Differential expression of genes in uninfected and rickettsia-infected Dermacentor variabilis ticks as assessed by differential-display PCR.
Ticks serve as both the vector and the reservoir for members of the spotted fever group rickettsiae. The molecular interaction(s) that results from this close relationship is largely unknown. To identify genetic factors associated with the tick response to rickettsial infection, we utilized differential-display PCR. The majority of upregulation appeared in the infected tissue. We cloned and sequenced 54 differentially expressed transcripts and compared the sequences to those in the GenBank database. Nine of the 54 clones were assigned putative identities and included a clathrin-coated vesicle ATPase, peroxisomal farnesylated protein, Ena/vasodilator-stimulated phosphoprotein-like protein, alpha-catenin, tubulin alpha-chain, copper-transporting ATPase, salivary gland protein SGS-3 precursor, glycine-rich protein, and Dreg-2 protein. Confirmation of the rickettsial influence on the differential expression in the ovaries for a number of these clones was demonstrated by semiquantitative reverse transcription-PCR and Northern blot analyses, resulting in confirmation of six out of nine and three out of four assessed clones, respectively. Further characterization of the clones identified tissue-dependent expression in the midguts and salivary glands. The potential roles of these molecules in the maintenance and transmission of rickettsiae are discussed.


Sufficient blood, safe blood: can we have both?
The decision in September 2011 in the UK to accept blood donations from non-practicing men who have sex with men (MSM) has received significant public attention. Will this rule change substantially boost the number of blood donations or will it make our blood less safe? Clearly, most European countries have a blood procurement problem. Fewer young people are donating, while the population is aging and more invasive therapies are requiring more blood. Yet if that was the reason for allowing non-practicing MSM to donate, clearly re-admission of some other, much larger populations that are currently deferred from donation should likewise be considered. As far as risks for blood safety are concerned, evidence has been provided that the current quality of infectious disease marker testing significantly mitigates against, although does not completely eradicate, risks associated with admission of donors with a high risk of carrying certain blood-transmissible agents. However, it could be argued that more effective recruitment of the non-donor pool, which is substantially larger than the group of currently ineligible donors, would be a better strategy. Recruitment of this group will benefit the availability of blood without jeopardizing the current excellent safety profile of blood.


Analysis of a novel field dilution method for testing samples that exceed the analytic range of point-of-care blood lead analyzers.
Investigators developed and evaluated a dilution method for the LeadCare II analyzer (LCII) for blood lead levels >65 μg/dL, the analyzer's maximum reporting value. Venous blood samples from lead-poisoned children were initially analyzed in the field using the dilution method. Split samples were analyzed at the US Centers for Disease Control and Prevention (CDC) laboratory using both the dilution method and inductively coupled plasma-mass spectrometry (ICP-MS). The concordance correlation coefficient of CDC LCII vs. ICP-MS values (N = 211) was 0.976 (95 % confidence interval (CI) 0.970-0.981); of Field LCII vs. ICP-MS (N = 68) was 0.910 (95% CI 0.861-0.942), and CDC LCII vs. Field LCII (N = 53) was 0.721 (95% CI 0.565-0.827). Sixty percent of CDC and 54% of Field LCII values were within ±10% of the ICP-MS value. Results from the dilution method approximated ICP-MS values and were useful for field-based decision-making. Specific recommendations for additional evaluation are provided.


Assessment of competitiveness of rhizobia infecting Galega orientalis on the basis of plant yield, nodulation, and strain identification by antibiotic resistance and PCR.
Competition between effective and ineffective Rhizobium galegae strains nodulating Galega orientalis was examined on the basis of plant growth, nodulation, antibiotic resistance, and PCR results. In a preliminary experiment in Leonard's jars, ineffective R. galegae strains HAMBI 1207 and HAMBI 1209 competed in similar manners with the effective strain R. galegae HAMBI 1174. In a pot experiment, soil was inoculated with 0 to 10(5) HAMBI 1207 cells per g before G. orientalis was sown. Seeds of G. orientalis were surface inoculated with 2 x 10(4) and 2 x 10(5) cells of HAMBI 1174 per seed (which represent half and fivefold the commercially recommended amount of inoculant, respectively). Plant yield and nodulation by the effective strain were significantly reduced, with as few as 10(2) ineffective rhizobia per g of soil, and the inoculation response was not improved by the 10-fold greater dose of the inoculant. Bacteria occupying the nodules were identified by antibiotic resistance and PCR with primers specific for R. galegae HAMBI 1174, R. galegae, and genes coding for bacterial 16S rRNA (bacterial 16S rDNA). Sixty-two large nodules examined were occupied by the effective strain HAMBI 1174, as proven by antibiotic resistance and amplification of the strain-specific fragment. From 20 small nodules, only the species-specific fragment could be amplified, and isolated bacteria had the same antibiotic resistance and 16S PCR restriction pattern as strain HAMBI 1207. PCR with our strain-specific and species-specific primers provides a powerful tool for strain identification of R. galegae directly from nodules without genetic modification of the bacteria.


Successful marrow transplantation for acute myelocytic leukemia following therapy for Hodgkin's disease.
Five patients with acute myelocytic leukemia (AML) after combined modality therapy for Hodgkin's disease (HD) were treated with cyclophosphamide and busulfan followed by bone marrow transplantation (BMT). Four patients received allogeneic transplants from histocompatibility locus antigen (HLA)-compatible siblings and the fifth patient received an autologous marrow treated with 4-hydroperoxycyclophosphamide. Two patients died of complications of acute graft-v-host disease (GVHD) despite prophylaxis with either low-dose cyclophosphamide or cyclosporine. The remaining three patients were alive and disease-free 382, 617, and 620 days after transplant. These initial results are encouraging and more patients with treatment-related AML need to be evaluated with both allogeneic and autologous BMT to fully elucidate the potentially curative role of this intensive therapy in an otherwise fatal hematologic malignancy.


Iron Pentapyridyl Complexes as Molecular Water Oxidation Catalysts: Strong Influence of a Chloride Ligand and pH in Altering the Mechanism.
The development of molecular water oxidation catalysts based on earth-abundant, non-noble metals is essential for artificial photosynthesis research. Iron, which is the most abundant transition metal in the earth's crust, is a prospective candidate for this purpose. Herein, we report two iron complexes based on the polypyridyl ligand Py5OH (Py5OH=pyridine-2,6-diylbis [di(pyridin-2-yl)methanol]) that can catalyse water oxidation to produce O2 in Ru(III) -induced (at pH 8, highest turnover number (TON)=26.5; turnover frequency (TOF)=2.2 s(-1) ), Ce(IV) -induced (at pH≈1.5 highest TON=16; TOF=0.75 s(-1) ) and photo-induced (at pH 8, highest TON=43.5; TOF=0.6 s(-1) ) reactions. A chloride ligand in one of the iron complexes is shown to affect the activity strongly, improve stability and, thereby, the performance at pH 8 but it inhibits oxygen evolution at pH≈1.5. The observations are consistent with a change in mechanism for catalytic water oxidation with the Fe(Py5OH) complexes between acidic (Ce(IV) ) and near-neutral pH (Ru(III) ).


Intestinal Barrier Function in Gluten-Related Disorders.
Gluten-related disorders include distinct disease entities, namely celiac disease, wheat-associated allergy and non-celiac gluten/wheat sensitivity. Despite having in common the contact of the gastrointestinal mucosa with components of wheat and other cereals as a causative factor, these clinical entities have distinct pathophysiological pathways. In celiac disease, a T-cell mediate immune reaction triggered by gluten ingestion is central in the pathogenesis of the enteropathy, while wheat allergy develops as a rapid immunoglobulin E- or non-immunoglobulin E-mediated immune response. In non-celiac wheat sensitivity, classical adaptive immune responses are not involved. Instead, recent research has revealed that an innate immune response to a yet-to-be-defined antigen, as well as the gut microbiota, are pivotal in the development in this disorder. Although impairment of the epithelial barrier has been described in all three clinical conditions, its role as a potential pathogenetic co-factor, specifically in celiac disease and non-celiac wheat sensitivity, is still a matter of investigation. This article gives a short overview of the mucosal barrier of the small intestine, summarizes the aspects of barrier dysfunction observed in all three gluten-related disorders and reviews literature data in favor of a primary involvement of the epithelial barrier in the development of celiac disease and non-celiac wheat sensitivity.


Effect of light-load resistance exercise on postprandial amino acid transporter expression in elderly men.
An impaired amino acid sensing is associated with age-related loss of skeletal muscle mass. We tested whether light-load resistance exercise (LL-RE) affects postprandial amino acid transporter (AAT) expression in aging skeletal muscle. Untrained, healthy men (age: +65 years) were subjected to 13 h of supine rest. After 2 1/2 h of rest, unilateral LL-RE was conducted (leg extensions, 10 sets of 36 repetitions) at 16% 1RM Thereafter, the subjects were randomized into groups that orally ingested 40 g of whey protein either as hourly drinks (4 g per drink) (PULSE, N = 10) or two boluses (28 g at 0 h and 12 g at 7 h) (BOLUS, N = 10), or hourly isocaloric maltodextrin drinks (placebo, N = 10). Quadriceps muscle biopsies were taken at 0, 3, 7, and 10 h postexercise from both the resting and exercised leg, from which the membrane protein and mRNA expression of select AATs were analyzed by Western Blot and RT-PCR, respectively. LAT1 and PAT1 protein expression increased in response to LL-RE in the PULSE group, and SNAT2 and PAT1 protein expression increased in the BOLUS group when plasma BCAA concentration was low. In all three groups, LL-RE increased LAT1 mRNA expression, whereas a time course decrease in SNAT2 mRNA expression was observed. LL-RE increased membrane-associated AAT protein expression and mRNA expression. Altered AAT protein expression was only seen in groups that ingested whey protein, with the greatest effect observed after hourly feeding. This points toward an importance of AATs in the anabolic response following LL-RE and protein intake.


VC dimensions of group convolutional neural networks.
We study the generalization capacity of group convolutional neural networks. We identify precise estimates for the VC dimensions of simple sets of group convolutional neural networks. In particular, we find that for infinite groups and appropriately chosen convolutional kernels, already two-parameter families of convolutional neural networks have an infinite VC dimension, despite being invariant to the action of an infinite group.


