Massive intra-abdominal recurrence after robotic-assisted radical cystectomy: A case report and critical appraisal of literature.
Over the years, the robotic surgery is gaining increasing importance in the treatment of bladder cancer. Some doubts remain about the oncological safety of robotic approach and alerts have been raised about the occurrence of atypical recurrences, including peritoneal carcinomatosis and port-site metastasis. The patient referred to our Emergency Department because of acute confusional state probably due to severe anemia and sepsis. A left nephroureterectomy, left hemicolectomy with end colostomy and the surgical excision of the huge mass was performed through a xipho-pubic incision associated to another left peri-stomal incision. The histological specimen analysis showed a high-grade sarcoma, not otherwise specified (sarcoma, NOS-type), measuring 29 cm × 8 cm × 5 cm in diameters. The left kidney and ureter were not infiltrated by the neoplasm while serosa membranes and muscular layers of left colon were infiltrated by the mass. The patient died because of a cardiac arrest 4 days after surgery. RARC is a safe and feasible alternative to open radical cystectomy (ORC) with satisfactory operative time, little blood loss, and low transfusion rates. Despite this, RARC is associated with a low, but not neglectable, risk of atypical metastases like peritoneal implants and port-site metastasis. Although a small amount (7%) of RARC were performed in non-urothelial variants of bladder cancer, the sarcomatoid one can be related to a greater risk of atypical recurrence and special precaution should be taken to reduce potential causes of tumor seeding.


Isolates of Colletotrichum truncatum with Resistance to Multiple Fungicides from Soybean in Northern Thailand.
In Thailand, four systemic fungicides-carbendazim (Car), azoxystrobin (Azo), difenoconazole (Dif), and penthiopyrad (Pen)-are commonly used to control soybean anthracnose caused by Colletotrichum truncatum; however, the pathogen has developed resistance. From 2019 to 2020, fungicide resistance in C. truncatum from fields in Chiang Rai and Chiang Mai was monitored. In tests of 85 C. truncatum isolates for resistance to multiple fungicides, 15.3% were CarRAzoR, 34.1% were triple resistant (CarRAzoRDifR or CarRAzoRPenR), and 50.6% were CarRAzoRDifRPenR. Surprisingly, all isolates tested had lost their sensitivity to one or more of the fungicides tested. The carbendazim-resistant isolates carried a point mutation in the β-tubulin gene at codon 198 (E198A) or 200 (F200Y), and all azoxystrobin-resistant isolates had a mutation in the cytochrome b gene at codon 143 (G143A) or 129 (F129L). Moreover, a novel mutation at codon 208 (S208Y) in the gene encoding succinate dehydrogenase subunit B was detected in all of the isolates highly resistant to penthiopyrad. No mutation linked with difenoconazole resistance was detected in the genes encoding cytochrome P450 sterol 14α-demethylase. To the best of our knowledge, this is the first report of C. truncatum isolates resistant to multiple fungicides and serves as a warning to take measures to prevent the occurrence and distribution of these multiple-fungicide-resistant populations in soybean fields.


Using human-centered design to improve the assault rifle.
The objective of the present study was to interview infantry soldiers to determine their preferences with respect to rifle design and to examine the effect of buttstocks on shooting performance. Factor analysis showed that seven main factors should be considered in rifle redesign including tactics necessary, interface design, saving weight, bullpup configuration, sight design, other devices, and bayonet lug. For the shooting experiment, a total of four shooting trials were performed with the T-91 rifle, with buttstock lengths of 26 mm, 34 mm, self-adjusting stock, and bullpup stock. The analysis revealed that buttstock length had a significant effect on shooting performance. The redesigned rifle weight and total length should be reduced to 3.2 kg and 750 mm, respectively. The rifle buttstock should be a non-adjustable bullpup style. The buttstock shape should be curved and the hand-guard type should be more deeply and density seams, while the trigger handle shape should be slanted.


Health Literacy and Change in Health-Related Quality of Life in Dialysed Patients.
Health-related quality of life (HRQoL) is likely to deteriorate with the progression of chronic kidney disease (CKD). This change may be worsened by low health literacy (HL). We performed a longitudinal study at over 20 dialysis clinics in Slovakia (n = 413; mean age = 64.8 years; males = 58.4%). We assessed the association of three HL groups with a change in HRQoL over two years using binary logistic regression adjusted for type of vascular access, dialysis effectiveness, comorbidity, age and gender. We found that patients with low HL had poorer HRQoL at baseline in comparison to high-HL patients. We did not find significant associations of lower HL with the deterioration of mental or physical HRQoL after two years. In the adjusted model, patients with lower HL were not more likely to have deteriorated physical (low-HL patients: odds ratio/95% confidence interval: 0.99/0.53-1.84; moderate-HL patients: 0.97/0.55-1.73) or mental HRQoL (low-HL patients: 1.00/0.53-1.87; moderate-HL patients: 0.95/0.53-1.70) in comparison to high-HL patients. The HRQoL of lower-HL patients is worse at baseline but develops similarly to that of high-HL patients during dialysis treatment. Their relative HRQoL, thus, does not worsen further, but it does not improve either. Tailoring care to their needs may help to decrease the burden of low HL in dialysed patients.


Toxic dilatation of the colon.
Toxic dilatation of the colon represents the most serious complication of colonic inflammatory diseases. It is an acute transmural extension of severe inflammation resulting in decreased tissue cohesion and an increased tendency for bowel perforation. The process is characterized by dilatation and apparent radiologic thickening and nodularity of the bowel wall. Cases of toxic megacolon without dilatation occur infrequently.


Clonal characterization of rat muscle satellite cells: proliferation, metabolism and differentiation define an intrinsic heterogeneity.
Satellite cells (SCs) represent a distinct lineage of myogenic progenitors responsible for the postnatal growth, repair and maintenance of skeletal muscle. Distinguished on the basis of their unique position in mature skeletal muscle, SCs were considered unipotent stem cells with the ability of generating a unique specialized phenotype. Subsequently, it was demonstrated in mice that opposite differentiation towards osteogenic and adipogenic pathways was also possible. Even though the pool of SCs is accepted as the major, and possibly the only, source of myonuclei in postnatal muscle, it is likely that SCs are not all multipotent stem cells and evidences for diversities within the myogenic compartment have been described both in vitro and in vivo. Here, by isolating single fibers from rat flexor digitorum brevis (FDB) muscle we were able to identify and clonally characterize two main subpopulations of SCs: the low proliferative clones (LPC) present in major proportion (approximately 75%) and the high proliferative clones (HPC), present instead in minor amount (approximately 25%). LPC spontaneously generate myotubes whilst HPC differentiate into adipocytes even though they may skip the adipogenic program if co-cultured with LPC. LPC and HPC differ also for mitochondrial membrane potential (DeltaPsi(m)), ATP balance and Reactive Oxygen Species (ROS) generation underlying diversities in metabolism that precede differentiation. Notably, SCs heterogeneity is retained in vivo. SCs may therefore be comprised of two distinct, though not irreversibly committed, populations of cells distinguishable for prominent differences in basal biological features such as proliferation, metabolism and differentiation. By these means, novel insights on SCs heterogeneity are provided and evidences for biological readouts potentially relevant for diagnostic purposes described.


Desynchrony of fear in phobic exposure.
Intuitively, phobic exposure would seem to be a very stressful experience. However, it is not clear whether the characteristic feature of a classic stress response, activation of the hypothalamic-pituitary-adrenal (HPA) axis, is present in phobic fear. Some instances of phobic fear have been found to be accompanied by robust increases in cortisol, whereas in other instances a dissociation between subjective-behavioural arousal and the HPA-axis has been found. The latter is referred to as desynchrony of fear. The aim of the current study was to test the hypothesis that phobic fear is similar to normal fear and, as such, is accompanied by a robust increase in cortisol values. In all, 16 spider phobic subjects and 16 healthy controls participated in the study. During and following a standardised stepwise exposure paradigm, saliva samples were collected for cortisol determination. In contrast to the controls, the spider phobics reacted with a strong fear reaction to the spiders. However, cortisol levels remained unaffected. The phobic response did not resemble the classic 'fight or flight' response. Some suggest that the HPA-axis response has become extinguished in modern man. Yet, it is possible that phobic fear is not a derivative of an ancient fear but rather a separate entity that relies on other neuroendocrinological systems.


Elevated corticosterone associated with food deprivation upregulates expression in rat skeletal muscle of the mTORC1 repressor, REDD1.
Food deprivation induces a repression of protein synthesis in skeletal muscle in part due to reduced signaling through the mammalian target of rapamycin complex 1 (mTORC1). Previous studies have identified upregulated expression of the protein Regulated in DNA Damage and Development (REDD1) as an important mechanism in the regulation of mTORC1 activity in response to a variety of stresses. Our goal in this investigation was to determine whether modulation of REDD1 expression occurs in response to food deprivation and refeeding, and, if it does, to ascertain if changes in REDD1 expression correlate with altered mTORC1 signaling. As expected, mTORC1 signaling was repressed after 18 h of food deprivation compared with freely-fed control rats and quickly recovered after refeeding for 45 min. Food deprivation caused a dramatic rise in REDD1 mRNA and protein expression; refeeding resulted in a reduction to baseline. Food deprivation is characterized by low-serum insulin and elevated glucocorticoid concentrations. Therefore, initially, alloxan-induced type I diabetes was used to minimize the food deprivation- and refeeding-induced changes in insulin. Although diabetic rats exhibited upregulated REDD1 expression compared with nondiabetic controls, there was no direct correlation between REDD1 mRNA expression and serum insulin levels, and insulin treatment of diabetic rats did not affect REDD1 expression. In contrast, serum corticosterone levels correlated directly with REDD1 mRNA expression (r = 0.68; P = 0.01). Moreover, inhibiting corticosterone-mediated signaling via administration of the glucocorticoid receptor antagonist RU486 blocked both the food deprivation- and diabetes-induced increase in REDD1 mRNA expression. Overall, the results demonstrate that changes in REDD1 expression likely contribute to the regulation of mTORC1 signaling during food deprivation and refeeding.


Pancreatic alpha cell autoantibodies and glucagon response to arginine.
The frequency and significance of cytoplasmic pancreatic alpha cell autoantibodies (ACA) were investigated in 2102 healthy controls, 879 patients with insulin-dependent diabetes mellitus (IDDM) who were negative for islet cell autoantibodies (ICA), and 1567 relatives of IDDM patients. ACA were found in approximately 1 in 200 people of all ages and were not significantly associated with IDDM, the IDDM-associated HLA phenotypes DR3 and DR4, or thyrogastric or adrenal autoantibodies. Of 11 ACA-positive patients studied by arginine stimulation tests, none had frank glucagon deficiency. Thus, ACA do not appear to be associated with defective alpha cell function or with IDDM.


