GFP-expressing influenza A virus for evaluation of the efficacy of antiviral agents.
To address its value as a screening tool in the development of antiviral drugs, a recombinant influenza virus expressing green fluorescent protein (rPR8-GFP virus) was investigated in vitro and in vivo. The inhibition of viral growth by a neuraminidase inhibitor in the cells or lower respiratory tracts of mice could be visualized by the level of fluorescence. In addition, the rPR8-GFP virus exhibited high pathogenicity in mice. Taken together, these results suggest that the rPR8-GFP virus can be a useful tool for the rapid identification of antiviral drugs active against influenza viruses.


Towards more efficient low-carbon agricultural technology extension in China: identifying lead smallholder farmers and their behavioral determinants.
In the transition to low-carbon agriculture, smallholder farmers face more constraints. Identifying lead smallholder farmers and leveraging their peer effects can accelerate low-carbon agricultural technology extension among smallholder farmers. Based on survey data from 643 rice farmers in Zhejiang Province, China, this study constructs a finite mixture model (FMM) to identify lead smallholder farmers and then uses a quantile regression model (QRM) to explore their behavioral determinants. The main conclusions are as follows. First, despite the homogeneity in the production mode and resource constraints, lead smallholder farmers are younger and more open to risk, and they have higher educational levels and more family laborers. Second, a higher use efficiency of heterogeneous information is the key to differentiating lead smallholder farmers from other smallholder farmers. Third, green agricultural producer services can effectively alleviate resource constraints and contribute to the low-carbon transition of all smallholder farmers. These results can help redesign targeted extension policies to incentivize lead smallholder farmers.


Prevention of defects of axonal transport and nerve conduction velocity by oral administration of myo-inositol or an aldose reductase inhibitor in streptozotocin-diabetic rats.
The effects of orally-administered myo-inositol have been compared with those of an aldose reductase inhibitor on acute neurological defects in experimentally diabetic rats. Three groups of streptozotocin-treated diabetic rats (50 mg/kg, IP) together with three groups of age-matched controls (saline, IP) were compared. One pair of groups (control and diabetic) were untreated for 3 weeks, another pair of groups received daily oral myo-inositol (667 mg/kg) and the third pair received an aldose reductase inhibitor (ICI 105 552; 50 mg . kg-1, day-1, orally). The untreated diabetic group showed statistically significant deficits in accumulation, proximal to 24 h sciatic nerve constrictions, of choline acetyltransferase activity by comparison with untreated controls (2.8 +/- 0.4 versus 5.1 +/- 0.4 nmol acetylcholine . h-1 . nerve-1; p less than 0.001). The untreated diabetic rats also showed a fall in motor nerve conduction velocity of 6.2 +/- 0.7 m/s which was statistically significant (p less than 0.001). Treatment of the diabetic group with myo-inositol prevented the development of both defects of axonal transport and conduction velocity; both measurements were similar to those of the myo-inositol treated control rats. Likewise the diabetic rats which received aldose reductase inhibitor showed prevention of both defects. Nerves from untreated diabetic rats showed marked sorbitol accumulation and a statistically significant reduction in myo-inositol content by comparison with the untreated controls (sorbitol, 1.56 +/- 0.22 versus 0.8 +/- 0.01 and myo-inositol, 1.47 +/- 0.10 versus 2.3 +/- 0.10 nmol/mg; p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


Infertility and psychiatric morbidity.
To assess the relationship between psychiatric disorders and infertility. Case-control study. Fertile and infertile volunteer couples in an academic research setting. Eighty-one infertile couples recruited from an infertility center before fertility treatment and 70 fertile controls recruited from an obstetrics and gynecology clinic. None. The presence of Axis 1 psychiatric disorders. The occurrence of current psychiatric disorders was significantly higher among infertile subjects than among fertile controls, especially for adjustment disorder with mixed anxiety and depressed mood (16% vs. 2%) and for binge eating disorder (8% vs. 0). Our data highlight that a percentage of infertile patients have already developed a psychiatric disorder at the time of their first contact with a specialized fertility service. Possible applications are discussed, including the recommendation that gynecologists screen for clinical or subclinical psychiatric disorders in infertility patients and offer treatment accordingly.


Glomerular filtration rate and cardiometabolic risk in an outpatient pediatric population with high prevalence of obesity.
To evaluate the relationship between estimated glomerular filtration rate (eGFR) and cardiometabolic risk factors (CMRF) in an outpatient pediatric population with high prevalence of obesity. eGFR was evaluated in 901 children unselected for chronic kidney disease of whom 694 were overweight/obese (6-16 years) and 207 were age- and sex-matched normal weight (NW). We generated three categories of eGFR: mild-low eGFR (< 20th percentile), high eGFR (>80th percentile) and intermediate eGFR (20-80th percentile), considered as the reference category Children with either mild-low or high eGFR category showed a 2-4 fold higher Odds ratio of high blood pressure, left ventricular hypertrophy, and microalbuminuria compared with children of the intermediate eGFR category. In addition, children with mild-low eGFR levels showed a 1.5-2 fold higher Odds ratio of impaired fasting glucose and high white blood cell count compared with children with intermediate eGFR levels. In outpatient children with high prevalence of obesity, children with either mildly reduced or high eGFR have an increased burden of CMRF. Children with eGFR < 97 mL/min/1.73 m² show a worse CMR profile. This finding supports the usefulness to assess eGFR to identify children with unfavorable CMR profile.


Risk Factors for Rescue Therapy in Crohn's Patients Maintained on Infliximab After Withdrawal of the Immunomodulator: A Long-Term Follow-Up.
Usefulness of thiopurine and scheduled infliximab combination therapy in non-immunomodulator (IM)-naïve Crohn's disease (CD) patients and the optimal length of dual therapy are still debated. To determine proportion of patients developing disease flare requiring rescue therapy and risk factors associated with disease flare after de-escalation of IM from combination therapy. Adult CD patients in clinical remission on combination therapy were identified from a large single-center database between 2002 and 2009. Patients who had their IM stopped in the absence of adverse events were included. Association between clinical and demographic variables and time until rescue therapy was analyzed using Cox-proportional hazard models. Forty-three CD patients on combination therapy in clinical remission at time of IM de-escalation were identified and followed up for a median duration of 61.6 months (range 5.4-129.5). Median duration of remission on combination therapy prior to IM de-escalation was 12.0 months (range 4-74). Thirty-one patients (72.1%) required rescue therapy during follow-up. On multivariable analysis, age at diagnosis < 16 years versus > 40 years (HR 4.55, 95% CI 1.18-17.62, p = 0.028), using methotrexate instead of azathioprine in combination with infliximab (HR 3.37, 95% CI 1.14, 9.96, p = 0.028), and duration of combination therapy < 6 months (HR 5.68, 95% CI 1.58, 20.36, p = 0.007) increased risk for rescue therapy. A large proportion of CD patients on combination therapy experienced a flare following IM withdrawal. Young age at diagnosis, short duration of combination therapy, and methotrexate use were independent predictors of the need for rescue therapy.


