Histologic features distinguish microsatellite-high from microsatellite-low and microsatellite-stable colorectal carcinomas, but do not differentiate germline mutations from methylation of the MLH1 promoter.
The detection of microsatellite-unstable (microsatellite instability [MSI]) colorectal carcinomas (CRCs) has prognostic value and can help screen for Lynch syndrome. We determined which histologic features are associated with MSI status and presence of germline mutation and/or methylation of MLH1 promoter. Patients diagnosed with CRC were offered participation in the Columbus-area hereditary nonpolyposis colorectal cancer syndrome study regardless of age or family history. Tumors were evaluated for MSI using a modified Bethesda panel of microsatellite markers. Methylation status of the MLH1 promoter was evaluated by methylation-specific polymerase chain reaction and bisulfite PCR followed by restriction digestion of tumor DNA. All patients with microsatellite-unstable tumors underwent mutation analysis of the MLH1, MSH2, and MSH6 genes by full sequencing of genomic DNA and by multiplex ligation probe assay of MLH1 and MSH2. Histologic end points were tumor type, grade, percentage of mucin, border, and lymphoid host response. Of the 482 CRCs, 87 were MSI with 69 MSI high (MSI-H), 18 MSI low (MSI-L), and 395 microsatellite stable (MSS). Of 87 MSI tumors, 12 had germline mutations and 34 had methylation of the MLH1 promoter. Younger age, but not histologic features, was significantly associated with a germline mutation. Percentage of mucin, histologic type, grade, and lymphoid host response differed significantly between MSI-H when compared with MSI-L or MSS. No difference was found between MSI-L versus MSS. Histologic features are associated with MSI-H CRC and are helpful to differentiate MSI-H from MSI-L and MSS. These features are not useful to distinguish MSI-L from MSS carcinomas, and those with a deleterious germline hereditary nonpolyposis colorectal cancer syndrome mutation from those with methylation of the MLH1 promoter region.


Sex differences in the development of malignancies among end-stage renal disease patients: a nationwide population-based follow-up study in Taiwan.
Increasing evidence indicates that end-stage renal disease (ESRD) is associated with the morbidity of cancer. However, whether different dialysis modality and sex effect modify the cancer risks in ESRD patients remains unclear. A total of 3,570 newly diagnosed ESRD patients and 14,280 controls matched for age, sex, index month, and index year were recruited from the National Health Insurance Research Database in Taiwan. The ESRD status was ascertained from the registry of catastrophic illness patients. The incidence of cancer was identified through cross-referencing with the National Cancer Registry System. The Cox proportional hazards model and the Kaplan-Meier method were used for analyses. A similar twofold increase in cancer risk was observed among ESRD patients undergoing hemodialysis (HD) or peritoneal dialysis (PD) after adjusting for other potential risk factors. Patients with the highest cancer risk, approximately fourfold increased risk, were those received renal transplants. Urothelial carcinoma (UC) had the highest incidence in HD and PD patients. However, renal cell carcinoma (RCC) had the highest incidence in the renal transplantation (RT) group. In addition, female patients undergoing RT or PD had a higher incidence of RCC and UC, respectively. Male patients under HD had both higher incidence of RCC and UC. In conclusion, different dialysis modality could modify the cancer risks in ESRD patients. We also found sex effect on genitourinary malignancy when they are under different dialysis modality.


[Spatiotemporal expression pattern of E-cadherin and P-cadherin during mouse tooth development].
To investigate the expression patterns of E-cadherin and P-cadherin in murine-tooth germs at early developmental stages. Mandible samples of CD1 mice from embryonic day 12.5 to postnatal day 3.5 were collected. The expressions of E-cadherin and P-cadherin in murine mandibular first molar germs were detected by immunofluorescence and observed under confocal fluorescence microscope. HE staining was performed for tissue morphology. Both E-cadherin and P-cadherin were widely expressed in the epithelial tissues through early developmental stages. The E-cadherin expression was increased in polarizing pre-ameloblasts, whereas the P-cadherin expression declined. The expression of the P-cadherin could be detected in epithelial tissues before bud stage, and expressed in mature ameloblasts at secretory stage. The E-cadherin and P-cadherin expressed in different spatiotemporal expression patterns, indicating their individual functions during tooth development. P-cadherin might function in the secretion and mineralization of enamel.


Remodeling of nasal mucosa by allergen exposure in guinea pigs is suppressed by steroid and pranlukast.
It is unclear whether remodeling exists in allergic rhinitis in man. The aim of this study was to establish a guinea pig model of allergic rhinitis with remodeling and to examine the effects of dexamethasone and pranlukast on nasal mucosa remodeling. In the first experiment, three groups of ovalbumin-sensitized Hartley guinea pigs received intranasal challenges with ovalbumin for 1, 8, 12 weeks, respectively. In the second experiment, to examine the effect of dexamethasone and pranlukast, the animals were divided into 4 groups: negative control group; ovalbumin-sensitized group; ovalbumin + dexamethasone group; and ovalbumin + pranlukast group. During 12 weeks of intranasal exposure to ovalbumin, the latter two groups received daily intraperitoneal injections of dexamethasone and pranlukast, respectively. In the first experiment, in contrast to the negative control group, the ovalbumin-sensitized group exhibited significant goblet cell hyperplasia, epithelial damage and deposition of extracellular matrix in the nasal septal mucosa and conchae. In the second experiment, these changes were significantly inhibited by dexamethasone and pranlukast, respectively. We have established a model of upper airway remodeling in guinea pigs. The tissue remodeling was inhibited by early intervention with the antiallergic-inflammatory agents dexamethasone and pranlukast.


Unilateral multicystic dysplastic kidney: long term outcomes.
To report the long term follow up of children with antenatally detected unilateral multicystic dysplastic kidney (MCDK) with documentation of complications, involution rate with time, and renal function at 10 years. Data were retrieved from a prospective regional registry of patients with MCDK between 1985 and 2004. Children were followed using a common protocol of investigation with follow up ultrasound scans (USS) at 2 (165 patients), 5 (117 patients), and 10 years (43 patients). Serial USS showed that 33% of the MCDK kidneys had completely involuted at 2 years of age, 47% at 5 years, and 59% at 10 years. No patients developed hypertension, significant proteinuria, or malignancy, but two developed pelviureteric junction obstruction in the contralateral kidney. Twenty seven of 143 children (19%) had vesicoureteric reflux (VUR) (96% mild to moderate VUR) into the contralateral kidney with no difference in the incidence of urinary tract infections or renal scarring between those with or without VUR. The mean estimated glomerular filtration rate (GFR) was 86.4 ml/min/1.73 m2 (range 48-125) in 31 of 43 patients followed to 10 years. Conservative management of unilateral MCDK is justified with clinical review and infrequent USS but longer term follow up continues in the 41% still with renal remnants at 10 years and those with impaired GFR. It is suggested that the initial micturating cystogram is deferred unless abnormal USS features are present in the contralateral kidney or ureter.


Evolution and Expansion of the Prokaryote-Like Lipoxygenase Family in the Brown Alga Saccharina japonica.
Lipoxygenase (LOX) plays important roles in fatty acid oxidation and lipid mediator biosynthesis. In this study, we give first insights into brown algal LOX evolution. Whole genome searches revealed four, three, and eleven LOXs in Ectocarpus siliculosus, Cladosiphon okamuranus, and Saccharina japonica, respectively. In phylogenetic analyses, LOXs from brown algae form a robust clade with those from prokaryotes, suggesting an ancestral origin and slow evolution. Brown algal LOXs were divided into two clades, C1 and C2 in a phylogenetic tree. Compared to the two species of Ectocarpales, LOX gene expansion occurred in the kelp S. japonica through tandem duplication and segmental duplication. Selection pressure analysis showed that LOX genes in brown algae have undergone strong purifying selection, while the selective constraint in the C2 clade was more relaxed than that in the C1 clade. Furthermore, within each clade, LOXs of S. japonica evolved under more relaxed selection constraints than E. siliculosus and C. okamuranus. Structural modeling showed that unlike LOXs of plants and animals, which contain a β barrel in the N-terminal part of the protein, LOXs in brown algae fold into a single domain. Analysis of previously published transcriptomic data showed that LOXs in E. siliculosus are responsive to hyposaline, hypersaline, oxidative, and copper stresses. Moreover, clear divergence of expression patterns was observed among different life stages, as well as between duplicate gene pairs. In E. siliculosus, all four LOXs are male-biased in immature gametophytes, and mature gametophytes showed significantly higher LOX mRNA levels than immature gametophytes and sporophytes. In S. japonica, however, our RNA-Seq data showed that most LOXs are highly expressed in sporophytes. Even the most recently duplicated gene pairs showed divergent expression patterns, suggesting that functional divergence has likely occurred since LOX genes duplicated, which potentially contributes to the production of various oxylipins in brown algae.


Changes in circulating miRNAs in healthy overweight and obese subjects: Effect of diet composition and weight loss.
MicroRNAs (miRNAs) are small non-coding RNA molecules that can play an important role in several chronic metabolic conditions, including obesity. However, to date little is known about how they are regulated. Weight loss induced by surgical procedures has been effective at modulating specific circulating miRNAs, but the effect of energy-restricted diets with different macronutrient compositions on circulating miRNAs is not well understood. The objective of the present analysis was to explore the effect of three energy-restricted diets of different macronutrient composition and carbohydrate quality on plasma miRNA levels. The GLYNDIET study is a 6-month, parallel, randomized clinical trial conducted on overweight and obese subjects who were randomized to one of three different dietary intervention groups: i) a moderate-carbohydrate and low glycemic index diet (LGI), ii) a moderate-carbohydrate and high glycemic index diet (HGI), and iii) a low-fat and high glycemic index diet (LF). We assessed the genome-wide circulating miRNA profile in a subsample of eight randomly selected participants. A total of 8 miRNAs (miR-411, miR-432, miR-99b, miR-340, miR-423, miR-361, let-7c) were differently quantified according to diet intervention, and were therefore longitudinally validated in 103 participants before and after the energy-restricted diets. Circulating miR-361 levels were lower in the LGI group than in the HGI group, even after adjusting for differences in weight loss. The intra-group analyses demonstrated a significant down-regulation of all miRNAs screened in our study subjects after the LGI intervention. Similarly, miR-139 and miR-340 were down-regulated after the HGI intervention, while miR-139, miR-432 and miR-423 were down-regulated after the low-fat diet. Changes in circulating miR-139 and let-7c were significantly associated with changes in lipid profile and insulin resistance. An energy-restricted low-glycemic index diet down-regulates circulating miRNA-361 more than an energy-restricted high-glycemic index, regardless of the magnitude of the weight loss.


