartment block for inguinal herniorrhaphy are its easiness to perform, little burden of anesthesia and high efficacy. Demerits are slow onset of block and necessity for prolonged patient monitoring. Further investigations on dose and patient position are required for psoas compartment block to become useful for inguinal herniorrhaphy.


Pharmacoepidemiology of drug use at two primary health care centres in northern India.
There is a consensus amongst most health care centers that limited drugs should be available to meet most requirements at the first tier of health care. However, few studies have addressed themselves to determining this. The objectives of this study were to provide a description of the availability and prescription of modern pharmaceuticals at 2 selected Primary Health Care Centres in Northern India. The main results of this study are: documentation of age/sex of patients on prescriptions was not kept in all cases; prescribing by both generic/brand names was observed with generics predominating; most patients received 0-2 drugs; dosages were not specified in 10% of the cases, duration not specified in 30% of the cases, and dosage form not indicated in 30% of all drugs prescribed; treatment was sought for problems pertaining to gastrointestinal, respiratory, dermatological and musculoskeletal systems; and the most prescribed drugs were analgesics, antipyretics, antimicrobials, hematinics and antihistaminics.


[Subdural tension pneumocephalus after trephination for chronic subdural hematoma].
Symptomatic tension pneumocephalus developed after an evacuation of chronic subdural hematoma is reported. In this 71 year-old man, a rapid deterioration of consciousness was noted 5 days after a trephination for bilateral chronic subdural hematomas. An evacuation of hematoma was performed under a local anesthesia, and the preoperative and immediate postoperative course was uneventful. Subdural tension pneumocephalus was confirmed by typical CT findings and treated by re-trephination and drainage. The literature on the tension subdural pneumocephalus was reviewed, and the mechanism of its development was briefly discussed.


Sleep disturbances in allergic diseases.
Sleep is fundamental for physical and mental health as well as daytime performance and represents a crucial aspect in the personal representations of well-being. Sleep disturbances in patients with chronic disorders constitute a factor that contributes to exacerbate symptoms, complicate management, and affect mood and quality of life (QoL). Allergic diseases are a global health problem of increasing prevalence that affects up to 15% of the population in Western countries. Sleep problems associated with allergic diseases may play a role in worsening the burden of illness, contributing to impairment of the QoL. The aim of this review was to describe the most common causes leading to sleep disturbance in allergic patients and their consequences on the QoL. The possible negative effects of treatment on sleep parameters has been also considered.


Locus control region elements HS1 and HS4 enhance the therapeutic efficacy of globin gene transfer in beta-thalassemic mice.
Globin gene transfer in autologous hematopoietic stem cells is a promising therapeutic option for subjects with beta-thalassemia major. In this approach, high level, erythroid-specific globin transgene expression should correct ineffective erythropoiesis and hemolytic anemia following the delivery of only 1 to 2 vector copies per cell. The generation of vectors that provide high-level globin expression and require low vector copy (VC) integration is therefore essential for both safety and efficacy. We show here the major roles played by 2 lesser-known locus control region elements, termed HS1 and HS4. Partial deletions within HS4 markedly reduce in vivo globin expression requiring multiple VC per cell to correct the anemia. Most strikingly, addition of HS1 to HS2-3-4 increases globin expression by 52%, yielding 9 g Hb/VC in beta-thalassemic mice. Thus, while vectors encoding HS2-3-4 provide curative levels of hemoglobin at 1 to 2 copies per cell, adding HS1 is a promising alternative strategy if upcoming clinical trials prove higher levels of expression to be necessary.


The Relationship Between Activity Pattern and Muscle Adaptation in Skeletal Muscle.
Muscle is highly plastic in terms of size (maximum force), speed, maximum power, and endurance. Well-controlled studies in animals have shown that the adult skeletal muscle fiber has a remarkable ability to modify its gene expression so that with long-term substantial changes in the daily activity pattern the contractile phenotype can be modified across the whole spectrum of fiber type found in control muscle. The contractile phenotype in this context includes the isoform content of myosin and therefore the maximum velocity of shortening, the mitochondrial content and therefore the specific force and aerobic capacity (endurance), and the calcium handling proteins and therefore the speed of activation and relaxation. With voluntary exercise in human subjects, similar responses are observed, although the degree of transformation is restricted by the practical limitations of exercise dosing to changes in mitochondrial activity and muscle size rather than the more profound changes in contractile protein isoform that can be induced with artificial activation over a substantial proportion of the day.


Acyl carrier protein (ACP) import into chloroplasts. Covalent modification by a stromal holoACP synthase is stimulated by exogenously added CoA and inhibited by adenosine 3',5'-bisphosphate.
During the import of the precursor for the acyl carrier protein (ACP) into chloroplasts, apoACP is converted to holoACP by the attachment of a phosphopantetheine group transferred from coenzyme A (CoA) by a chloroplast holoACP synthase [Fernandez, M. and Lamppa, G. (1990) Acyl carrier protein import into chloroplasts does not require the phosphopantetheine: evidence for a chloroplast holoACP synthase, Plant Cell 2, 195-206]. Here it is shown that exogenous addition of CoA to intact chloroplasts in the import assay stimulates the conversion of apoACP to holoACP. If adenosine 3',5'-bisphosphate [Ado(3',5')P2], the byproduct of the transfer reaction, was also included the extent of conversion was greatly reduced. CoA has its effect after ACP precursor (pre-ACP) import and proteolytic removal of the transit peptide, thus indicating that the chloroplast holoACP synthase resides in the stroma where fatty acid synthase is found. When Ado(3',5')P2 was added alone to the import assay, it inhibited the synthesis of holoACP. Inhibition of the conversion of apo- to holoACP with Ado(3',5')P2 made it possible to examine whether the holoform of preACP could be imported into chloroplasts. Pre-apoACP was synthesized in Escherichia coli and shown to be competent for import in an ATP- and temperature-dependent manner. A partially purified chloroplast holoACP synthase converted 60-90% of the pre-apoACP to pre-holoACP. Pre-holoACP incubated with chloroplasts in the presence of Ado(3',5')P2 yielded > 60% holoACP, whereas the control reaction with pre-apoACP gave primarily apoACP. Hence the phosphopantetheine prosthetic group of ACP does not block precursor movement through the translocation apparatus of the chloroplast envelope.


The effect of private insurance access on the choice of GP/specialist and public/private provider in Spain.
This paper sheds light into the investigation of differential patterns of utilisation of physician services by populations subgroups that is emerging in a number of studies. Using Spanish data from the National Health Survey of 1997 we try to explain the distinct role of the type of insurance on the choice between specialists and GPs and its intertwining with the choice between private and public providers. We estimate a two-stages probit to conclude that differences in insurance access is the main determinant of both, the choice of sector and the kind of physician contacted, giving rise to very different patterns of consumption of GP and specialist visits. People with only public insurance go 2.8 times to the GP per one time that they visit a specialist; individuals with duplicate coverage have a ratio of GP/specialist visits equal to 1.4 (the combination being public GP and private specialist) and people with only private insurance access actually have an 'inverted' pattern of visits: they contact specialists more often than GPs. Age, sex and health and public supply characteristics also have a distinct and interesting impact on these choices. Finally, equity concerns based on the implied assumption that specialists care is superior to general practitioner care are discussed.