Transition Readiness Assessment Questionnaire: Skill gaps and psychosocial predictors of transition readiness among adolescents and young adults with chronic medical conditions.
Transferring from paediatric to adult care can be challenging. Adolescents and young adults (AYAs) with chronic health conditions need to develop a specific set of skills to ensure lifelong medical follow-up due to the chronicity of their condition. The Transition Readiness Assessment Questionnaire-French version (TRAQ-FR) is a 19-item questionnaire measuring such skills. The aims of the study were to (1) describe participant characteristics and (2) identify constructs related to, and predictors of, having learned domain-specific transition readiness skills. Participants included 216 AYAs aged 14-20 years (M = 15.93; SD = 1.35; 54.1% male) recruited from five outpatient clinics in a Canadian tertiary hospital. AYAs completed the TRAQ-FR, the Pediatric Quality of Life Inventory 4.0 (PedsQL) and a sociodemographic questionnaire. Descriptive, bivariate and binary logistic regression analyses were conducted. Overall, participants reported significantly higher scores on the Talking with Providers, Managing Daily Activities and Managing Medications subscales than on the Appointment Keeping and Tracking Health Issues subscales (F[41075] = 168.970, p < .001). At the item level, median scores (on a 5-point Likert scale) suggest that AYAs had begun practising five of the 19 skills (median scores ≥4; 'Yes, I have started doing this'), while a median score of 1 ('No, I don't know how') was found for one item ('Do you get financial help with school or work?'). At the subscale level, TRAQ-FR skills and skill gaps were related to AYAs' age, sex and PedsQL scores (ps < .05). Older and female AYAs were more likely to have begun practising specific TRAQ-FR subscale skills. Better psychosocial functioning was also related to having learned specific transition readiness skills. AYAs show several gaps in transition readiness. Targeted intervention in transition readiness skill development could take into account AYAs' age, sex and psychosocial functioning for a successful transfer to adult care.


[Research on phytoestrogenic effects and its mechanism of psoralen].
To observe and evaluate the phytoestrogenic effects and its mechanism of psoralen in estrogen receptor (ER) alpha and beta positive T47D and ishikawa cells. The proliferation rate of T47D influenced by 1 x 10(-5) mol x L(-1) to 1 x 10(-9) mol x L(-1) psoralen and that of Ishikawa influenced by 1 x 10(-6) mol x L(-1) and 1 x 10(-7) mol x L(-1) psoralen were analyzed by MTT assay. PR mRNA expression in T47D was quantified by RT-PCR assay. Estrogen receptor antagonist ICI 182, 780 was employed as a tool. ER-alpha and ER-beta expression of T47D was measured by flow cytometry. The proliferation rates of T47D cells treated with 1 x 10(-5) mol x L(-1) to 1 x 10(-7) mol x L(-1) psoralen and ishikawa cells treated with 1 x 10(-6) mol x L(-1) to 1 x 10(-7) mol x L(-1) psoralen were increased significantly. The RT-PCR result showed that 1 x 10(-7) mol x L(-1) and 1 x 10(-6) mol x L(-1) psoralen could increase PR expression in T47D cells. The above effects could be blocked by ICI 182,780. Psoralen could also induce the augment of ER-alpha and ER-beta expression in T47D cells significantly. Psoralen has phytoestrogenic effects. The effects are attained through ER pathway.


Therapeutic effect of rebamipide on ammonia-induced gastric mucosal hemorrhagic lesion in rats.
Rebamipide, 2-(4-chlorobenzoylamino)-3-[2(1H)-quinolinone-4-yl]-propionic acid, a novel antipeptic ulcer agent, has been reported to prevent various acute experimental gastric mucosal lesions and to accelerate the healing of chronic ulcers. Therapeutic effect of rebamipide was investigated with regard to the inhibitory effect on xanthine oxidase activity and type conversion of the enzyme which play a profound role in oxygen radicals generation system. Intraperitoneal administration of rebamipide at 60 mg/kg body weight reduced the xanthine oxidase activity, lipid peroxide content in ammonia induced hemorrhagic lesion. These results suggest that the therapeutic effect of rebamipide on gastric mucosal lesion may be in part due to the inhibitory activity of xanthine oxidase and type conversion rate of the enzyme.


The resistance of Pteridium aquilinum (L.) Kuhn to insect attack by Trichoplusia ni (Hübn.).
Resistance of Pteridium aquilinum to insect attack was studied by incorporating air dried bracken leaf meal and extracts of bracken leaf meal into an artificial diet for Trichoplusia ni (Lepidoptera: Noctuidae). When larvae are reared on diets containing 6% bracken leaf meal, they do not mature past the second instar and after 16 days the average weight is approximately 1 mg compared to 70 mg for control larvae. Feeding initiation studies indicate that a feeding deterrent is present in bracken fern but feeding rates and food utilization efficiency studies suggest that either the deterrent or another compound also functions as a toxin. This toxin does not affect growth, feeding rates, or utilization efficiency for the first 4 days after third instar larvae are transferred to a diet containing the water extract of bracken leaf meal; thereafter growth is terminated and feeding is greatly reduced. The active factor is water soluble, heat labile, and non-volatile and this partial characterization indicates that neither the bracken ecdysones or the anti-thiamine factor of bracken is involved in the resistance of this fern to insect attack by T. ni.


[Use of PubMed to improve evidence-based medicine in routine urological practice].
Applying evidence-based medicine in daily clinical practice is the basis of patient-centered medicine and knowledge of accurate literature acquisition skills is necessary for informed clinical decision-making. PubMed is an easy accessible, free bibliographic database comprising over 21 million citations from the medical field, life-science journals and online books. The article summarizes the effective use of PubMed in routine urological clinical practice based on a common case scenario. This article explains the simple use of PubMed to obtain the best search results with the highest evidence. Accurate knowledge about the use of PubMed in routine clinical practice can improve evidence-based medicine and also patient treatment.


Crystal structures of the M1 and M4 muscarinic acetylcholine receptors.
Muscarinic M1-M5 acetylcholine receptors are G-protein-coupled receptors that regulate many vital functions of the central and peripheral nervous systems. In particular, the M1 and M4 receptor subtypes have emerged as attractive drug targets for treatments of neurological disorders, such as Alzheimer's disease and schizophrenia, but the high conservation of the acetylcholine-binding pocket has spurred current research into targeting allosteric sites on these receptors. Here we report the crystal structures of the M1 and M4 muscarinic receptors bound to the inverse agonist, tiotropium. Comparison of these structures with each other, as well as with the previously reported M2 and M3 receptor structures, reveals differences in the orthosteric and allosteric binding sites that contribute to a role in drug selectivity at this important receptor family. We also report identification of a cluster of residues that form a network linking the orthosteric and allosteric sites of the M4 receptor, which provides new insight into how allosteric modulation may be transmitted between the two spatially distinct domains.


Use of Framingham Risk Score as a Clinical Tool for the Assessment of Fitness for Work: Results From a Cohort Study.
The aim of this study is to validate the use of the Framingham Risk Score (FRS) as clinical tool to predict the risk of diagnosis of unsuitability for work in a cohort of Italian workers. A cohort of workers has been observed from January 2006 to March 2014. FRS was calculated at each visit. Health surveillance diagnosis of unsuitability for work was selected as outcome. Two thousand eight hundred fifty seven workers were observed, 58.9% were men, mean age was 51.6 (±6.7), the mean FRS was 15.1% (±10.7%). Increased values of FRS at baseline were associated with increased rate of diagnosis of unsuitability for work (Hazard ratio [HR], 11.2, 95%CI, 3.3 to 37.8). FRS is a strong predictor of diagnosis of unsuitability for work and should be used as a clinical tool for the assessment of fitness for work in health surveillance.