Cardiac dysrhythmias after transcatheter closure of ASD with Amplatzer device.
Transcatheter closure of atrial septal defect (ASD) has been used as an alternative to open heart surgery. Although transcatheter closure of ASD with the Amplatzer septal occluder is a safe and feasible method in pediatric patients, there is little published data on arrhythmia analysis following transcatheter device closure of secundum ASD. We evaluated cardiac dysrhythmias with 24-hour ambulatory electrocardiographic (ECG) monitoring after transcatheter closure of ASD with Amplatzer device. A total of 85 consecutive patients with ASD underwent transcatheter closure of secundum ASD with Amplatzer device between October 1998 and December 2003. The study involved 65 of these patients assessed by 24-hour ambulatory ECG monitoring. Seven patients were evaluated a second time by 24-hour monitorization. During the procedure, transient complete atrioventricular (AV) block was seen in two patients. One of them returned to normal sinus rhythm in catheterization lab and the other returned to normal sinus rhythm in two hours. Transient junctional rhythm was observed in another patient during the device placement. Twenty-four hour ambulatory ECG monitoring was performed on all patients after a mean four-month period (1-12 months). Holter recordings demonstrated rare supraventricular extrasystole in two patients, rare ventricular premature beats in two patients, and intermittent sinus arrest with sinus pause lasting <1.5 seconds in one patient, for a total of five patients (7.6%). In conclusion, dysrhythmias after transcatheter device closure of secundum ASD with Amplatzer device are rare and benign. We need further long-term follow-up to evaluate late dysrhythmias after the transcatheter device closure of secundum ASD.


Synthesis and biological evaluation of prostaglandin-F alkylphosphinic acid derivatives as bone anabolic agents for the treatment of osteoporosis.
A series of novel C(1) alkylphosphinic acid analogues of the prostaglandin-F family have been evaluated at the eight human prostaglandin receptors for potential use in the treatment of osteoporosis. Using molecular modeling as a tool for structure-based drug design, we have discovered that the phosphinic acid moiety (P(O)(OH)R) behaves as an isostere for the C(1) carboxylic acid in the human prostaglandin FP binding assay in vitro and possesses enhanced hFP receptor selectivity when compared to the parent carboxylic acid. When evaluated in vivo, the methyl phosphinic acid analogue (4b) produced a bone anabolic response in rats, returning bone mineral volume (BMV) [corrected], to intact levels in the distal femur in the ovariectomized rat (OVX) model. These results suggest that prostaglandins of this class may be useful agents in the treatment of diseases associated with bone loss.


Ten-Year Review of Gestational Trophoblastic Disease at Lady Reading Hospital, Peshawar.
To determine the frequency and management outcome of gestational trophoblastic disease (GTD) in Lady Reading Hospital. This was a retrospective observational study at Lady Reading Hospital, Peshawar, from January 2011 to December 2021. Hospital records of all patients with GTD were reviewed and all were included in this study except those with an incomplete record or unconfirmed histology. Treatment was analysed in terms of surgical, chemotherapy or no treatment, and outcomes were noted in terms of complete remission, disease persistence or death. In 10 years 353 patients were admitted with GTD, and the frequency of the disease was 3.72 cases per 1000 pregnancies. The most frequent lesions were complete mole 65.2% (n=230) followed by invasive mole 20.4% (n=72). Mortality rate was 0.56% (n=2). Maternal blood group analysis revealed that B positive 28.3% (n=100) was more frequent. O positive blood group was found more in the malignant form of the disease at 3.96% (n=14). GTD was most prevalent in 21 to 30 years of age (41.4%, n=146). Regarding treatment, in 69.97% (n=247) of cases, suction and evacuation were performed, in 4.2% (n=15) of cases hysterectomy was performed as primary therapy, and 4.8% (17) needed hysterectomy for chemoresistance. In this study 42.49% (n=150) were given single-agent chemotherapy and 4.8% (n=17) were given multi-agent therapy. We had 21.33% (32) patients with a risk score of 7-9. In the group with a risk score of 7-9, 15.62% (n=5) patients were directly started on multi-agent therapy because of evidence of metastasis or choriocarcinoma; the remaining 84.37% (n=27) of patients who had no evidence of metastasis, no prior chemotherapy, no choriocarcinoma and International Federation of Obstetrics and Gynecology (FIGO) stage 1 were given single-agent methotrexate with folinic acid (eight days) after informed consent. In 18.75% of patients (n=6) hysterectomy was performed as the primary treatment either for haemorrhage or with age > 40, family completed, or reluctance to undergo chemotherapy. They all had a complete cure. In 3.1% (n=1) of cases, resistance to single-agent therapy was found and multi-agent treatment was started. Overall, in 96.29% of patients, complete remission was achieved with single-agent therapy in patients with risk scores of 7-9. The frequency of GTD was 3.4/1000 pregnancies. Complete mole was the most frequent lesion, and single-agent chemotherapy had a good outcome in low- and high-risk patients with a risk score of 7-9 (with no evidence of metastasis, prior chemotherapy, or choriocarcinoma and FIGO stage 1).


Subtrochanteric fracture after fixation of slipped capital femoral epiphysis: a complication of unused drill holes.
Subtrochanteric fracture of the femur has been infrequently reported after in situ fixation of slipped capital femoral epiphysis, and this occurred in only 1.4% of our patients over a 10-year period. As this technique is used more frequently, however, the incidence of this complication is likely to rise. All four subtrochanteric fractures occurred through unused drill holes, and avoiding extraneous screw holes seems to be the best way to prevent postoperative subtrochanteric fracture. Once subtrochanteric fracture occurs, immediate open reduction and internal fixation with a compression hip screw device is the recommended treatment.


Site-directed mutagenesis of the simian virus 40 large T-antigen gene: replication-defective amino acid substitution mutants that retain the ability to induce morphological transformation.
We used a heteroduplex deletion loop mutagenesis procedure for directing sodium bisulfite-induced mutations to specific sites on viral or plasmid DNA to generate a series of SV40 large T-antigen point mutants. The mutations were directed to a region of the T-antigen gene, 0.5 map units, that is thought to be important for interaction of the protein with the viral origin of DNA replication. Of the 16 mutants reported here, 10 had lost the ability to replicate their DNA, and 3 others showed a reduced level of replication compared to wild type. All of the mutants tested were capable of transforming rat cells in culture by the dense focus assay. We conclude that the sequences of the early region around 0.5 map units are critical for the replication of viral DNA but not for the transformation function of T antigen.


Failure of hospital employees to comply with smoke-free policy is associated with nicotine dependence and motives for smoking: a descriptive cross-sectional study at a teaching hospital in the United Kingdom.
Smoke-free policy aims to protect the health of the population by reducing exposure to environmental tobacco smoke (ETS), and World Health Organisation (WHO) guidance notes that these policies are only successful if there is full and proper enforcement. We aimed to investigate the problem of resistance to smoking restrictions and specifically compliance with smoke-free policy. We hypothesised that an explanation for non-compliance would lie in a measurable difference between the smoking behaviours of compliant and non-compliant smokers, specifically that non-compliance would be associated with nicotine dependence and different reasons for smoking. We conducted a questionnaire-based, descriptive, cross-sectional study of hospital employees. Seven hundred and four members of staff at Addenbrooke's Hospital, Cambridge, UK, completed the questionnaire, of whom 101 were smokers. Comparison between compliant and non-compliant smokers was made based on calculated scores for the Fagerström test and the Horn-Waingrow scale, and level of agreement with questions about attitudes. For ordinal data we used a linear-by-linear association test. For non-parametric independent variables we used the Mann-Whitney test and for associations between categorical variables we used the chi-squared test. The demographic composition of respondents corresponded with the hospital's working population in gender, age, job profile and ethnicity. Sixty nine smokers reported they were compliant while 32 were non-compliant. Linear-by-linear association analysis of the compliant and non-compliant smokers' answers for the Fagerström test suggests association between compliance and nicotine dependence (p = 0.049). Mann-Whitney test analysis suggests there is a statistically significant difference between the reasons for smoking of the two groups: specifically that non-compliant smokers showed habitual smoking behaviour (p = 0.003). Overall, compliant and non-compliant smokers did not have significantly different attitudes towards the policy or their own health. We demonstrate that those who smoke in this setting in contravention to a smoke-free policy do so neither for pleasure (promotion of positive affect) nor to avoid feeling low (reduction of negative affect); instead it is a resistant habit, which has little or no influence on the smoker's mood, and is determined in part by chemical dependence.


Resveratrol Synergistically Promotes BMP9-Induced Osteogenic Differentiation of Mesenchymal Stem Cells.
Mesenchymal stem cells (MSCs) differentiate into osteocytes, adipocytes, and chondrocytes. Resveratrol and bone morphogenetic protein 9 (BMP9) are known osteogenic induction factors of MSCs, but the effect of both resveratrol and BMP9 on osteogenesis is unknown. Herein, we explored whether resveratrol cooperates with BMP9 to improve osteogenic induction. The osteogenic induction of resveratrol and BMP9 on C3H10T1/2 cells was evaluated by detecting the staining and activity of the early osteogenic marker alkaline phosphatase (ALP). In addition, the late osteogenic effect was measured by the mRNA and protein levels of osteogenic markers, such as osteopontin (OPN) and osteocalcin (OCN). To assess the bone formation function of resveratrol plus BMP9 in vivo, we transplanted BMP9-infected C3H10T1/2 cells into nude mice followed by intragastric injection of resveratrol. Western blot (WB) analysis was utilized to elucidate the mechanism of resveratrol plus BMP9. Resveratrol not only enhanced osteogenic induction alone but also improved BMP9-induced ALP at 3, 5, and 7 d postinduction. Both the early osteogenic markers (ALP, Runx2, and SP7) and the late osteogenic markers (OPN and OCN) were significantly increased when resveratrol was combined with BMP9. The fetal limb explant culture further verified these results. The in vivo bone formation experiment, which involved transplanting BMP9-overexpressing C3H10T1/2 cells into nude mice, also confirmed that resveratrol synergistically enhanced the BMP9-induced bone formation function. Resveratrol phosphorylated adenosine monophosphate- (AMP-) activated protein kinase (AMPK) and stimulated autophagy, but these effects were abolished by inhibiting AMPK and Beclin1 using an inhibitor or siRNA. Resveratrol combined with BMP9 significantly improves the osteogenic induction of C3H10T1/2 cells by activating AMPK and autophagy.