Identification of programmed cell death in normal human skin tissues by using specific labelling of fragmented DNA.
Programmed cell death (PCD) in normal human skin tissues was studied by using in situ specific labelling of fragmented DNA. This labelling method clearly stained the nuclei of Henle's layer in the bulb of the anagen hair follicle in serial sections, and the nuclei of the inner root sheath cuticle cells and Huxley's layer cells showed positive staining in the upper part of the hair follicles. This staining pattern was consistent with the sequence of keratinization in the three layers. The nuclei of differentiated cells located at the centre of the sebaceous glands, and those of the granular keratinocyte layer, were also stained. These findings suggest that PCD might play a key role in the terminal differentiation of the epidermis and epidermal appendages.


Novel proteins associated with MHC class I antigens in cells expressing the adenovirus protein E3/19K.
Assembly of histocompatibility class I antigens (MHC) with beta 2-microglobulin (beta 2m) and peptide takes place in the rough endoplasmic reticulum (ER). At present, it is unclear why peptides generated in the cytosol or ER by proteolysis are not further degraded. Also, it is an open question whether assembly and/or peptide binding is self-instructive or is promoted by additional molecules, for example, chaperones. We previously demonstrated that the adenovirus glycoprotein E3/19K binds to human and some mouse MHC molecules in the ER, interfering with their transport to the cell surface. Here we show that immunoprecipitates from human cells that express transfected E3/19K and murine MHC Kd molecules not only contain MHC heavy chain, beta 2m and E3/19K but also two additional proteins with apparent molecular weights of 100 kDa and 110 kDa. Biochemical characterization of these proteins, designated p100 and p110, demonstrates that they are transmembrane glycoproteins with a similar if not identical protein backbone. Consistent with a role as chaperones, we find that glucose starvation induces complex formation between p100/110 and MHC-E3/19K. Most interestingly, p100 and p110 are displaced from the complex by addition of Kd-specific peptides. Therefore, p100 and p110 might be chaperones that promote correct folding of MHC antigens and/or peptide binding to MHC.


Structural, optical, and photophysical properties of nickel(II) alkylthioporphyrins: insights from experimental and DFT/TDDFT studies.
The ground- and excited-state properties of a Ni(II) porphyrin bearing peripheral alkylthio group, NiOMTP (OMTP = 2,3,7,8,12,13,17,18-octakis methylthio porphyrinate) have been investigated by steady-state and time-resolved absorption spectrometry and DFT/TDDFT theoretical methods. Several conformations corresponding to different deformations of the porphyrin core and to different orientations of the alkylthio groups have been theoretically explored. The nearly degenerate, purely ruffled D(2d) and hybrid (ruffled with a modest degree of saddling) D(2) conformations, both characterized by an up-down (ud) orientation of the vicinal methylthio groups are by far the preferred conformations in the "gas phase". In contrast to NiOEP, it is the orientation of the peripheral substituents rather than the type and degree of distortions of the porphyrin core that determines the stability of the NiOMTP conformers. The ground-state electronic absorption spectra of NiOMTP exhibit significant changes compared to its parent NiP and beta-alkylated analogues, such as NiOEP, resulting in a considerable red shift of the B and the Q bands, intensification and broadening of the Q band, and additional weak absorptions in the region between the Q and B bands. These spectral changes can be understood in terms of the electronic effects of the methylthio groups with nonplanar distortions of the porphyrin ring playing a very minor role. Transient absorption measurements with sub-picosecond resolution performed in toluene and TDDFT calculations reveal that following photoexcitation, NiOMTP deactivates by the pathway 1(pi,pi) --> 3(d(z2),d(x2-y2))--> ground state. The (d,d) state exhibits complex spectral evolution over ca. 8 ps, interpreted in terms of vibrational relaxation and cooling. The cold ligand-field excited state decays with a lifetime of 320 ps. At variance with the highly distorted nickel porphyrins but similar to the planar analogues, the (d,d) spectrum of NiOMTP has transient absorption bands immediately to the red of the bleaching of the ground-state Q and B bands.


New furostanol glycosides from Polygonatum multiflorum (L.) All.
The phytochemical investigation of the whole plant of Polygonatum multiflorum resulted in the isolation of two new steroidal glycosides, polmultoside A (4) and polmultoside B (5), along with three known glycosides protobioside (1), protodeltonin (2) and huangjiangsu A (3). The structures of the isolated compounds have been elucidated by extensive 1D (1H, 13C) and 2D (COSY, HSQC, HMBC) NMR spectral data analysis, as well as high-resolution mass determinations.


Flavonol dioxygenation catalysed by cobalt(II) complexes supported with 3N(COO) and 4N donor ligands: a comparative study to assess the carboxylate effects on quercetin 2,4-dioxygenase-like reactivity.
Two new cobalt(II)-acetato complexes, [CoII(L3NCOO)(OAc)]·0.5H2O (1OAc·0.5H2O) and [CoII(L4N)(OAc)](PF6) (2OAc(PF6)), were synthesised using ligands L3NCOO- (Li+L3NCOO- = lithium 2-(benzyl((6'-methyl-[2,2'-bipyridin]-6-yl)methyl)amino)acetate) and L4N (N-benzyl-1-(6'-methyl-[2,2'-bipyridin]-6-yl)-N-(pyridin-2-ylmethyl)methanamine), respectively, to mimic the functional activity of cobalt(II)-quercetin-2,4-dioxygenase (CoII-2,4-QD). Additionally, Co(II)-flavonolato ternary complexes, [CoII(L3NCOO)(fla)]·H2O (1fla·H2O) and [CoII(L4N)(fla)](PF6) (2fla(PF6)), were synthesised as enzyme-substrate models. All four complexes were thoroughly characterised by elemental analyses and spectroscopic methods. Structural characterisation was performed for 1OAc·0.5H2O, 2OAc(PF6)·CH2Cl2 and 2fla+ with a perchlorate counter anion, 2fla(ClO4)·1.5H2O. Furthermore, density functional theory (DFT) calculations, time-dependent DFT (TD-DFT) and molecular orbital (MO) analysis were performed for the flavonolato adducts 1fla and 2fla+. The catalytic activities of complexes 1OAc·0.5H2O and 2OAc(PF6) in the oxygenative degradation of flavonol (multiple-turnover reactions) were investigated at 70 °C in DMF to determine the effect of the carboxylate substituent over a pyridyl donor residue on reactivity. Complex 1OAc·0.5H2O showed a higher catalytic rate than complex 2OAc(PF6). The same reactivity order was observed for single-turnover dioxygenation reactions with ternary complexes (1fla > 2fla+). The formation constants (Kf) of 1fla and 2fla+ species are comparable, implying that catalyst-substrate adduct formation occurs in similar amounts for both catalytic reactions. Therefore, the Kf values have a similar impact on reactivities. However, the oxidation potential of the bound fla-/fla˙ couple in 1fla is considerably lower than that in 2fla+. DFT calculations predicted that the negatively charged carboxylate group of ligand L3NCOO- determines the higher reactivity of 1fla with dioxygen by decreasing the oxidation potential of the bound fla-/fla˙ couple. During the dioxygenation process, the reactive Co(II)-bound flavonoxy radical was generated via single-electron transfer from the coordinated fla- to dioxygen, simultaneously forming a superoxide ion. The anionic carboxylate group improves the stability of the bound flavonoxy radical by providing substantial electron density to the electron-deficient fla˙ through the Co(II) centre, allowing the reactive fla˙ species to accumulate at an optimal concentration for effective catalysis. EPR spectroscopy successfully detected the cobalt-bound fla˙ species formed through the dioxygenation of 1fla. NBT2+ and EPR spin-trapping experiments confirmed superoxide formation during the dioxygenation process. So, the present work describes CoII-2,4-QD model studies and clarifies the function of carboxylate in quercetinase-like reactivity.