An evaluation of the protective effects of chlorogenic acid on broiler chickens in a dextran sodium sulfate model: a preliminary investigation.
This study was conducted to investigate the protective effects of chlorogenic acid (CGA) on broilers subjected to dextran sodium sulfate (DSS)-induced intestinal damage. One hundred and forty-four 1-day-old male Arbor Acres broiler chicks were allocated into one of 3 groups with 6 replicates of eight birds each for a 21-d trial. The treatments included: 1) Control group: normal birds fed a basal diet; 2) DSS group: DSS-treated birds fed a basal diet; and 3) CGA group: DSS-treated birds fed a CGA-supplemented control diet. An oral DSS administration via drinking water was performed from 15 to 21 d of age. Compared with the control group, DSS administration reduced 21-d body weight and weight gain from 15 to 21 d, but increased absolute weight of jejunum and absolute and relative weight of ileum (P < 0.05). DSS administration elevated circulating D-lactate concentration and diamine oxidase activity (P < 0.05), which were partially reversed when supplementing CGA (P < 0.05). The oral administration with DSS decreased villus height and villus height/crypt depth ratio, but increased crypt depth in jejunum and ileum (P < 0.05). Compared with the control group, DSS administration increased serum glutathione level and jejunal catalase activity and malonaldehyde accumulation, but decreased jejunal glutathione level (P < 0.05). In contrast, feeding a CGA-supplemented diet normalized serum glutathione and jejunal malonaldehyde levels, and increased jejunal glutathione concentration in DSS-administrated birds (P < 0.05). Additionally, CGA supplementation reduced ileal malonaldehyde accumulation in DSS-treated birds (P < 0.05). DSS challenge increased levels of serum interferon-γ and interleukin-6, jejunal interleukin-1β, tumor necrosis factor-α, and interleukin-6, and ileal interleukin-1β and interleukin-6 when compared with the control group (P < 0.05). The elevated serum interferon-γ and ileal interleukin-6 levels were normalized to control values when supplementing CGA (P < 0.05). The results suggested that CGA administration could partially prevent DSS-induced increased intestinal permeability, oxidative damage, and inflammation in broilers, although it did not improve their growth performance and intestinal morphology.


Prognostic significance of a complete pathological response after induction chemotherapy in operable breast cancer.
Only a few papers have been published concerning the incidence and outcome of patients with a pathological complete response after cytotoxic treatment in breast cancer. The purpose of this retrospective study was to assess the outcome of patients found to have a pathological complete response in both the breast and axillary lymph nodes after neoadjuvant chemotherapy for operable breast cancer. Our goal was also to determine whether the residual pathological size of the tumour in breast could be correlated with pathological node status. Between 1982 and 2000, 451 consecutive patients were registered into five prospective phase II trials. After six cycles, 396 patients underwent surgery with axillary dissection for 277 patients (69.9%). Pathological response was evaluated according to the Chevallier's classification. At a median follow-up of 8 years, survival was analysed as a function of pathological response. A pathological complete response rate was obtained in 60 patients (15.2%) after induction chemotherapy. Breast tumour persistence was significantly related to positive axillary nodes (P=5.10(-6)). At 15 years, overall survival and disease-free survival rates were significantly higher in the group who had a pathological complete response than in the group who had less than a pathological complete response (P=0.047 and P=0.024, respectively). In the absence of pathological complete response and furthermore when there is a notable remaining pathological disease, axillary dissection is still important to determine a major prognostic factor and subsequently, a second non cross resistant adjuvant regimen or high dose chemotherapy could lead to a survival benefit.


Bacteria and granulation tissue associated with Montgomery T-tubes.
Although complications (infection, development of granulation tissue) of silicone Montgomery T-tubes have been reported, the microbiological consequences and the origin of granulation tissue have not yet been evaluated. A prospective trial. Twenty-three Montgomery T-tubes from 10 patients were analyzed with regard to the development of granulation tissue, bacterial growth (including genotyping with polymerase chain reaction), and results of sensitivity testing. Furthermore, stent sterilization (n = 6) was investigated. Granulation tissue occurred with 74% of the stents, and all specimens showed signs of infection but no foreign body reaction. The predominant organisms were Staphylococcus aureus (35%) and Pseudomonas aeruginosa (17%). The differences between groups with and without granulation tissue were significant for P aeruginosa. Polymerase chain reaction fingerprinting of the S aureus obtained from 15 stents (n = 3 patients) revealed a total of seven different genotypes. Whereas two of these patients harbored six different genotypes of S aureus, the third patient was persistently colonized by S aureus over a 15-month period with the identical genotype. Susceptibility testing showed most commonly (65%) sensitivity to a combination of amoxicillin-clavulanate and ofloxacin. After sterilization, 92% of analyzed stent segments showed no bacterial growth. Granulation tissue commonly occurred next to the silicone (subglottic area, stoma) where S aureus and P aeruginosa were commonly isolated. A combination of mechanical irritation and bacterial infection seems to account for the development of granulation tissue. Polymerase chain reaction fingerprinting showed both prolonged persistence and a change of colonizing strains after multiple stent replacements. A combination of amoxicillin-clavulanate and ofloxacin is the most effective antibiotic therapy. Sterilization of the cost-intensive silicone stents is feasible, and reuse in the same patient is justifiable from economic aspects.


Mouse heart rate in a human: diagnostic mystery of an extreme tachyarrhythmia.
We report telemetry recording of an extreme non-fatal tachyarrhythmia noted in a hospitalized quadriplegic male with history of atrial fibrillation where the average ventricular conduction rate was found to be about 600 beats per minute and was associated with transient syncope. A medical literature review suggests that the fastest human ventricular conduction rate reported to date in a tachyarrhythmia is 480 beats per minute. We therefore report the fastest human heart rate noted in a tachyarrhythmia and the most probable mechanism of this arrhythmia being a rapid atrial fibrillation with 1:1 conduction in the setting of probable co-existing multiple bypass tracts.


A Comparative Study of Machine Learning Methods for Predicting Live Weight of Duroc, Landrace, and Yorkshire Pigs.
Live weight is an important indicator of livestock productivity and serves as an informative measure for the health, feeding, breeding, and selection of livestock. In this paper, the live weight of pig was estimated using six morphometric measurements, weight at birth, weight at weaning, and age at weaning. This study utilised a dataset including 340 pigs of the Duroc, Landrace, and Yorkshire breeds. In the present paper, we propose a comparative analysis of various machine learning methods using outlier detection, normalisation, hyperparameter optimisation, and stack generalisation to increase the accuracy of the predictions of the live weight of pigs. The performance of live weight prediction was assessed based on the evaluation criteria: the coefficient of determination, the root-mean-squared error, the mean absolute error, and the mean absolute percentage error. The performance measures in our experiments were also validated through 10-fold cross-validation to provide a robust model for predicting the pig live weight. The StackingRegressor model was found to provide the best results with an MAE of 4.331 and a MAPE of 4.296 on the test dataset.


Natural killer cell levels in adults living with type 2 diabetes: a systematic review and meta-analysis of clinical studies.
Chronic immune activation and hyperglycaemia are a hallmark of type 2 diabetes mellitus (T2D) while natural killer (NK) cells are involved in the pathogenesis of T2D. Dysregulated NK cell responses are associated with an increased risk of cardiovascular disease in patients living with T2D. To provide a comprehensive and systematic evidence-based estimate on the levels of NK cells in patients living with T2D. This systematic review and meta-analysis included 13 studies reporting on 491 adult patients with T2D and 1064 nondiabetic controls. The pooled effect estimates showed increased levels of NK cells in adult patients with T2D compared to controls (MD: 0.03 [- 3.20, 3.26], I2 = 97%, p < 0.00001). Overall, the evidence presented in this systematic review shows that the changes in NK cells in patients living with T2D are still unclear and further studies are needed.