Global burden of active smoking among people living with HIV on antiretroviral therapy: a systematic review and meta-analysis.
Although the high burden of both active smoking and human immunodeficiency virus (HIV) is clearly known, the relationship between them is still not well characterized. Therefore, we estimated the global prevalence of active smoking in people living with HIV (PLHIV) on antiretroviral therapy (ART) and investigated the association between exposure to active smoking and risk for suboptimal adherence to ART. Main text: We searched PubMed, Embase, and Web of Science to identify articles published until September 19, 2019. Eligible studies reported the prevalence of active smoking in PLHIV on ART or investigated the association between active smoking and ART adherence; or enough data to compute these estimates. We used a random-effects model to pool data and quantified heterogeneity (I2). The global prevalence of active smoking was 36.1% (95% CI: 33.7-37.2; 329 prevalence data; 462 104 participants) with substantial heterogeneity. The prevalence increased with level of country income; from 10.1% (95% CI: 6.8-14.1) in low-income to 45.2% (95% CI: 42.7-47.7) in high-income countries; P < 0.0001. With regards to the Joint United Nations Programme on HIV/AIDS (UNAIDS) regions, the prevalence was higher in West and Central Europe and North America 45.4% (42.7-48.1) and lowest in the two UNAIDS regions of sub-Saharan Africa: Eastern and Southern Africa 10.7% (95% CI: 7.8-14.0) and West and Central Africa 4.4% (2.9-6.3); P < 0.0001. Globally, we estimated that there were 4 110 669 PLHIV on ART who were active smokers, among which the highest number was from Eastern and Southern Africa (35.9%) followed by Asia and the Pacific (25.9%). Active smoking was significantly associated with suboptimal ART adherence: pooled odds ratio 1.57 (95% CI: 1.37-1.80; I2 = 56.8%; 19 studies; 48 450 participants); even after considering adjusted estimates: 1.67 (95% CI: 1.39-2.01; I2 = 53.0%; 14 studies). This study suggests a high prevalence of active smoking in PLHIV on ART and an association between active smoking and ART suboptimal adherence. As such, healthcare providers and policy makers should focus on adopting and implementing tobacco harm reduction strategies in HIV care, especially in sub-Saharan Africa known as epicenter of HIV pandemic with highest number of active tobacco smoking among PLHIV on ART.


Analysis of liquid-to-solid coupling and other performance parameters for microfluidically reconfigurable photonic systems.
In this paper, we analytically investigate the coupling of light from liquid-core waveguides to conventional solid-core waveguides and a series of other optical properties of liquid waveguides in order to gauge the practicality of such a system for use in microfluidically reconfigurable photonic systems. A finite element model of the system was constructed and relevant properties such as mode field diameter, attenuation, bending loss, and efficiency of evanescent and end-fire coupling were investigated as a function of the liquid waveguide Peclet number and the relative difference in refractive index. For pure liquid systems we show that the mode field diameter decreases monotonically with increasing Peclet number and that bending losses could be significantly reduced by increasing the Peclet number. More critically, we observed irreversible evanescent coupling, in which the light coupled in the solid waveguide is entrapped within the solid rather than coupled back into the liquid waveguide. This effect was caused by the lengthwise variation in the propagation constant of the liquid core due to downstream diffusion. We demonstrate that coupling efficiencies as high as 84% can be obtained for fluid based end-fire coupling by taking advantage of the tunable mode field diameter. By developing techniques for coupling light between liquid and solid states we hope to be able to overcome the drawbacks of solid waveguide systems (e.g. unchangeable structure and properties) and liquid waveguide systems (e.g. diversion and attenuation) yielding a new paradigm for reconfigurable photonics.


Usefulness of preoperative chemoradiation in locally advanced cervical carcinoma.
This study investigated response rate, toxicity, and influence on intra- and postoperative complications of neoadjuvant concurrent chemoradiation in locally advanced (Figo IB2-IIIB) cervical carcinomas. Twenty-eight patients (median age 46.5 years, age range 29-73 years), diagnosed as having squamous cell carcinomas (n=21), adenocarcinomas (n=6), and undifferentiated carcinoma (n=1), entered this study. The chemoradiation protocol included external radiotherapy to the pelvis (39.6 Gy), intra-arterial or intravenous infusion of cisplatin (70 mg/m2 on days 1 and 22), and 24-hour continuous intravenous infusion of 5-fluorouracil (700 mg/m2 on days 1-4 and 22-25). Two weeks after the end of chemoradiotherapy, the patients underwent restaging followed by suitable operation including pelvic lymphadenectomy. The median follow-up period in the surviving patients was 13.1 months. The response rate for chemoradiation was 93% (26/28). The 2-year disease free survival was 85.2%. Grade 3-4 neutropenia was observed in 14 patients (50%), 4 patients (14%) showed grade 3 anemia, and 5 patients (17.8%) showed grade 3-4 thrombocytopenia. Nineteen patients (67.8%) could undergo surgery after concurrent chemoradiation. Pathological examination revealed a complete response (pT0) in 7 patients, and 5 patients showed only microscopic residual disease (pTmic). In 4 patients, there were intraoperative complications: vesical lesions in 2 (10%) and small intestine lesions in 2 (10%). Neoadjuvant therapy with concurrent chemoradiation for locally advanced cervical carcinoma is effective, safe, and useful for increasing operability rates as well as decreasing intra- and postoperative complications. Therefore, neoadjuvant concurrent chemoradiation should be considered for treatment in patients with locally advanced cervical carcinomas who could not undergo primary operation.


Suppressor U1 snRNAs in Drosophila.
Although the role of U1 small nuclear RNAs (snRNAs) in 5' splice site recognition is well established, suppressor U1 snRNAs active in intact multicellular animals have been lacking. Here we describe suppression of a 5' splice site mutation in the Drosophila melanogaster white gene (wDR18) by compensatory changes in U1 snRNA. Mutation of positions -1 and +6 of the 5' splice site of the second intron (ACG[GTGAGT to ACC]GTGAGC) results in the accumulation of RNA retaining this 74-nucleotide intron in both transfected cells and transgenic flies. U1-3G, a suppressor U1 snRNA which restores base-pairing at position +6 of the mutant intron, increases the ratio of spliced to unspliced wDR18 RNA up to fivefold in transfected Schneider cells and increases eye pigmentation in wDR18 flies. U1-9G, which targets position -1, suppresses wDR18 in transfected cells less well. U1-3G,9G has the same effect as U1-3G although it accumulates to lower levels. Suppression of wDR18 has revealed that the U1b embryonic variant (G134 to U) is active in Schneider cells and pupal eye discs. However, the combination of 9G with 134U leads to reduced accumulation of both U1b-9G and U1b-3G,9G, possibly because nucleotides 9 and 134 both participate in a potential long-range intramolecular base-pairing interaction. High levels of functional U1-3G suppressor reduce both viability and fertility in transformed flies. These results show that, despite the difficulties inherent in stably altering splice site selection in multicellular organisms, it is possible to obtain suppressor U1 snRNAs in flies.


[Routine sonography in suspected appendicitis. A trial of 24-hour screening at a general hospital].
A prospective study was conducted to determine the diagnostic value of sonography in suspected acute appendicitis. From September 1987 until May 1988, the end of the study, 152 patients suspected of having "acute appendicitis" were admitted to hospital. In contrast to other recently published studies conducted by a few highly qualified examiners, we dimed at determining whether this method is useful as a 24-hour-screening performed ba y number of more or less qualified examiners. In the beginning, the patients were examined with a 3.5 MHz scanner, later on exclusively with 5 MHz linear and sector scanners. 150 cases could be evaluated, 96 of which underwent surgery. We found 35 sonographically correct results, 89 correctly negative, 4 false positive and 13 false negative diagnoses. This means a sensitivity of 73% and a specificity of 96% for the sonographical diagnostic method. In 20 cases (13%) sonography led to other diagnoses. A large majority of these findings can be detected by less experienced examiners. The goal of a 24-hour-screening could not be reached because there a competent examiner was not always available at night. It became clear that clinically obvious cases do not require sonographic conformation. The value of sonography lies with doubtful cases of appendicitis: if the ultrasound examination is negative, appendectomy can be postponed; if sonography leads to other differential diagnoses, appropriate therapy can be started immediately.


Nanopatterned aluminum nitride template for high efficiency light-emitting diodes.
Nanopatterned aluminum nitride (NP-AlN) templates were used to enhance the light extraction efficiency of the light-emitting diodes (LEDs). Here, the NP-AlN interlayer between the sapphire substrate and GaN-based LED was used as an effective light outcoupling layer at the direction of bottom side and as a buffer layer for growth of GaN LEDs. The cross-sectional transmission electron microscopy (TEM) analysis showed that the formation of stacking faults and voids could help reduce the threading dislocations. Micro Raman spectra also revealed that the GaN-based epilayer grown on the NP-AlN template had smaller residual stress than that grown on a planar sapphire substrate. The normalized electroluminescence (EL) spectra at the top and bottom sides of device revealed that the enhancement of the bottom side emission of the LED with the NP-AlN interlayer was more notable than a planar sapphire substrate due to the graded-refractive-index (GRIN) effect of the NP-AlN.