Preventive education for high-risk children: cognitive consequences of the Carolina Abecedarian Project.
Longitudinal mental test scores for 54 educationally treated disadvantaged preschool children at high-risk for nonbiologically based mild mental retardation and 53 control children were compared. The educationally treated children were in a child-centered prevention-oriented intervention program delivered in a daycare setting from infancy to age 5. Language, cognitive, perceptual-motor, and social development were stressed. Children were examined with age-appropriate tests of development at 6, 12, 18, 24, 30, 42, 48, and 54 months of age. Beginning at 18 months, and on every test occasion thereafter, educationally treated children significantly outscored control group children on mental tests; treated children consistently scored at the national average whereas control children's scores declined from the average level at 12 months to below average at 18 months and thereafter. Implications of the results for early intervention were discussed.


Periorbital necrotizing fasciitis: trivial facial injury resulting in cardiac arrest.
Facial injuries in the context of alcohol abuse are a common presentation to casualty departments. While the majority are self limiting, the presence of periorbital or facial oedema should alert physicians to the development of periorbital necrotizing fasciitis. Here, a case illustrates this unusual manifestation of streptococcal infection, and emphasises the rapid onset and devastating systemic and local cosmetic consequences of this condition.


[Characteristics of CO2 exchange at leaf and canopy levels in tropical seasonal rain forest of Xishuangbanna].
By the methods of chamber-based and eddy covariance measurements, the CO2 exchange of dominant tree species Pometia tomentosa and Gironniera subaequalis at leaf and canopy levels in the tropical seasonal rain forest of Xishuangbanna was measured in different seasons of 2004. The results showed that for the two tree species, the maximum net photosynthesis (P(max A)) of canopy based on chamber-based measurement ranked in the order of rainy season (RS) > end of rainy season (ERS) > foggy-cool season (FS) > dry-hot season (DS), and the dark respiration rate (Rd) of leaf was RS > ERS > DS > FS. The P(max B) based on eddy covariance measurement was in the same order as that based on chamber-based measurement, while the canopy respiration rate (Re) was RS > DS > FS > ERS. The maximum net photosynthetic rate of canopy in different seasons measured by the two methods had a comparatively small difference, ranging from 0.9 to 2.0 micromol m(-2) s(-1).


Crystal growth, observation, and characterization of the low-temperature structure of the fluorite-related ruthenates: Sm(3)RuO(7) and Eu(3)RuO(7).
The compounds Sm(3)RuO(7) and Eu(3)RuO(7) were grown as single crystals from molten hydroxide fluxes. They crystallize in the orthorhombic space group Cmcm and are part of a well-known family of fluorite-related oxides of stoichiometry Ln(3)MO(7). This structure contains rare earth cations in two different coordination environments, 8-fold pseudocubic and 7-fold pentagonal bipyramidal, and contains Ru(V) cations that are octahedrally coordinated. The RuO(6) octahedra are trans vertex-sharing to yield chains oriented along the c-axis. Upon cooling, single crystals of Sm(3)RuO(7) and Eu(3)RuO(7) undergo a structural transition at 190 and 280 K, respectively, from space group Cmcm to P2(1)nb. The structure transition results in a loss of lattice centering, a doubling of the b-axis, a distortion of the vertex-shared Ru-O chains, and a reduction in the coordination of one of the rare earth cations from 8-fold to 7-fold. Accompanying this structural transition are anomalies in the magnetic susceptibility at about 190 and 280 K for Sm(3)RuO(7) and Eu(3)RuO(7), respectively. The structures of these low-temperature phases of Ln(3)RuO(7) have been determined for the first time and are described.


Resveratrol inhibits collagen-induced platelet stimulation through suppressing NADPH oxidase and oxidative inactivation of SH2 domain-containing protein tyrosine phosphatase-2.
Reactive oxygen species (ROS) produced upon collagen stimulation are implicated in propagating various platelet-activating pathways. Among ROS-producing enzymes, NADPH oxidase (NOX) is largely responsible for collagen receptor-dependent ROS production. Therefore, NOX has been proposed as a novel target for the development of antiplatelet agent. We here investigate whether resveratrol inhibits collagen-induced NOX activation and further examine the effects of resveratrol on ROS-dependent signaling pathways in collagen-stimulated platelets. Collagen-induced superoxide anion production in platelets was inhibited by resveratrol. Resveratrol suppressed collagen-induced phosphorylation of p47(phox), a major regulatory subunit of NOX. Correlated with the inhibitory effects on NOX, resveratrol protected SH2 domain-containing protein tyrosine phosphatase-2 (SHP-2) from ROS-mediated inactivation and subsequently attenuated the specific tyrosine phosphorylation of key components (spleen tyrosine kinase, Vav1, Bruton's tyrosine kinase, and phospholipase Cγ2) for collagen receptor signaling cascades. Resveratrol also inhibited downstream responses such as cytosolic calcium elevation, P-selectin surface exposure, and integrin-αIIbβ3 activation. Furthermore, resveratrol inhibited platelet aggregation and adhesion in response to collagen. The antiplatelet effects of resveratrol through the inhibition of NOX-derived ROS production and subsequent oxidative inactivation of SHP-2 suggest that resveratrol is a potential compound for prevention and treatment of thrombovascular diseases.