Complement-mediated killing of Borrelia burgdorferi by nonimmune sera from sika deer.
Various species of cervid deer are the preferred hosts for adult, black-legged ticks (Ixodes scapularis and Ixodes pacificus) in the United States. Although frequently exposed to the agent of Lyme disease (Borrelia burgdorferi), these animals, for the most part, are incompetent as transmission reservoirs. We examined the borreliacidal activity of normal and B. burgdorferi-immune sera from sika deer (Cervus nippon) maintained in a laboratory setting and compared it to that of similar sera from reservoir-competent mice and rabbits. All normal deer sera (NDS) tested killed > 90% of B. burgdorferi cells. In contrast, normal mouse and rabbit sera killed < or = 22% of the Borrelia. Anti-B. burgdorferi antibodies could not be detected in any normal sera by indirect fluorescent antibody assay (IFA). Sera collected from deer 6 wk after exposure to B. burgdorferi by tick feeding exhibited IFA titers of 1:256, whereas sera from mice and rabbits similarly exposed had titers of > 1:1,024. Heat treatment (56 C, 30 min) of NDS reduced borreliacidal activity, with < 20% of the B. burgdorferi cells killed, suggesting complement-mediated killing. The chelators EGTA and EDTA were used to block the classical or both the classical and alternative complement pathways, respectively. Addition of 10 mM EGTA to NDS had a negligible effect on borreliacidal activity, with > 90% of the cells killed. Addition of 10 mM EDTA reduced the killing to approximately 30%, whereas the addition of Mg2+ (10 mM) restored borreliacidal activity to NDS. The addition of zymosan A, an activator of the alternative pathway, increased the survival of B. burgdorferi cells to approximately 80% in NDS. These data suggest that the alternative complement activation pathway plays a major role in the borreliacidal activity of NDS. Additionally, 10 mM EGTA had almost no effect on the killing activity of B. burgdorferi-exposed deer sera, suggesting that the classical pathway is not involved in Borrelia killing, even in sera from B. burgdorferi-exposed deer.


Cardiac Complications of Cancer Therapy: Pathophysiology, Identification, Prevention, Treatment, and Future Directions.
Cardiotoxicity is an important complication of cancer therapy. With a significant improvement in the overall survival and prognosis of patients undergoing cancer therapy, cardiovascular toxicity of cancer therapy has become an important public health issue. Several well-established as well as newer anticancer therapies such as anthracyclines, trastuzumab, and other HER2 receptor blockers, antimetabolites, alkylating agents, tyrosine kinase inhibitors, angiogenesis inhibitors, checkpoint inhibitors, and thoracic irradiation are associated with significant cardiotoxicity. Cardiovascular imaging employing radionuclide imaging, echocardiography, and magnetic resonance imaging is helpful in early detection of the cardiotoxicity and prevention of overt heart failure. These techniques also provide important tools for understanding the mechanism of cardiotoxicity of these modalities, which would help develop strategies for the prevention of cardiac morbidity and mortality related to the use of these agents. An understanding of the mechanism of the cardiotoxicity of cancer therapies can help prevent and treat their adverse cardiovascular consequences. Clinical implementation of algorithms based upon cardiac imaging and several non-imaging biomarkers can prevent cardiac morbidity and mortality associated with the use of cardiotoxic cancer therapies.


Novel Xanthomonas Species From the Perennial Ryegrass Seed Microbiome - Assessing the Bioprotection Activity of Non-pathogenic Relatives of Pathogens.
The productivity of the Australian dairy industry is underpinned by pasture grasses, and importantly perennial ryegrass. The performance of these pasture grasses is supported by the fungal endophyte Epichloë spp. that has bioprotection activities, however, the broader microbiome is not well characterized. In this study, we characterized a novel bioprotectant Xanthomonas species isolated from perennial ryegrass (Lolium perenne L. cv. Alto). In vitro and in planta bioassays against key fungal pathogens of grasses (Sclerotium rolfsii, Drechslera brizae and Microdochium nivale) indicated strong bioprotection activities. A complete circular chromosome of ∼5.2 Mb was generated for three strains of the novel Xanthomonas sp. Based on the 16S ribosomal RNA gene, the strains were closely related to the plant pathogen Xanthomonas translucens, however, comparative genomics of 22 closely related xanthomonad strains indicated that these strains were a novel species. The comparative genomics analysis also identified two unique gene clusters associated with the production of bioprotectant secondary metabolites including one associated with a novel nonribosomal peptide synthetase and another with a siderophore. The analysis also identified genes associated with an endophytic lifestyle (e.g., Type VI secretion system), while no genes associated with pathogenicity were identified (e.g., Type III secretion system and effectors). Overall, these results indicate that these strains represent a novel, bioactive, non-pathogenic species of the genus Xanthomonas. Strain GW was the designated type strain of this novel Xanthomonas sp.


Metabolic syndrome and major depression.
Major depression is associated with a 4-fold increased risk for premature death, largely accounted by cardiovascular disease (CVD). The relationship between depression and CVD is thought to be mediated by the so-called metabolic syndrome (MeS). Epidemiological studies have consistently demonstrated a co-occurrence of depression with MeS components, ie, visceral obesity, dyslipidemia, insulin resistance, and hypertension. Although the exact mechanisms linking MeS to depression are unclear, different hypotheses have been put forward. On the one hand, MeS could be the hallmark of the unhealthy lifestyle habits of depressed patients. On the other, MeS and depression might share common alterations of the stress system, including the hypothalamus-pituitary-adrenal (HPA) axis, the autonomic nervous system, the immune system, and platelet and endothelial function. Both the conditions induce a low grade chronic inflammatory state that, in turn, leads to increased oxidative and nitrosative (O&NS) damage of neurons, pancreatic cells, and endothelium. Recently, neurobiological research revealed that peripheral hormones, such as leptin and ghrelin, which are classically involved in homeostatic energy balance, may play a role in mood regulation. Metabolic risk should be routinely assessed in depressed patients and taken into account in therapeutic decisions. Alternative targets should be considered for innovative antidepressant agents, including cytokines and their receptors, intracellular inflammatory mediators, glucocorticoids receptors, O&NS pathways, and peripheral mediators.


A dual-modality probe utilizing intravascular ultrasound and optical coherence tomography for intravascular imaging applications.
We have developed a dual-modality biomedical imaging probe utilizing intravascular ultrasound (IVUS) and optical coherence tomography (OCT). It consists of an OCT probe, a miniature ultrasonic transducer and a fixed mirror. The mirror was mounted at the head of the hybrid probe 45° relative to the light and the ultrasound beams to change their propagation directions. The probe was designed to be able to cover a larger area in blood vessel by IVUS and then visualize a specific point at a much finer image resolution using OCT. To demonstrate both its feasibility and potential clinical applications, we used this ultrasound-guide OCT probe to image a rabbit aorta in vitro. The results offer convincing evidence that the complementary natures of these two modalities may yield beneficial results that could not have otherwise been obtained.


Hidden Markov model-based activity recognition for toddlers.
Physical activity has been shown to impact future health outcomes in adults, but little is known about the long-term impact of physical activity in toddlers. Accurately measuring the specific types and amounts of physical activity in toddlers will help us to understand, predict, and better affect their future health outcomes. Although activity recognition has been extensively developed for adults as well as older children, toddlers move in ways that are significantly different from older children, indicating the need for a more tailored approach. In this study, 22 toddlers wore Actigraph waist-worn accelerometers which recorded their movements during guided play. The toddlers were videotaped and their activities were later annotated for the following eight distinct activity classes: lying down, being carried, riding in a stroller, sitting, standing, running/walking, crawling, and climbing up/down. Accelerometer data were extracted in 2 s signal windows and paired with the activities the toddlers were performing during that time interval. A variety of classifiers were tuned to a validation set. A random forest classifier was found to achieve the highest accuracy of 63.8% in a test set. To improve the accuracy, a hidden Markov model (HMM) was applied by providing the predictions of the static classifiers as observations. The HMM was able to improve the accuracy to 64.8% with all five classifiers increasing the accuracy an average of 1.3% points (95% confidence interval = 0.7-1.9, p < 0.01). When the three most misclassified activities (sitting, standing, and riding in a stroller) were collapsed together, the accuracy increased to 79.3%. Further refinement of the toddler activity recognition classifier will enable more accurate measurements of toddler activity and improve future health outcomes of toddlers.


Ataluren Pharmacokinetics in Healthy Japanese and Caucasian Subjects.
To evaluate the potential for ethnicity-related differences in ataluren pharmacokinetics (PK) and safety, a phase 1 single-dose study was conducted in 48 healthy (24 Japanese and 24 Caucasian subjects), nonsmoking male volunteers who were equally divided into 3 cohorts of oral doses at 5, 10, and 20 mg/kg. Blood samples were collected until 48 hours postdose. PK results demonstrated rapid absorption of ataluren, with peak plasma levels (Cmax ) being attained between 0.875 and 2.5 hours after dosing. The mean Cmax and area under the concentration-time curve (AUC(0-last) ) increased with each increasing dose level in both Japanese and Caucasian subjects. Although the Cmax was similar across all subjects at each dose regardless of ethnicity, Japanese subjects had a mean AUC(0-last) approximately 14% to 34% lower than that of Caucasian subjects across the 3 dose levels. This difference was likely due to the higher variability of AUC values in Caucasian subjects and the relatively small study population. In conclusion, similar ataluren PK profiles were observed in healthy Japanese and Caucasian subjects following single oral administration of ataluren at all dose levels.


[Problems of nephroblastoma in children].
An evaluation of 388 cases of Wilms tumor within 13 years established a high proportion of advanced cases among children: stage II - 30.6; III - 35.4; IV - 27.6 and V (bilateral) - 6.4%. Main homeostatic indices remained unchanged until stages III-IV. Primary tumor and metastases can be detected by a complex of clinical, X-ray and radioisotope methods the effectiveness of which is determined by degree of tumor extension. Dissemination of tumor at different sites were detected by primary examination in 40.5% and at later stages in 15% of patients. Most metastases (96%) were detected within the first 12 months of therapy. They were significantly more frequent at stage III and their frequency was in correlation with the patient's age at the time of tumor detection. Out of 388 cases, 112 (28.9%) survived for more than 3 years, stage II - 57.1; stage III - 25.4 and stage IV - 4.6% included. 1.5 - 2 year-long courses of pre- and postoperative polychemotherapy proved to be effective, whereas in cases of stage III tumor they should be supplemented with irradiation of primary tumor, its bed, regional lymph collectors and metastases. These measures ensured a 3-year survival in 69.5% of stage II patients and in 32.3% of stage III patients.