The chrondoprotective actions of a natural product are associated with the activation of IGF-1 production by human chondrocytes despite the presence of IL-1beta.
Cartilage loss is a hallmark of arthritis and follows activation of catabolic processes concomitant with a disruption of anabolic pathways like insulin-like growth factor 1 (IGF-1). We hypothesized that two natural products of South American origin, would limit cartilage degradation by respectively suppressing catabolism and activating local IGF-1 anabolic pathways. One extract, derived from cat's claw (Uncaria guianensis, vincaria), is a well-described inhibitor of NF-kappaB. The other extract, derived from the vegetable Lepidium meyenii (RNI 249), possessed an uncertain mechanism of action but with defined ethnomedical applications for fertility and vitality. Human cartilage samples were procured from surgical specimens with consent, and were evaluated either as explants or as primary chondrocytes prepared after enzymatic digestion of cartilage matrix. Assessments included IGF-1 gene expression, IGF-1 production (ELISA), cartilage matrix degradation and nitric oxide (NO) production, under basal conditions and in the presence of IL-1beta. RNI 249 enhanced basal IGF-1 mRNA levels in human chondrocytes by 2.7 fold, an effect that was further enhanced to 3.8 fold by co-administration with vincaria. Enhanced basal IGF-1 production by RNI 249 alone and together with vincaria, was confirmed in both explants and in primary chondrocytes (P < 0.05). As expected, IL-1beta exposure completely silenced IGF-1 production by chondrocytes. However, in the presence of IL-1beta both RNI 249 and vincaria protected IGF-1 production in an additive manner (P < 0.01) with the combination restoring chondrocyte IGF-1 production to normal levels. Cartilage NO production was dramatically enhanced by IL-1beta. Both vincaria and RNI 249 partially attenuated NO production in an additive manner (p < 0.05). IL-1beta - induced degradation of cartilage matrix was quantified as glycosaminoglycan release. Individually RNI 249 or vincaria, prevented this catabolic action of IL-1beta. The identification of agents that activate the autocrine production of IGF-1 in cartilage, even in the face of suppressive pro-inflammatory, catabolic cytokines like IL-1beta, represents a novel therapeutic approach to cartilage biology. Chondroprotection associated with prevention of the catabolic events and the potential for sustained anabolic activity with this natural product suggests that it holds significant promise in the treatment of debilitating joint diseases.


Machine learning and phone data can improve targeting of humanitarian aid.
The COVID-19 pandemic has devastated many low- and middle-income countries, causing widespread food insecurity and a sharp decline in living standards1. In response to this crisis, governments and humanitarian organizations worldwide have distributed social assistance to more than 1.5 billion people2. Targeting is a central challenge in administering these programmes: it remains a difficult task to rapidly identify those with the greatest need given available data3,4. Here we show that data from mobile phone networks can improve the targeting of humanitarian assistance. Our approach uses traditional survey data to train machine-learning algorithms to recognize patterns of poverty in mobile phone data; the trained algorithms can then prioritize aid to the poorest mobile subscribers. We evaluate this approach by studying a flagship emergency cash transfer program in Togo, which used these algorithms to disburse millions of US dollars worth of COVID-19 relief aid. Our analysis compares outcomes-including exclusion errors, total social welfare and measures of fairness-under different targeting regimes. Relative to the geographic targeting options considered by the Government of Togo, the machine-learning approach reduces errors of exclusion by 4-21%. Relative to methods requiring a comprehensive social registry (a hypothetical exercise; no such registry exists in Togo), the machine-learning approach increases exclusion errors by 9-35%. These results highlight the potential for new data sources to complement traditional methods for targeting humanitarian assistance, particularly in crisis settings in which traditional data are missing or out of date.


Enhancers with cooperative Notch binding sites are more resistant to regulation by the Hairless co-repressor.
Notch signaling controls many developmental processes by regulating gene expression. Notch-dependent enhancers recruit activation complexes consisting of the Notch intracellular domain, the Cbf/Su(H)/Lag1 (CSL) transcription factor (TF), and the Mastermind co-factor via two types of DNA sites: monomeric CSL sites and cooperative dimer sites called Su(H) paired sites (SPS). Intriguingly, the CSL TF can also bind co-repressors to negatively regulate transcription via these same sites. Here, we tested how synthetic enhancers with monomeric CSL sites versus dimeric SPSs bind Drosophila Su(H) complexes in vitro and mediate transcriptional outcomes in vivo. Our findings reveal that while the Su(H)/Hairless co-repressor complex similarly binds SPS and CSL sites in an additive manner, the Notch activation complex binds SPSs, but not CSL sites, in a cooperative manner. Moreover, transgenic reporters with SPSs mediate stronger, more consistent transcription and are more resistant to increased Hairless co-repressor expression compared to reporters with the same number of CSL sites. These findings support a model in which SPS containing enhancers preferentially recruit cooperative Notch activation complexes over Hairless repression complexes to ensure consistent target gene activation.


Deactivation rate and shortening velocity as determinants of contractile frequency.
The kinetic properties of muscle that could influence locomotor frequency include rate of activation, rate of cross-bridge "attachment", intrinsic shortening velocity, and rate of deactivation. The latter two mechanisms are examined using examples from high-speed running in lizards and escape swimming in scallops. During running, inertial loading and elastic energy storage probably mitigate the effects of thermal alterations in intrinsic muscle shortening velocity. The result is a rather low thermal dependence of stride frequency over a 15-20 degree C temperature range. However, at lower temperatures, the longer times required for deactivation cause the thermal dependence of frequency to increase greatly. Scallops use a single muscle to swim by jet propulsion. In vivo shortening velocity in these animals also shows a low thermal dependence. As with high-speed running, the mechanics of jet propulsion may limit the effects of thermally induced changes in intrinsic shortening velocity. The largest thermal effect during swimming is on the initial phase of valve opening. The effects of temperature on the rate of deactivation of the adductor muscle could play an important role in limiting reextension of the muscle, which is dependent on elastic energy storage in the hinge ligament. These examples illustrate that the relative importance of various intrinsic contractile properties in controlling locomotor performance depends on the mechanics of the movements.


Social Determinants of Health: Underreported Heterogeneity in Systematic Reviews of Caregiver Interventions.
Although most people have some experience as caregivers, the nature and context of care are highly variable. Caregiving, socioeconomic factors, and health are all interrelated. For these reasons, caregiver interventions must consider these factors. This review examines the degree to which caregiver intervention research has reported and considered social determinants of health. We examined published systematic reviews and meta-analyses of interventions for older adults with age-related chronic conditions using the PRISMA and AMSTAR 2 checklists. From 2,707 papers meeting search criteria, we identified 197 potentially relevant systematic reviews, and selected 33 for the final analysis. We found scant information on the inclusion of social determinants; the papers lacked specificity regarding race/ethnicity, gender, sexual identity, socioeconomic status, and geographic location. The majority of studies focused on dementia, with other conditions common in later life vastly underrepresented. Significant gaps in evidence persist, particularly for interventions targeting diverse conditions and populations. To advance health equity and improve the effectiveness of interventions, research should address caregiver heterogeneity and improve assessment, support, and instruction for diverse populations. Research must identify aspects of heterogeneity that matter in intervention design, while recognizing opportunities for common elements and strategies.


Percutaneous endoscopic transforaminal lumbar discectomy: a critical appraisal.
Percutaneous endoscopic transforaminal lumbar discectomy (PETD) is one of the surgical techniques for the treatment of the lumbar disc herniations. Fenestration of the annulus and partial resection of the nuclear substance are achieved percutaneously by a posterolateral approach under local anaesthesia. The results of the first 42 patients are evaluated regarding the learning curve and indications for this procedure. The mean follow-up time was 15 months. Excellent and good results were evaluated as successful and the overall success rate is 77 %. All six patients with foraminal disc herniations in whom a free fragment could be removed had excellent results. Military personnel can return to work quickly without deficits with the use of this technique.


Preparation and cellular uptake of pH-dependent fluorescent single-wall carbon nanotubes.
Fluorescent single-wall carbon nanotubes (SWCNTs) were prepared by mixing cut SWCNTs with acridine orange (AO). The optical absorbance and fluorescence characteristics of AO-SWCNT conjugates display interesting pH-dependent properties. Fluorescence microscopy in combination with transmission electron microscopy proves that AO-SWCNTs can enter HeLa cells and are located inside lysosomes. The endocytosis-inhibiting tests show that the clathrin-mediated endocytosis is a key step in the internalization process. The internalized AO-SWCNTs remain inside lysosomes for more than a week and have little effect on cell proliferation. These findings may be useful in understanding the SWCNT-based intracellular drug delivery mechanism and help to develop new intracellular drug transporters.


Novel Bacillus subtilis Spore-Displayed Tyrosinase Kit for Rapid Detection of Tyrosine in Urine: Pharmaceutical Applications for the Early Diagnosis of Kidney-Related Diseases.
Purpose: Simple and cheap diagnostic kit development is one of the important aims of pharmaceutical developers and companies focused on public health improvement. The Bacillus subtilis spore surface-display technique is a genetic engineering method that is used to develop new-generation diagnostic kits applicable for the early detection of various types of diseases. In this study, we developed a novel simple, rapid, and inexpensive diagnostic paper-based kit to detect tyrosine in urine samples of humans and animals that is applicable for home or laboratory use. Methods: The B. subtilis spore-displayed tyrosinase system developed by genetic engineering methods was used to prepare a paper-based kit to detect tyrosine in urine samples of different groups of patients (i.e., patients with diabetes, diabetes with chronic kidney disease (CKD), and chronic kidney disease) for the detection of tyrosine during the acute disease phase. To confirm the sensitivity and specificity of the kit, tyrosine was also detected in urine samples using conventional liquid chromatography/mass spectroscopy. Results: Different concentrations of tyrosine (0.1-1 mM) were detected in urine samples based on visible changes of color from bright brownish-gray to dark brownish-gray within 1 hour. The kit could screen samples to distinguish the three groups of patients based on formation of a broad spectrum of colors reflecting the concentration of tyrosine. Conclusion: To the best of our knowledge, this is the first diagnostic kit with potential to rapidly diagnose various diseases related to the production of tyrosine in biological samples. This kit is not only widely applicable, including for personal use in the home, but is also appropriate as a part of standard screening tests and health protection programs in countries with limited resources.


Single-fulcrum laparoscopic cholecystectomy in uncomplicated gallbladder diseases: a retrospective comparative analysis with conventional laparoscopic cholecystectomy.
Single-fulcrum laparoscopic cholecystectomy (SFLC) is a variant type of single incision and multi-port technique that does not use specialized one-port devices or articulating instruments. We retrospectively compared perioperative outcomes of SFLC with those of conventional laparoscopic cholecystectomy (CLC). Between March 2009 and December 2010, SFLC was performed in 130 patients. Among them, 105 patients with uncomplicated gallbladder disease (no inflammation or no clinical symptoms) and another 105 patients who underwent CLC were selected for this study. There was no open conversion. In comparison with CLC, SFLC was performed more often in young (46.4±12.2 years vs. 52.5±13.6 years, p=0.001) female patients (80/25 vs. 62/43, p=0.008). The total operation time was longer in SFLC (56.7±14.1 min vs. 47.5±17.1 min, p<0.001), but pain scores immediately after operation and at discharge time were lower for SFLC than for CLC (3.1±1.3 vs. 4.0±1.9, p<0.001, 2.0±0.9 vs. 2.4±0.8, p=0.002). Total cost was lower for SFLC than for CLC (US $ 1801±289.9 vs. US $ 2003±617.4, p=0.004). There were no differences in hospital stay or complication rates. SFLC showed greater technical feasibility and cost benefits in treating uncomplicated benign gallbladder disease than CLC.