Transitional wall pressure fluctuations on axisymmetric bodies.
This paper reports the characteristics of wall pressure fluctuations in the transition region of the flow over axisymmetric bodies. Measurements are conducted in a low noise wind tunnel to obtain the boundary-layer flow field, the local static pressures, and the wall pressure fluctuations using hot wires, micromanometers and flush-mounted microphones. The spatial and the temporal developments of Tollmien-Schlichting (T-S) waves were first observed in the time domain. The wall pressure fluctuations in the transitional boundary-layer flows were of intermittent pulses and intensified as they were convected downstream at typically 63% of the upstream velocity. The Wigner-Ville distributions are then obtained to examine the energy evolution jointly in time and in frequency. The center frequency of the T-S wave is decreased with increases of the boundary-layer thickness and the bandwidth of the energy distribution is broadened as the local Reynolds number is increased. Finally the nondimensional spectra of the transitional wall pressure fluctuations scaled on the outer variables was obtained and it was then found that the characteristic frequency of the T-S wave was related to the outer variables as omegadelta( *)/U(infinity) approximately 0.2. The peak level of the pressure fluctuations during late transition at the characteristic frequency is about 10 dB higher than that of the fully developed flow.


Cardiopulmonary Profiling of Athletes with Post-Exertional Malaise after COVID-19 Infection-A Single-Center Experience.
(1) Background: Cardiopulmonary exercise testing (CPET) has been suggested by the European Society of Cardiology (ESC) for assessing the exercise limitations of apparently healthy individuals, but data on elite athletes regarding this test are scarce. (2) Methods: We analyzed CPET in elite (n = 43, 21.9 ± 3.7 years) and recreational (n = 40, 34.7 ± 13.0 years) athletes with persistent subjective exercise intolerance and post-exertional malaise (PEM) after COVID-19 infection. The primary outcome was the point prevalence of the adequate cardiopulmonary response (ACPR), defined by the presence of all of the following ESC criteria for apparently healthy individuals: (1) >100% of predicted peak oxygen consumption (predVO2peak), (2) VE/VCO2 < 30, (3) no exercise oscillatory ventilation (EOV), and (4) heart rate recovery of ≥12 beats/minute 1 min after exercise termination (HRR1). Results: ACPR occurred more frequently in elite athletes than in recreational athletes (70.0% vs. 39.5%; p = 0.005), mainly driven by the lower VE/VCO2 (<30: 97.7% vs. 65%, p < 0.001). Elite (11.6%) and recreational athletes (22.5%) showing a plateau of O2 pulse did not display ACPR. Conclusions: ACPR was not observed in all recreational and elite athletes with PEM. In particular, perturbed VE/VCO2 and the plateauing of O2 pulse are suitable for quantifying exercise limitations and may identify a high-risk population with long-COVID-19 syndrome who require their training intensities to be adapted.


Labor unions: a public health institution.
Using a social-ecological framework, we drew on a targeted literature review and historical and contemporary cases from the US labor movement to illustrate how unions address physical and psychosocial conditions of work and the underlying inequalities and social determinants of health. We reviewed labor involvement in tobacco cessation, hypertension control, and asthma, limiting articles to those in English published in peer-reviewed public health or medical journals from 1970 to 2013. More rigorous research is needed on potential pathways from union membership to health outcomes and the facilitators of and barriers to union-public health collaboration. Despite occasional challenges, public health professionals should increase their efforts to engage with unions as critical partners.


Exploration of 2-((Pyridin-4-ylmethyl)amino)nicotinamide Derivatives as Potent Reversal Agents against P-Glycoprotein-Mediated Multidrug Resistance.
Overexpression of the ATP-binding cassette (ABC) transport proteins, like ABCB1, commonly referred to as P-glycoprotein (P-gp), initiates active efflux of a broad spectrum of unrelated chemotherapeutic drugs in structure and function, leading to chemotherapy failure. A series of 2-((pyridin-4-ylmethyl)amino)nicotinamide derivatives as potent reversal agents against P-glycoprotein-mediated multidrug resistance (MDR) were designed and synthesized. The majority of target compounds displayed great reversal potency, especially 9n. In-depth studies demonstrated 9n has high potency (EC50 = 119.6 ± 6.9 nM), low cytotoxicity, and long duration (>24 h) in reversing adriamycin (ADM) resistance in K562/A02 cells. 9n also improved the effects of other cytotoxic agents related to MDR, increased accumulation of ADM, interrupted P-gp-mediated Rh123 efflux function, and suppressed P-gp ATPase activity in K562/A02 MDR cells. The Western blot analysis indicated that the MDR reversal by 9n was not due to a decrease in protein expression. Besides, the effect of CYP3A4 was not influenced by 9n, avoiding the toxicity caused by drug interactions. The study yielded 9n with superior properties compared to the classical inhibitor verapamil (VRP) and leading compound apatinib.


Plasma Peptidylarginine Deiminase IV Promotes VWF-Platelet String Formation and Accelerates Thrombosis After Vessel Injury.
PAD4 (peptidylarginine deiminase type IV), an enzyme essential for neutrophil extracellular trap formation (NETosis), is released together with neutrophil extracellular traps into the extracellular milieu. It citrullinates histones and holds the potential to citrullinate other protein targets. While NETosis is implicated in thrombosis, the impact of the released PAD4 is unknown. This study tests the hypothesis that extracellular PAD4, released during inflammatory responses, citrullinates plasma proteins, thus affecting thrombus formation. Here, we show that injection of r-huPAD4 in vivo induces the formation of VWF (von Willebrand factor)-platelet strings in mesenteric venules and that this is dependent on PAD4 enzymatic activity. VWF-platelet strings are naturally cleaved by ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type-1 motif-13). We detected a reduction of endogenous ADAMTS13 activity in the plasma of wild-type mice injected with r-huPAD4. Using mass spectrometry and in vitro studies, we found that r-huPAD4 citrullinates ADAMTS13 on specific arginine residues and that this modification dramatically inhibits ADAMTS13 enzymatic activity. Elevated citrullination of ADAMTS13 was observed in plasma samples of patients with sepsis or noninfected patients who were elderly (eg, age >65 years) and had underlying comorbidities (eg, diabetes mellitus and hypertension) as compared with healthy donors. This shows that ADAMTS13 is citrullinated in vivo. VWF-platelet strings that form on venules of Adamts13-/- mice were immediately cleared after injection of r-huADAMTS13, while they persisted in vessels of mice injected with citrullinated r-huADAMTS13. Next, we assessed the effect of extracellular PAD4 on platelet-plug formation after ferric chloride-induced injury of mesenteric venules. Administration of r-huPAD4 decreased time to vessel occlusion and significantly reduced thrombus embolization. Our data indicate that PAD4 in circulation reduces VWF-platelet string clearance and accelerates the formation of a stable platelet plug after vessel injury. We propose that this effect is, at least in part, due to ADAMTS13 inhibition.


microRNA deregulation in keloids: an opportunity for clinical intervention?
Keloids are defined as benign dermal scars invading adjacent healthy tissue, characterized by aberrant fibroblast dynamics and overproduction of extracellular matrix. However, the aetiology and molecular mechanism of keloid production remain poorly understood. Recent discoveries have shed new light on the involvement of a class of non-coding RNAs known as microRNAs (miRNA), in keloid formation. A number of miRNAs have differential expression in keloid tissues and keloid-derived fibroblasts. These miRNAs have been characterized as novel regulators of cellular processes pertinent to wound healing, including extracellular matrix deposition and fibroblast proliferation. Delineating the functional significance of miRNA deregulation may help us better understand pathogenesis of keloids, and promote development of miRNA-directed therapeutics against this condition.