High intensity exercise decreases global brain glucose uptake in humans.
Physiological activation increases glucose uptake locally in the brain. However, it is not known how high intensity exercise affects regional and global brain glucose uptake. The effect of exercise intensity and exercise capacity on brain glucose uptake was directly measured using positron emission tomography (PET) and [18F]fluoro-deoxy-glucose ([18F]FDG). Fourteen healthy, right-handed men were studied after 35 min of bicycle exercise at exercise intensities corresponding to 30, 55 and 75% of on three separate days. [18F]FDG was injected 10 min after the start of the exercise. Thereafter exercise was continued for another 25 min. PET scanning of the brain was conducted after completion of the exercise. Regional glucose metabolic rate (rGMR) decreased in all measured cortical regions as exercise intensity increased. The mean decrease between the highest and lowest exercise intensity was 32% globally in the brain (38.6+/-4.6 versus 26.1+/-5.0 micromol (100 g)-1 min-1, P<0.001). Lactate availability during exercise tended to correlate negatively with the observed brain glucose uptake. In addition, the decrease in glucose uptake in the dorsal part of the anterior cingulate cortex (37% versus 20%, P<0.05 between 30% and 75% of VO2max) was significantly more pronounced in subjects with higher exercise capacity. These results demonstrate that brain glucose uptake decreases with increase in exercise intensity. Therefore substrates other than glucose, most likely lactate, are utilized by the brain in order to compensate the increased energy needed to maintain neuronal activity during high intensity exercise. Moreover, it seems that exercise training could be related to adaptive metabolic changes locally in the frontal cortical regions.


Selective autoretroperfusion preserves myocardial function during coronary artery ligation in swine.
External pumps have been previously used to minimize edema and hemorrhage caused by coronary retroperfusion. The objective of this study was to use a pump-less approach (selective autoretroperfusion, SARP) to preserve myocardial function after acute coronary artery ligation. In five experimental pigs, the LAD artery was ligated distal to the first diagonal and retroperfusion was instituted for three hours from a brachiocephalic artery at 50 mmHg pressure through an adjustable occluder on the cannula. In eight control pigs, the LAD artery was ligated distal to the second diagonal for the same duration with no SARP. ECG showed more prominent S-T segment elevation in the untreated control group despite the more distal ligation. The degree of myocardial contraction was significantly attenuated in the control group but was largely preserved in the SARP treated group. The myocytes were well preserved in the SARP group with no rupture of venous microvessels. Myocyte edema and disruption was observed in the control group with only mild extracellular edema in the SARP treated group. SARP preserved myocardial function with no damage to the myocyte and venules during three hours of acute LAD ligation.


Pleomorphic intimal sarcoma of pulmonary artery. A case report.
A rare case of pleomorphic intimal sarcoma of pulmonary trunk is reported. The patient presented with symptoms of right ventricular out flow tract (RVOT) obstruction. Metastatic deposits were seen in lungs, diaphragm and thyroid. Bronchial mucosal involvement was also seen. The tumour showed multicentric origin and on electron microscopic examination in a particular cell line was seen.


Only lower limb controlled interactive computer gaming enables an effective increase in energy expenditure.
Limited research documents if new and existing interactive computer gaming "exergaming" increase energy expenditure and cardio-respiratory costs comparable to common exercise modalities. To address this, healthy male volunteers (n = 15) completed six by 12-min test elements: PlayStation2 (PS2(hand)), Nintendo Wii boxing, walk at 5.6 km h(-1), cycle at 120 W, playing an adapted lower limb controlled PS2 (PS2(limb)) and run at 9.6 km h(-1). In addition, they played PS2(limb) for 30 min and performed an incremental treadmill test to exhaustion. Data were analysed using repeated measures ANOVA with post hoc Tukey tests, P < 0.05 inferred significance. PS2(limb) increased energy expenditure (EE) and post-exercise blood lactate (BLa) significantly higher (P < 0.001) than PS2(hand), Wii gaming or walking at 5.6 km h(-1) (EE: 30.3 ± 4.9 vs. 4.7 ± 1.1, 22.0 ± 6.1 and 17.9 ± 1.9 kJ h(-1) kg(-1); BLa: 2.4 ± 1.5 vs. 1.0 ± 0.3, 1.8 ± 0.8 and 0.9 ± 0.2 mmol L(-1)), playing the PS2(limb) raised mean EE over six times greater than PS2(hand). Mean fat and carbohydrate oxidation rates during the 9- to 12-min period playing the PS2(limb) were five and ten times greater than PS2(hand) (0.25 ± 0.10 vs. 0.05 ± 0.10, 1.69 ± 0.52 vs. 0.15 ± 0.14 g min(-1), respectively). PS2(limb) met ACSM guidelines for cardiovascular fitness; however, current Wii technology failed. In conclusion, gaming interactive technology must be adapted or designed to include the lower limbs in order to provide a significant exercise stimulus.


Thyroid hormones directly alter human hair follicle functions: anagen prolongation and stimulation of both hair matrix keratinocyte proliferation and hair pigmentation.
Both insufficient and excess levels of thyroid hormones (T3 and T4) can result in altered hair/skin structure and function (e.g. effluvium). However, it is still unclear whether T3 and T4 exert any direct effects on human hair follicles (HFs), and if so, how exactly human HFs respond to T3/T4 stimulation. Our objective was to asses the impact of T3/T4 on human HF in vitro. Human anagen HFs were isolated from skin obtained from females undergoing facelift surgery. HFs from euthyroid females between 40 and 69 yr (average, 56 yr) were cultured and treated with T3/T4. Studying microdissected, organ-cultured normal human scalp HFs, we show here that T4 up-regulates the proliferation of hair matrix keratinocytes, whereas their apoptosis is down-regulated by T3 and T4. T4 also prolongs the duration of the hair growth phase (anagen) in vitro, possibly due to the down-regulation of TGF-beta2, the key anagen-inhibitory growth factor. Because we show here that human HFs transcribe deiodinase genes (D2 and D3), they may be capable of converting T4 to T3. Intrafollicular immunoreactivity for the recognized thyroid hormone-responsive keratins cytokeratin (CK) 6 and CK14 is significantly modulated by T3 and T4 (CK6 is enhanced, CK14 down-regulated). Both T3 and T4 also significantly stimulate intrafollicular melanin synthesis. Thus, we present the first evidence that human HFs are direct targets of thyroid hormones and demonstrate that T3 and/or T4 modulate multiple hair biology parameters, ranging from HF cycling to pigmentation.


Systematic investigation of the hydrothermal syntheses of Pr(III)-PDA (PDA = pyridine-2,6-dicarboxylate anion) metal-organic frameworks.
A series of novel two-dimensional (2D) and three-dimensional (3D) praseodymium coordination polymers, namely, {[Pr3(PDA)4(HPDA)(H2O)8] x 8H2O}n (2), {[Pr2(PDA)3(H2O)3] x H2O}n (3), {[Pr(PDA)(H2O)4] x ClO4}n (4), and { [Pr2(PDA)2(H2O)5SO4] x 2H2O}n (5) (PDA = pyridine-2,6-dicarboxylic anion), was designed and synthesized under hydrothermal conditions. Complexes 1-3 (chainlike polymer, {[Pr(PDA)(HPDA)(H2O)2] x 4H2O}n (1) was also obtained independently by us, although it has been reported recently by Ghosh et al.) were fabricated successfully by simply tuning the Pr/PDA ratio and exhibited various and intriguing topological structures from a 1D chain to a 3D network. While the synthetic strategy of 5 was triggered and further performed only after 1 was structurally characterized. The complexes were characterized by X-ray single-crystal determination, spectroscopic, and variable-temperature magnetic susceptibility analyses. In complex 2 an unusual nanosized square motif as a building block constructed by eight Pr ions was further assembled into a highly ordered 2D grid compound. In complex 3 the decanuclear Pr metal-based structure as a repeat unit interpenetrated to form a novel 3D polymer. Complex 4 was a 3D network polymer fabricated through a hexanuclear Pr ring as a building block, and ClO4- anions as guests were trapped in the cavity. In complex 5 six Pr atoms, two SO4(2-) anions, and carboxylic oxygen bridges constructed an intriguing rectangle structure as a repeat unit in the grid to form a 2D coordination polymer in which the unique bi-bidentate coordination mode of SO4(2-) anion was observed.


Cefadroxil in skin and skin-structure foot infections: a retrospective review.
The efficacy and safety of cefadroxil in eradicating localized skin and skin-structure infections of the foot were investigated in a retrospective chart review of 222 consecutive patients from two private practices seen over a 10-year period. Of the 189 patients for whom follow-up data were available, 187 (99%) received cefadroxil 500 mg twice daily, and 2 patients (1%) received 250 mg twice daily. The duration of therapy was 2 weeks or less in 87% of patients, with a median duration of therapy of 11.4 days (range, 5 to 35 days). Of the 189 clinically evaluable patients, 179 (95%) achieved a favorable clinical response to treatment; of the 57 patients with microbiologic cultures, 54 (95%) experienced a satisfactory bacteriologic response to therapy; no adverse events related to cefadroxil therapy were identified during the review. The overall results from this retrospective study suggest that cefadroxil is an effective agent with a favorable safety profile for the treatment of skin and skin-structure infections of the foot.


Protein Profiling of WERI-RB1 and Etoposide-Resistant WERI-ETOR Reveals New Insights into Topoisomerase Inhibitor Resistance in Retinoblastoma.
Chemotherapy resistance is one of the reasons for eye loss in patients with retinoblastoma (RB). RB chemotherapy resistance has been studied in different cell culture models, such as WERI-RB1. In addition, chemotherapy-resistant RB subclones, such as the etoposide-resistant WERI-ETOR cell line have been established to improve the understanding of chemotherapy resistance in RB. The objective of this study was to characterize cell line models of an etoposide-sensitive WERI-RB1 and its etoposide-resistant subclone, WERI-ETOR, by proteomic analysis. Subsequently, quantitative proteomics data served for correlation analysis with known drug perturbation profiles. Methodically, WERI-RB1 and WERI-ETOR were cultured, and prepared for quantitative mass spectrometry (MS). This was carried out in a data-independent acquisition (DIA) mode. The raw SWATH (sequential window acquisition of all theoretical mass spectra) files were processed using neural networks in a library-free mode along with machine-learning algorithms. Pathway-enrichment analysis was performed using the REACTOME-pathway resource, and correlated to the molecular signature database (MSigDB) hallmark gene set collections for functional annotation. Furthermore, a drug-connectivity analysis using the L1000 database was carried out to associate the mechanism of action (MOA) for different anticancer reagents to WERI-RB1/WERI-ETOR signatures. A total of 4756 proteins were identified across all samples, showing a distinct clustering between the groups. Of these proteins, 64 were significantly altered (q < 0.05 & log2FC |>2|, 22 higher in WERI-ETOR). Pathway analysis revealed the "retinoid metabolism and transport" pathway as an enriched metabolic pathway in WERI-ETOR cells, while the "sphingolipid de novo biosynthesis" pathway was identified in the WERI-RB1 cell line. In addition, this study revealed similar protein signatures of topoisomerase inhibitors in WERI-ETOR cells as well as ATPase inhibitors, acetylcholine receptor antagonists, and vascular endothelial growth factor receptor (VEGFR) inhibitors in the WERI-RB1 cell line. In this study, WERI-RB1 and WERI-ETOR were analyzed as a cell line model for chemotherapy resistance in RB using data-independent MS. Analysis of the global proteome identified activation of "sphingolipid de novo biosynthesis" in WERI-RB1, and revealed future potential treatment options for etoposide resistance in RB.