Ursolic acid inhibits acid sphingomyelinase in intestinal cells.
Ursolic acid (UA) has antiinflammatory and anticancer effects on mammalian cells. Increase in acid sphingomyelinase (SMase) is associated with several inflammatory diseases including inflammatory bowel diseases. The enzyme has become a target for drug discovery. The present study examined the roles of UA on acid SMase in intestinal cells. We found that UA specifically inhibited acid SMase activity in both human colon cancer Caco-2 cells and rat nontransformed IEC-6 intestinal cells in a dose-dependent manner, with 50% inhibition occurred at 30 μM for Caco-2 cells and less than 20 μM for IEC-6 cells. In comparison with some chemicals known to inhibit acid SMase, UA appeared most effective. The decreased acid SMase activity was not associated with significant accumulation of cellular sphingomyelin but significant increase in phosphatidylcholine, the donor of choline for sphingomyelin synthesis. Western blot analysis showed a decreased enzyme levels in the cells after UA stimulation, but real time quantitative polymerase chain reaction (qPCR) failed to show a parallel reduction of acid SMase mRNA after UA stimulation. Finally, UA had no direct effect on acid SMase activity in cell-free extracts. In conclusion, UA has inhibitory effects on acid SMase synthesis and the effect occurs presumably at posttranslational levels.


In vitro effects of HgCl2 on murine lymphocytes. I. Preferable activation of CD4+ T cells in a responder strain.
Mercury-induced autoimmune disorders have been demonstrated in rats and mice injected with HgCl2. We have studied the ability of HgCl2 to activate murine lymphocytes in vitro and found that it induced increased DNA synthesis, which peaked at days 4 to 6. Other metal ions, such as Mg2+ and Zn2+, had no or much less effect. Consistent with the in vivo studies, there were strain differences, and the most significant increase in thymidine uptake was induced in A.SW and BALB/c spleen cells. Both T lymphocytes and adherent cells were required for activation, and anti-CD4 Ab completely abrogated HgCl2-induced proliferation, suggesting the involvement of T helper cells. Cell phenotype analysis demonstrated that HgCl2 induced both proliferation and transformation of CD4+ and CD8+ T cells from BALB/c mice (responder strain). In contrast, only CD8+ T cells from the nonresponder DBA/2 mice were transformed. These findings indicate that helper T cells play a crucial role for the immunologic effects caused by HgCl2 and determine the ability of different mouse strains to respond to HgCl2.


Pathogenic mechanisms in sporadic amyotrophic lateral sclerosis.
In recognition of the 100th anniversary of Charcot's death we have reviewed possible pathogenic mechanisms in amyotrophic lateral sclerosis (ALS). Advances in the last 5 years in molecular biology and genetics have identified mutations in the cytosolic dismutase (SOD1) gene in some patients with familial ALS raising the possibility that oxidative stress may be involved in the pathogenesis. An excitotoxic pathogenesis has been implicated based on elevated plasma and CSF levels of amino acids and altered contents of amino acids in the nervous system of ALS patients and changes in the number of excitatory amino acid receptors. ALS sera containing antibodies to L-type calcium channels and the development of immune mediated lower and upper and lower motor neuron models have revitalized research efforts focusing on an immune basis for ALS. Other pathogenic mechanisms which have been the subject of recent research include elemental toxicity, apoptosis and programmed cell death and possibly a deficiency or abnormality in growth factors. Pathogenic processes for ALS must account for an increasing incidence of ALS, male preponderance, and the selective vulnerability of the cortico-motoneuronal system.


Secondary Bone Defect in Neuromuscular Diseases in Childhood: A Longitudinal "Muscle-Bone Unit" Analysis.
To evaluate the potential bone defect in neuromuscular diseases, we conducted a longitudinal study including three groups of patients: 14 Duchenne muscular dystrophies (DMD) and 2 limb-girdle muscular dystrophies (LGMD); 3 Becker muscular dystrophies (BeMD) and 7 spinal muscular atrophies (SMA). Yearly osteodensitometries assessed body composition and bone mineral density (BMD) associated with bone markers and leptin. Along the 7-year study, 107 osteodensitometries showed that bone status evolved to osteopenia in most patients except BeMD. When analyzing the crude values, BMD improved with age in BeMD and SMA but not in DMD/LGMD. The correlation using the Z-scores displayed a decrease in BMD with age in DMD/LGMD for all regions, in SMA at total body less head, whereas BMD increased in BeMD at lumbar spine. As observed in healthy persons, muscular mass and bone tissue were significantly correlated. Glucocorticoids were deleterious on trabecular and cortical bone. Leptin was high in most patients and correlated to fat mass and bone parameters. This study confirms a secondary bone defect in neuromuscular diseases, further confirming the functional relationship between bone and muscle and arguing for regular bone follow-up in patients to prevent fracture risk. Adipose tissue seems to interfere with bone remodeling in neuromuscular diseases.


Neonatal mortality risks among preterm births in a rural Bangladeshi cohort.
Preterm birth leads to an estimated 35% of neonatal deaths worldwide. Our study analyses neonatal mortality risks among preterm births in rural Bangladesh. Trained community health workers (CHW) prospectively collected data between June 2007 and September 2009. Among 32 126 livebirths, 22.3% were preterm (delivered at <37 weeks gestation) and almost half (46.4%) of all neonatal deaths occurred among preterm babies. Preterm babies who were born as the first child {[risk ratio (RR) 1.4; 95% confidence interval (CI) 1.1, 1.8]} and in the poorest households [RR 1.7; 95% CI 1.2, 2.4] were at higher mortality risk. Birth and newborn care preparedness was associated with lower risk of mortality [RR 0.3; 95% CI 0.2, 0.4] while preterm infants who had symptoms of infection [RR 5.6; 95% CI 4.3, 7.1] or whose mother suffered antenatal complications [RR 1.4; 95% CI 1.1, 1.8] were at higher mortality risk. Elimination of excess neonatal deaths caused by preterm would decrease population-level neonatal mortality rate by 31.0% [95% CI 27.60%, 34.5%]. Given that 87% of preterm births and 60% of preterm deaths were in late or moderate preterm infants, and that 87% preterm babies received a visit from CHW within third day of life, a home-based essential care package delivered by CHWs for sick preterm infants, specifically focused on birth preparedness, skin-to-skin care, immediate breast feeding, early recognition of danger signs, and linked through referral to intensive and quality care in health facilities, could be an effective approach in low resource settings.


Single-channel characteristics of wild-type IKs channels and channels formed with two minK mutants that cause long QT syndrome.
IKs channels are voltage dependent and K+ selective. They influence cardiac action potential duration through their contribution to myocyte repolarization. Assembled from minK and KvLQT1 subunits, IKs channels are notable for a heteromeric ion conduction pathway in which both subunit types contribute to pore formation. This study was undertaken to assess the effects of minK on pore function. We first characterized the properties of wild-type human IKs channels and channels formed only of KvLQT1 subunits. Channels were expressed in Xenopus laevis oocytes or Chinese hamster ovary cells and currents recorded in excised membrane patches or whole-cell mode. Unitary conductance estimates were dependent on bandwidth due to rapid channel "flicker." At 25 kHz in symmetrical 100-mM KCl, the single-channel conductance of IKs channels was approximately 16 pS (corresponding to approximately 0.8 pA at 50 mV) as judged by noise-variance analysis; this was fourfold greater than the estimated conductance of homomeric KvLQT1 channels. Mutant IKs channels formed with D76N and S74L minK subunits are associated with long QT syndrome. When compared with wild type, mutant channels showed lower unitary currents and diminished open probabilities with only minor changes in ion permeabilities. Apparently, the mutations altered single-channel currents at a site in the pore distinct from the ion selectivity apparatus. Patients carrying these mutant minK genes are expected to manifest decreased K+ flux through IKs channels due to lowered single-channel conductance and altered gating.


Longer duration of operative time enhances healing metabolites and improves patient outcome after Achilles tendon rupture surgery.
The relationship between the duration of operative time (DOT), healing response and patient outcome has not been previously investigated. An enhanced healing response related to DOT may potentiate repair processes, especially in hypovascular and sparsely metabolized musculoskeletal tissues such as tendons. This study aimed to investigate the association between DOT and the metabolic healing response, patient-reported outcome and the rate of post-operative complications after acute Achilles tendon injury. Observational cohort, cross-sectional study with observers blinded to patient grouping. A total of two-hundred and fifty-six prospectively randomized patients (210 men, 46 women; mean age 41 years) with an acute total Achilles tendon rupture all operated on with uniform anaesthetic and surgical technique were retrospectively assessed. At 2 weeks post-operatively, six metabolites were quantified using microdialysis. At 3, 6 and 12 months, patient-reported pain, walking ability and physical activity were examined using self-reported questionnaires, Achilles tendon total rupture score, foot and ankle outcome score and physical activity scale. At 12 months, functional outcome was assessed using the heel-rise test. Complications, such as deep venous thrombosis, infections and re-operations, were recorded throughout the study. Patients who underwent longer DOT exhibited higher levels of glutamate (p = 0.026) and glycerol (p = 0.023) at 2 weeks. At the 1-year follow-up, longer DOT was associated with significantly less loss in physical activity (p = 0.003), less pain (p = 0.009), less walking limitations (p = 0.022) and better functional outcome (p = 0.014). DOT did not significantly correlate with the rate of adverse events, such as deep venous thrombosis, infections or re-ruptures. Higher glutamate levels were associated with less loss in physical activity (p = 0.017). All correlations were confirmed by multiple linear regressions taking confounding factors into consideration. The results from this study suggest a previously unknown mechanism, increased metabolic response associated with longer DOT, which may improve patient outcome after Achilles tendon rupture surgery. Allowing for a higher amount of traumatized tissue, as reflected by up-regulation of glycerol in patients with longer DOT, may prove to be an important surgical tip for stimulation of repair of hypometabolic soft tissue injuries, such as Achilles tendon ruptures. II.