It's Groundhog Day! What Can the History of Science Say About the Crisis in Alzheimer's Disease Research?
For years now, Alzheimer's disease (AD) research has been stuck in a Groundhog-Day scenario: an endless time loop with no breakthrough in sight. Disagreement about the validity of the field's dominant approach, based on the Amyloid Cascade Hypothesis, has led to a seemingly unresolvable trench war between proponents and critics. Our paper evaluates the recent scientific literature on AD from a historical and philosophical perspective. We show that AD research is a classic example of the boundary work at play in a field in crisis: both parties deploy historical and philosophical references to illustrate what counts as good and bad science, as proper scientific method and appropriate scientific conduct. We also show that boundary work has proved unable to point a way out of the deadlock and argue that the science system's tools for establishing scientific quality, such as peer review and the grant system, are unlikely to resolve the crisis. Rather, they consolidate the dominant model's position even more. In conclusion, we suggest that some kind of reverse boundary-work is needed that reopens the discussion on the nature of AD, an issue that has never been settled scientifically. Drawing on historical and philosophical work, we make clear that the definition of AD as a biomedical disease for which a cure can be found has consequences, not only for funding opportunities, but also for patients and their lives. A reconsideration of the desirability of these consequences may lead to different choices with respect to research priorities and patient care.


[Malaria and the work of two writer-physicians: Louis Ferdinand Celine and Carlo Levi].
Malaria rarely mentioned in literary works has been dealt with by the two writers, both physicians, studied here. Louis-Ferdinand Destouches (1894-1961) alias Celine, studied medicine in Rennes and Paris from 1921 to 1924. He had been previously engaged by the Rockefeller Foundation to lecture on tuberculosis in Brittany. He later worked for the Commission of Hygiene of the League of Nations in Geneva and took part in 1925 to a long voyage in America (Cuba, United States) and Italy where malaria was still occurring. In 1926 he published a book: La quinine en therapeutique and participated to another voyage in West Africa where he contracted malaria. Numerous hints to this disease inspired to Celine by his first trip to Africa (Cameroun) in 1916-17 are found in Voyage au bout de la nuit (1932). Carlo Levi (1902-1975) studied medicine in Torino from 1919 to 1924. Painter and writer he wrote there his masterpiece: Christ has stopped in Eboli (1945). Malaria which then decimated a very poor population of peasants abandoned to themselves represents the background of the book and it is as a competent and human physician that C. Levi gained the friendship of the inhabitants of this remote and inhospitable province.


Progress in global measles control and mortality reduction, 2000-2007.
Despite the availability of a safe and effective vaccine since 1963, measles has been a major killer of children in developing countries (causing an estimated 750,000 deaths as recently as 2000), primarily because of underutilization of the vaccine. At the World Health Assembly in 2008, all World Health Organization (WHO) member states reaffirmed their commitment to achieving a 90% reduction in measles mortality by 2010 compared with 2000, a goal that was established in 2005 as part of the Global Immunization Vision and Strategy (2). This WHO-UNICEF comprehensive strategy for measles mortality reduction (1) focuses on 47 priority countries. The strategy's components include 1) achieving and maintaining high coverage (>90%) with the routinely scheduled first dose of measles-containing vaccine (MCV1) among children aged 1 year; 2) ensuring that all children receive a second opportunity for measles immunization (either through a second routine dose or through periodic supplementary immunization activities [SIAs]); 3) implementing effective laboratory-supported disease surveillance; and 4) providing appropriate clinical management for measles cases. This report updates previously published reports and describes immunization and surveillance activities implemented during 2007. Increased routine measles vaccine coverage and SIAs implemented during 2000--2007 resulted in a 74% decrease in the estimated number of measles deaths globally. An estimated 197,000 deaths from measles occurred in 2007; of these, 136,000 (69%) occurred in the WHO South-East Asian Region. Achievement of the 2010 goal will require full implementation of measles mortality reduction strategies, especially in the WHO South-East Asian Region.


Regulation of major histocompatibility (MHC) class II human leukocyte antigen-DR alpha gene expression in thyrocytes by single strand binding protein-1, a transcription factor that also regulates thyrotropin receptor and MHC class I gene expression.
The single strand binding protein (SSBP-1) is a positive regulator of TSH receptor gene expression and binds to an element with a GXXXXG motif. The S box of the mouse major histocompatibility class II gene has multiple GXXXXG motifs and can also bind SSBP-1. The S box is one of four highly conserved elements on the 5'-flanking region of class II genes that are necessary for interferon-gamma (IFNgamma) to overcome the normally suppressed state of the gene and induce aberrant class II expression. In this report we show that SSBP-1, when overexpressed in FRTL-5 thyroid cells, is a positive regulator of human leukocyte antigen (HLA)-DR alpha class II gene expression, as is IFNgamma or the class II trans-activator (CIITA). This is evidenced by increased exogenous promoter activity, increased endogenous RNA levels, and increased endogenous antigen expression after transfecting full-length SSBP-1 complementary DNA together with a HLA-DR alpha promoter-reporter gene chimera into TSH-treated FRTL-5 thyroid cells whose endogenous SSBP-1 levels are low. IFNgamma reverses the ability of TSH to decrease endogenous SSBP-1 RNA levels. Also, whereas SSBP-1 transfection does not cause any increase in IFNgamma-induced exogenous promoter activity, transfection of SSBP-1 and CIITA additively increases endogenous class II RNA levels to levels measured in cells treated with IFNgamma. Further, competition studies show that SSBP-1 binding is necessary for formation of the double strand protein/DNA complexes that are seen in electrophoretic mobility shift assays when the class II 5'-flanking region is incubated with extracts from IFNgamma-treated FRTL-5 cells and that have been previously associated with IFNgamma-induced aberrant class II expression. These data suggest that SSBP-1 is involved in the action of IFNgamma to overcome the normally suppressed state of the class II gene; it functions together with CIITA, whose expression is independently increased by IFNgamma. The effect of SSBP-1 as a positive regulator of class II promoter activity is lost in cells maintained without TSH, in which endogenous SSBP-1 RNA levels are already high in the absence of aberrant class II gene expression. These data suggest that high levels of endogenous SSBP-1 are insufficient to cause aberrant class II expression, but, rather, TSH or IFNgamma treatment additionally modulates the cell, albeit differently, such that transfected or endogenous SSBP-1, respectively, can express its positive regulatory activity. The effect of TSH is consistent with reports indicating that TSH enhances the ability of IFNgamma to increase class II gene expression despite the fact IFNgamma increases endogenous SSBP-1 to only the same levels as in cells untreated with TSH. Finally, the effect of SSBP-1 as a positive regulator is lost when GXXXXG motifs, which exist on both the coding and noncoding strands of the S box, are mutated. Consistent with this, mutation and oligonucleotide competition studies show that GXXXXG motifs are necessary for either strand of the S box to bind protein/DNA complexes containing SSBP-1 in FRTL-5 cell extracts or to bind to recombinant SSBP-1. They also suggest that the SSBP-1-binding sites on either strand of the HLA-DR alpha S box are functionally distinct. We conclude from these data that the positive regulatory action of SSBP-1 on class II gene expression involves GXXXXG motifs on each strand of the highly conserved S box of the class II 5'-flanking region. As SSBP-1 is modulated by IFNgamma and is involved in class I and TSH receptor as well as class II gene expression in FRTL-5 cells, the sum of the data supports the hypotheses that common transcription factors regulate all three genes, and their altered activities may contribute to the development of autoimmunity.