The human connexin gene family of gap junction proteins: distinct chromosomal locations but similar structures.
Connexins are protein subunits that constitute gap junction channels. Two members of this gene family, connexin43 (Cx43) and connexin32 (Cx32), are abundantly expressed in the heart and liver, respectively. Human genomic DNA analysis revealed the presence of two loci for Cx43: an expressed gene and a processed pseudogene. The expressed gene (GJA1) was mapped to human chromosome 6 and the pseudogene (GJA1P) to chromosome 5. To determine whether Cx32 was linked to Cx43, somatic cell hybrids were analyzed by polymerase chain reaction and hybridization, resulting in the assignment of the gene for Cx32 (GJB1) to the X chromosome at Xp11----q22. Comparison of the structures of connexin genes suggests that members of this multigene family arose from a single precursor, but evolved to distinct chromosomal locations.


Duloxetine in the treatment of chronic pain due to fibromyalgia and diabetic neuropathy.
Duloxetine is a serotonin-norepinephrine reuptake inhibitor approved by the US Food and Drug Administration for the treatment of fibromyalgia and painful diabetic neuropathy at doses of 60 mg daily. Duloxetine has been shown to significantly improve the symptoms of chronic pain associated with these disorders, as measured by the Fibromyalgia Impact Questionnaire, Brief Pain Inventory scores, the Clinical Global Impressions Scale, and other various outcome measures in several placebo-controlled, randomized, double-blind, multicenter studies. Symptom improvement generally began within the first few weeks, and continued for the duration of the study. In addition, the efficacy of duloxetine was found to be due to direct effects on pain symptoms rather than secondary to improvements in depression or anxiety. Adverse events including nausea, constipation, dry mouth, and insomnia, were mild and transient and occurred at relatively low rates. In conclusion, duloxetine, a selective inhibitor for the serotonin and norepinephrine transporters, is efficacious in the treatment of chronic pain associated with fibromyalgia or diabetic neuropathy, and has a predictable tolerability profile, with adverse events generally being mild to moderate.


NPM/ALK mutants resistant to ASP3026 display variable sensitivity to alternative ALK inhibitors but succumb to the novel compound PF-06463922.
ALK is involved in the onset of several tumors. Crizotinib (XalkoriTM), a potent ALK inhibitor, represents the current front-line treatment for ALK+ NSCLC and shows great clinical efficacy. However, resistant disease often develops after initial response. ASP3026 is a novel second-generation ALK inhibitor with activity on crizotinib-resistant ALK-L1196M gatekeeper mutant. As resistance is likely to be a relevant hurdle for any drug, we sought to determine the resistance profile of ASP3026 in the context of NPM/ALK+ ALCL. We selected six ASP3026-resistant cell lines by culturing human ALCL cells in the presence of increasing concentrations of drug. The established resistant cell lines carry several point mutations in the ALK kinase domain (G1128S, C1156F, I1171N/T, F1174I, N1178H, E1210K and C1156F/D1203N were the most frequent) that are shown to confer resistance to ASP3026 in the Ba/F3 cell model. All mutants were profiled for cross-resistance against a panel of clinically relevant inhibitors including ceritinib, alectinib, crizotinib, AP26113 and PF-06463922. Finally, a genetically heterogeneous ASP3026-resistant cell line was exposed to second-line treatment simulations with all inhibitors. The population evolved according to relative sensitivity of its mutant subclones to the various drugs. Compound PF-06463922 did not allow the outgrowth of any resistant clone, at non-toxic doses.


Identification and monitoring of metabolite markers of dry bean consumption in parallel human and mouse studies.
Aim of the study was to identify and monitor metabolite markers of dry bean consumption in parallel human and mouse studies that each had shown chemopreventive effects of dry bean consumption on colorectal neoplasia risk. Using LC/mass spectroscopy ± ESI and GC/mass spectroscopy, serum metabolites of dry beans were measured in 46 men before and after a 4-week dry bean enriched diet (250 g/day) and 12 mice that received a standardized diet containing either 0 or 10% navy bean ethanol extract for 6 weeks; we also investigated fecal metabolites in the mice. The serum metabolites identified in these controlled feeding studies were then investigated in 212 polyp-free participants from the Polyp Prevention Trial who self-reported either increased (≥+31 g/day from baseline), high dry bean intake of ≥42 g/day in year 3 or low, unchanged dry bean consumption of <8 g/day; serum was analyzed from baseline and year 3. Serum pipecolic acid and S-methyl cysteine were elevated after dry bean consumption in human and mouse studies and reflected dry bean consumption in the Polyp Prevention Trial. Serum levels of pipecolic acid and S-methyl cysteine are useful biomarkers of dry bean consumption.


Adjuvant atezolizumab versus observation in muscle-invasive urothelial carcinoma (IMvigor010): a multicentre, open-label, randomised, phase 3 trial.
Despite standard curative-intent treatment with neoadjuvant cisplatin-based chemotherapy, followed by radical surgery in eligible patients, muscle-invasive urothelial carcinoma has a high recurrence rate and no level 1 evidence for adjuvant therapy. We aimed to evaluate atezolizumab as adjuvant therapy in patients with high-risk muscle-invasive urothelial carcinoma. In the IMvigor010 study, a multicentre, open-label, randomised, phase 3 trial done in 192 hospitals, academic centres, and community oncology practices across 24 countries or regions, patients aged 18 years and older with histologically confirmed muscle-invasive urothelial carcinoma and an Eastern Cooperative Oncology Group performance status of 0, 1, or 2 were enrolled within 14 weeks after radical cystectomy or nephroureterectomy with lymph node dissection. Patients had ypT2-4a or ypN+ tumours following neoadjuvant chemotherapy or pT3-4a or pN+ tumours if no neoadjuvant chemotherapy was received. Patients not treated with neoadjuvant chemotherapy must have been ineligible for or declined cisplatin-based adjuvant chemotherapy. No post-surgical radiotherapy or previous adjuvant chemotherapy was allowed. Patients were randomly assigned (1:1) using a permuted block (block size of four) method and interactive voice-web response system to receive 1200 mg atezolizumab given intravenously every 3 weeks for 16 cycles or up to 1 year, whichever occurred first, or to observation. Randomisation was stratified by previous neoadjuvant chemotherapy use, number of lymph nodes resected, pathological nodal status, tumour stage, and PD-L1 expression on tumour-infiltrating immune cells. The primary endpoint was disease-free survival in the intention-to-treat population. Safety was assessed in patients who either received at least one dose of atezolizumab or had at least one post-baseline safety assessment. This trial is registered with ClinicalTrials.gov, NCT02450331, and is ongoing but not recruiting patients. Between Oct 5, 2015, and July 30, 2018, we enrolled 809 patients, of whom 406 were assigned to the atezolizumab group and 403 were assigned to the observation group. Median follow-up was 21·9 months (IQR 13·2-29·8). Median disease-free survival was 19·4 months (95% CI 15·9-24·8) with atezolizumab and 16·6 months (11·2-24·8) with observation (stratified hazard ratio 0·89 [95% CI 0·74-1·08]; p=0·24). The most common grade 3 or 4 adverse events were urinary tract infection (31 [8%] of 390 patients in the atezolizumab group vs 20 [5%] of 397 patients in the observation group), pyelonephritis (12 [3%]) vs 14 [4%]), and anaemia (eight [2%] vs seven [2%]). Serious adverse events occurred in 122 (31%) patients who received atezolizumab and 71 (18%) who underwent observation. 63 (16%) patients who received atezolizumab had a treatment-related grade 3 or 4 adverse event. One treatment-related death, due to acute respiratory distress syndrome, occurred in the atezolizumab group. To our knowledge, IMvigor010 is the largest, first-completed phase 3 adjuvant study to evaluate the role of a checkpoint inhibitor in muscle-invasive urothelial carcinoma. The trial did not meet its primary endpoint of improved disease-free survival in the atezolizumab group over observation. Atezolizumab was generally tolerable, with no new safety signals; however, higher frequencies of adverse events leading to discontinuation were reported than in metastatic urothelial carcinoma studies. These data do not support the use of adjuvant checkpoint inhibitor therapy in the setting evaluated in IMvigor010 at this time. F Hoffmann-La Roche/Genentech.


Measurement of the Nucleon F_{2}^{n}/F_{2}^{p} Structure Function Ratio by the Jefferson Lab MARATHON Tritium/Helium-3 Deep Inelastic Scattering Experiment.
The ratio of the nucleon F_{2} structure functions, F_{2}^{n}/F_{2}^{p}, is determined by the MARATHON experiment from measurements of deep inelastic scattering of electrons from ^{3}H and ^{3}He nuclei. The experiment was performed in the Hall A Facility of Jefferson Lab using two high-resolution spectrometers for electron detection, and a cryogenic target system which included a low-activity tritium cell. The data analysis used a novel technique exploiting the mirror symmetry of the two nuclei, which essentially eliminates many theoretical uncertainties in the extraction of the ratio. The results, which cover the Bjorken scaling variable range 0.19<x<0.83, represent a significant improvement compared to previous SLAC and Jefferson Lab measurements for the ratio. They are compared to recent theoretical calculations and empirical determinations of the F_{2}^{n}/F_{2}^{p} ratio.


T1TAdb: the database of type I toxin-antitoxin systems.
Type I toxin-antitoxin (T1TA) systems constitute a large class of genetic modules with antisense RNA (asRNA)-mediated regulation of gene expression. They are widespread in bacteria and consist of an mRNA coding for a toxic protein and a noncoding asRNA that acts as an antitoxin preventing the synthesis of the toxin by directly base-pairing to its cognate mRNA. The co- and post-transcriptional regulation of T1TA systems is intimately linked to RNA sequence and structure, therefore it is essential to have an accurate annotation of the mRNA and asRNA molecules to understand this regulation. However, most T1TA systems have been identified by means of bioinformatic analyses solely based on the toxin protein sequences, and there is no central repository of information on their specific RNA features. Here we present the first database dedicated to type I TA systems, named T1TAdb. It is an open-access web database (https://d-lab.arna.cnrs.fr/t1tadb) with a collection of ∼1900 loci in ∼500 bacterial strains in which a toxin-coding sequence has been previously identified. RNA molecules were annotated with a bioinformatic procedure based on key determinants of the mRNA structure and the genetic organization of the T1TA loci. Besides RNA and protein secondary structure predictions, T1TAdb also identifies promoter, ribosome-binding, and mRNA-asRNA interaction sites. It also includes tools for comparative analysis, such as sequence similarity search and computation of structural multiple alignments, which are annotated with covariation information. To our knowledge, T1TAdb represents the largest collection of features, sequences, and structural annotations on this class of genetic modules.