Simplified laparoscopic radical cystectomy with orthotopic ileal neobladder creation in a porcine model.
Laparoscopic radical cystectomy with orthotopic ileal neobladder creation is a technically challenging and lengthy surgical procedure. We present our experience with a simplified technique for laparoscopic cystectomy and neobladder creation in the porcine model. Ten female minipigs underwent a purely laparoscopic radical cystectomy with orthotopic ileal neobladder creation. Nine ureterointestinal anastomoses were performed using a simplified "dunk" technique, where the ureter was prolapsed 5 mm into the afferent limb and the periureteral tissue was secured to the bowel serosa with three superficial sutures. Six ureters were not stented, and three had indwelling stents inserted. In 11 ureters, the anastomosis was performed using a running mucosa-to-mucosa technique (three with stents, eight without stents). The Lapra-Ty suture clip (Ethicon Endosurgery, Cincinnati, OH) was used to secure the running sutures on the urethra, ureters, and neobladder. Animals were harvested at 3 to 8 weeks (mean 6.5 weeks) after surgery. Serology, static cystogram, intravenous urography, and gross and histopathologic evaluations were performed. Of six unstented dunked ureterointestinal anastomoses, two (33%) were widely patent, two were strictured but patent, and two were completely obstructed. In the three stented ureters implanted using the dunk technique, one (33%) was widely patent, one was strictured, and one was completely obstructed. All ureterointestinal anastomoses performed with a mucosa-to-mucosa running anastomosis, whether stented (three ureters) or not stented (eight ureters), were widely patent. Lapra-Ty clip migration into the neobladder pouch caused urethral obstruction resulting in delayed bladder perforation in two animals. Laparoscopic cystectomy and ileal neobladder creation is technically feasible. Attempts to simplify the ureterointestinal anastomosis require further evaluation and modification. Stent placement appears to be unnecessary in the laparoscopic ureterointestinal anastomosis. Laparoscopic creation of the ileal neobladder remains a technically challenging procedure.


The Thoracic Inlet Length as a Reference Point to Radiographically Assess Cardiac Enlargement in Dogs with Myxomatous Mitral Valve Disease.
The diagnostic value of the vertebral heart size (VHS) in dogs with mitral valve degeneration (MVD) is compromised when middle thoracic vertebral anomalies are present. The objective of this study was to assess the use of the thoracic inlet heart score (TIHS) to identify left heart enlargement (LHE) secondary to MVD. The cardiac silhouette of 50 clinically healthy dogs and 106 MVD dogs in different stages was assessed on a right lateral chest radiograph. The TIHS and VHS value were calculated for each patient and compared. The TIHS was significantly different between the control dogs and the dogs with MMVD, increasing with disease stage, control 2.91 ± 0.23, Stage B1 2.98 ± 0.36, B2 3.25 ± 0.34, and C 3.53 ± 0.36, p < 0.05. A THIS ≥3.3 showed 69% sensitivity and 81% specificity to identify LHE. The TIHS showed moderate correlation with the VHS, LA/Ao, and LVIDDN 0.59, 0.42, and 0.62, respectively. The intraobserver and interobserver agreement were almost perfect, 0.96, and substantial, 0.73. The TIHS method can be used to identify LHE secondary to MMVD on dogs' thoracic radiographs.


Diagnosis of breast cancer: MDCT versus MRI.
In Japan, helical CT has been applied to breast cancers for the accurate determination of cancer extent before surgery. In recent years, multidetector CT (MDCT) has become commercially available. Compared with mammography and ultrasonography, MDCT of the breast is thought to be useful for preoperative assessment of breast-conserving surgery. In contrast, magnetic resonance imaging (MRI) is extremely accurate in the diagnosis of intraductal carcinoma and multicentricity of breast cancer. A real problem related to the use of CT is X-ray exposure. Therefore, it is controversial whether the use of CT for the patients with breast cancer is appropriate or not. Recent studies that compared the use of MDCT and MRI in the same patients concluded that MRI was more accurate for the detection and evaluation of the extent of intraductal carcinoma. We should consider the pitfalls and limitations of CT for assessing the breast cancer extent.


Interaction of 70-kDA heat shock cognate protein with peptides and myo-inositol monophosphatase.
Fluorescence techniques have been used to investigate the interaction of bovine 70-kDa heat shock cognate protein (Hsc 70) with small molecular weight peptides and myo-inositol monophosphatase. The emission properties of Hsc 70 remain invariant upon addition of ATP. The results of steady-state fluorescence indicate that the tryptophan residues of Hsc 70 are exposed to the rapidly relaxing aqueous solvent. Binding of residues 1-20 of ribonuclease A (RNase S-peptide) to Hsc 70 causes protein fluorescence quenching which was used to determine a dissociation constant Kd = 2.7 microM for the binary Hsc 70.RNase S-peptide complex. The octapeptide corresponding to the NH2-terminal portion of sickle cell hemoglobin recognizes Hsc 70 and binds with a Kd = 9.3 microM. Binding of RNase S-peptide to Hsc 70 produces a small enhancement of ATPase activity. Unfolded myo-inositol monophosphatase, tagged with the fluorescent probe 5-[2-(2-iodoacetamido)ethylamino]-1-naphthalenesulfonic acid, recognizes Hsc 70; the formation of a stable complex was detected by steady-state emission anisotropy measurements. The rate and extent of recovery of catalytic activity of unfolded myo-inositol monophosphatase is not influenced by Hsc 70. It is suggested that interaction of Hsc 70 with unfolded proteins in the cell may be able to delay the formation of misfolded structures.


Limy 'milky' bile: without apparent gall-bladder obstruction by a gallstone.
Limy bile, characterized by excessive precipitation of calcium carbonate, is an uncommon complication of gallstone disease, and is a condition that seems to follow gall-bladder obstruction. Limy bile was observed at cholecystectomy in a 57 year old woman who presented with biliary colic. At operation a mass of precipitated cohesive crystals, that had coalesced into a whitish paste of calcium carbonate, was observed obstructing the outlet of the gall-bladder. This case report is unique in that calcium carbonate precipitation appears to have preceded gall-bladder obstruction.


Diversity of Fe2+ entry and oxidation in ferritins.
The essential metal iron presents two major problems for life: it is potentially highly toxic due to its redox activity, and its extremely low solubility in aqueous solution in the presence of O2 can make it hard to acquire and store safely. Ferritins are part of nature's answer to these problems, as they store iron in a safe but accessible form in all types of cells. How they achieve this has been the subject of intense research for several decades. Here, we highlight recent progress in elucidating the routes by which Fe2+ ions access the catalytic ferroxidase centers, and the mechanisms by which Fe2+ is oxidized. Emerging from this is a picture of diversity, both in terms of Fe2+ entry pathways and the roles played by the structurally distinct diiron ferroxidase centers.


Differential effects of age and cerebral atrophy upon span of immediate recall and paired-associate learning in older patients suspected of dementia.
Seventy-eight patients suspected of dementia, 50 years of age or older and without evidence of focal cerebral lesion or systemic illness, were administered the Digit-Span Forward and Paired-Associate Learning subtests of the Wechsler Memory Scale. Computerized axial tomography, allowing a visualization and measurement of the lateral ventricles and cortical sulci, was employed to measure cerebral atrophy. Four groups were defined as follows: (1) age 50-69, without atrophy (N = 27), (2) age 50-69, with atrophy (N = 17), (3) age 70-89, without atrophy (N = 18), (4) age 70-89, with atrophy (N = 16). Results of analyses of variance indicate that Digit-Span is affected by cerebral atrophy but not by age, while Paired-Associate Learning is affected by both age and cerebral atrophy. No interaction effect between age and cerebral atrophy was found for either task. These results are discussed within the context of previous research, and implications for both models of memory and theories of the effects of cerebral atrophy in different ag groups are suggested.