[Neurosurgical embryology. Part 8: Post-natal angiogenesis and remodeling].
Post-natal angiogenesis (microvascular proliferation) and remodeling (modifications of diameter and wall thickness) occur in various physiological circumstances including adaptation to exercise or wound healing. The formation of a new vessel is submitted to the combinative action of growth factors. New endothelial cells migrate, proliferate, differentiate and attract pericytes and future smooth muscle cells to create the new vessel. Powerful stimuli leading to remodeling and to the creation of new vessels are mainly represented by hemodynamic forces generated by pressure and blood flow, and by hypoxia. Our purpose is to overview the molecular mechanisms and the stimuli giving rise to these processes.


Five successful pregnancies with antenatal anakinra exposure.
Our aim is to add to the limited existing prospective data on IL-1 inhibitor use in pregnancy. Data were obtained from the Organization of Teratology Information Specialists Autoimmune Disease in Pregnancy Project, a prospective cohort study of pregnancy outcomes in the USA and Canada. Eligible women were enrolled prior to 19 weeks' gestation between 2004 and 2017. Outcomes were obtained by maternal interview and medical record abstraction. Five pregnancies with anakinra exposure were identified, all resulting in full-term singleton live births with no major or long-term complications. Three maternal subjects used anakinra for adult-onset Still's disease and two for systemic JIA. For all individuals who discontinued anakinra, some amount of steroid medication was necessary for treatment of disease flare. Two maternal subjects developed oligohydramnios, one also with pregnancy-induced hypertension. Two women had Caesarian sections, one medically indicated and one scheduled. One infant had low birth weight, but follow-up records indicated normal adjusted weight at 1 year. Three women successfully breastfed their infants, at least two of whom continued anakinra while breastfeeding. Anakinra was used successfully in five full-term pregnancies; however, two subjects developed oligohydramnios, a process that can be linked to fetal renal anomalies. Given previously reported cases of congenital renal anomalies associated with both antenatal anakinra use and maternal hyperthermia, the relationship between maternal IL-1 inhibitor use, uncontrolled maternal febrile disease and fetal outcomes should be further explored.


Prevalence and severity of vestibular recession in mandibular incisors after orthodontic treatment.
To assess the prevalence and severity of vestibular gingival recession of mandibular incisors after orthodontic treatment and to evaluate possible contributing factors. From the record pool of patients who completed orthodontic treatment from 1999-2006 at the Department of Orthodontics, University of Oslo, Norway, 588 patients fulfilled the inclusion criteria. Intraoral color slides were used for the evaluation of gingival recessions (based on Miller classification), presence of visible plaque, and gingival inflammation. Cephalometric radiographs were used to assess the sagittal intermaxillary relation, mandibular and intermaxillary angles, and the position of the lower incisors. A control group was drawn from the same pool of 588 patients. All statistical analyses were performed using SPSS. The prevalence of gingival recessions after orthodontic treatment was 10.3%. Most (8.6%) were classified as Miller Class I, and 1.7% were classified as Miller Class II. Gingival recession was predominantly found on central incisors. Reduction of the sagittal intermaxillary angle and retroclination of the lower incisors was correlated with the development of a more severe gingival recession. The present study indicates that vestibular gingival recession of mandibular incisors after orthodontic treatment is of minor prevalence and severity. The presence of gingival recession or retroclination of the incisors with mesial basal relations increases the risk of more severe gingival recession.


Reflecting on the Germanwings Disaster: A Systematic Review of Depression and Suicide in Commercial Airline Pilots.
The 2015 Germanwings Flight 9525 disaster, in which 150 people were killed after the co-pilot may have intentionally crashed the plane in a suicide attempt, highlights the importance of better understanding the mental health of commercial airline pilots. However, there have been few systematic reviews investigating the topic of mental health in commercial aviation. This systematic review aims to identify the types and prevalence of mental health disorders that commercial airline pilots experience with a focus on mood disorders and suicide risk. A systematic literature search was performed using PubMed, EMBASE, and PsycINFO databases. Eligible studies were assessed and data was extracted and analyzed. 20 studies were identified. The prevalence of depression experienced by commercial airline pilots in this review ranged from 1.9% to 12.6%. Factors that negatively impacted the mental health of pilots included substance abuse, experiencing verbal or sexual abuse, disruption in sleep circadian rhythms and fatigue. This systematic review identifies that commercial airline pilots may experience depression at least as frequently as the general population. Commercial airline pilots experience occupational stressors, such as disrupted circadian rhythms and fatigue which may increase risks of developing mood disorders. Most studies identified in this review were cross-sectional in nature with substantial limitations. There is a clear need for further higher quality longitudinal studies to better understand the mental health of commercial airline pilots.


Role of receptor proteins for enterobactin and 2,3-dihydroxybenzoylserine in virulence of Salmonella enterica.
Single, double, and triple mutants of an enterobactin-deficient mutant strain of Salmonella enterica serovar Typhimurium were constructed that were defective in the expression of the iron-regulated outer membrane proteins (IROMPs) FepA, IroN, and Cir, which are proposed to function as catecholate receptors. Uptake of naturally occurring and chemically synthesized catecholate molecules by these mutants was assessed in standard growth promotion assays. Unique patterns of uptake were identified for each IROMP; specifically, FepA and IroN were confirmed to be required for transport of enterobactin, and all three proteins were shown to function as receptors for the enterobactin breakdown product 2,3-dihydroxybenzoylserine. The fepA, iroN, and cir alleles were transduced to enterobactin-proficient strains of S. enterica serovar Typhimurium and S. enterica serovar Enteritidis, and the resulting phenotypes were confirmed by analysis of outer membrane protein profiles, by sensitivity to KP-736, a catecholate-cephalosporin conjugate, and by growth promotion tests on egg white agar. Intragastric infections of mice with the S. enterica serovar Typhimurium strains indicated that the parental strain and the fepA iroN double mutant were similarly virulent but that the fepA iroN cir triple mutant was significantly attenuated. Moreover, in mixed infections, the fepA iroN mutant showed similar cecal colonization and invasion of the liver to the parental strain, while the triple mutant showed significantly reduced cecal colonization and no measurable spread to the liver. Infections of 4-day-old chicks with S. enterica serovar Enteritidis strains also indicated that mutation of the fepA iroN genes did not significantly reduce cecal colonization and systemic spread compared with those of the parental strain. The results indicate that, while enterobactin uptake is not essential for the virulence of S. enterica serovars in mouse and chicken infection models, the ability to take up 2,3-dihydroxybenzoylserine via any of the three catecholate siderophore receptors appears to play an important role, since the S. enterica serovar Typhimurium triple mutant was significantly attenuated in the mouse model. Salmochelins appear not to be involved in the virulence of S. enterica.


The level of endotoxins in hemodialysis water and dialysate in Lithuanian hemodialysis centers.
The composition and quality of the dialysis fluid play an important role in the modulation of dialysis-related complications. During hemodialysis, patient's blood has a contact with dialysate through a semipermeable membrane. Bacterial endotoxins can pass through the membrane pores into the patient's blood and cause a silent chronic microinflammation. The aim of this study was to determine the level of endotoxins in hemodialysis water and dialysate in Lithuanian hemodialysis centers. Dialysis water (n=50) and dialysate (n=50) were collected from 91% (n=50) of all hemodialysis centers. The presence of bacterial endotoxins was evaluated using a sensitive Limulus amebocyte lysate test, which detects intact lipopolysaccharides. The level of endotoxins was lower than 0.25 EU/mL in 43 (86%) dialysis water samples and in 46 (92%) dialysate samples, and complied with the recommendations of the European Pharmacopoeia and the European Best Practice Guidelines for pure dialysis fluid. The dialysate of 39 (78%) Lithuanian hemodialysis centers complied with the definition of an ultrapure dialysis fluid. The water and dialysate were of insufficient quality in 14% and in 8% of Lithuanian hemodialysis centers, respectively, and this could be improved by the establishment of routine investigation of endotoxins.


Twenty-five-year survival of children with birth defects in New York State: a population-based study.
Few studies have been conducted on long-term survival of children with major birth defects because of a lack of longitudinal birth defects surveillance data. The objective of this study was to conduct a 25-year survival analysis among children in New York born with major defects by survival age, birth defect category, and other possible contributing factors. A cohort was constructed containing children born in 1983 to 2006 with selected major birth defects. Deaths among the study cohort were identified by matching the children to their death certificates. The survival probability was estimated by Kaplan-Meier methods. Cox proportional hazards regression was used to examine the effect of the risk factors on survival. A total of 9112 deaths were identified among 57,002 live births with selected birth defects between 1983 and 2006. The overall 25-year survival probability of the study cohort was 82.51% (95% confidence interval, 82.11-82.89%). The estimated survival probability was comparable to that reported from previous studies regarding individual defects including spina bifida, encephalocele, atrioventricular septal defects, tracheoesophageal fistula and esophageal atresia or stenosis, renal agenesis or dysgenesis, lower limb reduction, diaphragmatic hernia, abdominal wall defects, and Down syndrome. Sex, low birth weight for gestational age, existence of multiple birth defects (nonisolated), and maternal age and nativity were identified as risk factors. Using the statewide, population-based birth defects surveillance data in New York State, the survival experience of the study cohort was examined across all survival time periods by individual birth defect of interest. Several risk factors that affect survival were identified.


Bone mineral and matrix. Are they altered in osteoporosis?
Osteoporosis is a major health threat whose etiology remains elusive. Disease heterogeneity and until recently the absence of appropriate techniques prevented detailed analyses of the underlying defects in this disease. Among the questions that must be asked are: (1) Are the cells in the osteoporotic bone functioning normally? (2) Is the matrix produced by these cells normal? (3) Is the pattern of mineral deposition normal? If not, how do each of these differ from normal, what is the effect of that abnormality, and how can it be prevented or corrected? Through answering these questions the scientific community will learn the function of many of the matrix proteins and may gain insight into revolutionary new therapies. The tools of human bone cell culture, new imaging techniques, analyses in biopsy samples, and other aspects of modern biotechnology are now available to start answering these questions.


[The Italian contribution to studies on the biological rhythms implicated in psychiatric disorders].
This editorial summarizes the main studies, carried out in the last 10 years, by various Italian research groups, on the alterations of circadian rhythms in psychiatric disorders. The results of these researches, as well as those obtained in various international contexts, encourage to teach in the medical schools for psychiatry, about the new chronoterapeutic interventions and the implementation of combined therapies for increasingly personalized psychiatric therapies.