Average glandular dose coefficients for pendant-geometry breast CT using realistic breast phantoms.
To design volume-specific breast phantoms from breast CT (bCT) data sets and estimate the associated normalized mean glandular dose coefficients for breast CT using Monte Carlo methods. A large cohort of bCT data sets (N = 215) was used to evaluate breast volume into quintiles (plus the top 5%). The average radius profile was then determined for each of the six volume-specific groups and used to both fabricate physical phantoms and generate mathematical phantoms (V1-V6; "V" denotes classification by volume). The MCNP6 Monte Carlo code was used to model a prototype bCT system fabricated at our institution; and this model was validated against physical measurements in the fabricated phantoms. The mathematical phantoms were used to simulate normalized mean glandular dose coefficients for both monoenergetic source photons "DgNCT (E)" (8-70 keV in 1 keV intervals) and polyenergetic x-ray beams "pDgNCT " (35-70 kV in 1 kV intervals). The Monte Carlo code was used to study the influence of breast size (V1 vs. V5) and glandular fraction (6.4% vs. 45.8%) on glandular dose. The pDgNCT coefficients estimated for the V1, V3, and V5 phantoms were also compared to those generated using simple, cylindrical phantoms with equivalent volume and two geometrical constraints including; (a) cylinder radius determined at the breast phantom chest wall "Rcw "; and (b) cylinder radius determined at the breast phantom center-of-mass "RCOM ". Satisfactory agreement was observed for dose estimations using MCNP6 compared with both physical measurements in the V1, V3, and V5 phantoms (R2 = 0.995) and reference bCT dose coefficients using simple phantoms (R2 = 0.999). For a 49 kV spectrum with 1.5 mm Al filtration, differences in glandular fraction [6.5% (5th percentile) vs. 45.8% (95th percentile)] had a 13.2% influence on pDgNCT for the V3 phantom, and differences in breast size (V1 vs. V5) had a 16.6% influence on pDgNCT for a breast composed of 17% (median) fibroglandular tissue. For cylindrical phantoms with a radius of RCOM , the differences were 1.5%, 0.1%, and 2.1% compared with the V1, V3, and V5 phantoms, respectively. Breast phantoms were designed using a large cohort of bCT data sets across a range of six breast sizes. These phantoms were then fabricated and used for the estimation of glandular dose in breast CT. The mathematical phantoms and associated glandular dose coefficients for a range of breast sizes (V1-V6) and glandular fractions (5th to 95th percentiles) are available for interested users.


Imaging integrin αvβ3 on blood vessels with 111In-RGD2 in head and neck tumor xenografts.
Arginine-glycine-aspartic acid (RGD)-based imaging tracers allow specific imaging of integrin αvβ3, a protein overexpressed during angiogenesis, leading to the possibility that it might serve as a tool to stratify patients for antiangiogenic treatment. However, these tracers have generally been characterized in xenograft models in which integrin αvβ3 was constitutively expressed by the tumor cells themselves. In the studies presented here, the use of (111)In-RGD2 as a tracer to image only integrin αvβ3 expression on blood vessels in the tumor was determined using tumor xenografts in which tumor cells were integrin αvβ3-negative. DOTA-E-[c(RGDfK)]2 was radiolabeled with (111)In ((111)In-RGD2), and biodistribution studies were performed in squamous cell carcinoma of the head and neck (HNSCC) xenograft mouse models to determine the optimal peptide dose to image angiogenesis. Next, biodistribution and imaging studies were performed at the optimal peptide dose in 3 HNSCC mouse models, FaDu, SCCNij3, and SCCNij202. Immunohistochemical analysis of tumor vascular and cell surface expression of integrin αvβ3 and correlation analysis of vascular integrin αvβ3 and autoradiography were completed. All 3 HNSCC xenografts expressed integrin αvβ3 on the vessels only. The optimal peptide dose of (111)In-RGD2 was 1 μg or less for specific integrin αvβ3-mediated uptake of the tracer. SPECT/CT imaging showed clear uptake of the tracer in the periphery of the tumors, corresponding with well-vascularized areas of the tumor. Within the tumor, (111)In-RGD2 autoradiography coincided with vascular integrin αvβ3 expression, as determined immunohistochemically. Integrin αvβ3-mediated uptake was also detected in nontumor tissues, which, through immunohistochemical analysis, proved positive for integrin αvβ3. (111)In-RGD2 allows the visualization of integrin αvβ3 in xenograft models in which integrin αvβ3 is expressed only on the neovasculature, such as in the HNSCC tumors. Thus, (111)In-RGD2 allows specific visualization of angiogenesis in tumor models lacking constitutive tumoral integrin αvβ3 expression but may be less useful for this purpose in many tumors in which tumor cells express integrin αvβ3.


Checkmate: Metabolic flexibility with a STING in its tail.
Inhibiting a key metabolic enzyme, ACLY, in cancer cells impacts T cell function in immunotherapy-resistant tumors and may offer a target for therapeutic treatment.


Drosophila Mechanosensory Transduction.
Mechanosensation in Drosophila relies on sensory neurons transducing mechanical stimuli into ionic currents. The molecular mechanisms of this transduction are in the process of being revealed. Transduction relies on mechanogated ion channels that are activated by membrane stretch or the tension of force-conveying tethers. NOMPC (no-mechanoreceptor potential C) and DmPiezo were put forward as bona fide mechanoelectrical transduction (MET) channels, providing insights into MET channel architecture and the structural basis of mechanogating. Various additional channels were implicated in Drosophila mechanosensory neuron functions, and parallels between fly and vertebrate mechanotransduction were delineated. Collectively, these advances put forward Drosophila mechanosensory neurons as cellular paradigms for mechanotransduction and mechanogated ion channel function in the context of proprio- and nociception as well as the detection of substrate vibrations, touch, gravity, and sound.


Optical coherence tomography evaluation of tissue prolapse after carotid artery stenting using closed cell design stents for unstable plaque.
During carotid artery stenting (CAS) with the use of closed cell design stents for unstable plaques, tissue prolapse between stent struts was evaluated by optical coherence tomography (OCT). 14 carotid stenosis lesions diagnosed as unstable plaques by MRI were evaluated by OCT imaging during CAS using closed cell stents. Cross sectional OCT images within the stented segment were evaluated at 1 mm intervals. The slice rate for the presence of tissue prolapse between the struts was calculated. No intra-procedural complications occurred. After single stent placement, plaque prolapse was observed in all cases. Slices with any and >500 µm tissue prolapse were seen in 30% and 7.8% of cases, respectively. In 5 of 7 lesions with tissue prolapse >500 µm, additional stents were overlapped. In cases with overlapping stents, slices with any tissue prolapse were significantly decreased from 26% to 16% (p=0.008); in particular, the occurrence of tissue prolapse >500 µm was significantly decreased from 15% to 2.3% (p<0.001). In one case of >500 µm tissue prolapse without an overlapping stent, delayed embolization due to an in-stent thrombus occurred 9 months after the procedure. OCT during CAS using closed cell stent for unstable plaques frequently revealed tissue prolapse between struts. Placement of overlapping stents significantly reduced tissue prolapse, particularly tissue prolapse >500 µm. However, closed cell stents used for unstable plaques may not solve the problem of tissue prolapse.


DNA repair mechanisms in mammalian germ cells.
Mammalian germ cells encounter several types of DNA damage. This damage is almost completely repaired in a short period of time to provide the maintenance of genomic integrity. The main repair mechanisms operating in mammalian germline cells are: nucleotide excision repair (NER), base excision repair (BER), mismatch repair (MMR), DNA double strand break repair (DSBR), and post replication repair (PRR). Currently, there are relatively few publications that summarize basic information and new findings on DNA repair mechanisms used in mammalian germ cells. In the present article, we review the studies that discuss repair mechanisms operating in the female and male germ cells. We then survey some of the recent discoveries made in this field.


Isolation of Saccharomyces Cerevisiae from Pineapple and Orange and Study of Metal's Effectiveness on Ethanol Production.
In view of the anticipated shortage of the traditional supplies of fossil fuels, there is a great deal of interest in the production of ethanol as an alternative biofuel in recent years. The main objective of this research work was to isolate and characterize stress tolerant, high potential ethanol producing yeast strains from various fruit peel. Two yeast isolates from pineapple (Pa) and orange (Or) have been isolated, characterized on the basis of morphological and physic-chemical characters and optimized on ethanol producing capability using sugarcane molasses as substrate. Ethanol production percentage was estimated by Conway method. Isolates were thermotolerant, pH tolerant, ethanol tolerant as well as osmotolerant. They were resistant to Chloramphenicol (30 μg/disc) and Nalidixic acid (30 μg/disc). The isolates showed no killer toxin activity against E. coli. The highest production capacity of the yeasts was found to be 7.39% and 5.02% for Pa and Or, respectively, at pH 5.0, 30 °C temperature in media with an initial reducing sugar concentration of 6.5% for Pa and 5.5% for Or (shaking). Addition of metal ions increased the rate of ethanol production highest to 10.61% by KH2PO4. This study revealed that indigenous yeast isolates could be used to benefit the fuel demand and industrial alcohol industries.


RNA Interference Mediated Interleukin-1β Silencing in Inflamed Chondrocytes Decreases Target and Downstream Catabolic Responses.
Posttraumatic activation of the catabolic cascade plays a major role in degradation of cartilage. Interleukin-1β (IL-1β), a primary instigator in the catabolic axis, is upregulated in chondrocytes following injury. IL-1β activates key degradative enzymes, including MMPs and aggrecanases, and other proinflammatory mediators such as PGE2 which contribute to ECM breakdown. Posttranscriptional silencing of IL-1β by RNA interference (RNAi) may drive a reduction in IL-1β. We hypothesized that transduction of chondrocytes using rAAV2 expressing a short hairpin RNAi motif targeting IL-1β (shIL-1β) would significantly decrease IL-1β expression and, in turn, decrease expression of other catabolic enzymes. Chondrocyte cultures were transduced with rAAV2-tdT-shIL-1β in serum-free media. The fluorescent protein, tdTomato, was used to determine transduction efficiency via flow cytometry and fluorescent microscopy. Cells were stimulated with lipopolysaccharide (LPS) 48 hours following transduction. After 24-hour stimulation, supernatants were collected for cytokine analysis, and cells lysed for gene expression analysis. IL-1β knockdown led to significantly decreased expression of IL-1β, TNF-α, and ADAMTS5. PGE2 synthesis was also significantly downregulated. Overall, effective silencing of IL-1β using rAAV2 vector expressing a short hairpin IL-1β knockdown sequence was shown. Additionally, significant downstream effects were evident, including decreased expression of TNF-α and ADAMTS5. Targeted silencing of catabolic cytokines may provide a promising treatment avenue for osteoarthritic (OA) joints.