An incidental and uncommon pulmonary sequestration with an uncommon feeding artery.
Pulmonary sequestration is a rare congenital anomaly and most intralobar sequestrations were located in lower lobes. There is little information on middle lobe intralobar sequestration. A 44 year-old man with right middle lobe syndrome was referred for surgical management. He underwent video-assisted thoracoscopic right middle lobectomy and his postoperative course was uneventful. Pathology is notable for an incidental intralobar sequestration. Our case was unique in that the location of incidental pulmonary sequestration is uncommon and in that its feeding artery was uncommonly located in the fissure. We report an extremely rare case of right middle lobe intralobar sequestration with a feeding artery in the fissure.


Synthesis, anti-tubercular activity and 3D-QSAR study of coumarin-4-acetic acid benzylidene hydrazides.
A set of 25 coumarin-4-acetic acid benzylidene hydrazides were synthesized and characterized by NMR, IR and mass spectroscopic techniques. The compounds were evaluated for their anti-tubercular activity against Mycobacterium tuberculosis H(37)Rv strain using the BACTEC 460 system to determine percentage inhibition. To understand the relationship between structure and activity, a 3D-QSAR analysis has been carried out by Comparative Molecular Field Analysis (CoMFA). Several statistically significant CoMFA models were generated. The CoMFA model generated with database alignment was the best in terms of overall statistics. The CoMFA contours provide a good insight into the structure activity relationships of the compounds reported herein.


Positive peritoneal cytology is highly predictive of prognosis and relapse patterns in stage III (FIGO 2009) endometrial cancer.
According to the updated FIGO system, positive peritoneal cytology does not affect endometrial cancer stage. This revision may reduce rates of obtaining cytology, with unclear implications in advanced disease. This study evaluates the significance of positive cytology in stage III (FIGO 2009) endometrial cancer. Eligible patients received treatment for stage III endometrial cancer at a single institution and had peritoneal cytology performed. Of 196 patients, 58% were ≥ 60 years old, 48% had deep myometrial invasion, 71% lymphovascular invasion, 25% cervical invasion, 37% adnexal involvement, 79% nodal involvement, and 46% aggressive histology. Positive cytology was present in 23% (45/196) and significantly associated with cervical stromal invasion, adnexal involvement, and aggressive histology (P ≤ 0.03). There was no significant difference in rates of lymphadenectomy, chemotherapy, or radiation between negative and positive cytology groups. At a median follow-up of 47 months, the 5-year freedom from relapse was 39% for positive cytology vs. 69% for negative, disease-specific survival 42% vs. 77%, and overall survival 34% vs. 72% (P < 0.001). Positive cytology correlated with higher recurrence rates in the para-aortic nodes and peritoneum (30% vs. 9%, 23% vs. 4%; P ≤ 0.008). When controlling for adverse features including aggressive histology, positive cytology was associated with an increased hazard for relapse (HR 2.3; P = 0.002) and death (HR 2.9; P < 0.001). In stage III endometrial cancer, positive cytology independently predicts outcome and is associated with distinct relapse patterns. Obtaining peritoneal cytology in stage III endometrial cancer is critical.


Synthesis of unequally-spaced arrays using the fractional Fourier series.
This paper introduces an algebraic method to synthesize the radiation pattern of unequally-spaced arrays. In this method, first, the characteristic matrix, containing the sampled data of the desired pattern, is established. Then, the fractional Fourier series is calculated using the distinct eigenvalues and eigenvectors of the characteristic matrix. In the following, the location of the array elements is estimated using the eigenvalues by considering the mutual coupling effect. The magnitude and phase of the excitation currents are computed using the least square method. Also, it is proved that the number of elements can be reduced using the eigenvalue decomposition and differential spectrum method. Several important patterns are investigated to verify the performance of the proposed method, and then a comprehensive discussion is expressed for all cases. It is shown that using the proposed method, a greater average spacing is achieved which allows bettering mitigate the phenomenon of mutual coupling. Furthermore, the various results show that the proposed method offers a better approximation of the desired array factor especially for beam-shaped patterns. This method also has a noble ability to reduce the number of array elements compared to a generic reference array, while still retaining a good ability to approximate the desired pattern fairly faithfully.


Platelet adhesion to cultured bovine cerebral microvascular endothelial cells by stimulation of platelet activating factor and antagonism of drugs.
To study platelet activating factor (PAF) stimulating the platelets to adhere to cultured bovine cerebral microvascular endothelial cells (CMEC) and the inhibitory effect of triazelodiazepine (WEB), 1,5-bis-(3,4-dimethoxyphenyl)-tetrahydro-(4H)-pyran (DMPP), tetrandrine (Tet). The platelets adhesion to CMEC and the inhibitory effect of drugs were investigated by [3H]adenine labeling of rabbit blood platelet. The platelet adhesion to CMEC was increased by 36% vs control after CMEC was stimulated with PAF 10 nmol L-1 for 25 min. WEB 0.1, 1, 10 mmol L-1 or DMPP 0.1, 1, 10 mmol L-1 or Tet 0.1, 1, 10 mmol L-1 inhibited the PAF stimulating platelet adhesion to CMEC by 5.4%, 16.3%, 20.1%; 13.7%, 19.4%, 22.4%; and 5.5%, 23.1%, 32.6%, respectively. DMPP and Tet inhibited the PAF action in cerebral vascular system.


cDNA array analysis of pineal gene expression reveals circadian rhythmicity of the dominant negative helix-loop-helix protein-encoding gene, Id-1.
The pineal gland is a major output of the endogenous vertebrate circadian clock, with melatonin serving as the output signal. In many species, elevated nocturnal melatonin production is associated with changes in pineal gene expression. In the current study, cDNA array analysis was used in an attempt to identify additional genes that exhibit day/night differential expression in the rat pineal gland. This revealed 38 candidate genes, including Id-1 (inhibitor of DNA binding and differentiation). Id-1 encodes a helix-loop-helix (HLH) protein that lacks a basic DNA binding domain and could affect pineal physiology via a dominant negative trans-acting regulatory activity. For this reason Id-1 was selected for further analysis. Id-1 was expressed in a major population of pineal cells and the Id-1 protein was associated with a nuclear complex. The levels of Id-1 mRNA and protein exhibit approximately six-fold day/night rhythms. In contrast, the related genes Id-2 and Id-3 do not exhibit marked day/night differences in pineal expression. Rhythmic Id-1 expression is primarily limited to a C-terminally extended splice variant of Id-1, which would restrict the functional output of the rhythm to protein binding partners of this isoform of Id-1. Our findings add to the body of evidence indicating that transcriptional regulators play a role in neuroendocrine rhythms, and extend this by introducing the concept of a dominant negative HLH involvement. The rhythm in Id-1 in the pineal gland provides an experimental opportunity to identify Id-1-binding partners which may also be involved in Id-1 activity in other functional contexts.