[Personalization in the medicine of the future : Opportunities and risks].
Personalized medicine is not a new concept. The renaissance of the term is due to the enormous progress in gene sequencing technology and functional imaging, as well as the development of targeted therapies. Application of these technologies in clinical medicine will necessitate infrastructural as well as organizational and educational changes in the healthcare system. An important change required already in the short-term is the introduction of centralized structures, preferably in university clinics, which adopt these innovations and incorporate them into clinical care. Simultaneously, the collation and use of large quantities of relevant data from highly variable sources must be successfully mastered, in order to pave the way for disruptive technologies such as artificial intelligence.


Vitamin-mediated regulation of intestinal immunity.
The intestine is exposed continuously to complex environments created by numerous injurious and beneficial non-self antigens. The unique mucosal immune system in the intestine maintains the immunologic homeostasis between the host and the external environment. Crosstalk between immunocompetent cells and endogenous (e.g., cytokines and chemokines) as well as exogenous factors (e.g., commensal bacteria and dietary materials) achieves the vast diversity of intestinal immune functions. In addition to their vital roles as nutrients, vitamins now also are known to have immunologically crucial functions, specifically in regulating host immune responses. In this review, we focus on the immunologic functions of vitamins in regulating intestinal immune responses and their roles in moderating the fine balance between physiologic and pathologic conditions of the intestine.


Ultrastructure of kidney renal proximal convoluted tubules of experimental animals after Cladribine (2-CdA) administration.
The proximal convoluted tubules of the kidney of white Wistar rats were examined. The animals were given Cladribine (2-CdA) subcutaneously at dosages of 0.07 mg/kg b.m./24 h for 7 days and 0.1 mg/kg b.m./24 h for 6 days in three courses with 5 weeks' break between each. The animals were decapitated 24 hours after the last dose of the drug and 4 weeks after the last dose. The kidney samples were taken for ultrastructural examination. Giving Cladribine at the dose of 0.1 mg/kg b.m./24 h for 7 successive days does not lead to instant changes in the ultrastructure of the proximal convoluted tubules. Changes noticed 4 weeks after Cladribine administration are the following: decrease in amount of mitochondria, the presence of numerous vacuoles, changes in the structure of the brush border, presence of numerous glycogen granules, and the presence of a diluted cytoplasm around the nucleus. Giving 2-CdA at the dose of 0.07 mg/kg b.m./24 h for 6 days in three courses leads to similar changes in proximal convoluted tubules with more extensive damages of the brush border. These were no more intensive after 4 weeks' break in Cladribine administration.


Mucopolysaccharidoses - Clinical Spectrum and Frequency of Different Types.
To determine the relative frequency and clinical features of different varieties of mucopolysaccharidosis. Descriptive study. Department of Pediatric Gastroenterology, Hepatology and Nutrition, The Children's Hospital and The Institute of Child Health, Lahore, from January 2013 to December 2015. All patients who had any feature suggestive of mucopolysaccharidosis were screened with detailed history, clinical examination and skeletal survey. Urine samples for glycosaminoglycan (GAGs) levels and dried blood samples for enzyme analysis were sent. Patients who were confirmed to be suffering from mucopolysaccharidosis were included in the study. The data was analysed using SSPS version 20. A total of 90 confirmed MPS cases, 52 males and 38 females, median age 42 months, were included. Hurler/Hurler-Scheie syndrome was the most frequent (75, 83.33%) followed by Morquio (6, 6.67%), Sanfilippo (5, 5.56%), Maroteaux-Lamy (3, 3.33%) and Hunter (1, 1.11%) syndromes. Consanguinity was present in 79 (87.78%) cases. Common features were hepatomegaly (80, 88.89%), coarse facies (70, 77.78%), splenomegaly (67, 74.44%), and bone disease (48, 53.33%). Most common variety of mucopolysaccharidosis was Hurler/Hurler Scheie followed by Morquio syndrome. Most of the patients were born to consanguineous parents. Common clinical features were coarse facies, hepatosplenomegaly and dysostosis multiplex.


Lactational performance of cows fed low or high ruminally undegradable protein prepartum and supplemental methionine and lysine postpartum.
Multiparous Holstein cows (n = 24) were fed diets containing 34 or 41% ruminally undegradable protein (RUP) for 30 d before parturition; then each group was fed a basal diet supplemented with or without ruminally stable Met (10.6 g/d) and Lys (15.2 g/d) for 75 d in the subsequent lactation. Supplementation of Met and Lys increased the milk yield of cows previously fed the low RUP diet, but milk yields before and after amino acid (AA) supplementation were similar for cows previously fed the high RUP diet. Milk protein content (percentage) increased from 2.83 to 2.96 for cows previously fed the high RUP diet. Milk protein yield increased from 1.13 to 1.21 kg/d when Met and Lys were fed. Data on AA concentration in plasma and AA extraction by the mammary gland suggest that the supplementation of Met and Lys corrected a Met limitation. According to the Cornell Net Carbohydrate and Protein System, the lactation diet was limiting for Met for maximum milk yield and was corrected by AA supplementation. Based on limiting AA, allowable milk yield was 42.5 kg/d, and the observed yield was 40.9 kg/d averaged across treatments. The group with the greatest allowable milk yield (45.2 kg/d) had the greatest actual milk yield (43.0 kg/d). The regression equation of observed milk yield on allowable milk yield was Y = 3.4 + 0.8805X.