Prosthetic Disuse Leads to Lower Balance Confidence in a Long-Term User of a Transtibial Prosthesis.
Residual limb wounds or ulcers are one of the most frequent skin problems reported by lower extremity prosthesis users. Healing often requires prosthesis disuse, which can logically impair physical functioning. However, there are limited data available to support this idea. We report the impact of prosthesis disuse by presenting assessments of balance, gait, physical activity, and balance confidence obtained on a case subject before experiencing a wound and following reintroduction to a well-fit prosthesis after wound-related prosthesis disuse. The case subject was a 76-year-old male who suffered a unilateral, transtibial amputation due to synovial sarcoma 13 years before. After presenting with a history of pain in the area of a chronic skin plaque, he received a punch biopsy, which resulted in 4 weeks of prosthesis disuse followed by 12 weeks of limited use before a final well-fitting socket was received. The following data were collected 24 weeks before the biopsy and 4 weeks after receiving the final well-fitting socket: Berg Balance Scale, L-test of walking, quantitative gait analysis, Activity-specific Balance Confidence Scale, and 1 week of community-based activity. Balance confidence decreased nearly 19%, walking speed decreased by 12%, and steps/day decreased by 19% following ∼4 months of prosthesis disuse/limited use; functional measures were not impacted. Lower balance confidence is not trivial as it can lead to activity avoidance and increased fall risk. Interventions to target balance confidence changes following prosthesis disuses may be important to minimize the impact of disuse on physical and mental well-being.


Postoperative impedance-pH testing is unreliable after Nissen fundoplication with or without giant hiatal hernia repair.
Combined 24-h multichannel intralumenal impedance-pH monitoring (MII-pH) is gaining popularity as a diagnostic tool for gastroesophageal reflux. Since the surgical reduction of hiatal hernias and creation of a fundoplication anatomically restores the gastroesophageal reflux barrier, one would assume that it effectively stops all reflux regardless of composition. Our aim is to evaluate the results of routine MII-pH testing in successful Nissen fundoplication patients. Sixty-two patients with normal acid exposure, confirmed by 24-h pH testing, after Nissen fundoplication were evaluated with symptomatic questionnaire, esophageal manometry and MII-pH testing more than 6 months after surgery. Patients were grouped into normal and abnormal based on postoperative impedance results. Patients with Nissen alone were separately compared to patients with Nissen + giant hiatal hernia (GHH). Twenty-nine (47%) patients exhibited abnormal impedance after successful Nissen fundoplication. Abnormal impedance was associated with GHH repair, lower bolus pressures, and lower distal esophageal contraction amplitudes. Postoperative testing with the standard MII-pH catheters using published normative values seems to be clinically irrelevant. Clinicians should analyze the results of routine MII-pH testing in the setting of a fundoplication critically as the current technology is associated with a high false positive rate.


Phase-shifted Bragg microstructured optical fiber gratings utilizing infiltrated ferrofluids.
Results are presented on the efficient spectral manipulation of uniform and chirped Bragg reflectors inscribed in microstructured optical fibers utilizing short lengths of ferrofluids infiltrated in their capillaries. The infiltrated ferrofluidic defects can generate either parasitic reflection notch features in uniform Bragg reflectors of up to 80% visibility and ~0.1 nm spectral shift or tunability of the bandwidth and strength reflection up to 100% when introduced into chirped gratings. Spectra are presented for different spatial positions and optical characteristics of the ferrofluidic section.


Blood volume measurements using an integrated fiberoptic monitoring system in a porcine septic shock model.
To compare the accuracy of an integrated fiberoptic monitoring system using transpulmonary thermo-dye dilution technique to measure blood volume (BV) with standard method using chromium-51-tagged erythrocytes in septic shock. Prospective blinded animal laboratory study. University department of anesthesiology. Thirty-five anesthetized and mechanically ventilated pigs (21.4 +/- 2.2 kg) were investigated over a period of 6 hrs. Septic shock was induced with fecal peritonitis (0.75 g . kg per body weight autologous feces). A central venous catheter was used for injection of the indicator dyes. BV was measured by detecting indocyanine green using a 4-Fr aortic catheter with an integrated fiberoptic and thermistor connected to a computer system for calculation of transpulmonary indicator dilution BV (BVTPID). Cr-tagged erythrocytes were used as standard method of BV measurement (BV-Cr). Hemodynamic treatment scheme was aimed at maintenance of a central venous pressure of 12 mm Hg. Data were analyzed using Bland-Altman analyses. One hundred and five data pairs of simultaneous BV measurements were yielded during hemodynamic stability with a mean BVTPID of 64.2 +/- 17.8 mL . kg. Mean BV-Cr was 83.1 +/- 17.0 mL . kg. Linear regression equation was BVTPID = 0.58 x BV-Cr + 15.8 (r = .56, p < .01). Mean bias was 18.9 mL . kg (95% confidence interval, 15.7-22.1 mL . kg), with limits of agreement of -13.9 to 51.7 mL . kg. Transpulmonary indicator dilution for blood volume measurement agrees moderately with standard method using Cr-tagged erythrocytes in porcine septic shock.


Effect of reducing the posted speed limit to 30 km per hour on pedestrian motor vehicle collisions in Toronto, Canada - a quasi experimental, pre-post study.
Pedestrian related deaths have recently been on the rise in Canada. The effect of changing posted speeds on the frequency and severity of pedestrian motor vehicle collisions (PMVC) is not well studied using controlled quasi-experimental designs. The objective of this study was to examine the effect of lowering speed limits from 40 km/h to 30 km/h on PMVC on local roads in Toronto, Canada. A 30 km/h speed limit on local roads in Toronto was implemented between January 2015 and December 2016. Streets that remained at a 40 km/h speed limit throughout the study period were selected as comparators. A quasi-experimental, pre-post study with a comparator group was used to evaluate the effect of the intervention on PMVC rates before and after the speed limit change using repeated measures Poisson regression. PMVC data were obtained from police reports for a minimum of two years pre- and post-intervention (2013 to 2018). Speed limit reductions from 40 km/h to 30 km/h were associated with a 28% decrease in the PMVC incidence rate in the City of Toronto (IRR = 0.72, 95% CI: 0.58-0.89). A non-significant 7% decrease in PMVC incidence rates were observed on comparator streets that remained at 40 km/h speed limits (IRR = 0.93, 95% CI: 0.70-1.25). Speed limit reduction also influenced injury severity, with a significant 67% decrease in major and fatal injuries in the post intervention period on streets with speed limit reductions (IRR = 0.33, 95% CI: 0.13-0.85) compared with a 31% not statistically significant decrease in major and fatal injuries on comparator streets (IRR = 0.69, 95% CI: 0.37-1.31). The interaction term for group and pre-post comparisons was not statistically significant (p = 0.14) indicating that there was no evidence to suggest a pre-post difference in IRRs between the intervention and comparator streets. Declines in the rate of PMVC were observed on roads with posted speed limit reductions from 40 km/h to 30 km/h, although this effect was not statistically greater than reductions on comparator streets.


CBXC: a set of recombinant inbred strains between CBA/Ca and BALB/c.
A new set of nine recombinant inbred strains designated CBXC-1 to CBXC-9 has been developed from a cross between CBA/Ca female and BALB/c male mice. All of the strains have been brother x sister mated for more than 20 generations, and have been characterized at eight electrophoretic, one immunological and two coat colour loci at which the progenitor strains differ. This set of strains should be useful in investigating the genetics of any characters which differ between the two progenitor strains.


Pelvic and extremity immobilization after bladder exstrophy closure: complications and impact on success.
A successful initial bladder closure is critical to the development of adequate bladder capacity and ultimate continence among patients with bladder exstrophy. Although secure pelvic fixation is essential to success, the various methods of pelvic and extremity immobilization and their impact on surgical outcome have not previously been examined. A review of the approved bladder exstrophy database at our institution identified 194 patients for whom data were available regarding initial bladder closure and the method of pelvic immobilization. The records were reviewed with respect to age at bladder closure, use/type of osteotomy, method of pelvic immobilization, complications related to the immobilization technique, and surgical outcome. Of 194 patients identified, initial exstrophy closure failed in 86. Of these 86 patients, the initial closure failed in 80 at another hospital who were referred to our institution for additional treatment. Of the 114 initially closed at this institution, 6 failed. The success rates for initial and second closures were highest after osteotomy (75% versus 38% and 81.5% versus 27%, respectively), and when the patients were immobilized with an external fixator and 6 to 8 weeks of modified Buck's traction with osteotomy (96%) or 4 to 6 weeks of modified Bryant's traction without osteotomy (61%). The spica cast and "mummy wrapping" immobilization techniques were less effective and were associated with significant complications. These results demonstrate that a successful exstrophy bladder closure depends on the proper use of the osteotomy and effective postoperative immobilization of the pelvis. Our best outcomes have been achieved using lower extremity traction (with or without external fixation of the pelvis).


Assessing the impact of the heart of New Ulm Project on cardiovascular disease risk factors: A population-based program to reduce cardiovascular disease.
The Heart of New Ulm Project (HONU), is a population-based project designed to reduce modifiable cardiovascular disease (CVD) risk factors in the rural community of New Ulm, MN. HONU interventions address multiple levels of the social-ecological model. The community is served by one health system, enabling the use of electronic health record (EHR) data for surveillance. The purpose of this study was to assess if trends in CVD risk factors and healthcare utilization differed between a cohort of New Ulm residents age 40-79 and matched controls selected from a similar community, using EHR data from baseline (2008-2009) through three follow up time periods (2010-2011, 2012-2013, 2014-2015). Matching, using covariate balance sparse technique, yielded a sample of 4077 New Ulm residents and 4077 controls. We used mixed effects longitudinal models to examine trends over time between the two groups. Blood pressure, total cholesterol, low-density lipoprotein-cholesterol, and triglycerides showed better management in New Ulm over time compared to the controls. The proportion of residents in New Ulm with controlled blood pressure increased by 6.2 percentage points compared to an increase of 2 points in controls (p < 0.0001). As the cohort aged, 10-year ASCVD risk scores increased less in New Ulm (5.1) than the comparison community (5.9). The intervention and control community did not differ with regard to inpatient stays, smoking, or glucose. Findings suggest efficacy for the HONU project interventions for some outcomes.