Phase Modulation by Vacuum Poling toward Enhanced Performance of Quasi-Two-Dimensional Bromide Perovskite-Based Light-Emitting Diodes.
As a new kind of highly efficient luminescent materials, quasi-two-dimensional (quasi-2D) perovskites show great potential in industrial display applications. In this work, the poling methods are used in modulating the phase arrangement in quasi-2D perovskite light-emitting diodes (PeLEDs). We find the effective modulation of different phase components in uniform arrangement can enhance both brightness and current efficiency to 30 810 cd/m2 and 8.3 cd/A, about 2 times in comparison to results for the control devices. Upon further investigation by optical and impedance spectroscopy, we find the phase modulation can improve the balance of charge carrier injection without sacrificing the transport ability. We believe our studies can provide new inspiration on the influence of phase arrangement in PeLED devices.


Sleep cyclicity in infants during the earliest postnatal weeks.
Cyclicity within sleep was investigated in 20 fullterm infants who were monitored in the home for 24-hour periods when they were 2-5 weeks old. Using a pressure sensitive mattress in the crib, analog signals from the baby's respiration and body movements were computer scored for: Quiet Sleep, Active Sleep, Active-Quiet Transitional Sleep, Sleep-Wake Transition, and Wakefulness. Sleep episodes with at least four recurrences of Quiet Sleep were analyzed for cyclicity. Significant cyclicity of Quiet Sleep was found in 49 of the 55 sleep episodes analyzed. The mean Quiet Sleep recurrence period was 63.2 minutes. Eighteen of the 20 infants exhibited cyclicity on one or more weeks. Thus, sleep during the neonatal period is generally characterized by the presence of cyclicity. Cyclicity was not related to other parameters of the sleep cycle, but was correlated with stability of state patterning over weeks. The results provide new evidence for neurobehavioral competence of the neonate in terms of sleep organization.


Stereoselective synthesis of spirocyclopentanones via N-heterocyclic carbene-catalyzed reactions of enals and dienones.
Homoenolates generated from enals by nucleophilic heterocyclic carbene (NHC) catalysis undergo a conjugate addition/cyclization sequence with cyclic dienones, culminating in the efficient synthesis of spirocyclopentanones.


Management of an invasive and metastatic Sertoli cell tumor with associated myelotoxicosis in a dog.
We describe the surgical and post-operative management of a large, invasive, and metastatic functional Sertoli cell tumor in a 9-year-old cryptorchid male Labrador retriever dog. Despite residual disease after surgery, bone marrow recovery occurred without administration of bone marrow stimulants and serum estradiol accurately predicted tumor recurrence.


The phospholipid- and calcium-dependent protein kinase as a target in tumor chemotherapy.
Evidence for a constitutive activation of protein kinase C (EC 22.214.171.124) in Ha-ras transformed 3T3 cells is presented. Several compounds which inhibit protein kinase C in vitro have been studied with regard to their antiproliferative activity in cultured tumor cells. The following agents were investigated: 3-hexadecyl-mercapto-2-methoxy-methyl-propyl-1- phosphocholine (BM 41440); 1-octadecyl-2-methyl-sn-glycero-3-phosphocholine (ET-18-OCH3); quercetin, tamoxifen and staurosporine. All compounds decrease protein kinase C activity in vitro as well as in intact cells and inhibit cell multiplication within the same dose range. The results suggest a causal relation between the antiproliferative effects and the inhibition of protein kinase C. All inhibitors of protein kinase C synergistically enhance the antiproliferative activity of cis-diamminedichloroplatinum(II). Available data suggest that the effects of protein kinase C inhibitors should be exploitable for tumor chemotherapy.


Research progress on risk factors of delirium in burn patients: A narrative review.
Delirium, an acute brain dysfunction, is a common and serious complication in burn patients. The occurrence of delirium increases the difficulty of patient treatment, is associated with various adverse outcomes, and increases the burden on the patient's family. Many scholars have studied the factors that cause delirium, but the causes, pathogenesis, and treatment of delirium in burn patients have not been fully revealed. There is no effective pharmacological treatment for delirium, but active preventive measures can effectively reduce the incidence of delirium in burn patients. Therefore, it is necessary to study the relevant factors affecting the occurrence of delirium in burn patients. This study was conducted on December 20, 2021 by searching the PubMed database for a narrative review of published studies. The search strategy included keywords related to "burns," "delirium," and "risk factors." We reviewed the characteristics of delirium occurrence in burn patients and various delirium assessment tools, and summarized the risk factors for the development of delirium in burn patients in terms of personal, clinical, and environmental factors, and we found that although many risk factors act on the development of delirium in burn patients, some of them, such as clinical and environmental factors, are modifiable, suggesting that we can estimate the exposure of burn patients to risk factors by assessing their likelihood of delirium occurring and to make targeted interventions that provide a theoretical basis for the prevention and treatment of burn delirium.


Projected Estimates of African American Medical Graduates of Closed Historically Black Medical Schools.
There continue to be low numbers of underrepresented minorities, including African Americans, in academic medicine. Historically Black medical colleges and universities are major sources of training for medical school graduates who are African American or who belong to other underrepresented minority groups. Several historically Black medical schools were closed during the period surrounding the 1910 Flexner report. The implications of these school closures with regard to the number of African American medical school graduates have not been fully examined. To examine the consequences associated with the closure of historically Black medical schools for the number of African American medical school graduates. This observational economic evaluation used steady expansion and rapid expansion models to estimate the consequences associated with the closure of historically Black medical schools for the number of African American medical school graduates. The numbers of graduates from 13 historically Black medical schools that are now closed were obtained through historical records. Data on historically Black medical schools that are currently open were obtained from school-specific reports and reports published by the Association of American Medical Colleges. The study focused on projected estimates of outcomes from the hypothetical continued operation and expansion of 5 closed historically Black medical schools that were included in the Flexner report: Flint Medical College of New Orleans University, Knoxville Medical College, Leonard Medical School of Shaw University, Louisville National Medical College, and the University of West Tennessee College of Medicine and Surgery-Memphis. The main outcome was the estimate of the number of African American students who would have graduated from historically Black medical schools that were closed during the period surrounding the 1910 Flexner report. Among the 5 historically Black medical schools that were closed, the estimated mean number of graduates per year was 5.27 students at Flint Medical College, 2.60 students at Knoxville Medical College, 11.06 students at Leonard Medical School, 4.17 students at Louisville National Medical College, and 6.74 students at the University of West Tennessee. If the 5 closed historically Black medical schools had remained open, the steady expansion and rapid expansion models indicated that these schools might have collectively provided training to an additional 27 773 graduates and 35 315 graduates, respectively, between their year of closure and 2019. In the analysis of Leonard Medical School and the University of West Tennessee only, the steady expansion and rapid expansion models indicated that these 2 schools would have provided training to an additional 10 587 graduates and 13 403 graduates, respectively, between their year of closure and 2019. An extrapolation based on the racial and ethnic self-identification of current graduates of historically Black medical schools indicated that if these closed schools had remained open, the number of graduating African American physicians might have increased by 355 individuals (29%) in 2019 alone. To increase the number of African American medical school graduates, consideration should be given to creating medical education programs at historically Black colleges and universities. Such programs may start with small enrollment but could have positive consequences for the diversity of the physician workforce.