A COMPARISON OF CHANGE IN 3D SCAPULAR KINEMATICS WITH MAXIMAL CONTRACTIONS AND FORCE PRODUCTION WITH SCAPULAR MUSCLE TESTS BETWEEN ASYMPTOMATIC OVERHEAD ATHLETES WITH AND WITHOUT SCAPULAR DYSKINESIS.
The significance of scapular dyskinesis is being challenged due to a lack of the association with pain and ability to predict injury in athletic populations. However, it is unknown whether asymptomatic overhead athletes with dyskinesis cope by normalizing scapular position with higher demand activities. The purpose of this study was to compare change in scapular kinematics from an active unweighted contraction to a maximal isometric contraction in asymptomatic overhead athletes with and without scapular dyskinesis. Secondarily, force generated with manual muscle tests were explored for differences and relationships with kinematics. Cross-sectional laboratory study. Twenty-five matched asymptomatic overhead athletes with (n=14) and without (n=11) scapular dyskinesis, defined with a reliable and validated clinical method, participated in this study. Three-dimensional scapular kinematics were evaluated in an active unweighted condition, and during maximal isometric contractions at 90 ° of shoulder flexion. Isometric force produced with lower trapezius and serratus anterior manual muscle tests were assessed with a dynamometer. Changes in scapular kinematics were compared between groups. Differences in force generated with manual muscle tests between groups and relationships with kinematics were explored. Athletes with dyskinesis demonstrated greater deficits in scapular upward rotation with maximal contraction (p=<0.001), less external rotation (p=0.036) and weaker lower trapezius manual muscle test strength (p=0.031). Lower trapezius (p=0.003;r=0.57) and serratus anterior (p=0.042;r=0.41) manual muscle test strength deficits were fair to moderately associated with a lack of scapular upward rotation during maximal contraction. Small to moderate changes in scapular kinematics are normal responses to a maximal contraction, but with scapular dyskinesis this response is accentuated. Athletes with dyskinesis generate less force with lower trapezius manual muscle testing compared to athletes without dyskinesis. Decreased strength with lower trapezius and serratus anterior manual muscle testing was also related to a lack of upward rotation in all athletes. 3.


Multianalyte microspot immunoassay--microanalytical "compact disk" of the future.
Throughout the 1970s, controversy centered both on immunoassay "sensitivity" per se and on the relative sensitivities of labeled antibody (Ab) and labeled analyte methods. Our theoretical studies revealed that RIA sensitivities could be surpassed only by the use of very high-specificity nonisotopic labels in "noncompetitive" designs, preferably with monoclonal antibodies. The time-resolved fluorescence methodology known as DELFIA--developed in collaboration with LKB/Wallac--represented the first commercial "ultrasensitive" nonisotopic technique based on these theoretical insights, the same concepts being subsequently adopted in comparable methodologies relying on the use of chemiluminescent and enzyme labels. However, high-specific-activity labels also permit the development of "multianalyte" immunoassay systems combining ultrasensitivity with the simultaneous measurement of tens, hundreds, or thousands of analytes in a small biological sample. This possibility relies on simple, albeit hitherto-unexploited, physicochemical concepts. The first is that all immunoassays rely on the measurement of Ab occupancy by analyte. The second is that, provided the Ab concentration used is "vanishingly small," fractional Ab occupancy is independent of both Ab concentration and sample volume. This leads to the notion of "ratiometric" immunoassay, involving measurement of the ratio of signals (e.g., fluorescent signals) emitted by two labeled Abs, the first (a "sensor" Ab) deposited as a microspot on a solid support, the second (a "developing" Ab) directed against either occupied or unoccupied binding sites of the sensor Ab. Our preliminary studies of this approach have relied on a dual-channel scanning-laser confocal microscope, permitting microspots of area 100 microns 2 or less to be analyzed, and implying that an array of 10(6) Ab-containing microspots, each directed against a different analyte, could, in principle, be accommodated on an area of 1 cm2. Although measurement of such analyte numbers is unlikely ever to be required, the ability to analyze biological fluids for a wide spectrum of analystes is likely to transform immunodiagnostics in the next decade.


Polydopamine and eumelanin molecular structures investigated with ab initio calculations.
A set of computational methods that contains a brute-force algorithmic generation of chemical isomers, molecular dynamics (MD) simulations, and density functional theory (DFT) calculations is reported and applied to investigate nearly 3000 probable molecular structures of polydopamine (PDA) and eumelanin. All probable early-polymerized 5,6-dihydroxyindole (DHI) oligomers, ranging from dimers to tetramers, have been systematically analyzed to find the most stable geometry connections as well as to propose a set of molecular models that represents the chemically diverse nature of PDA and eumelanin. Our results indicate that more planar oligomers have a tendency to be more stable. This finding is in good agreement with recent experimental observations, which suggested that PDA and eumelanin are composed of nearly planar oligomers that appear to be stacked together via π-π interactions to form graphite-like layered aggregates. We also show that there is a group of tetramers notably more stable than the others, implying that even though there is an inherent chemical diversity in PDA and eumelanin, the molecular structures of the majority of the species are quite repetitive. Our results also suggest that larger oligomers are less likely to form. This observation is also consistent with experimental measurements, supporting the existence of small oligomers instead of large polymers as main components of PDA and eumelanin. In summary, this work brings an insight into the controversial structure of PDA and eumelanin, explaining some of the most important structural features, and providing a set of molecular models for more accurate modeling of eumelanin-like materials.


Serum CD109 levels reflect the node metastasis status in head and neck squamous cell carcinoma.
Various biomarkers are being developed for the early diagnosis of cancer and for predicting its prognosis. The aim of this study is to evaluate the diagnostic significance of serum CD109 in head and neck squamous cell carcinoma (HNSCC). The serum CD109 levels in a total of 112 serum samples collected before and after surgery from 56 HNSCC patients were analyzed with an enzyme-linked immunosorbent assay (ELISA). The clinical factor that showed a statistically significant association with both the preoperative serum CD109 level, and the CD109 index: which was defined as the ratio of the preoperative serum CD109 level to the postoperative serum CD109 level, were assessed. The correlations between the serum CD109 levels and lymph node density (LND), pathological features such as lymphatic invasion, and serum SCC antigen levels were also assessed. The ELISA measurement revealed that preoperative serum CD109 levels were elevated in patients with node metastasis-positive and stage IV disease, in comparison to those with node metastasis-negative and Stage I+II+III disease, respectively. A multiple regression analysis indicated that serum CD109 level was significantly associated with the node metastasis status. A Spearman's rank correlation analysis also revealed a positive correlation between the preoperative serum CD109 level and LND. Furthermore, the probabilities of the overall and relapse-free survival were significantly lower in patients with a preoperative serum CD109 level of ≥38.0 ng/ml and a CD109 index of ≥1.6, respectively, than in others. There was no significant correlation between the serum CD109 and SCC antigen levels. The serum CD109 levels were elevated in patients with advanced stage disease, reflecting the node metastasis status. CD109 in sera could be a novel prognostic marker for HNSCC involving lymph node metastasis.


Organ weights and histopathology of double-crested cormorants (Phalacrocorax auritus) dosed orally or dermally with artificially weathered Mississippi Canyon 252 crude oil.
A series of toxicity tests were conducted to assess the effects of low to moderate exposure to artificially weathered Deepwater Horizon Mississippi Canyon 252 crude oil on representative avian species as part of the Natural Resource Damage Assessment. The present report summarizes effects of oral exposure (n=26) of double-crested cormorants (DCCO; Phalacrocorax auritus) to 5 or 10ml oil kg-1 day-1 for up to 21 days or dermal application (n=25) of 13ml oil to breast and back feathers every three days totaling 6 applications in 21 days on organ weights and histopathology. Absolute and relative kidney and liver weights were increased in birds exposed to oil. Additionally, gross and/or histopathologic lesions occurred in the kidney, heart, pancreas and thyroid. Clinically significant renal lesions in the orally dosed birds included squamous metaplasia and increased epithelial hypertrophy of the collecting ducts and renal tubules and mineralization in comparison to controls. Gross cardiac lesions including thin walls and flaccid musculature were documented in both orally and dermally dosed birds and myocardial fibrosis was found in low numbers of dermally dosed birds only. Cytoplasmic vacuolation of the exocrine pancreas was noted in orally dosed birds only. Thyroid follicular hyperplasia was increased in dermally dosed birds only possibly due to increased metabolism required to compensate damaged feather integrity and thermoregulate. Gastrointestinal ulceration was found in orally dosed birds only. There were no significant hepatic histopathologic lesions induced by either exposure route. Therefore, hepatic histopathology is likely not a good representation of oil-induced damage. Taken together, the results suggest that oral or dermal exposure of DCCOs to artificially weathered MC252 crude oil induced organ damage that could potentially affect survivability.


RBE of X rays of different energies: a cytogenetic evaluation by FISH.
Mammography using 26-30 kVp X rays is routinely used in breast cancer screening. Discussion about the radiation-related risk associated with this methodology is ongoing. For radioprotection purposes, a quality factor of 1 has been assigned for all photon energies. However, the relative biological effectiveness (RBE) could increase as the photon energy decreases. Analyzing different biological parameters, for 30 kVp X rays, RBE values from 1 to 8 have been estimated. In the present study, a cytogenetic FISH evaluation of the RBE of 30, 80 and 120 kVp X rays has been done. Blood samples were irradiated with 10 doses from 0.05 to 3 Gy for each energy studied. The yields of translocations and dicentrics were determined by fluorescence in situ hybridization (FISH) using whole chromosome probes for chromosomes 1, 4 and 11 together with a pancentromeric probe. The alpha coefficients of the dose-effect curves for dicentrics, minimum number of breaks needed to produce exchange-type aberrations, and apparently simple translocations were used to estimate the RBE. Using the curves obtained for 120 kVp as a reference, the RBE values for dicentrics were 1.08+/-0.43 and 1.73+/-0.59 for 80 and 30 kVp X rays, respectively; for minimum number of breaks these values were 1.38+/-0.39 and 1.42+/-0.41, and for apparently simple translocations they were 1.26+/-0.40 and 1.51+/-0.47, respectively. Moreover, the induction of complex aberrations by these energies was compared. The percentage of complex aberrations relative to total aberrations showed a significant tendency to increase as X-ray energy decreased: 7.8+/-1.19, 9.8+/-1.6 and 14.1+/-1.9 for 120, 80 and 30 kVp, respectively (P<0.02).