Localization of arabinogalactan proteins in egg cells, zygotes, and two-celled proembryos and effects of beta-D-glucosyl Yariv reagent on egg cell fertilization and zygote division in Nicotiana tabacum L.
Arabinogalactan proteins (AGPs) have been implicated in a variety of plant development processes including sexual plant reproduction. As a crucial developmental event, plant sexual reproduction generally occurs inside an ovule embedded in an ovary. The inaccessibility of the egg cells, zygotes, and embryos has hindered our understanding of the importance of AGPs in the early events involving fertilization, zygotic division, and early embryogenesis. In this study, the well-established in vitro zygote and ovary culture systems, together with immunofluorescence and immunogold labelling techniques, were employed to investigate the role of AGPs in the early events of sexual reproduction in Nicotiana tabacum. Dramatic changes in AGP content during ovule development were evidenced by western blotting. Subcellular localization revealed that AGPs are localized in the plasma membrane, cell wall, and cytoplasm of pre- and post-fertilized egg cells, and cytoplasm and vacuoles of two-celled proembryos. Abundant AGPs were detected in unfertilized egg cells; however, the level of AGPs substantially decreased in fertilized egg cells. Polar distribution of AGPs in elongated zygotes was observed. The early two-celled proembryos just from zygote division displayed accumulation of AGPs at a low level, while in the elongated two-celled proembryos at the late stage, the AGP content clearly increased. Provision of betaGlcY, a synthetic phenylglycoside that specifically binds AGPs, to the in vitro cultures of isolated zygote and fertilized ovaries increased abnormal symmetrical division of zygotes. In the culture of pollinated but unfertilized ovaries, addition of betaGlcY resulted in arrest of fertilization of the egg cells, but had no effect on fertilization of the central cells. The possible roles of AGPs in fertilization, zygotic division, and proembryo development are discussed.


Improved accuracy for radiation damping in coupled finite element/equivalent source computations.
In coupled structural-acoustic computations, radiation damping is due to the resistive component of the surface pressure created by structural vibrations. Equivalent sources using tripole sources as basis functions can be used to compute the surface pressure forces for exterior radiation problems. This technique is similar to the Burton and Miller method for eliminating numerical difficulties due to interior acoustic resonances in boundary element computations and has been proven to yield unique solutions. However, numerical computations presented here will show that for the specific equivalent source formulation under investigation, tripole sources overpredict the resistive component of the surface impedance, especially in the mid-to-high frequency range. It will also be shown that for frequency domain calculations, an accurate representation for the resistive component of the pressure forces can be derived from an analytical representation for the source radiation resistance. Unfortunately, this technique is not applicable to time domain computations. It is also shown that more accurate results can be obtained by allowing both the simple and dipole source amplitudes to be independent variables and enforcing boundary conditions in both the exterior and interior directions simultaneously to reduce the magnitude of the interior acoustic field.


Analysis of operating room allocations to optimize scheduling of specialty rotations for anesthesia trainees.
Because specialty workloads and corresponding operating room (OR) allocations vary among days of the week, anesthesia residents and student nurse anesthetists are sometimes assigned to cases off rotation (e.g., scheduled for cardiac surgery but assigned to urology for the day). We describe a method to create hybrid rotations of two specialties (e.g., cardiac and vascular surgery), thereby reducing the numbers of days that trainees are "pulled" from their scheduled rotations. Raw data were the number of hours of OR time used by each surgical specialty on each workday for the preceding 9 months. These OR workloads were converted to the number of ORs to be allocated to each specialty for each day of the week on the basis of maximization of the efficiency of use of OR time. We considered all potential hybrid rotations of pairwise combinations of specialties to which trainees could be assigned. Integer linear programming was used to calculate the maximum number of trainees who could be scheduled to hybrid rotations and receive daily assignments matching those rotations. Validity of the results was shown by using data from a small facility for which the optimal solution could be discerned by inspection. Validity (appropriateness) of the constraints was demonstrated by the exclusion of each constraint, resulting in answers that are obviously incorrect. Novelty and usefulness of the method was evidenced by its choosing from among hundreds of thousands of potential combinations of specialties and its identifying appropriate assignments that were substantively different from current rotations. We developed a methodology to determine rotations consisting of combinations of specialties to be paired for purposes of trainee scheduling to reduce the incidence of daily assignments off rotation. Practically, with this method, anesthesia residents and student nurse anesthetists can be assigned cases within their scheduled rotations as often as possible.


The effect of pedometer position and normal gait asymmetry on step count accuracy.
Manufacturers of pedometers recommend wearing a pedometer on the midline of the right thigh and this recommendation is used in research. However, there is conflicting evidence regarding the effect of pedometer position on accuracy. The purpose of this study was to systematically evaluate the effect of pedometer position on accuracy for 3 modes of gait. The Yamax SW200 pedometer was evaluated in 20 subjects in 5 different positions simultaneously: left mid-axillary, left mid-thigh, umbilical, right mid-thigh, and right mid-axillary. Each subject was asked to walk on a treadmill (54, 80, and 107 m.min(-1)), to walk overground (slow, preferred, and fast speeds), and to ascend and descend stairs. Pedometer steps were recorded and compared with actual steps observed. Using the same protocol, instantaneous triaxial accelerometry was used to explain position-dependent differences in accuracy. At slow speeds, the left mid-axillary position demonstrated the lowest net mean (+/-SD) error across all modes of gait (7.7% (+/-11.6%) overground;-0.6% (+/-2.2%) stairs). The right mid-thigh position had significantly (p<0.01) higher error for treadmill (18.1% (+/-17.3%)), overground (12.9% (+/-15.2%)), and stairs (2.9% (+/-3.9%)). Pedometer position dependent error was demonstrated, with the left mid-axillary position superior to the recommended position of right mid-thigh. The greater accuracy on the left side was wholly explained by gait asymmetry evident in step-induced accelerations recorded at right and left pedometer positions. A model of absolute error based on the ratio of steps at different gait speeds was generated to demonstrate the importance of this finding in pedometer-based lifestyle intervention studies.


An assessment of intermediate coronary care.
The effect of intermediate coronary care, with and without ECG monitoring, was compared with general medical ward care on the basis of mortality, resuscitation, and detection and treatment of arrhythmias from days 3 to 14 after admission in 2,095 cases of acute coronary heart disease. Mortality was significantly reduced (P less than .02), and number of successful resuscitations for ventricular fibrillation was increased (P less than .05) but only in the unit with monitoring. Number of arrhythmias detected was significantly increased, particularly incidence of ventricular ectopics and heart block (P less than .02). Number of arrhythmias corrected to sinus rhythm was increased, but not significantly. Death from pulmonary embolism fell (P less than .01). Review of causes of death and autopsies showed an increased proportion of deaths due to intractable heart failure and cardiogenic shock. Not only specially trained nurses, but also ECG monitoring, were necessary to obtain the benefits of this treatment.


The Historic Built Environment As a Long-Term Geochemical Archive: Telling the Time on the Urban "Pollution Clock".
This study introduces a novel methodology for utilizing historic built environments as reliable long-term geochemical archives, addressing a gap in the reconstruction of past anthropogenic pollution levels in urban settings. For the first time, we employ high-resolution laser ablation mass spectrometry for lead isotope (206Pb/207Pb and 208Pb/206Pb) analysis on 350-year-old black crust stratigraphies found on historic built structures, providing insights into past air pollution signatures. Our findings reveal a gradual shift in the crust stratigraphy toward lower 206Pb/207Pb and higher 208Pb/206Pb isotope ratios from the older to the younger layers, indicating changes in lead sources over time. Mass balance analysis of the isotope data shows black crust layers formed since 1669 primarily contain over 90% Pb from coal burning, while other lead sources from a set of modern pollution including but not limited to leaded gasoline (introduced after 1920) become dominant (up to 60%) from 1875 onward. In contrast to global archives such as ice cores that provide integrated signals of long-distance pollution, our study contributes to a deeper understanding of localized pollution levels, specifically in urban settings. Our approach complements multiple sources of evidence, enhancing our understanding of air pollution dynamics and trends, and the impact of human activities on urban environments.


Eradication of high viable loads of Listeria monocytogenes contaminating food-contact surfaces.
This study demonstrates the efficacy of cold gaseous ozone treatments at low concentrations in the eradication of high Listeria monocytogenes viable cell loads from glass, polypropylene, stainless steel, and expanded polystyrene food-contact surfaces. Using a step by step approach, involving the selection of the most resistant strain-surface combinations, 11 Listeria sp. strains resulted inactivated by a continuous ozone flow at 1.07 mg m(-3) after 24 or 48 h of cold incubation, depending on both strain and surface evaluated. Increasing the inoculum level to 9 log CFU coupon(-1), the best inactivation rate was obtained after 48 h of treatment at 3.21 mg m(-3) ozone concentration when cells were deposited onto stainless steel and expanded polystyrene coupons, resulted the most resistant food-contact surfaces in the previous assays. The addition of naturally contaminated meat extract to a high load of L. monocytogenes LMG 23775 cells, the most resistant strain out of the 11 assayed Listeria sp. strains, led to its complete inactivation after 4 days of treatment. To the best of our knowledge, this is the first report describing the survival of L. monocytogenes and the effect of ozone treatment under cold storage conditions on expanded polystyrene, a commonly used material in food packaging. The results of this study could be useful for reducing pathogen cross-contamination phenomena during cold food storage.


PhamClust: a phage genome clustering tool using proteomic equivalence.
Bacteriophage genomes are pervasively mosaic, presenting challenges to describing phage relatedness. Here, we describe PhamClust, a bioinformatic approach for phage genome comparisons that uses a new metric of proteomic equivalence quotient for comparative genomics. PhamClust reliably assorts genomes into groups or clusters of related phages and can subdivide clusters into subclusters. PhamClust is computationally efficient and can readily process thousands of phage genomes. It is also a useful analytic tool for exploring the different types of inter-genome relatedness characteristic of phages in different clusters.


Soap-film flow induced by electric fields in asymmetric frames.
Net fluid flow of soap films induced by (ac or dc) electric fields in asymmetric frames is presented. Previous experiments of controllable soap film flow required the simultaneous use of an electrical current passing through the film and an external electric field or the use of nonuniform ac electric fields. Here a single voltage difference generates both the electrical current going through the film and the electric field that actuates on the charge induced on the film. The film is set into global motion due to the broken symmetry that appears by the use of asymmetric frames. If symmetric frames are used, the film flow is not steady but time dependent and irregular. Finally, we study numerically these film flows by employing the model of charge induction in ohmic liquids.