Radiation Dose Survey for Common Computed Tomography Exams: 2013 British Columbia Results.
In 2013 Health Canada conducted a national survey of computed tomography (CT) radiation usage. We analysed contributions from all 7 public health authorities in the province of British Columbia, which covered scanner age, number of slices, and common adult protocols (≥ 19 years: 70 ± 20 kg, head, chest, abdomen/pelvis, and trunk). Patient doses were recorded for common protocols. Diagnostic reference levels (DRLs) was calculated using scanner data with >10 patient doses recorded for each protocol. Data was analysed based on image reconstruction (filtered backprojection vs iterative reconstruction [IR] vs IR available but not in use). Provincial response was 92%, with 59 of 64 CT data used for analysis. The average scanner age was 5.5 years old, with 39% of scanners installed between 2008-2013; 78.5% of scanners were multislice (>64 slices), and 44% of scanners had IR available. Overall British Columbia DRLs were: head = 1305, chest = 529, abdomen/pelvis = 819, and trunk = 1225. DRLs were consistent with Health Canada recommendations and other Canadian published values, but above international standards. For sites with IR available, less than 50% used this technology routinely for head, chest and trunk exams. Overall, use of IR reduced radiation usage between 11%-32% compared to filtered backprojection, while sites using IR vs IR available used 30%/43% less radiation for head/chest exams (P < .05). No significant difference was observed for abdomen/pelvis exams (P = .385). With the fast pace of CT technical advancement, DRLs should reflect the technology used, instead of just globally applied to anatomical regions. Federal guidelines should be updated at a higher frequency to reflect new technology. In addition, new technologies must be utilised to optimize image quality vs radiation usage.


Role of nano-lipid formulation of CARP-1 mimetic, CFM-4.17 to improve systemic exposure and response in osimertinib resistant non-small cell lung cancer.
EGFR mutated NSCLCs have been shown to employ the use of CARP-1 in overriding the signaling inhibition of tyrosine kinase inhibitors (such as Osimertinib). CFM 4.17 is a CARP-1 inhibitor which has a promising role in overcoming Tyrosine Kinase Inhibitor (TKI) resistance when used as a pre-treatment through promoting apoptosis. Lack of solubility, hydrophobicity leading to poor systemic exposure are the limitations of CFM 4.17. This can be overcome by nano lipid-based formulation (NLPF) of CFM 4.17 which can enhance systemic exposure in preclinical animal models as well as improve therapeutic efficacy in drug-resistant cancer cell lines. Molecular docking simulation studies were performed for CFM 4.17. CFM 4.17-NLPF was formulated by melt dispersion technique and optimized using a Box-Behnken designed surface response methodology approach using Design Expert and MATLAB. In vitro, CFM 4.17 release studies were performed in simulated gastric fluids (SGF-pH-1.2) and simulated intestinal fluids (SIF- pH-6.8). Cell viability assays were performed with HCC827 and H1975 Osimertinib resistant and non-resistant cells in 2D and 3D culture models of Non-small cell lung cancer to determine the effects of CFM 4.17 pre-treatment in Osimertinib response. In vivo pharmacokinetics in rats were performed measuring the effects of NLPF on CFM 4.17 to improve the systemic exposure. CFM 4.17 was well accommodated in the active pocket of the active site of human EGFR tyrosine kinase. CFM 4.17 NLPF was optimized with robust experimental design with particle size less than 300 nm and % entrapment efficiency of 92.3 ± 1.23. Sustained diffusion-based release of CFM 4.17 was observed from NLPF in SGF and SIFs with Peppas and Higuchi based release kinetics, respectively. CFM 4.17 pretreatment improved response by decreasing IC50 value by 2-fold when compared to single treatment Osimertinib in both 2D monolayer and 3D spheroid assays in HCC827 and H1975 Osimertinib resistant and non-resistant cells of Non-small cell lung cancer. There were no differences between CFM 4.17 NLPF and suspension in 2D monolayer culture pretreatments; however, The 3D culture assays showed that CFM 4.17 NLPF improved combination sensitivity. Pharmacokinetic analysis showed that CFM 4.17 NLPF displayed higher AUCtot (2.9-fold) and Cmax (1.18-fold) as compared to free CFM 4.17. In contrast, the animal groups administered CFM 4.17 NLPF showed a 4.73-fold (in half-life) and a 3.07-fold increase (in MRT) when compared to equivalent dosed suspension. We have successfully formulated CFM 4.17 NLPFs by robust RSM design approach displaying improved response through sensitizing cells to Osimertinib treatment as well as improving the oral bioavailability of CFM 4.17.


Prediction of clinically relevant postoperative pancreatic fistula using radiomic features and preoperative data.
Clinically relevant postoperative pancreatic fistula (CR-POPF) can significantly affect the treatment course and outcome in pancreatic cancer patients. Preoperative prediction of CR-POPF can aid the surgical decision-making process and lead to better perioperative management of patients. In this retrospective study of 108 pancreatic head resection patients, we present risk models for the prediction of CR-POPF that use combinations of preoperative computed tomography (CT)-based radiomic features, mesh-based volumes of annotated intra- and peripancreatic structures and preoperative clinical data. The risk signatures were evaluated and analysed in detail by visualising feature expression maps and by comparing significant features to the established CR-POPF risk measures. Out of the risk models that were developed in this study, the combined radiomic and clinical signature performed best with an average area under receiver operating characteristic curve (AUC) of 0.86 and a balanced accuracy score of 0.76 on validation data. The following pre-operative features showed significant correlation with outcome in this signature ([Formula: see text]) - texture and morphology of the healthy pancreatic segment, intensity volume histogram-based feature of the pancreatic duct segment, morphology of the combined segment, and BMI. The predictions of this pre-operative signature showed strong correlation (Spearman correlation co-efficient, [Formula: see text]) with the intraoperative updated alternative fistula risk score (ua-FRS), which is the clinical gold standard for intraoperative CR-POPF risk stratification. These results indicate that the proposed combined radiomic and clinical signature developed solely based on preoperatively available clinical and routine imaging data can perform on par with the current state-of-the-art intraoperative models for CR-POPF risk stratification.


Case reports - reversal of sensory deficit associated with pain relief after treatment with gabapentin.
Many patients with neuropathic pain have coexistent sensory deficits. Neuropathic pain may be alleviated by a variety of drugs but sensory deficits are assumed to be permanent. In an audit of the effects of gabapentin therapy on patients with neuropathic pain, monthly detailed sensory examinations were performed during the first three months of treatment. Of five patients with sensory deficits who tolerated gabapentin therapy, three showed marked improvement of their sensory deficits associated with pain alleviation. The cases are presented and possible explanations for the observed sensory improvements are discussed. These findings raised exciting neurophysiological questions in addition to being of potential importance to the clinical problem of neurotrophic tissue injury.


Parental decision regret among Australian parents after consenting to or refusing hypospadias repair for their son: Results of a survey with controls.
Parental decision regret in hypospadias surgery is a recognised source of long-lasting psycho-social morbidity. High parental decision regret after their child's hypospadias repair is reported. The aim of this study is to report on decision regret in Australian parents, who accepted and declined surgery for their son and explore underlying factors for decision-making, satisfaction, and regret. An online anonymous survey was administered to three groups of parents: 1) parents who consented for hypospadias repair, 2) parents who declined repair and 3) a control group who requested circumcision for their child. Operations occurred between 2010 and 2020 in two paediatric hospitals in New South Wales, Australia. The survey included a validated decision regret assessment tool and additional questions to explore the possible basis of the opinions. One hundred and eighteen parents (invited - 381, completed - 116, response rate - 31%) participated in the survey. Decision regret was present in group 1 (n = 89) - 55% (moderate-to-severe 15%), in group 2 (n = 14) - 71% (moderate-to-severe 57%), and in the control group (n = 15) - 15% (moderate-to-severe 8%) of parents. There was a significant difference in the median decision regret score between all three groups. Parents who chose hypospadias repair were mostly concerned about function. The prevalence of decision regret among Australian parents who consented for their son's hypospadias repair was lower compared with the mean decision regret reported in the literature to date (55% vs 65%). Decision regret and its severity were highest among parents who declined hypospadias repair. New strategies are needed to reduce decision regret in parents whether they elect for surgery or not.