A structural rationale for SV40 Vp1 temperature-sensitive mutants and their complementation.
Two groups of temperature-sensitive (ts) mutants, termed ts B and ts C, have mutations in the major capsid protein of SV40, Vp1. These mutants have virion assembly defects at the nonpermissive temperature, but can complement one another when two mutants, one from each group, coinfect a cell. A third group of mutants, termed ts BC, have related phenotypes, but do not complement other mutants. We found that the mutations fall into two structural and functional classes. All ts C and one ts BC mutations map to the region close to the Ca2+ binding sites, and are predicted to disrupt the insertion of the distal part of the C-terminal invading arm (C-arm) into the receiving clamp. They share a severe defect in assembly at the nonpermissive temperature, with few capsid proteins attached to the viral minichromosome. By contrast, all ts B and most ts BC mutations map to a contiguous region including acceptor sites for the proximal part of the C-arm and intrapentamer contacts. These mutants form assembly intermediates that carry substantial capsid proteins on the minichromosome. Thus, accurate virion assembly is prevented by mutations that disrupt interactions between the receiving pentamer and both the proximal and distal parts of the C-arms, with the latter having a greater effect. The distinct spatial localization and assembly defects of the two classes of mutants provide a rationale for their intracistronic complementation and suggest models of capsid assembly.


Role of testosterone in stimulating seasonal changes in a potential avian chemosignal.
Songbird preen oil contains volatile and semivolatile compounds that may contain information about species, sex, individual identity, and season. We examined the relationship between testosterone (T) and the amounts of preen oil volatile and semivolatile compounds in wild and captive dark-eyed juncos (Junco hyemalis). In wild males and females, we observed an increase in volatile compound relative concentration early in the breeding season. This increase mirrored previously described seasonal elevation in T levels in wild males and females, suggesting a positive relationship between hormone levels and preen gland secretions, and a possible role for these secretions in signaling receptivity. In females, the greatest relative concentrations of most compounds were observed close to egg laying, a time when steroid hormones are high and also the only time that females respond to an injection of gonadotropin-releasing hormone with a short-term increase in T. In a study of captive juncos held on short days, we asked whether the seasonal increases observed in the wild could be induced with experimental elevation of T alone. We found that exogenous T stimulated the production of some volatile compounds in non-breeding individuals of both sexes. However, of the 15 compounds known to increase during the breeding season, only four showed an increase in relative concentration in birds that received T implants. Our results suggest that testosterone levels likely interact with other seasonally induced physiological changes to affect volatile compound amounts in preen oil.


Lipid deposition in human tendon xanthoma.
Tendon xanthomas often accompany the rapidly progressive atherosclerosis that develops in patients with familial hypercholesterolemia. This report describes lipid accumulation, as studied histochemically, in an Achilles tendon xanthoma from a patient whose death was secondary to complications of familial hypercholesterolemia. Lipid was stained with oil red O and filipin dyes for detection of esterified and unesterified cholesterol, respectively. As in human atherosclerosis, unesterified cholesterol accumulated in the tendon predominantly in the extracellular space but separately from oil-red-O-stained lipid that accumulated both intra- and extracellularly. Deposition of unesterified cholesterol in human atherosclerotic lesions and tendon xanthomas is an interesting but as yet unexplained phenomenon.


Acute Kidney Injury Masked by Malnutrition: A Case Report and the Problem of Protein.
Acute kidney injury (AKI) is common among critically ill patients. There are a number of nutrition considerations in the management of AKI, including fluid balance, electrolyte and acid-base disturbances, protein provision, and management of comorbid conditions. The optimal amount of protein provision for patients with AKI who are not on renal replacement therapy (RRT) has been a topic of debate for years. Excessive protein provision may contribute to azotemia in these patients, but inadequate protein intake may harm nutrition status and result in poorer clinical outcomes. This review discusses a patient case of AKI masked by malnutrition and muscle loss and reviews the current literature on optimal protein intake in AKI (not on RRT). Based on a structured search strategy, 4 articles were reviewed. We conclude that the available evidence suggests that significant restrictions in protein intake are not necessary for those critically ill patients with AKI. However, the studies reviewed here showed significant heterogeneity in protein dose and delivery, estimation of protein needs, patient population, and definition of AKI, and thus further research is needed to systematically determine the optimal dose of protein for critically ill adults with AKI.


An updated review of microsatellite instability in the era of next-generation sequencing and precision medicine.
Deficient DNA mismatch repair causes a robust mutator phenotype known as microsatellite instability (MSI). MSI is a feature of Lynch syndrome-related cancers and is also found in approximately 15% of sporadic gastric, colorectal, and endometrial cancers. Epigenetic inactivation of the MLH1 gene is often associated with sporadic MSI cancers. Recent next-generation sequencing (NGS)-based analyses have comprehensively characterized MSI-positive (MSI+) cancers, and several approaches for detecting the MSI phenotype of tumors using NGS have been developed. The FDA has recently granted accelerated approval to an anti-PD-1 antibody, pembrolizumab, for use in pediatric and adult patients with MSI+ solid tumors. Genome-wide analyses using NGS have revealed that hypermutation defined as >10 somatic mutations per megabase appears to be more prevalent than previously estimated, affecting approximately 17% of adult cancers. These results potentially expand the use of immunotherapy, which is thought to be effective in cancers with an increased mutational burden. Therefore, evaluation of MSI and MSI-associated molecular changes in tumors has emerged as clinically important. MSI is a valuable diagnostic marker of Lynch syndrome and a potential predictive marker for chemotherapy and immunotherapy efficacy. Here, we provide an update on MSI-associated cancers, focusing on findings obtained by genome-wide analyses using NGS, and the predictive role of MSI in immune checkpoint immunotherapy.


The Effect of the Affordable Care Act Medicaid Expansions on Financial Wellbeing.
We examine the effect of the Medicaid expansions under the 2010 Patient Protection and Affordable Care Act (ACA) on consumer financial outcomes using data from a major credit reporting agency for a large, national sample of adults. We employ the synthetic control method to compare individuals living in states that expanded Medicaid to those that did not. We find that the Medicaid expansions significantly reduced the number of unpaid bills and the amount of debt sent to third-party collection agencies among those residing in zip codes with the highest share of low-income, uninsured individuals. Our estimates imply a reduction in collection balances of approximately $1,140 among those who gain Medicaid coverage due to the ACA. Our findings suggest that the ACA Medicaid expansions had important financial impacts beyond increasing health care use.


Development and Validation of a Prognostic Risk Score System for COVID-19 Inpatients: A Multi-Center Retrospective Study in China.
Coronavirus disease 2019 (COVID-19) has become a worldwide pandemic. Hospitalized patients of COVID-19 suffer from a high mortality rate, motivating the development of convenient and practical methods that allow clinicians to promptly identify high-risk patients. Here, we have developed a risk score using clinical data from 1479 inpatients admitted to Tongji Hospital, Wuhan, China (development cohort) and externally validated with data from two other centers: 141 inpatients from Jinyintan Hospital, Wuhan, China (validation cohort 1) and 432 inpatients from The Third People's Hospital of Shenzhen, Shenzhen, China (validation cohort 2). The risk score is based on three biomarkers that are readily available in routine blood samples and can easily be translated into a probability of death. The risk score can predict the mortality of individual patients more than 12 d in advance with more than 90% accuracy across all cohorts. Moreover, the Kaplan-Meier score shows that patients can be clearly differentiated upon admission as low, intermediate, or high risk, with an area under the curve (AUC) score of 0.9551. In summary, a simple risk score has been validated to predict death in patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2); it has also been validated in independent cohorts.


The Plant Parasitic Nematodes Database: A Comprehensive Genomic Data Platform for Plant Parasitic Nematode Research.
Plant parasitic nematodes are important phytopathogens that greatly affect the growth of agricultural and forestry plants. Scientists have conducted several studies to prevent and treat the diseases they cause. With the advent of the genomics era, the genome sequencing of plant parasitic nematodes has been considerably accelerated, and a large amount of data has been generated. This study developed the Plant Parasitic Nematodes Database (PPND), a platform to combine these data. The PPND contains genomic, transcriptomic, protein, and functional annotation data, allowing users to conduct BLAST searches and genome browser analyses and download bioinformatics data for in-depth research. PPND will be continuously updated, and new data will be integrated. PPND is anticipated to become a comprehensive genomics data platform for plant parasitic nematode research.


Selective renal angiography: its value in renal transplantation.
To date angiography has been the most precise diagnostic modality for the evaluation of a transplanted kidney. In the immediate postoperative period angiography can clearly demonstrate the integrity of the vascular anastomoses, and in most cases helps to differentiate between acute tubular necrosis and acute rejection. It is also helpful in the evaluation of the transplant kidney later in the postoperative period, since it can provide some estimate of the vascular effects of chronic rejection and distinguish between hypertension secondary to chronic rejection or renal artery stenosis. Serial isotope techniques may play a more important role in the evaluation of the transplanted kidney in the future, but at present angiography is the diagnostic modality of choice at the Cleveland Clinic.


The Effect of Different Storage Media on Color Stability of Self-Adhesive Composite Resin Cements for up to One Year.
The aim of this study was to analyze the long-term color stability of eight self-adhesive composite resin cements (SACRCs) after storage in diverse media for up to one year. 480 discs (diameter: 12 mm/thickness: 1.0 ± 0.05 mm) were fabricated (n = 60/SACRC): (1) BeautyCem (BEA); (2) Bifix SE (BIF); (3) Clearfil SA Cement Automix (CLE); (4) RelyX Unicem 2 Automix (RXU); (5) SeT (SET); (6) SmartCem 2 (SMC); (7) SoloCem (SOC); and (8) SpeedCEM (SPC). After polishing, specimens were immersed in (a) red wine (RW); (b) curry-solution (CU); (c) cress-solution (CR); and (d) distilled water (DW) at 37 °C and measured after 7, 28, 90, 180, and 365 days for color differences (ΔE) and water absorption (WA). Non-aged specimens were used as baselines. After 365 days, all of the discs were polished and their ΔE was measured. Data were analyzed using Kolmogorov-Smirnov, partial-eta-squared/ηP², 3-/1-way ANOVA with Tukey-HSD post-hoc test (α = 0.05). Significant differences occurred between all SACRCs for WA (p ≤ 0.003), except in RXU and in SET and in ΔE (p ≤ 0.002), except in SET and SPC. The significantly highest WA presented in SOC; the lowest showed in BEA. Significant ΔE differences and a decrease after polishing between all storage media were found (p < 0.001) with highest values for RW, followed by CU, CR, and DW. The lowest ΔE was measured for CLE, followed by SOC, BIF, RXU, BEA, SPC, SET, and SMC (p < 0.001) and increased significantly during aging. The highest ΔE decrease presented in BEA. SACRCs showed an increase in WA/ΔE within total aging time. Discoloration could not be removed completely by polishing. SACRCs need to be carefully selected for restorations in the esthetical zone with visible restoration margins. Polishing can significantly reduce the marginal discoloration.