Heating Induced by Therapeutic Ultrasound in the Presence of Magnetic Nanoparticles.
The efficiency of ultrasound hyperthermia for anti-cancer treatments such as radiotherapy or chemotherapy can be improved by using sonosensitizers, which are materials that enhance the attenuation and dissipation of acoustic energy. We propose the use of magnetic nanoparticles as sonosensitizers because of their biocompatibility, nontoxicity, and common use in several medical applications. A magnetic material was synthetized and then incorporated in the form of a magnetic fluid in agar tissue-mimicking phantoms. Ultrasound hyperthermia studies were conducted at various ultrasound frequencies and concentrations of magnetic nanoparticles in the phantoms. The theoretical modeling based on a heat transfer equation and the experimental results show good agreement and confirm that the temperature rise during ultrasound heating in tissue-mimicking phantoms doped with sonosensitizers is greater than that in a pure agar phantom. Furthermore, on the basis of Pennes' bio-heat equation, which takes into consideration the blood perfusion and metabolic heat, the thermal dose and lesion shapes after sonication were determined for a hypothetical tissue.


[Cystic tumors of the pancreas].
Cystic tumors of the pancreas are diagnosed with increasing frequency. The main problems associated with their diagnosis are to determine if they are benign or malignant and to decide their surgical excision or clinical follow-up, since these tumors can be benign, potentially malignant or malignant. Imaging techniques such as CT scan, magnetic resonance and endoscopic ultrasonography are the key procedures that must be performed before adopting a final therapeutic decision. However, 10% of cystic lesions are unable to be categorized even after thorough imaging evaluation. We analyze the characteristics of the more common cystic pancreatic lesions and the means to recognize them.


Gallic acid increases atrial natriuretic peptide secretion and mechanical dynamics through activation of PKC.
Gallic acid (GA) protects against myocardial ischemia-reperfusion (I/R) injury, prevents cardiac hypertrophy and fibrosis, and has anti-inflammatory activity in the heart. However, its effects in regulating atrial natriuretic peptide (ANP) secretion are unknown. The aim of this study was to determine the function of GA in regulating ANP secretion and atrial dynamics in rat atria. GA (0.01, 0.05, and 0.1μmol/L) significantly increased atrial ANP secretion and induced positive inotropy dose-dependently. GA (0.1μmol/L) also increased plasma level of ANP and hemodynamics in rats. These effects were accompanied by upregulation of atrial protein kinase C subtypes β and ε (PKCβ and PKCε), which was completely blocked by LY333531 and EAVSLKPT, antagonists of protein PKCβ and PKCε, respectively. GA-induced ANP secretion was also attenuated by Gö6983 but not rottlerin, antagonists of PKCα and PKCδ, and the positive inotropy was reversed by Gö6983. U-73122, a phospholipase C (PLC) antagonist, mitigated the effects of GA on ANP secretion and mechanical dynamics and downregulated Phospho-PLCβ at Ser537 (pPLCβ S537), Phospho-PLCβ at Ser1105 (pPLCβ S1105), PKCβ and PKCε levels, whereas KN62, an inhibitor of Ca2+/calmodulin-dependent kinase II, was not modified the GA-induced ANP secretion and suppressed GA-induced mechanical dynamics. GA promotes ANP secretion and effects positive inotropy with regard to mechanical dynamics through the activation of PLC-PKC signaling in rat atria.


pKa determination of verapamil by liquid-liquid partition.
The pKa of verapamil was determined by measuring the partition coefficient of verapamil between n-heptane and aqueous buffer solution at various pH values. The magnitude of the effects of ionic strength or temperature on the pKa of verapamil was in agreement with those reported previously. The estimated pKa of verapamil in human plasma was 8.75. The pKa of 2,4,6-trimethylpyridine measured by means of the present partition method was in good agreement with that determined by UV spectrophotometry.


Evaluation of mutagenicity and antimutagenicity of cashew stem bark methanolic extract in vitro.
Plants remain to be a great source of medicines for humanity. Among them is the Anacardium occidentale L. species, popularly known as the cashew, which has several therapeutic indications, such as cicatrizing, antihypertensive, hypoglycemic and antitumoral properties. The objective of the present work was to assess the mutagenicity and antimutagenicity of cashew stem bark methanolic extract (CSBME) on cell cultures of Chinese hamster lung fibroblasts (V79). The cultures were treated with different concentrations of the CSBME (500, 1000 and 2000 microg/mL) or with the extract associated with doxorubicin (DXR, 0.75 microg/mL) during the cell cycle phases G1, S, and G2 and throughout continuous treatment. Apart from these treatments, the cell cultures were also treated with PBS (negative-control) and DXR (positive-control). The data obtained in the chromosome aberrations (CA) test showed a significant reduction in CA frequency in the cultures treated with DXR and extract in comparison with those that received only DXR during the cell cycle phases G1 and S and throughout the entire cycle, as well as the absence of mutagenicity in all the treatments realized. The antimutagenic effect observed in this work reinforces the presence of the previously described therapeutic properties of cashew and indicates the safe use of this extract.


Long non-coding RNA CASC2 in human cancer.
Long non-coding RNAs cover large part of the non-coding information of the human DNA, which represents more than 90% of the whole genome. They constitute a wide and complex group of molecules with more than 200 nucleotides, which generally lack an open reading frame, and are involved in various ways in the pathophysiology of cancer. Their roles in the regulation of gene expression, imprinting, transcription, and post-translational processing have been described in several types of cancer. CASC2 was discovered in 2004 in patients with endometrial carcinoma as a potential tumor suppressor. Since then, additional studies in other types of neoplasia have been carried out, and both mechanisms and interactions of CASC2 in cancer have been better elucidated. In this review, we summarize the current knowledge on the role of CASC2 in the genesis, progression, and clinical management of human cancer.


Systemic sporotrichosis with osteomyelitis.
We report a patient with systemic sporotrichosis and osteomyelitis without concomitant suppurative arthritis. To our knowledge this is the first such documented occurrence of hematogenous osteomyelitis in the systemic form of the disease. It was diagnosed by radionuclide bone scintigraphy and radiography and confirmed by biopsy.


[Temporal arteritis].
Temporal arteritis or giant cell arteritis occurs most commonly in elderly individuals. The lesion is usually restricted to the temporal arteries, but rarely, those elsewhere in the body may be involved. The symptoms include a pulsatile headache, usually in the temporal regions, together with anorexia, fever, jaw claudication and muscle pain, known as polymyalgia rheumatica. Histopathological study shows a granulomatous inflammatory lesion with mono-nuclear cell infiltration, associated with Langhans type giant cells, involving mainly the tunica media. Temporal arteritis is not considered to be a life-threatening disorder, however, visual disturbance, the most serious complication, may appear in those with affection of the ophthalmic artery, resulting in blindness in approximately 26% of the untreated cases. Therefore, early diagnosis by temporal artery biopsy and immediate steroid administration should be the keynote of successful therapy for preventing such a critical complication and for relief of the symptoms.