FACTORS IN THE TESTING AND APPLICATION OF ALGICIDES.
A review is presented of some of the factors affecting the laboratory testing and practical applications of chemicals toxic to algae. The basic factor demonstrated is that the amount of chemical required to inhibit the growth of algae is dependent on the amount of algae present and not on the volume of water in which the algae are dispersed. It is shown how a chemical can be tested for algistatic or algicidal properties, thus enabling one to decide how best to apply a particular chemical. The selectivity of chemicals and the development of resistance in algae towards certain chemicals is demonstrated. Also, it is shown how certain algae can appear to be resistant to chemical treatments because of their growth habit or their production of extracellular products which affect the toxicity of added chemicals. With a better understanding of how various factors can influence the effectiveness of toxic chemicals, it is hoped that the selection of a chemical and method of application to a particular problem will be more successful.


Surface phenotype and Ig heavy-chain gene usage in chronic B-cell leukemias: expression of myelomonocytic surface markers in CD5- chronic B-cell leukemia.
We investigated the surface expression of leukocyte differentiation antigens and the Ig heavy-chain variable region (VH) gene family use in leukemic cells from 26 Japanese patients with chronic B-cell leukemias with special reference to CD5 antigen expression. CD5 was expressed on leukemic cells in 21 of 26 cases (CD5+) but not in 5 cases (CD5-). Myelomonocytic marker, CD13 antigen was expressed on the leukemic cells in all 5 CD5- cases but in none of CD5+ cases. Leukemic cells in CD5- cases also expressed CD11b antigen more frequently than those in CD5+ cases (80% v 11%; P < .01). Another myeloid marker, CD33, was expressed neither on CD5+ nor CD5- leukemic cells. CD22, a restricted B-cell marker, was expressed more frequently on CD5- leukemic cells than on CD5+ leukemic cells (80% v 33%; P < .05). Another restricted B-cell activation marker, CD23, was expressed at similar frequency in both the CD5+ and CD5- groups (67% v 60%). Although CD45RA was expressed on the majority of leukemic B cells, the CD45RA expression level was significantly higher among CD5- cases than CD5+ cases (P < .01). In the analysis of VH gene expressed in chronic B-cell leukemias by polymerase chain reaction amplification, CD5+ cases preferentially used VH4 family members (48%; 10 of 21). CD5- cases, on the other hand, mainly used VH3 family (80%; 4 of 5). Thus, from our present observation of an albeit limited patient population, we have found an association between VH gene family use and CD5 antigen expression in chronic B-cell leukemias. We have also shown the differential expression of myelomonocytic markers in the CD5+ and CD5- chronic B-cell leukemias. These result are in agreement with previous suggestions that CD5 positivity is the hallmark for distinct clinical entity commonly referred to in the literature as chronic lymphocytic leukemia.


Facile Synthesis of Kwakhurin, a Marker Compound of Pueraria mirifica and Its Quantitative NMR Analysis for Standardization as a Reagent.
The side effects of kwao keur dietary supplements (obtained from the tuberous root of Pueraria mirifica) have recently been reported by the Ministry of Health, Labour and Welfare, Japan. To control the quality of kwao keur products, its ingredients need to be maintained by characteristic marker compounds, such as miroestrol, deoxymiroestrol, and kwakhurin (KWA). In this study, we described the facile synthesis of KWA, a marker compound of P. mirifica. Our revised synthetic method produced KWA with shorter steps and higher yield than the reported method. Furthermore, the absolute purity of KWA was determined by quantitative NMR analysis for standardization as a reagent, and its purity was 92.62 ± 0.12%.


Interferon-Free Sofosbuvir-Based Anti-HCV Therapy After Liver Transplantation.
Therapy for HCV-infected patients after orthotopic liver transplantation (OLT) is based on interferon (IFN) as the gold standard, but sustained virologic response (SVR) and safety profiles of the IFN-based therapies are very unsatisfactory. The aim of this continuing analysis is evaluation of the impact of an IFN-free sofosbuvir (SOF)-based therapy in HCV-infected liver transplant recipients. Post-OLT patients with a proven recurrence of HCV were treated with SOF and ribavirin (RBV) for 24 weeks (n=10). Laboratory parameters and FibroScan® are continuously evaluated at weeks 0, 12, 24, and 36. A retrospectively analyzed HCV patient cohort who received antiviral therapy with pegylated INF and RBV± telaprevir (TLV) were used as a control group. All patients who finished their treatment with SOF/RBV at least 12 weeks ago showed an SVR. The SOF-based therapy showed a significantly higher rate of rapid virologic response (RVR) and sustained virologic response (SVR) compared to the IFN-based therapies (RVR: p=0.007; SVR: p=0.009). According to temporary data on FibroScan® analysis, regression of fibrosis was observed in 8 patients treated with SOF/RBV. No premature termination of SOF became necessary. In this small group of patients, the preliminary results indicate that a regression of fibrosis is achievable within 24 weeks of therapy with SOF after OLT. SOF seems to be effective and safe in the treatment of OLT patients infected with HCV and will likely improve patient and transplant survival.


A structured viroid RNA serves as a substrate for dicer-like cleavage to produce biologically active small RNAs but is resistant to RNA-induced silencing complex-mediated degradation.
RNA silencing is a potent means of antiviral defense in plants and animals. A hallmark of this defense response is the production of 21- to 24-nucleotide viral small RNAs via mechanisms that remain to be fully understood. Many viruses encode suppressors of RNA silencing, and some viral RNAs function directly as silencing suppressors as counterdefense. The occurrence of viroid-specific small RNAs in infected plants suggests that viroids can trigger RNA silencing in a host, raising the question of how these noncoding and unencapsidated RNAs survive cellular RNA-silencing systems. We address this question by characterizing the production of small RNAs of Potato spindle tuber viroid (srPSTVds) and investigating how PSTVd responds to RNA silencing. Our molecular and biochemical studies provide evidence that srPSTVds were derived mostly from the secondary structure of viroid RNAs. Replication of PSTVd was resistant to RNA silencing, although the srPSTVds were biologically active in guiding RNA-induced silencing complex (RISC)-mediated cleavage, as shown with a sensor system. Further analyses showed that without possessing or triggering silencing suppressor activities, the PSTVd secondary structure played a critical role in resistance to RISC-mediated cleavage. These findings support the hypothesis that some infectious RNAs may have evolved specific secondary structures as an effective means to evade RNA silencing in addition to encoding silencing suppressor activities. Our results should have important implications in further studies on RNA-based mechanisms of host-pathogen interactions and the biological constraints that shape the evolution of infectious RNA structures.


Association of accelerated body mass index gain with repeated measures of blood pressure in early childhood.
We examined the association for rates of age- and sex-standardized body mass index (zBMI) gain between 0-3, 3-18, and 18-36 months with BP in children at 36-72 months of age. We collected repeated measures of zBMI and BP in 2502 children. zBMI was calculated using the World Health Organization standards. Each child's zBMI at birth and rates of zBMI gain in each period from birth to 36 months were estimated using linear spline multilevel models. Generalized estimating equations were used to determine whether zBMI at birth and zBMI gain between 0-3, 3-18, and 18-36 months were each associated with repeated measures of BP at 36-72 months of age. We sequentially conditioned on zBMI at birth and zBMI gain in each period prior to each period tested, as covariates, and adjusted for important socio-demographic, familial, and study design covariates. We examined whether these associations were modified by birthweight or maternal obesity, by including interaction terms. After adjusting for all covariates and conditioning on prior zBMI gains, a 1 standard deviation unit faster rate of zBMI gain during 0-3 months, (β = 0.59 mmHg; 95% CI 0.31, 0.86) and 3-18 months (β = 0.74 mmHg; 95% CI 0.46, 1.03) were each associated with higher systolic BP at 36-72 months. No significant associations were observed, however, for zBMI at birth or zBMI gain in the 18-36 month growth period. zBMI gains from 0-3 and 3-18 months were also associated with diastolic BP. Birthweight significantly modified the relationship during the 3-18 month period (p = 0.02), with the low birthweight group exhibiting the strongest association for faster rate of zBMI gain with higher systolic BP (β = 1.31 mmHg; 95% CI 0.14, 2.48). Given that long-term exposure to small elevations in BP are associated with subclinical cardiovascular disease, promoting interventions targeting healthy growth in infancy may be important.


Glycerol kinase activities in muscles from vertebrates and invertebrates.
1. Glycerol kinase (EC 18.104.22.168) activity was measured in crude extracts of skeletal muscles by a radiochemical method. The properties of the enzyme from a number of different muscles are very similar to those of the enzyme from rat liver. Glycerol kinase from locust flight muscle was inhibited competitively by l-3-glycerophosphate with a K(i) of 4.0x10(-4)m. 2. The activity of glycerol kinase was measured in a variety of muscles from vertebrates and invertebrates in an attempt to explain the large variation in the activity of this enzyme in different muscles. 3. In vertebrates glycerol kinase activities were generally higher in red muscle than in white muscle; the highest activities (approx. 0.2mumole/min./g. fresh wt.) were found in the red breast muscle of some birds (e.g. pigeon, duck, blue tit) whereas the activities in the white breast muscle of the pheasant and domestic fowl were very low (approx. 0.02mumole/min./g.). 4. On the basis of glycerol kinase activities, muscles from insects can be classified into three groups: muscles that have a low enzyme activity, i.e. <0.3mumole/min./g. (leg muscles of all insects studied and the flight muscles of cockroaches and the tsetse fly); muscles that have an intermediate enzyme activity, i.e. 0.3-1.5mumoles/min./g. (e.g. locusts, cockchafers, moths, water-bugs); and muscles that have a high enzyme activity, i.e. >1.5mumoles/min./g. (e.g. bees, wasps, some blowflies). 5. The function of glycerol kinase in vertebrate and insect muscles that possess a low or intermediate activity is considered to be the removal of glycerol that is produced from lipolysis of triglyceride or diglyceride by the muscle. Therefore in these muscles the activity of glycerol kinase is related to the metabolism of fat, which is used to support sustained muscular activity. A possible regulatory role of glycerol kinase in the initiation of triglyceride or diglyceride lipolysis is discussed. 6. The function of glycerol kinase in the insect muscles that possess a high activity of the enzyme is considered to be related to the high rates of glycolysis that these muscles can perform. The oxidation of extramitochondrial NADH, and therefore the maintenance of glycolysis, is dependent on the functioning of the glycerophosphate cycle; if at any stage of flight (e.g. at the start) the rate of mitochondrial oxidation of l-3-glycerophosphate was less than the activity of the extramitochondrial glycerophosphate dehydrogenase, this compound would accumulate, inhibit the latter enzyme and inhibit glycolysis. It is suggested that such excessive accumulation of l-3-glycerophosphate is prevented by hydrolysis of this compound to glycerol; the latter would have to be removed from the muscle when the accumulation of l-3-glycerophosphate had stopped, and this would explain the presence of glycerol kinase in these muscles and its inhibition by l-3-glycerophosphate.