Patch test dose-response study of p-phenylenediamine: thresholds and anatomical regional differences.
The elicitation response in allergic contact dermatitis for the hair dye substance p-phenylenediamine (PPD) is dose dependent, but threshold concentrations have not previously been investigated. 15 PPD-sensitive patients participated in a serial dilution 48-hr patch test with PPD using 8 concentrations of PPD ranging from 1 to 10 000 on the upper back. Petrolatum was applied as control. Three concentrations (50, 100 and 500 p.p.m. PPD) were also applied to the retroauricular area and on the lateral aspects of the upper arms. 14 of the 15 participants reacted to one or more of the test samples. The threshold value for 10% of the tested persons (ED10) based on+or stronger reactions for PPD on the back was 38 p.p.m. (CI: 4.3-100). There were no statistically significant differences in the sensitivity of the three anatomical regions. The upper back is a suitable region for patch testing patients with hair dye dermatitis.


Characterization of a retrovirus shuttle vector capable of either proviral integration or extrachromosomal replication in mouse cells.
A retrovirus shuttle vector is described that contains the dominant selectable neo gene which confers resistance to kanamycin in bacteria and to the drug G418 in animal cells. The bacterial supF gene and the origins of DNA replication from polyomavirus and the ColE1 replicon also have been included in this vector. Infection of normal rodent cells results in single-copy proviral integration, whereas infection of mouse (MOP) cells expressing polyoma large T antigen results in extrachromosomal replication of the DNA form of the virus. The copy number of the extrachromosomal circles in MOP cells varies from 0 to 100 copies per cell. G418-resistant MOP cells lose their drug-resistant phenotype after passage under nonselective conditions, suggesting that maintenance of the extrachromosomal circles is unstable. The extrachromosomal form of the virus can be recovered as plasmids in Escherichia coli. Two-thirds of the circles analyzed were found to be structurally intact. The others have undergone rearrangements including deletions and insertions. The bacterial supF gene was found to be intact in the majority of recovered plasmids. The data presented here suggest that these retroviruses should be useful as gene transfer vectors for animal cells in culture or in vivo.


In silico simulations suggest that Th-cell development is regulated by both selective and instructive mechanisms.
Th-cell differentiation is highly influenced by the local cytokine environment. Although cytokines such as IL-12 and IL-4 are known to polarize the Th-cell response towards Th1 or Th2, respectively, it is not known whether these cytokines instruct the developmental fate of uncommitted Th cells or select cells that have already been committed through a stochastic process. We present an individual based model that accommodates both stochastic and deterministic processes to simulate the dynamic behaviour of selective versus instructive Th-cell development. The predictions made by each model show distinct behaviours, which are compared with experimental observations. The simulations show that the instructive model generates an exclusive Th1 or Th2 response in the absence of an external cytokine source, whereas the selective model favours coexistence of the phenotypes. A hybrid model, including both instructive and selective development, shows behaviour similar to either the selective or the instructive model dependent on the strength of activation. The hybrid model shows the closest qualitative agreement with a number of well-established experimental observations. The predictions by each model suggest that neither pure selective nor instructive Th development is likely to be functional as exclusive mechanisms in Th1/Th2 development.


Responsiveness to erythropoiesis-stimulating agents and renal survival in patients with chronic kidney disease.
Renal anemia of chronic kidney disease (CKD) is generally treated by erythropoiesis-stimulating agents (ESAs). However, there are individual differences in patients' responsiveness to ESA, which may affect the prognosis of CKD. The effect of ESAs on hemoglobin was followed in 297 CKD patients with renal anemia. Three types of ESA, epoetin alfa or beta, darbepoetin alfa, and epoetin beta pegol, were used in this study and dose of ESA was converted to that of epoetin using a dose conversion ratio (epoetin:darbepoetin alfa:epoetin beta pegol = 200:1:0.93). After initial 12-week administration of ESAs, the patients were divided into three groups: poor, intermediate, and good responders based on ΔHb/week/epoetin dose as an index. Hemoglobin values were followed for 144 weeks. Initial patient characteristics--including age, body mass index, hemoglobin, estimated glomerular filtration rate, transferrin saturation, ferritin, albumin, calcium, parathyroid hormone, C-reactive protein, and urine protein--were similar in the three responder groups, except phosphate in the poor responder group was significantly higher than in the other two groups. The period from ESA use to renal death (RD) was significantly shortest in the poor responder group, and the number of RD patients was fewer in the good responder group. Multivariate Cox regression revealed that low final ΔHb(ΔHb from ESA use to just before dialysis)/week/epoetin dose, and low Hb after 12-week ESA use were significant factors related to responsiveness to ESA, suggesting that hyporesponsiveness to ESA was a risk factor for RD. Cox regression also found that hyperphosphatemia and diabetic nephropathy were risks for RD as well. The study results suggest that hyporesponsiveness to ESA after the first 12-week administration as well as after 12 weeks is a risk for RD in pre-dialysis CKD patients. Furthermore, hyperphosphatemia and diabetic nephropathy are risk factors for RD.


Summary of cases of adventitial cystic disease of the popliteal artery.
Adventitial cystic disease of the popliteal artery is explored. The results of correspondence with authors reporting this condition are elaborated upon. This has provided an opportunity to discuss the history of the condition, the findings in 115 cases which have come to the attention of the Correspondence Office dealing with this entity, and the results of treatment. A discussion of the suspected etiology of the condition is presented. The condition remains one of unknown etiology which can be treated by cyst evacuation or aspiration when the popliteal artery is patent and which is best treated by arterial reconstruction when the artery is occluded. The results of such treatment are good but are dependent upon technical excellence of the operative procedure.


Aggressive Behavior in Silent Subtype III Pituitary Adenomas May Depend on Suppression of Local Immune Response: A Whole Transcriptome Analysis.
Silent subtype III pituitary adenomas (SS-3) are clinically nonfunctional adenomas that are more aggressive in terms of invasion and risk of recurrence than their conventional null cell counterparts. We previously showed that these tumors can be distinguished by immunohistochemistry based on the identification of a markedly enlarged and fragmented Golgi apparatus. To understand the molecular correlates of differential aggressiveness, we performed whole transcriptome sequencing (RNAseq) on 4 SS-3 and 4 conventional null cell adenomas. The genes that were highly upregulated in all the SS-3 adenomas included 2 secreted proteins involved in the suppression of T-lymphocyte activity, i.e., ARG2 (multiple testing adjusted padj = 1.5 × 10-3) and SEMA3A (padj = 3.3 × 10-3). Highly downregulated genes in all the SS-3 adenomas included HLA-B (padj = 3.3 × 10-6), suggesting reduced antigen presentation by the adenoma to cytotoxic T-cells. Quantitative RT-PCR of these genes performed on the adenoma samples supported the RNAseq results. We also found a relative decrease in the overall concentration of T-lymphocytes in the SS-3 tumors. These results suggest that SS-3 adenomas actively suppress the immune system and raise the possibility that they may be treatable with immune checkpoint inhibitors or nonspecific cancer immunotherapies.


An update on human papillomavirus infection and Papanicolaou smears in adolescents.
Human papillomavirus infection is one of the most common sexually transmitted infections in young women, and may lead to clinical sequelae such as anogenital condylomata and cervical squamous cell carcinoma. Recent data on the biology and natural history of HPV infection in adolescents will have important implications for the development of adolescent-specific protocols for cervical cancer screening and for follow-up of abnormal cytology.


Pancytopenia after low dose methotrexate therapy in a hemodialysis patient: case report and review of literature.
Methotrexate (MTX) is widely used in the treatment of rheumatoid arthritis (RA) with a side effect of pancytopenia. However, only a few cases of severe pancytopenia caused by low-dose MTX therapy have been reported, and the condition is rarely reported in uremic patients on dialysis therapy. We thereby report a hemodialysis patient who developed severe pancytopenia after oral treatment with low-dose MTX for RA. A 55-year-old woman who had been on regular hemodialysis treatment for 7 yr suffered from RA for 10 yr. She was regularly treated with celecoxib, prednisolone, and sulfasalazine in the past year. Because of the increasing arthralgia, 7.5 mg per week MTX was prescribed 3 months before admission. Stomatitis, fever, general fatigue, multiple skin carbuncles, and easy bruising developed after a cumulative dose of 90 mg. Pancytopenia was found at admission and the nadir of white blood cell count was 250/microL with 28% neutrophils, hematocrit was 22%, and platelet count was 6000/microL. Eosinophil counts increased from 11.5% initially to 26.1% on the sixth admission day. Transfusion with red blood cells and platelets, and appropriate antibiotics and folic acid were prescribed. She continued receiving regular hemodialysis and eventually recovered within 3 weeks.


Technology for Augmenting Type 1 Diabetes Mellitus Management.
Type 1 diabetes mellitus has witnessed significant progress in its management over the past several decades. This review highlights technologic advancements in type 1 diabetes management. Continuous glucose monitoring systems are now available at various functionality and cost levels, addressing diverse patient needs, including a recently US Food and Drug Administration (FDA)-approved implantable continuous glucose monitoring system (CGMS). Another dimension to these state-of-the-art technologies is CGMS and insulin pump integration. These integrations have allowed for CGMS-based adjustments to basal insulin delivery rates and suspension of insulin delivery when a low blood glucose event is predicted. This review also includes a brief discussion of upcoming technologies such as patch-based CGMS and insulin-glucagon dual-hormonal delivery.


Structural and Dynamics Perspectives on the Binding of Substrate and Inhibitors in Mycobacterium tuberculosis DHFR.
Dihydrofolate reductase (DHFR), an essential enzyme in the folate pathway, is a potential target for new anti-tuberculosis drugs. Fifteen crystal structures of Mycobacterium tuberculosis DHFR complexed with NADPH and various inhibitors are available in the RCSB Protein Data Bank, but none of them is a substrate binding structure. Therefore, we performed molecular dynamics simulations on ternary complexes of M. tuberculosis DHFR:NADPH with a substrate (dihydrofolate) and each of three competitive inhibitors the in 2,4-diaminopyrimidine series (P1, P157, and P169), in order to gain insight into the inhibition-mechanism of DHFR in the folate pathway. The binding energy and thermodynamics values of each system were calculated by the Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) method. The dynamics of the enzyme and the motion of each amino acid residue at the active site were examined. The key factors that promote the binding of P157 and P169 on M. tuberculosis DHFR (mtbDHFR) reveal opportunities for using these compounds as novel anti-tuberculosis drugs.