Reducing within-subject variation in chewing cycle kinematics--a statistical approach.
High levels of within-subject variability have limited the use of chewing cycle kinematics in the experimental and clinical context. The purpose of this study was to validate a new strategy for reducing within-subject variability in chewing cycle kinematics, based on the 10 most representative cycles from a chewing sequence. This prospective study included 25 young subjects, with normal class I occlusions. An optoelectronic recording system was used to track chin movements of subjects chewing gum (2.5 g). Computer programs provided estimates for duration and movement as well as the 3D coordinates of the chin point. The total output files were further processed for selection of 10 representative cycles based on standard scores for total duration, excursive ranges in the lateral, vertical and antero-posterior directions. Multilevel modelling procedures were used to test for significant differences. There were no significant differences in cycle duration or excursions between the estimates for all cycles versus the 10 most representative cycles. Cycle shapes were very similar. There were no statistically significant differences in between-subject variances. All within-subject variances were smaller when using data from the 10 most representative cycles. The reduction of variance was approximately 33% for total duration and 75% for total 3D excursion. The results validate the pre-processing strategy that selects the 10 most representative cycles from a sequence without altering cycle duration, excursions or shape or affecting between-subject variation but reducing within-subject variation substantially.


Evaluation of chromosomal damage by flow cytometry in turbot (Scophthalmus maximus L.) exposed to fuel oil.
Flatfishes, turbots (Scophthalmus maximus), were injected intraperitoneally with two doses of fuel oil number 2. Biliary metabolites were evaluated by fixed fluorescence to verify the efficiency of intoxication. Ethoxyresorufin-O-deethylase (EROD) activity was compared with chromosomal damage measured by flow cytometry. The analysis of biliary metabolites showed a good dose-response relation and constitutes a clear reference for the subsequent measurements. Comparing flow cytometry and EROD results, a shorter delay of response for EROD activity was obtained, but chromosomal damage was significant only after one week. The persistence of the EROD response was shorter, while the genotoxic signal still persisted after one month. The measurement of chromosomal damage allowed a good differentiation between the two tested doses. In the case of EROD activity, the results were less clear. The results suggest that within a few weeks after exposure to fuel oil number 2, the measurements of chromosomal damage by flow cytometry can be used to detect a dose-dependent genotoxic response in fish.


Activity of a new antiemetic agent: alizapride. A randomized double-blind crossover controlled trial.
Alizapride is a methoxy-2-benzamide derivative three times more potent than its parent compound, metoclopramide, as an antagonist of apomorphine-induced emesis in dogs. The antiemetic activity of alizapride plus dexamethasone (DXM) was compared with that of placebo plus DXM in a randomized, double-blind, crossover study in cancer patients receiving cisplatin (DDP). Alizapride, given at the maximally tolerated dose of 4 mg/kg x 5, or placebo was given in a sequence determined by randomization during two successive, identical courses of antitumor chemotherapy. The antiemetic treatment was given 30 min before and 1.5, 3.5, 5.5, and 7.5 h after starting. DXM, in a dose of 12 mg, was given IV with the first administration of alizapride or placebo. A total of 39 patients completed the two courses of chemotherapy. The severity of gastrointestinal symptoms was influenced by previous treatment but not by the treatment sequence. Although our overall results suggest that alizapride does not add to the activity of DXM against DDP-induced amesis, a statistically significant difference favoring alizapride plus DXM was found among patients with the lowest gastrointestinal tolerance to DDP: women, patients under 50 years of age, and patients pretreated with chemotherapy including DDP and non-DDP agents. Side effects consisted of orthostatic hypotension, which was symptomatic in two patients, and a single occurrence of severe extrapyramidal syndrome. We conclude that alizapride is more active than placebo when combined with DXM for DDP-induced emesis in patients at high risk of severe nausea and vomiting. The severity of the side effects in this study indicates that a dose reduction of alizapride might be appropriate for further studies.


The association between cardiopulmonary exercise testing and postoperative outcomes in patients with lung cancer undergoing lung resection surgery: A systematic review and meta-analysis.
Exercise capacity should be determined in all patients undergoing lung resection for lung cancer surgery and cardiopulmonary exercise testing (CPET) remains the gold standard. The purpose of this study was to investigate associations between preoperative CPET and postoperative outcomes in patients undergoing lung resection surgery for lung cancer through a review of the existing literature. A search was conducted on PubMed, Scopus, Cochrane Library and CINAHL from inception until December 2022. Studies investigating associations between preoperative CPET and postoperative outcomes were included. Risk of bias was assessed using the QUIPS tool. A random effect model meta-analysis was performed. I2 > 40% indicated a high level of heterogeneity. Thirty-seven studies were included with 6450 patients. Twenty-eight studies had low risk of bias. [Formula: see text] peak is the oxygen consumption at peak exercise and serves as a marker of cardiopulmonary fitness. Higher estimates of [Formula: see text] peak, measured and as a percentagege of predicted, showed significant associations with a lower risk of mortality [MD: 3.66, 95% CI: 0.88; 6.43 and MD: 16.49, 95% CI: 6.92; 26.07] and fewer complications [MD: 2.06, 95% CI: 1.12; 3.00 and MD: 9.82, 95% CI: 5.88; 13.76]. Using a previously defined cutoff value of > 15mL/kg/min for [Formula: see text] peak, showed evidence of decreased odds of mortality [OR: 0.55, 95% CI: 0.28-0.81] and but not decreased odds of postoperative morbidity [OR: 0.82, 95% CI: 0.64-1.00]. There was no relationship between [Formula: see text] slope, which depicts ventilatory efficiency, with mortality [MD: -9.60, 95% CI: -27.74; 8.54] however, patients without postoperative complications had a lower preoperative [Formula: see text] [MD: -2.36, 95% CI: -3.01; -1.71]. Exercise load and anaerobic threshold did not correlate with morbidity or mortality. There was significant heterogeneity between studies. Estimates of cardiopulmonary fitness as evidenced by higher [Formula: see text] peak, measured and as a percentage of predicted, were associated with decreased morbidity and mortality. A cutoff value of [Formula: see text] peak > 15mL/kg/min was consistent with improved survival but not with fewer complications. Ventilatory efficiency was associated with decreased postoperative morbidity but not with improved survival. The heterogeneity in literature could be remedied with large scale, prospective, blinded, standardised research to improve preoperative risk stratification in patients with lung cancer scheduled for lung resection surgery.


Clinical relevance of routine semen analysis and controversies surrounding the 2010 World Health Organization criteria for semen examination.
Semen analysis is the corner stone of infertility evaluation as it provides information on the functional status of the seminiferous tubules, epididymis and accessory sex glands. The methods on how the human semen should be evaluated are provided by the World Health Organization, which periodically releases manuals that include specific protocols and reference standards. In 2010, the WHO published new criteria for human semen characteristics that were markedly lower than those previously reported. In this review initially it is discussed the limitations of semen analysis as a surrogate measure of a man's ability to father a pregnancy. Secondly, it is analyzed methodology issues that could explain why the newly released reference values were different from those earlier reported. Thirdly, it is speculated on the likely effects of the 2010 WHO criteria in the management of male infertility. Due to the several inherent limitations of semen analysis as a surrogate marker of male infertility, physicians should exercise caution when interpreting results. A template for semen analysis reports that incorporates the distribution of the semen characteristics of recent fathers in centiles rather than solely the minimum thresholds could aid clinicians to better understand how a given patient results compare with the reference population. Importantly, a male infertility evaluation must go far beyond a simple semen analysis, as it has to be complemented with a proper physical examination, a comprehensive history taking, and relevant endocrine, genetic, and other investigations.


Numerical simulation and experimental validation of blood flow in arteries with structured-tree outflow conditions.
Blood flow in the large systemic arteries is modeled using one-dimensional equations derived from the axisymmetric Navier-Stokes equations for flow in compliant and tapering vessels. The arterial tree is truncated after the first few generations of large arteries with the remaining small arteries and arterioles providing outflow boundary conditions for the large arteries. By modeling the small arteries and arterioles as a structured tree, a semi-analytical approach based on a linearized version of the governing equations can be used to derive an expression for the root impedance of the structured tree in the frequency domain. In the time domain, this provides the proper outflow boundary condition. The structured tree is a binary asymmetric tree in which the radii of the daughter vessels are scaled linearly with the radius of the parent vessel. Blood flow and pressure in the large vessels are computed as functions of time and axial distance within each of the arteries. Comparison between the simulations and magnetic resonance measurements in the ascending aorta and nine peripheral locations in one individual shows excellent agreement between the two.


Detection of strain variation in isolates of Rhodococcus equi from an affected foal using repetitive sequence-based polymerase chain reaction.
Rhodococcus equi is an important pathogen of foals aged 1-6 months. Evidence exists that foals are exposed to a wide diversity of R. equi strains in their environment. However, limited data are available regarding the extent to which genotypic variation exists among isolates infecting individual foals. Therefore, electrophoresis of repetitive sequence-based polymerase chain reaction (rep-PCR) amplicons in an automated microfluidics chip format was used to genotype 9 virulent R. equi isolates obtained from distinct anatomic locations in a single foal. Four of the isolates were obtained from different regions of the lung, and 5 were from abscessed intra-abdominal lymph nodes (LNs). Six distinct genotypes were identified among the 9 isolates. None of the pulmonary isolates was identical; however, a pulmonary isolate was found to be identical to an isolate recovered from a small intestinal LN, and another pulmonary isolate was identical to an isolate from a mesenteric LN. These results indicate that foals can be infected with multiple strains of virulent R. equi. Furthermore, identical strains can be found in multiple, remote anatomic locations in an infected foal, and this can occur for >1 strain in the same foal. The automated system used in the current study provided a rapid, reproducible, and discriminating method for typing R. equi isolates.


CIRRHOTIC PATIENTS WITH ACUTE KIDNEY INJURY (AKI) HAVE HIGHER MORTALITY AFTER ABDOMINAL HERNIA SURGERY.
The incidence of abdominal hernia in cirrhotic patients is as higher as 20%; in cases of major ascites the incidence may increase up to 40%. One of the main and most serious complications in cirrhotic postoperative period (PO) is acute kidney injury (AKI). To analyze the renal function of cirrhotic patients undergoing to hernia surgery and evaluate the factors related to AKI. Follow-up of 174 cirrhotic patients who underwent hernia surgery. Laboratory tests including the renal function were collected in the PO.AKI was defined based on the consensus of the ascite´s club. They were divided into two groups: with (AKI PO) and without AKI . All 174 patients were enrolled and AKI occurred in 58 (34.9%). In the AKI PO group, 74.1% had emergency surgery, whereas in the group without AKI PO it was only 34.6%.In the group with AKI PO, 90.4% presented complications, whereas in the group without AKI PO they occurred only in 29.9%. Variables age, baseline MELD, baseline creatinine, creatinine in immediate postoperative (POI), AKI and the presence of ascites were statistically significant for survival. There is association between AKI PO and emergency surgery and, also, between AKI PO and complications after surgery. The factors related to higher occurrence were initial MELD, basal Cr, Cr POI. The patients with postoperative AKI had a higher rate of complications and higher mortality.