Evaluating risk for alcohol use disorder: Polygenic risk scores and family history.
Early identification of individuals at high risk for alcohol use disorder (AUD) coupled with prompt interventions could reduce the incidence of AUD. In this study, we investigated whether Polygenic Risk Scores (PRS) can be used to evaluate the risk for AUD and AUD severity (as measured by the number of DSM-5 AUD diagnostic criteria met) and compared their performance with a measure of family history of AUD. We studied individuals of European ancestry from the Collaborative Study on the Genetics of Alcoholism (COGA). DSM-5 diagnostic criteria were available for 7203 individuals, of whom 3451 met criteria for DSM-IV alcohol dependence or DSM-5 AUD and 1616 were alcohol-exposed controls aged ≥21 years with no history of AUD or drug dependence. Further, 4842 individuals had a positive first-degree family history of AUD (FH+), 2722 had an unknown family history (FH?), and 336 had a negative family history (FH-). PRS were derived from a meta-analysis of a genome-wide association study of AUD from the Million Veteran Program and scores from the problem subscale of the Alcohol Use Disorders Identification Test in the UK Biobank. We used mixed models to test the association between PRS and risk for AUD and AUD severity. AUD cases had higher PRS than controls with PRS increasing as the number of DSM-5 diagnostic criteria increased (p-values ≤ 1.85E-05 ) in the full COGA sample, the FH+ subsample, and the FH? subsample. Individuals in the top decile of PRS had odds ratios (OR) for developing AUD of 1.96 (95% CI: 1.54 to 2.51, p-value = 7.57E-08 ) and 1.86 (95% CI: 1.35 to 2.56, p-value = 1.32E-04 ) in the full sample and the FH+ subsample, respectively. These values are comparable to previously reported ORs for a first-degree family history (1.91 to 2.38) estimated from national surveys. PRS were also significantly associated with the DSM-5 AUD diagnostic criterion count in the full sample, the FH+ subsample, and the FH? subsample (p-values ≤6.7E-11 ). PRS remained significantly associated with AUD and AUD severity after accounting for a family history of AUD (p-values ≤6.8E-10 ). Both PRS and family history were associated with AUD and AUD severity, indicating that these risk measures assess distinct aspects of liability to AUD traits.


A research feasibility assessment of regulatory information systems capacities for investigation of epidemiologic factors in pseudorabies.
Pseudorabies (PR) outbreak-investigation forms from 10 states having the most PR-infected herds were evaluated for agreement in question response-data type, information intent, and outbreak information categories. A question randomly selected from an investigation form had 0.6304 probability of being unique to a single state, and 0.0062 probability of being common to all states. Analysis of outbreak forms, on the basis of information intent, revealed that the probability of a randomly selected question being derived from an information category unique to a single state was 0.0323, whereas the probability of a question being derived from an information category shared by all states was 0.1935. A telephone survey revealed that state PR control officials did not believe additional research on between-herd spread of PR was necessary to successfully complete the eradication program. However, officials believed a better understanding of PR risk factors would enhance program effectiveness and build producer confidence.


Melatonin alleviates lung injury in H1N1-infected mice by mast cell inactivation and cytokine storm suppression.
Influenza A virus (IAV) H1N1 infection is a constant threat to human health and it remains so due to the lack of an effective treatment. Since melatonin is a potent antioxidant and anti-inflammatory molecule with anti-viral action, in the present study we used melatonin to protect against H1N1 infection under in vitro and in vivo conditions. The death rate of the H1N1-infected mice was negatively associated with the nose and lung tissue local melatonin levels but not with serum melatonin concentrations. The H1N1-infected AANAT-/- melatonin-deficient mice had a significantly higher death rate than that of the WT mice and melatonin administration significantly reduced the death rate. All evidence confirmed the protective effects of melatonin against H1N1 infection. Further study identified that the mast cells were the primary targets of melatonin action, i.e., melatonin suppresses the mast cell activation caused by H1N1 infection. The molecular mechanisms involved melatonin down-regulation of gene expression for the HIF-1 pathway and inhibition of proinflammatory cytokine release from mast cells; this resulted in a reduction in the migration and activation of the macrophages and neutrophils in the lung tissue. This pathway was mediated by melatonin receptor 2 (MT2) since the MT2 specific antagonist 4P-PDOT significantly blocked the effects of melatonin on mast cell activation. Via targeting mast cells, melatonin suppressed apoptosis of alveolar epithelial cells and the lung injury caused by H1N1 infection. The findings provide a novel mechanism to protect against the H1N1-induced pulmonary injury, which may better facilitate the progress of new strategies to fight H1N1 infection or other IAV viral infections.


Involvement of c-Jun NH2-terminal kinase activation and c-Jun in the induction of apoptosis by the ether phospholipid 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine.
The ether phospholipid 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH3; edelfosine) is a potent inducer of apoptosis in human tumor cells. We show that ET-18-OCH3-induced apoptosis is associated with activation of the c-Jun NH2-terminal kinase (JNK) signaling. The addition of ET-18-OCH3 to distinct human leukemic cells (HL-60, U937, and Jurkat), which undergo rapid apoptosis on treatment with ET-18-OCH3, induced a dramatic and sustained increase in the of c-jun mRNA level that was associated with activation of activator protein-1 transcription factor. We found that ET-18-OCH3 induced a persistent activation of JNK in HL-60 cells that was detected before the onset of apoptosis, the latter being assessed by DNA fragmentation and by the appearance of phosphatidylserine on the external leaflet of the plasma membrane. The inductions of JNK after HL-60 monocyte/macrophage differentiation and ET-18-OCH3-mediated apoptosis were distinguished by the different activation patterns, transient versus persistent, respectively. ET-18-OCH3 analogues unable to induce apoptosis failed to activate JNK. ET-18-OCH3-dependent JNK activation was not detected in K562 cells, which did not undergo apoptosis on treatment with ET-18-OCH3. Phorbol myristate acetate inhibited both ET-18-OCH3-induced apoptosis and sustained JNK activation; thus, persistent JNK activation by ET-18-OCH3 is associated with the capacity of this ether phospholipid to induce apoptosis. Furthermore, antisense oligonucleotides directed against c-jun blocked ET-18-OCH3-induced apoptosis, indicating a role for c-Jun in this apoptotic response. These data indicate that JNK activation and c-Jun are involved in the induction of apoptosis by ET-18-OCH3.


Understanding the Subjective Experience of Long-term Remote Measurement Technology Use for Symptom Tracking in People With Depression: Multisite Longitudinal Qualitative Analysis.
Remote measurement technologies (RMTs) have the potential to revolutionize major depressive disorder (MDD) disease management by offering the ability to assess, monitor, and predict symptom changes. However, the promise of RMT data depends heavily on sustained user engagement over extended periods. In this paper, we report a longitudinal qualitative study of the subjective experience of people with MDD engaging with RMTs to provide insight into system usability and user experience and to provide the basis for future promotion of RMT use in research and clinical practice. We aimed to understand the subjective experience of long-term engagement with RMTs using qualitative data collected in a longitudinal study of RMTs for monitoring MDD. The objectives were to explore the key themes associated with long-term RMT use and to identify recommendations for future system engagement. In this multisite, longitudinal qualitative research study, 124 semistructured interviews were conducted with 99 participants across the United Kingdom, Spain, and the Netherlands at 3-month, 12-month, and 24-month time points during a study exploring RMT use (the Remote Assessment of Disease and Relapse-Major Depressive Disorder study). Data were analyzed using thematic analysis, and interviews were audio recorded, transcribed, and coded in the native language, with the resulting quotes translated into English. There were 5 main themes regarding the subjective experience of long-term RMT use: research-related factors, the utility of RMTs for self-management, technology-related factors, clinical factors, and system amendments and additions. The subjective experience of long-term RMT use can be considered from 2 main perspectives: experiential factors (how participants construct their experience of engaging with RMTs) and system-related factors (direct engagement with the technologies). A set of recommendations based on these strands are proposed for both future research and the real-world implementation of RMTs into clinical practice. Future exploration of experiential engagement with RMTs will be key to the successful use of RMTs in clinical care.


Assessment and management of acute coronary syndromes (ACS): a Canadian perspective on current guideline-recommended treatment--part 1: non-ST-segment elevation ACS.
Despite the reduction of coronary heart disease mortality over the past 40 years, hospital admissions for acute coronary syndromes (ACS) continue to increase. The goal of this 2-part article is to review the issues at each stage of assessment and management of the ACS patient, and to propose an optimal treatment strategy for the individual patient in the context of the realities, culture, and delivery of healthcare in Canada. ACS patients are categorized as either ST segment elevation myocardial infarction (STEMI) or non-ST-elevation ACS (NSTE-ACS). For the patients with NSTE-ACS, prevention of recurrent ischemic events is the primary goal. Assessment of risk for recurrent ischemic and bleeding events helps to determine the net benefit of early cardiac catheterization and percutaneous coronary intervention (PCI) and intensive antiplatelet and anticoagulant treatment. Those with higher ischemic risk features should be considered for an early invasive strategy and receive both dual antiplatelet therapy and an anticoagulant at the time of first medical assessment. Patients without high-risk features could be considered for medical treatment and a selectively invasive strategy; with coronary angiography and revascularization only if high-risk features become apparent. Long-term vascular protection with lifestyle modification (especially smoking cessation), lipid lowering, blood pressure and glycemic control, and the use of renin angiotensin aldosterone system (RAAS) blockade to prevent recurrent ischemic events, is important in all patients with ACS.


Inhaled tobacco sterols: uptake by the lungs and disposition to selected organs of rats.
Tobacco sterols (cholesterol, beta-sitosterol, campesterol, and stigmasterol) are present in tobacco smoke and appear in plasma of mammals exposed to cigarette smoke. Because tobacco sterols may be important in the pathogenesis of smoking-induced lung and vascular diseases, we studied the pattern of deposition of cigarette sterols in the lungs and appearance of cigarette sterols in plasma and body organs of rats. After exposure to twenty 5 ml "puffs" of smoke from tobacco labeled with [4-14C]cholesterol or beta-[4-14C]sitosterol, rats were killed just after exposure (day 0) and on days 2, 5, 8, 11, 15, and 30, and the lungs and selected body organs analyzed for activity. We found that cigarette sterols are associated with particulates in cigarette smoke, deposited mostly in distal airspaces and parenchyma of the lungs, and appear in plasma and several body organs for more than 30 days after this single exposure to cigarette smoke. Bronchoalveolar lavage fluid contained relatively small amounts of radiolabel for only the first few days, suggesting that most of the sterols were rapidly incorporated in lung parenchyma. Because disorders of sterol metabolism have been implicated in a variety of diseases including atherosclerosis and cancer, the significance of tobacco sterols to human smoking-induced diseases deserves further study.