Effects of continuous positive airway pressure on energy balance regulation: a systematic review.
Obesity is both a cause and a possible consequence of obstructive sleep apnoea (OSA), as OSA seems to affect parameters involved in energy balance regulation, including food intake, hormonal regulation of hunger/satiety, energy metabolism and physical activity. It is known that weight loss improves OSA, yet it remains unclear why continuous positive airway pressure (CPAP) often results in weight gain.The goal of this systematic review is to explore if and how CPAP affects the behaviour and/or metabolism involved in regulating energy balance.CPAP appears to correct for a hormonal profile characterised by abnormally high leptin and ghrelin levels in OSA, by reducing the circulating levels of each. This is expected to reduce excess food intake. However, reliable measures of food intake are lacking, and not yet sufficient to make conclusions. Although studies are limited and inconsistent, CPAP may alter energy metabolism, with reports of reductions in resting metabolic rate or sleeping metabolic rate. CPAP appears to not have an appreciable effect on altering physical activity levels. More work is needed to characterise how CPAP affects energy balance regulation.It is clear that promoting CPAP in conjunction with other weight loss approaches should be used to encourage optimal outcomes in OSA patients.


Phloem unloading follows an extensive apoplasmic pathway in cucumber (Cucumis sativus L.) fruit from anthesis to marketable maturing stage.
The phloem unloading pathway remains unclear in fruits of Cucurbitaceae, a classical stachyose-transporting species with bicollateral phloem. Using a combination of electron microscopy, transport of phloem-mobile symplasmic tracer carboxyfluorescein, assays of acid invertase and sucrose transporter, and [(14)C]sugar uptake, the phloem unloading pathway was studied in cucumber (Cucumis sativus) fruit from anthesis to the marketable maturing stage. Structural investigations showed that the sieve element-companion cell (SE-CC) complex of the vascular bundles feeding fruit flesh is apparently symplasmically restricted. Imaging of carboxyfluorescein unloading showed that the dye remained confined to the phloem strands of the vascular bundles in the whole fruit throughout the stages examined. A 37 kDa acid invertase was located predominantly in the cell walls of SE-CC complexes and parenchyma cells. Studies of [(14)C]sugar uptake suggested that energy-driven transporters may be functional in sugar trans-membrane transport within symplasmically restricted SE-CC complex, which was further confirmed by the existence of a functional plasma membrane sucrose transporter (CsSUT4) in cucumber fruit. These data provide a clear evidence for an apoplasmic phloem unloading pathway in cucumber fruit. A presumption that putative raffinose or stachyose transporters may be involved in soluble sugars unloading was discussed.


[Discussion on the electromagnetic compatibility testing and evaluation of radio frequency ablation catheter].
With the enforcement of YY 0505-2012, the testing items and evaluation points of radio frequency ablation catheter in electromagnetic compatibility field should be studied and discussed. Based on the requirements of relevant standards, this paper discusses on the testing items that should be applied and the evaluation points that should be focused on by analyzing the intended use and the structure of radio frequency ablation catheter, when it intends to apply registration individually with the basic knowledge of electromagnetic compatibility field.


A case of Type A insulin resistance syndrome in a 14-year-old girl without common clinical features.
Type A insulin resistance syndrome is a rare congenital disorder caused by insulin receptor dysfunction arising from heterozygous mutations in the insulin receptor (INSR) gene. It is usually diagnosed in non-obese female adolescents with insulin resistance, acanthosis nigricans, or ovarian hyperandrogenism. We present the case of a 14-year-old adolescent girl with non-obesity diabetes and insulin resistance; however, she had no acanthosis nigricans, or menstrual abnormalities. The laboratory results were as follows: fasting plasma glucose 179 mg/dL; C-peptide 1.8 ng/mL; insulin 34.8 lU/mL; HbA1c 10.6%. Islet cell autoantibodies, insulin autoantibodies, and glutamic acid decarboxylase antibodies were negative. We performed a next-generation sequencing-based targeted gene panel for the known maturity-onset diabetes of the young and found a heterozygous mutation in the INSR gene c.3602G>A (p.Arg1201Gln). Her mother carried the same mutation as her and also did not present with acanthosis nigricans or menstruation abnormalities. Type A insulin resistance syndrome has diverse clinical phenotypes even with the same type of mutation at the same site in the INSR gene. Clinicians should consider genetic testing for early detection and adequate treatment approaches.


Investigating Stress Response during Vaginal Delivery and Elective Cesarean Section through Assessment of Levels of Cortisol, Interleukin 6 (IL-6), Growth Hormone (GH) and Insulin-Like Growth Factor 1 (IGF-1).
How do stress related phenomena during labor differ between vaginal delivery (VD) and elective cesarean section (CS), remains of heightened interest. The purpose of this study is to investigate discrepancies regarding the stress response during VD and CS. Cortisol, interleukin 6 (IL-6), growth hormone (GH) and insulin-like growth factor 1 (IGF-1) levels from parturients' peripheral blood were evaluated on three time-points, namely during the first stage of labor (TP1), two hours post labor (TP2) and 48 h post labor (TP3). Levels were also evaluated from the umbilical cord blood. A total of 50 women were enrolled in this prospective cohort study, with 24 and 26 subjected to CS and VD, respectively. No statistically significant differences were observed between the two groups at TP1. Only GH levels presented the same pattern during the three time-points among both groups. In the umbilical cord blood, the CS group presented statistically significant higher IGF-1 and GH levels. In the umbilical cord blood, IGF-1 and GH levels were positively correlated, while GH and cortisol levels were negatively correlated. CS is a less stressful procedure than VD and is further associated with less intense inflammation, albeit with a longer inflammatory response period. Labor physiology during CS differs considerably regarding respective observations during VD. This merits extensive investigation in order to decipher these data for optimal clinical practice and guidelines.


Interest in geriatric medicine in Canada: how can we secure a next generation of geriatricians?
In Canada, there is minimal training of geriatrics for physicians, a shortage of geriatricians, and extremely low numbers of students entering geriatrics. This study explored student interest in and barriers and enticements to geriatric medicine as a career choice. Medical students attending a university in Ontario, Canada, were surveyed in their first year (N=121), after a geriatric education session, and again in their second year (N=118) about their interest in a career in geriatrics. In the first year, less than 20% of students were interested in geriatrics; in the second year this decreased to 16%. In both years, female students were more interested than male students. Those students interested in geriatrics had higher hopes that their practice would involve primarily adults and seniors. Students not interested in geriatrics rated performing procedures and technical skills, not wanting to work with chronically ill patients, and caring for younger patients as important practice characteristics. Although the importance of prestige was low for all students, it was significantly higher for those not interested in geriatrics. Although changes to prestige, income, lifestyle, and length of residency training were identified as potential enticements to geriatrics, they were not major deterrents to a career in geriatrics. The findings suggest strategies that may affect student interest in geriatrics, such as increased and early student exposure to geriatrics with emphasis on fostering and nurturing student interest, consideration of various enticements to this specialty, and the development of health system-specific solutions to this problem. Knowledge of student and practice characteristics that increase the likelihood of selecting geriatrics as a specialty may allow for early identification and support of future geriatricians.


Immune Control of γ-Herpesviruses.
Vaccination against γ-herpesviruses has been hampered by our limited understanding of their normal control. Epstein-Barr virus (EBV)-transformed B cells are killed by viral latency antigen-specific CD8+ T cells in vitro, but attempts to block B cell infection with antibody or to prime anti-viral CD8+ T cells have protected poorly in vivo. The Doherty laboratory used Murid Herpesvirus-4 (MuHV-4) to analyze γ-herpesvirus control in mice and found CD4+ T cell dependence, with viral evasion limiting CD8+ T cell function. MuHV-4 colonizes germinal center (GC) B cells via lytic transfer from myeloid cells, and CD4+ T cells control myeloid infection. GC colonization and protective, lytic antigen-specific CD4+ T cells are now evident also for EBV. Subunit vaccines have protected only transiently against MuHV-4, but whole virus vaccines give long-term protection, via CD4+ T cells and antibody. They block infection transfer to B cells, and need include no known viral latency gene, nor any MuHV-4-specific gene. Thus, the Doherty approach of in vivo murine analysis has led to a plausible vaccine strategy for EBV and, perhaps, some insight into what CD8+ T cells really do.


Influence of Scanning Speed on Microstructure and Properties of Laser Cladded Fe-Based Amorphous Coatings.
Fe-based amorphous alloys with excellent mechanical properties are suitable for preparing wear resistant coatings by laser cladding. In this study, a novel Fe-based amorphous coating was prepared by laser cladding on 3Cr13 stainless steel substrates. The influence of scanning speeds on the microstructures and properties of the coatings was investigated. The microstructure compositions and phases were analyzed by scanning electron microscope, electron probe microanalyzer, and x-ray diffraction respectively. Results showed that the microstructures of the coatings changed significantly with the increase of scanning speeds. For a scanning speed of 6 mm/s, the cladding layer was a mixture of amorphous and crystalline regions. For a scanning speed of 8 mm/s, the cladding layer was mainly composed of block grain structures. For a scanning speed of 10 mm/s, the cladding layer was composed entirely of dendrites. Different dilution rates at the bonding zones were the main reasons for the microstructure change for different claddings. For all three scanning speeds, the coatings had higher hardness and wear resistance when compared with the substrate; as the scanning speed increased, the hardness and wear resistance of the coatings gradually decreased due to the change in microstructure.


A gender-balanced approach to the study of peer victimization and aggression subtypes in early childhood.
A short-term longitudinal study during early childhood (N = 301; 155 girls; M = 44.76 months old, SD = 8.20) investigated the prospective associations between peer victimization and aggression subtypes. Specifically, observations of relational and physical victimization as well as teacher reports of the forms (i.e., relational and physical) and functions (i.e., proactive and reactive) of aggression were collected at two time points during an academic year. Within- and between-group gender differences were examined as part of the preliminary analyses. In order to address key study questions, both directions of effect between peer victimization and aggression subtypes were examined. We found that teacher-reported proactive relational aggression predicted decreases in observed relational victimization over time, whereas reactive relational aggression predicted increases in observed relational victimization over time. Ways in which these and other findings extend the literature are discussed.