Growth of the genetically engineered strain Cupriavidus necator RW112 with chlorobenzoates and technical chlorobiphenyls.
Cupriavidus necator (formerly Ralstonia eutropha) strain H850 is known to grow on biphenyl, and to co-oxidize congeners of polychlorinated biphenyls (PCBs). Using a Tn5-based minitransposon shuttle system and the TOL plasmid, the rational construction of hybrids of H850 was achieved by subsequent introduction of three distinct elements carrying 11 catabolic loci from three other biodegrading bacteria into the parent strain, finally yielding C. necator RW112. The new genetic elements introduced into H850 and its derivatives were tcbRCDEF, which encode the catabolic enzymes needed for chlorocatechol biodegradation under the control of a transcriptional regulator, followed by cbdABC, encoding a 2-halobenzoate dioxygenase, and xylXYZ, encoding a broad-spectrum toluate dioxygenase. The expression of the introduced genes was demonstrated by measuring the corresponding enzymic activities. The engineered strain RW112 gained the ability to grow on all isomeric monochlorobenzoates and 3,5-dichlorobenzoate, all monochlorobiphenyls, and 3,5-dichloro-, 2,3'-dichloro- and 2,4'-dichlorobiphenyl, without accumulation of chlorobenzoates. It also grew and utilized two commercial PCB formulations, Aroclor 1221 and Aroclor 1232, as sole carbon and energy sources for growth. This is the first report on the aerobic growth of a genetically improved bacterial strain at the expense of technical Aroclor mixtures.


Identification of enzymes acting on alpha-glycated amino acids in Bacillus subtilis.
We have characterized the Bacillus subtilis homologs of fructoselysine 6-kinase and fructoselysine-6-phosphate deglycase, two enzymes that specifically metabolize the Amadori compound fructose-epsilon-lysine in Escherichia coli. The B. subtilis enzymes also catalyzed the phosphorylation of fructosamines to fructosamine 6-phosphates (YurL) and the conversion of the latter to glucose 6-phosphate and a free amino acid (YurP). However, their specificity was totally different from that of the E. coli enzymes, since they acted on fructoseglycine, fructosevaline (YurL) or their 6-phosphoderivatives (YurP) with more than 30-fold higher catalytic efficiencies than on fructose-alpha-lysine (6-phosphate). These enzymes are therefore involved in the metabolism of alpha-glycated amino acids.


Targeting ETS1 with RNAi-based supramolecular nanoassemblies for multidrug-resistant breast cancer therapy.
Overexpression of erythroblastosis virus E26 oncogene homolog 1 (ETS1) gene is correlated with both tumor progression and poor response to chemotherapy in cancer treatment, and the exploitation of RNA interference (RNAi) technology to downregulate ETS1 seems to be a promising approach to reverse multidrug-resistant cancer cells to chemotherapy. Hence, the RNAi-based nanomedicine which is able to simultaneously downregulate ETS1 expression and to deliver chemotherapeutic agents may improve multidrug-resistant cancer therapy synergistically. In this study, we developed a supramolecular nanoassembly that could deliver siRNA targeting ETS1 (siETS1) and doxorubicin (DOX) as an effective nanomedicine to achieve successful chemotherapy towards multidrug-resistant breast cancer. The nanotherapeutic system was prepared by loading adamantane-conjugated doxorubicin (AD) into polyethyleneimine-modified (2-hydroxypropyl)-γ-cyclodextrin (HP) through the supramolecular assembly to form AD-loaded HP (HPAD), followed by electrostatically-driven self-assembly between siETS1 and HPAD. When the HPAD/siETS1 nanoassemblies were delivered into drug-resistant MCF-7/ADR cells, the drug efflux was significantly reduced as a result of simultaneous silencing of ETS1 and MDR1 genes. Importantly, the HPAD/siETS1 nanoassembly could enhance drug residence time at tumor site, and effectively inhibit drug-resistant tumor growth due to the inhibition of angiogenesis and necrosis in tumor tissues. Western blot analysis indicated that the gene expression of both ETS1 and MDR1 in vivo was considerably downregulated after the drug-resistant tumor-bearing mouse was treated with HPAD/siETS1 nanoassemblies. This study offers a new therapeutic delivery strategy targeting ETS1 for the effective multidrug-resistant chemotherapy.


First evidence of the presence of microtriches in the Gyrocotylidea.
The Gyrocotylidea, a small and enigmatic group of intestinal parasites of chimaeras, has been considered to be related either to the Monogenea, or, more frequently, to the most primitive monozoic tapeworms (Cestoda), i.e., the Amphilinidea and Caryophyllidea. The present study, based on transmission electron microscopical observations of a species of Gyrocotyle from the rabbit fish, Chimaera monstrosa, in the North Atlantic, demonstrates for the first time the presence of microtriches as surface structures of gyrocotylideans. Because microtriches are considered to be an autapomorphy of tapeworms (Cestoda), in which they differ from other Neodermata (Monogenea and Trematoda), the present data represent another source of evidence in support of a close relationship between the gyrocotylideans and the tapeworms sensu stricto (Eucestoda). Simple morphology, small size, and shape uniformity of the microtriches of Gyrocotyle sp. may indicate they represent an original (plesiomorphic) form that then evolved in more derived cestode groups into a variety of types present mainly on the scolex. The microtriches of Gyrocotyle sp. resemble those found in caryophyllidean, spathebothriidean, pseudophyllidean, and trypanorhynch cestodes, which are considered to represent the most basal groups of the Eucestoda.


Breathlessness limiting exertion in very old adults: findings from the Newcastle 85+ study.
Long-term breathlessness is more common with age. However, in the oldest old (>85 years), little is known about the prevalence, or impact of breathlessness. We estimated breathlessness limiting exertion prevalence and explored (i) associated characteristics; and (ii) whether breathlessness limiting exertion explains clinical and social/functional outcomes. Health and socio-demographic characteristics were extracted from the Newcastle 85+ Study cohort. Phase 1 (baseline) and follow-up data (18 months, Phase 2; 36 months, Phase 3; 60 months, Phase 4 after baseline) were examined using descriptive statistics and cross-sectional regression models. Eight hundred seventeen participants provided baseline breathlessness data (38.2% men; mean 84.5 years; SD 0.4). The proportions with any limitation of exertion, or severe limitation by breathlessness were 23% (95% confidence intervals (CIs) 20-25%) and 9% (95%CIs 7-11%) at baseline; 20% (16-25%) and 5% (3-8%) at Phase 4. Having more co-morbidities (odds ratio (OR) 1.34, 1.18-1.54; P < 0.001), or self-reported respiratory (OR 1.88, 1.25-2.82; P = 0.003) or cardiovascular disease (OR 2.38, 1.58-3.58; P < 0.001) were associated with breathlessness limiting exertion. Breathlessness severely limiting exertion was associated with poorer self-rated health (OR 0.50, 029-0.86; P = 0.012), depression (beta-coefficient 0.11, P = 0.001), increased primary care contacts (beta-co-efficient 0.13, P = 0.001) and number of nights in hospital (OR 1.81; 1.02-3.20; P = 0.042). Breathlessness limiting exertion appears to become less prevalent over time due to death or withdrawal of participants with cardio-respiratory illness. Breathlessness severely limiting exertion had a wide range of service utilisation and wellbeing impacts.