Polybromo-1: the chromatin targeting subunit of the PBAF complex.
The human Polybromo-1 protein (Pb1) was recently identified as a unique subunit of the PBAF (Polybromo, Brg1-Associated Factors) chromatin-remodeling complex required for kinetochore localization during mitosis and the transcription of estrogen-responsive genes. Pb1 coordinates key features common to all remodeling complexes, including chromatin localization, recruitment of protein subunits and alteration of chromatin architecture. A comprehensive analysis of individual domains composing Pb1 is used to propose new information regarding the function of Pb1 in the PBAF chromatin-remodeling complex. The newly identified regulatory role of this important protein is also examined to explain both native function and the emerging role of Pb1 as a tumor suppressor found to be mutated in breast cancer.


Individualized Medicine in Gastroenterology and Hepatology.
After the completion of the Human Genome Project, there has been an acceleration in methodologies on sequencing nucleic acids (DNA and RNA) at a high precision and with ever-decreasing turnaround time and cost. Collectively, these approaches are termed next-generation sequencing and are already affecting the transformation of medical practice. In this symposium article, we highlight the current knowledge of the genetics of selected gastrointestinal tract and liver diseases, namely, inflammatory bowel disease, hereditary cholestatic liver disease, and familial colon cancer syndromes. In addition, we provide a stepwise approach to use next-generation sequencing methodologies for clinical practice with the goal to improve the diagnosis as well as management of and/or therapy of the chosen digestive diseases. This early experience of applying next-generation sequencing in the practice of gastroenterology and hepatology will delineate future best practices in the field, ultimately for the benefit of our patients.


Increased mitogenic activity of scleroderma serum: inhibitory effect of human recombinant interferon-gamma.
To investigate the role of platelet activation in the development of systemic sclerosis and the role of interferon-gamma (IFN gamma) in the inhibition of mitogenic activity induced by whole blood serum of patients with systemic sclerosis. The mitogenic activity of whole blood serum in the absence or presence of different concentrations of IFN gamma (a potent inhibitor of induced collagen synthesis in dermal fibroblasts) and platelet-poor plasma derived serum were tested on human dermal fibroblasts by measuring incorporation of [3H]thymidine. Platelet activation was determined by quantification of plasma beta-thromboglobulin (beta-TG) using a beta-TG radioimmunoassay kit. The mitogenic activity was significantly increased in whole blood serum and in platelet-poor plasma derived serum of the patients compared with controls. In contrast, no significant increase in beta-TG concentration was observed in scleroderma platelet-poor plasma compared with control. Recombinant human IFN gamma had a greater inhibitory effect on the mitogenic activity induced by whole blood serum of patients than on that produced with control sera, at any concentration of IFN gamma tested. Our results suggest that mitogenic activity observed in the plasma of sclerodermic patients could originate from cells other than platelets and could be involved in the development of fibrosis. The potent inhibitory effect of IFN gamma on this proliferative activity may account for the beneficial effect of this cytokine in the treatment of progressive systemic sclerosis.


Symptoms associated with inflammatory arthritis are common in the primary care population: results from the joint symptoms survey.
To describe the prevalence of self-reported inflammatory joint symptoms, such as joint pain, stiffness and swelling, in UK primary care patients consulting for both musculoskeletal (MSK) and non-musculoskeletal (non-MSK) complaints. A joint symptoms questionnaire survey was sent to 10 161 individuals, of whom 5050 had consulted for MSK problems. These were matched by age, gender and general practice to non-MSK consulters. Participants provided data on relevant symptoms such as joint pain, stiffness and swelling. The prevalence of these symptoms, their severity and impact were compared between MSK and non-MSK consulters. A total of 4549 adults responded to the survey (adjusted response 45.8%) of whom 52.3% consulted for a MSK problem. The mean (s.d.) age was 61.6 (14.8) years and 58.9% were female. Persistent (on at least half of the days in the last month) inflammatory symptoms were common even in non-MSK consulters, with 42% reporting joint pain, 36% reporting joint stiffness and 18% reporting joint swelling. This is in comparison with 62% reporting joint pain, 50% stiffness and 24% swelling among MSK consulters. Although symptoms such as persistent joint pain, swelling and stiffness are predictive of inflammatory arthritis, large numbers of people consulting primary care for non-MSK reasons report these symptoms when asked by questionnaire. This compounds the challenges of diagnosing inflammatory arthritis in a non-specialist setting where new approaches are needed to ensure accurate, early diagnosis, facilitating a treat-to-target approach.


Epidemiology of human papillomavirus-associated anogenital cancers in Granada: a three-decade population-based study.
HPV infection is a common risk factor for all anogenital cancers. However, there are important differences in the epidemiology of anogenital cancers and these have not been compared considering diverse epidemiological indicators over a long period of time. To fill this gap, we investigated incidence, mortality, and survival trends of anogenital cancers over a period of three decades. We conducted an observational registry-based study using data from the population-based cancer registry of Granada in southern Spain. We collected data on all incident cases of anogenital cancer (cervical, anal, penile, vulvar, and vaginal cancer) diagnosed between 1985 and 2017. We calculated crude and age-standardized incidence and mortality rates, and 1, 3, and 5-year overall and net survival. We further conducted time-trend analysis calculating annual percent changes (APC) for each cancer site. The incidence of anogenital cancers decreased slightly during the past 30 years, with the exception of vulvar cancer, where a slight increase was observed. Mortality decreased significantly for cervical cancer over the study period but increased non-significantly for the remaining cancer sites. Survival rates were similar to those reported in comparable countries and increased for cervical and vulvar cancer. Cervical cancer was the greatest contributor to the burden of anogenital cancers and showed a marked improvement in all indicators in comparison to the remaining cancer sites.


Involvement of proteases in apoptosis.
Genetically programmed (apoptotic) cell death plays a key role in cell and tissue homeostasis and in pathogenesis of various diseases. However, the mechanisms involved in apoptotic cell death are poorly understood. At present, the role of proteases in key events of apoptosis is intensively studied and discussed and the involvement of various proteolytic enzymes in the induction and development of the cell death is well-recognized. Proteases of various classes participating in apoptosis have been identified as well as some substrates of these proteases whose cleavage is critical to cell viability; specific protease inhibitors which prevent the cell death have been synthesized. This review summarizes new data on proteolytic enzymes involved in apoptosis and considers the mechanisms of activation of proteases upon induction of apoptosis and the pathways of their involvement in the cell death. The participation of nuclear proteolytic enzymes in the destabilization of chromatin structure and regulation of DNA fragmentation by endonucleases in apoptotic cells is discussed.


Natural history and predictors of obesity after orthotopic heart transplantation.
Excessive weight gain resulting in obesity is commonly seen after orthotopic heart transplantation. Obesity increases the risk for the development of many significant health problems and the associated morbidity and mortality. The purpose of this study was to define the occurrence, magnitude, and predictors of overweight/obesity in this group. We followed 47 consecutive patients for 1 year after orthotopic heart transplantation for changes in weight. Weight gain from baseline was significant at months 2, 4, 6, 8, and 10, with significant weight increments between each measurement interval (p < 0.05). The mean weight gain was 10.1 kg +/- 1.6 (standard error [SE]) at 12 months. Based on entry body mass index patients were divided into two groups: group I, overweight/obese, body mass index more than 27 kg/m2 (n = 11); group II, not overweight/obese, body mass index 27 kg/m2 or less (n = 36). No significant difference was found in the amount of weight gained between the two groups (group I, 10.3 kg +/- 1.2 [SE]; group II, 10.1 kg +/- 1.4 [SE]). Only age predicted weight gain, with younger patients gaining more than older patients after transplantation (14.2 +/- 13.2 kg versus 8.8 +/- 7.9 kg; p < 0.05). We found no relationship between the observed weight gain and any of the other measured predictors, that is, history of overweight, age, diabetes, family history of overweight, sex, patient's participation in cardiac rehabilitation. The universal nature of this weight gain and the lack of markers predicting patients at greatest risk for obesity underscores the seriousness of this problem.


Incidence and outcome of early Candida peritonitis after liver and pancreas transplantation.
Candida peritonitis is a potentially life-threatening infection after abdominal transplantation, although there is scant information regarding its incidence and outcome. We analysed the incidence rate and outcome of Candida peritonitis in 717 liver or pancreas transplant recipients. Five cases of Candida peritonitis were diagnosed, representing the second most frequent cause of invasive fungal infection in the cohort. The incidence rate of Candida peritonitis during the first 30 days after transplantation was 6.5 cases/10 000 transplant days in pancreas recipients and 1.2 cases/10 000 transplant days in liver recipients (P = 0.035). Four of the five patients received an echinocandin in combination with other antifungal. All patients were alive and with good graft function at 1-year follow-up. In our series, Candida peritonitis in liver and pancreas transplant recipients was not uncommon and had a good prognosis.


DHX33 Transcriptionally Controls Genes Involved in the Cell Cycle.
The RNA helicase DHX33 has been shown to be a critical regulator of cell proliferation and growth. However, the underlying mechanisms behind DHX33 function remain incompletely understood. We present original evidence in multiple cell lines that DHX33 transcriptionally controls the expression of genes involved in the cell cycle, notably cyclin, E2F1, cell division cycle (CDC), and minichromosome maintenance (MCM) genes. DHX33 physically associates with the promoters of these genes and controls the loading of active RNA polymerase II onto these promoters. DHX33 deficiency abrogates cell cycle progression and DNA replication and leads to cell apoptosis. In zebrafish, CRISPR-mediated knockout of DHX33 results in downregulation of cyclin A2, cyclin B2, cyclin D1, cyclin E2, cdc6, cdc20, E2F1, and MCM complexes in DHX33 knockout embryos. Additionally, we found the overexpression of DHX33 in a subset of non-small-cell lung cancers and in Ras-mutated human lung cancer cell lines. Forced reduction of DHX33 in these cancer cells abolished tumor formation in vivo Our study demonstrates for the first time that DHX33 acts as a direct transcriptional regulator to promote cell cycle progression and plays an important role in driving cell proliferation during both embryo development and tumorigenesis.