Steady shearing flows of deformable, inelastic spheres.
We extend models for granular flows based on the kinetic theory beyond the critical volume fraction at which a rate-independent contribution to the stresses develops. This involves the incorporation of a measure of the duration of the particle interaction before and after this volume fraction. At volume fractions less than the critical, the stress components contain contributions from momentum exchanged in collisions that are influenced by the particle elasticity. At volume fractions greater than the critical, the stress components contain both static contributions from particle elasticity and dynamic contributions from the momentum transfer associated with the release of elastic energy by the breaking of force chains. A simple expression for the duration of a collision before and after the critical volume fraction permits a smooth transition between the two regimes and predictions for the components of the stress in steady, homogeneous shearing that are in good agreement with the results of numerical simulations. Application of the theory to steady, inhomogeneous flows reproduces the features of such flows seen in numerical simulations and physical experiments.


Dynamic lumbrical muscle transfer for correction of posttraumatic extensor tendon subluxation.
Posttraumatic ulnar subluxation of the extensor tendon over the metacarpal head results from rupture of the radial sagittal fibers. The patient will complain of limited digital extension and pain. Various techniques have been described to correct the disorder by either reefing the sagittal fibers or using a slip of extensor tendon around the radial collateral ligament. Unfortunately, these techniques are either technically not feasible, reefing of the sagittal fibers, or result in significant stiffness, using a strip of extensor tendon. The author describes an extraarticular technique which uses a dynamic muscle transfer that is synergistic with metacarpophalangeal flexion.


Two-dimensional lamellar polyimide/cardanol-based benzoxazine copper polymer composite coatings with excellent anti-corrosion performance.
The economic loss and environmental damage caused by metal corrosion is irreversible. Thus, effective methods, such as coating technologies are used to protect metal surfaces from corrosion. In this work, cardanol-based benzoxazine (CB) was synthesized by a solvent-free method using cardanol, paraformaldehyde and n-octylamine. A cardanol-based benzoxazine copper polymer (CBCP) with good mechanical properties was then prepared by CuCl2 catalysis and can be cured at room temperature. Subsequently, polyimide corrosion inhibitors with a two-dimensional sheet structure (pyromellitic dianhydride polyimide (PDPI) and 1,4,5,8-naphthalene tetracarboxylic dianhydride polyimide (NDPI)) were designed and prepared. Lastly, PDPI or NDPI was mixed with CBCP to obtain two-dimensional lamellar polyimide/cardanol-based benzoxazine copper polymer composite coatings. The Tafel curves and electrochemical impedance spectroscopy (EIS) measurements showed composite coatings with good corrosion resistance in different corrosive media. Compared to CBCP coating, the anticorrosion performance of the composite coatings improved obviously, especially the coating obtained with 0.5 wt% PDPI. It exhibits a high polarization resistance (3.874 × 109 Ω), a high protection efficiency (99.99% and 97.98%) and low corrosion rate (3.376 × 10-6 mm year-1). This work suggested a facile and eco-friendly strategy for preparing bio-based anticorrosive composite coatings from low cost and abundant cardanol and polyimide corrosion inhibitors, which will significantly promote their application in metal anticorrosion.


Dose-dependent effects of tryptophan on learning and memory.
The concentration of 5-hydroxytryptamine (5-HT, Serotonin) varies as a result of physiological changes in the availability of its precursor tryptophan to the serotonergic neurons in the brain. Increase in brain tryptophan occurs following an increase in plasma tryptophan concentration. Tryptophan intake increases brain serotonin metabolism and enhances memory. The Present study was designed to investigate the effects of oral administration of tryptophan (TRP) at different doses (100, 300 and 500mg/kg) for two weeks on learning and memory functions and Neurochemical changes in rats. Control rats were given drinking water. Assessment of memory in rats was done by using the water Maze. on the 14th day trail training of water Maze was given to rats and after 1h of this 2nd trial of these rats were done. On the next day (After 24h of trail) long-term memories of these rats were monitored. After 1 hour of this all rats were killed by decapitation using guillotine. Brain and blood was collected and stored at -70°C. Neurochemical estimations of Plasma and brain tryptophan, 5-HT and 5-HIAA in brain were made by HPLC-EC. Result showed that administration of tryptophan enhanced performance on water Maze test. Tryptophan treated animals exhibited higher level of Plasma as well as brain tryptophan. 5-HT and 5-HIAA levels were also increased in tryptophan treated rats. Findings are discussed in context with the role of 5-HT metabolism in learning and memory process in rats. Results may help to understand the 5-HT changes following long term TRP administration in a dose dependent manner and will help to suggest the use of TRP in serotonin related illnesses.


Accuracy of magnetic resonance imaging of the knee and unjustified surgery.
Magnetic resonance imaging of the knee is greater than 90% accurate in detecting intraarticular disease when performed and interpreted by musculoskeletal magnetic resonance imaging specialists in specialized medical centers. However, independent imaging institutions often offer less expensive services to health insurers. We wondered if the magnetic resonance imaging performed in our community is of equivalent quality and accuracy. We studied a homogenous group of healthy, young, and fit military recruits to represent a cross section of our country's population. We analyzed all knee magnetic resonance images of soldiers who subsequently had primary arthroscopic knee surgery within a 3-month period from 1997-1998. The results were compared with surgical findings of four structures: medial meniscus, lateral meniscus, anterior cruciate ligament, and articular cartilage. Of the 1185 arthroscopies and 633 magnetic resonance images of the knee performed in 14 institutions, 139 paired magnetic resonance imaging arthroscopic reports met our inclusion criteria. The results showed a false positive rate of 65% for the medial meniscus, 43% for the lateral meniscus, 47.2% for the anterior cruciate ligament, and 41.7% for articular cartilage disease when compared with surgical findings. Accuracy rates were 52%, 82%, 80%, and 77%, respectively. Thirty-seven percent of the operations supported by a significant disorder on magnetic resonance imaging were unjustified. Our findings highlight the consequences that may occur when basing medical care on cost rather than quality of care.


Negative viscosity can enhance learning of inertial dynamics.
We investigated how learning of inertial load manipulation is influenced by movement amplification with negative viscosity. Using a force-feedback device, subjects trained on anisotropic loads (5 orientations) with free movements in one of three conditions (inertia only, negative viscosity only, or combined), prior to common evaluation conditions (prescribed circular pattern with inertia only). Training with Combined-Load resulted in lower error (6.89±3.25%) compared to Inertia-Only (8.40±4.32%) and Viscosity-Only (8.17±4.13%) according to radial deviation analysis (% of trial mean radius). Combined-Load and Inertia-Only groups exhibited similar unexpected no-load trials (8.38±4.31% versus 8.91±4.70% of trial mean radius), which suggests comparable low-impedance strategies. These findings are remarkable since negative viscosity, only available during training, evidently enhanced learning when combined with inertia. Modeling analysis suggests that a feedforward after-effect of negative viscosity cannot predict such performance gains. Instead, results from Combined-Load training are consistent with greater feedforward inertia compensation along with a small increase in impedance control. The capability of the nervous system to generalize learning from negative viscosity suggests an intriguing new method for enhancing sensorimotor adaptation.


Risk factors for saddle-related skin lesions on elephants used in the tourism industry in Thailand.
Lesions related to working conditions and improper saddle design are a concern for a variety of working animals including elephants. The objectives of the present study were to determine the prevalence of cutaneous lesions in anatomic regions (i.e., neck, girth, back, tail) in contact with saddle-related equipment among elephants in Thailand working in the tourism industry, and to identify potential risk factors associated with these lesions. Data for this cross-sectional study were collected between May 2007 and July 2007 on 194 elephants from 18 tourism camps across Thailand. There was a high prevalence (64.4 %; 95 % CI 57.3 - 71.2) of active lesions, most often located on the back region. Using multilevel multivariable logistic regression modelling containing a random intercept for camp we identified the following risk factors: increasing elephant age, the use of rice sacks as padding material in contact with the skin, and the provision of a break for the elephants. Working hours had a quadratic relationship with the log odds of an active lesion where the probability of an active lesion initially increased with the number of working hours per day and then declined possibly reflecting a "healthy worker" bias where only animals without lesions continue to be able to work these longer hours. While we recognize that the cross-sectional nature of the study posed some inferential limitations, our results offer several potential intervention points for the prevention of these lesions. Specifically, we recommend the following until longitudinal studies can be conducted: increased monitoring of older elephants and the back region of all elephants, working less than 6 hours per day, and the avoidance of rice sacks as padding material in contact with skin.


[Endonasal endoscopic chiasmapexy for secondary empty sella syndrome following fenestration of a Rathke's cleft cyst].
Drainage by fenestration of the cyst wall via the transsphenoidal apporach is the most commonly used treatment for symptomatic Rathke's cleft cyst (RCC). The same procedure is usually adopted for recurrence of RCC. We have encountered a case of secondary empty sella syndrome presented with visual field defects after repeated surgery for RCC. Secondary empty sella syndrome following the surgery of RCC is rare. The condition was explained by the mechanism that the optic nerve adhered to the cyst wall and it was tethered downward as the cyst shrank after the surgery. We treated the patient via the endonasal endoscopic transsphenoidal approach by placing holed silicone plates under the sellar floor to elevate the sellar contents and the optic nerve. Silicone plate is hard enough to support the sella and the small holes on it would facilitate drainage of the cyst contents. This method has proved useful as chiasmapexy for secondary empty sella syndrome after the surgery of RCC.


Network analysis of mild traumatic brain injury, persistent neurobehavioral and psychiatric symptoms, and functional disability among recent-era United States veterans.
Recent-era U.S. veterans are clinically complex, with a high prevalence of co-occurring mild traumatic brain injury (mTBI), psychiatric conditions, and behavioral dysfunction. The current study examined the direct and indirect associations between mTBI and persistent neurobehavioral, psychiatric, and functional disability symptoms among recent-era U.S. veterans and service members (n = 648). We evaluated the postconcussive syndrome (PCS) potential causal model with two network analysis modeling approaches. Separate analyses were conducted for military mTBI and lifetime mTBI. An exploratory factor analysis was conducted to limit topological overlap in the network analysis. The most influential symptoms (i.e., the unique variables most strongly associated with the rest of the network) in the military mTBI network were behavioral disengagement, expected influence (EI) = 1.10; cognitive difficulties, EI = 1.08; agitation/irritability, EI = 1.05; and PTSD-related reexperiencing and avoidance symptoms, EI = 0.98. After accounting for other symptoms, mTBI was only minimally informative, EI = 0.34. Additionally, military mTBI did not moderate the association between symptoms or the overall connectivity of the network. The results for lifetime mTBI were consistent with those for military mTBI. The present analyses identified a variety of behavioral, cognitive, and emotional symptoms that play an important role in understanding comorbidity and daily functioning among recent-era U.S. veterans. Associations between cumulative mTBI that occurred in civilian or military settings were indirect and relatively small in magnitude. The current results add to a growing literature raising doubts about the PCS model.


Vasoinhibin generation and effect on neuronal apoptosis in the hippocampus of late mouse embryos.
Morphological and behavioral evidence suggests that vasoinhibin is present in the central nervous system (CNS), triggering neuroendocrine and behavioral responses to stress. Moreover, vasoinhibin reduces neuronal survival and differentiation of primary sensory neurons of the peripheral nervous system. To address the functional role played by vasoinhibin at the CNS, and to better understand the underlying mechanisms involved in its actions, we treated primary cultured hippocampal neurons obtained from embryonic day 16 (E16) mice with a human recombinant vasoinhibin. We examined the resulting cellular changes, focusing on neuronal cell death, and explored the local generation of vasoinhibin within the hippocampus. Our results show that vasoinhibin significantly reduced neuronal cell density and increased immunoreactive activated caspase-3 and TUNEL-positive staining at 72, 16, and 24 h, respectively. Furthermore, vasoinhibin increased the expression of proapoptotic genes BAX, BAD, BIM, and PUMA and decreased that of the antiapoptotic gene BCL-2 at 24 h, as assessed by quantitative real-time reverse transcription-polymerase chain reaction. Vasoinhibin effects were blocked by coincubation with a vasoinhibin antibody or with prolactin. Immunoreactive bands consistent with vasoinhibin were observed in hippocampal extracts by Western blot analysis, and a prolactin standard was cleaved to vasoinhibin by a hippocampal lysate in a heat- and cathepsin D inhibitor pepstatin A-dependent fashion. Taken together, these data support the notion that vasoinhibin is locally produced by cathepsin D within the embryonic mouse hippocampus, a brain region that plays a critical role in emotional regulation, resulting in decreased neuronal cell viability via the activation of the intrinsic apoptosis pathway.


Acute short-term hyperoxia followed by mild hypoxia does not increase EPO production: resolving the "normobaric oxygen paradox".
Recent findings suggest that besides renal tissue hypoxia, relative decrements in tissue oxygenation, using a transition of the breathing mixture from hyperoxic to normoxic, can also stimulate erythropoietin (EPO) production. To further clarify the importance of the relative change in tissue oxygenation on plasma EPO concentration [EPO], we investigated the effect of a consecutive hyperoxic and hypoxic breathing intervention. Eighteen healthy male subjects were assigned to either IHH (N = 10) or CON (N = 8) group. The IHH group breathed pure oxygen (F(i)O(2) ~ 1.0) for 1 h, followed by a 1-h period of breathing a hypoxic gas mixture (F(i)O(2) ~ 0.15). The CON group breathed a normoxic gas mixture (F(i)O(2) ~ 0.21) for the same duration (2 h). Blood samples were taken just before, after 60 min, and immediately after the 2-h exposure period. Thereafter, samples were taken at 3, 5, 8, 24, 32, and 48 h after the exposure. During the breathing interventions, subjects remained in supine position. There were significant increases in absolute [EPO] within groups at 8 and 32 h in the CON and at 32 h only in the IHH group. No significant differences in absolute [EPO] were observed between groups following the intervention. Relative (∆[EPO]) levels were significantly lower in the IHH than in the CON group, 5 and 8 h following exposure. The tested protocol of consecutive hyperoxic-hypoxic gas mixture breathing did not induce [EPO] synthesis stimulation. Moreover, the transient attenuation in ∆[EPO] in the IHH group was most likely due to a hyperoxic suppression. Hence, our findings provide further evidence against the "normobaric O(2) paradox" theory.


Dysregulation of the PDGFRA gene causes inflow tract anomalies including TAPVR: integrating evidence from human genetics and model organisms.
Total anomalous pulmonary venous return (TAPVR) is a congenital heart defect inherited via complex genetic and/or environmental factors. We report detailed mapping in extended TAPVR kindreds and mutation analysis in TAPVR patients that implicate the PDGFRA gene in the development of TAPVR. Gene expression studies in mouse and chick embryos for both the Pdgfra receptor and its ligand Pdgf-a show temporal and spatial patterns consistent with a role in pulmonary vein (PV) development. We used an in ovo function blocking assay in chick and a conditional knockout approach in mouse to knock down Pdgfra expression in the developing venous pole during the period of PV formation. We observed that loss of PDGFRA function in both organisms causes TAPVR with low penetrance (approximately 7%) reminiscent of that observed in our human TAPVR kindreds. Intermediate inflow tract anomalies occurred in a higher percentage of embryos (approximately 30%), suggesting that TAPVR occurs at one end of a spectrum of defects. We show that the anomalous pulmonary venous connection seen in chick and mouse is highly similar to TAPVR discovered in an abnormal early stage embryo from the Kyoto human embryo collection. Whereas the embryology of the normal venous pole and PV is becoming understood, little is known about the embryogenesis or molecular pathogenesis of TAPVR. These models of TAPVR provide important insight into the pathogenesis of PV defects. Taken together, these data from human genetics and animal models support a role for PDGF-signaling in normal PV development, and in the pathogenesis of TAPVR.


Construction of a recombinase-deficient mutant recA protein that retains single-stranded DNA-dependent ATPase activity.
The recA1 mutation is a single point mutation that replaces glycine 160 of the recA polypeptide with an aspartic acid residue. The mutant recA1 protein has a greatly reduced single-stranded DNA-dependent ATPase activity at pH 7.5 compared to the wild-type protein. Interestingly, the recA1 protein does exhibit a vigorous ATPase activity at pH 6.2. To explore the molecular basis of this pH effect, we used site-directed mutagenesis to replace aspartic acid 160 of the recA1 polypeptide with an isosteric, but nonionizing, asparagine residue. The new [Asn160]recA protein catalyzes ATP hydrolysis at pH 7.5 with the same turnover number as the wild-type protein. This result suggests that the activation of the recA1 protein ATPase activity that occurs at pH 6.2 may be due, in part, to neutralization of the negatively charged aspartic acid 160 side chain. Although it is an active single-stranded DNA-dependent ATPase, the [Asn160]recA protein is unable to complement a recA deletion in vivo and is unable to carry out the three-strand exchange reaction in vitro. Further examination of ATP hydrolysis (under strand exchange conditions) revealed that the ATPase activity of the [Asn160]recA protein is strongly suppressed in the presence of Escherichia coli single-stranded DNA-binding protein (a component of the strand exchange assay), whereas the ATPase activity of the wild-type recA protein is stimulated by the E. coli protein. To account for these results, we speculate that ATP may induce specific conformational changes in the wild-type recA protein that are essential to the DNA pairing process and that these conformational changes may not occur with the [Asn160]recA protein.


The 14-3-3 cerebrospinal fluid immunoassay lacks utility in the diagnosis of clinical scrapie.
This study determined whether the immunoassay for cerebrospinal fluid 14-3-3 protein concentration was sensitive and specific in the diagnosis of naturally occurring clinical scrapie in sheep. Cerebrospinal fluid was collected from 9 sheep with the confirmed diagnosis of scrapie. Additionally, cerebrospinal fluid was collected from 13 clinically normal sheep, which originated from a closely monitored flock with no history of scrapie. Sensitivity and specificity were calculated using standard epidemiological methods. Cerebrospinal fluid immunoassay results did not differ significantly between positive and negative sheep. Test sensitivity varied from 0.55 to 0.66, depending on the choice of test endpoint. Test specificity varied from 0.30 to 0.77, depending on the choice of test endpoint. The 14-3-3 cerebrospinal fluid immunoassay appears to have no value in the diagnosis of clinical scrapie in sheep.


Cell-Penetrating Peptide Conjugated SERS Nanosensor for in Situ Intracellular pH Imaging of Single Living Cells during Cell Cycle.
Intracellular pH is an important modulator of cell functions, and its subtle change may dramatically affect the cellular activities and cause diseases. A reliable imaging of the intracellular pH is still a great challenge. We imaged the intracellular pH during the cell cycle at the single living cell level using newly designed cell-penetrating peptide conjugated pH nanosensors on a home-built in situ microscopic cell culture platform. The conjugated cell-penetrating peptide greatly enhanced the uptake of nanosensors without sacrificing the pH response. We observed a gradual alkalization from interphase to prophase and rapid acidification from prometaphase to telophase, reflecting variation and consumption of the species related to the energy storage during cell cycle. We realized SERS-based pH and fluorescence dual-mode imaging when the pH sensor was further modified with fluorescence dye. The integration of SERS imaging with in situ microscopic cell culture system offers great opportunity for revealing the intracellular pH-related biological and pathological processes.


Searching for unknown transfusion-transmitted hepatitis viruses: a binational cohort study of 1.5 million transfused patients.
Both hepatitis B and C viruses were transmitted through blood transfusion before implementation of donor screening. The existence of additional, yet unknown transfusion transmittable agents causing liver disease could have important public health implications. Analyses were based on the Scandinavian Donations and Transfusions (SCANDAT2) database. Cox regression models were used to estimate the hazard ratio (HR) of developing chronic liver disease in recipients of blood from donors who later developed any chronic liver disease compared to recipients who received blood transfusion from healthy donors. We also studied whether the risk of liver disease was increased in patients who received units from 'high-risk' donors, defined as donors who had a higher than expected occurrence of liver disease amongst their previous recipients. All analyses were stratified before and after 1992 to account for the effect of screening for hepatitis C virus. A total of 1 482 922 transfused patients were included in the analyses. Analyses showed evidence of transfusion transmission of liver diseases before, but not after the implementation of hepatitis C virus screening in 1992, with HRs for any liver disease of 1.38 [95% confidence interval (CI), 1.30-1.46] and 0.99 (95% CI, 0.91-1.07), before and after 1992, respectively. Similarly, blood components from 'high-risk' donors conferred increased risks before, but not after 1992. Our data provide no evidence for transfusion transmission of agents causing liver disease after the implementation of screening for hepatitis B and C, and suggest that if such transmission does occur, it is rare.


Rapid colorimetric sensing of gadolinium by EGCG-derived AgNPs: the development of a nanohybrid bioimaging probe.
Polyphenol functionalized silver nanoparticles (AgNPs) have been developed and demonstrated as colorimetric sensors for the selective detection of gadolinium. The newly obtained AgNP-Gd3+ conjugates exhibit high aqueous dispersibility and excitation dependent fluorescence emission. The conjugates offer multicolor bioimaging potential owing to their excellent luminescence properties.