Matrix metalloproteinase-like activity from hemocytes of the eastern oyster, Crassostrea virginica.
Investigation of oyster blood cell lysate revealed one prominent band of proteolytic activity when analyzed using gelatin and collagen impregnated polyacrylamide gel electrophoresis. The proteolytic activity was inhibited by 1,10 phenanthroline and EDTA, but not by other proteinase inhibitors. Maximal activity was shown at pH 8.2 and the molecular weight of the protein responsible for the activity was estimated to be 68 kDa. Proteolytic activity was also measured by fluorescence assays containing hemocyte lysate and fluorescein-labeled gelatin, type I or type IV collagen. Characteristics of this proteolytic activity suggest that an invertebrate matrix metalloproteinase is responsible.


Segregation of age-related skin microbiome characteristics by functionality.
Although physiological changes are the most evident indicators of skin aging by alteration of the skin's structure and function, we question whether skin aging is also affected by the structure and assembly process of the skin microbiome. We analysed the skin microbiomes of 73 healthy Chinese women in two age groups (25-35 years old and 56-63 years old) using 16S rRNA gene amplicon sequencing; the overall microbiome structure was significantly different between the two age groups. An analysis using ecological theory to evaluate the process of microbial community assembly processes revealed that the microbiomes of the older group were formed under a greater influence of the niche-based process, with the network of microbes being more collapsed than that of the younger group. Inferred metagenomic functional pathways associated with replication and repair were relatively more predominant in the younger group whereas, among the various metabolism-related pathways, those associated with biodegradation were more predominant in the older group. Interestingly, we found two segregated sub-typing patterns in the younger group which were also observed in the skin microbiomes of young Chinese women living in four other cities in China. The results of our study highlights candidate microbes and functional pathways that are important for future research into preventing skin aging and which could lead to a comprehensive understanding of age-related skin microbiome characteristics.


Insulin-dependent ketosis-resistant diabetes in Ethiopia.
Anthropometric, clinical and biochemical findings were compared in 30 rural (group A), 18 urban insulin-requiring (group B) and 45 urban oral-agent-responsive (group C) newly diagnosed diabetics. Mean ages at onset were 28.3 +/- 12.0, 25.6 +/- 14.5 and 42.1 +/- 10.5 years respectively. The differences between A and C and between B and C were significant. Group A were poor and malnourished, with body mass index (BMI) 15.9 +/- 1.9 and 17.2 +/- 3.7 kg/m2 for males and females respectively, presented with a long history of classical diabetes without ketoacidosis and required insulin in modest doses. 3 of 10 cases had excess stool fat but none of 13 unselected cases had pancreatic calcification. Group C were better nourished, with BMI 22.6 +/- 2.8 and 22.4 +/- 4.5 kg/m2, and responded to oral agents. Group B, with BMI 17.2 +/- 2.6 and 18.6 +/- 3.1 kg/m2, required insulin for control but had C-peptide levels above 0.02 nmol/1 in 10 of 15 cases. Anthropometric indices for males, but not for females, were significantly lower in group A than in group B or C. There were significant differences in levels of glucose between A and B and A and C, free fatty acids between A and C and B and C, insulin between A and B and A and C and C-peptide between A and C and B and C. Of the 3 groups the rural type most closely resembled the tropical variants.


Characterization of an N-terminal Nav1.5 channel variant - a potential risk factor for arrhythmias and sudden death?
Alterations in the SCN5A gene encoding the cardiac sodium channel Nav1.5 have been linked to a number of arrhythmia syndromes and diseases including long-QT syndrome (LQTS), Brugada syndrome (BrS) and dilative cardiomyopathy (DCM), which may predispose to fatal arrhythmias and sudden death. We identified the heterozygous variant c.316A > G, p.(Ser106Gly) in a 35-year-old patient with survived cardiac arrest. In the present study, we aimed to investigate the functional impact of the variant to clarify the medical relevance. Mutant as well as wild type GFP tagged Nav1.5 channels were expressed in HEK293 cells. We performed functional characterization experiments using patch-clamp technique. Electrophysiological measurements indicated, that the detected missense variant alters Nav1.5 channel functionality leading to a gain-of-function effect. Cells expressing S106G channels show an increase in Nav1.5 current over the entire voltage window. The results support the assumption that the detected sequence aberration alters Nav1.5 channel function and may predispose to cardiac arrhythmias and sudden cardiac death.


Analysis of intracellular expressed proteins of Mycobacterium tuberculosis clinical isolates.
Tuberculosis (TB) is the most threatening infectious disease globally. Although progress has been made to reduce global incidence of TB, emergence of multidrug resistant (MDR) TB threatens to undermine these advances. To combat the disease, novel intervention strategies effective against drug resistant and sensitive subpopulations of M. tuberculosis are urgently required as adducts in the present treatment regimen. Using THP-1 cells we have analyzed and compared the global protein expression profile of broth-cultured and intraphagosomally grown drug resistant and sensitive M.tuberculosis clinical isolates. On comparing the two dimensional (2-DE) gels, many proteins were found to be upregulated/expressed during intracellular state which were identified by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). Four proteins (adenosylhomocysteinase, aspartate carbomyltransferase, putatitive thiosulfate sulfurtransferase and universal stress protein) were present in both intracellular MDR and sensitive isolates and three of these belonged to intermediary metabolism and respiration category. Two proteins (alanine dehydrogenase and adenosine kinase) of intracellular MDR isolate and two (glucose-6-phosphate isomerase and ATP synthase epsilon chain) of intracellular sensitive isolate belonged to intermediary metabolism and respiration category. One protein (Peroxidase/Catalase) of intracellular MDR and three (HSPX, 14 kDa antigen and 10 kDa chaperonin) of sensitive isolate belonged to virulence, detoxification and adaptation category. ESAT-6 of intracellular MDR belonged to cell wall and cell processes category. Two proteins (Antigen 85-C and Antigen 85-A) of intracellular sensitive isolate were involved in lipid metabolism while probable peptidyl-prolyl cis-trans isomerase A was involved in information pathways. Four (Rv0635, Rv1827, Rv0036c and Rv2032) of intracellular MDR and two proteins (Rv2896c and Rv2558c) of sensitive isolate were hypothetical proteins which were functionally characterized using bioinformatic tools. Bioinformatic findings revealed that the proteins encoded by Rv0036, Rv2032c, Rv0635, Rv1827 and Rv2896c genes are involved in cellular metabolism and help in intracellular survival. Mass spectrometry and bioinformatic analysis of both MDR and sensitive isolates of M. tuberculosis during intraphagosomal growth showed that majority of commonly upregulated/expressed proteins belonged to the cellular metabolism and respiration category. Inhibitors of the metabolic enzymes/intermediate can therefore serve as suitable drug targets against drug-resistant and sensitive subpopulations of M. tuberculosis.


Perceived Discrimination, Coping Styles, and Internalizing Symptoms Among a Community Sample of Hispanic and Somali Adolescents.
Perceived discrimination, perceptions of receiving differential treatment due to negative attitudes, and stereotypes about one's racial/ethnic group can increase vulnerability to depression and anxiety. Although ethnic minority youth now represent over half of the U.S. youth population, few studies have investigated potential protective factors in the relationship between perceived discrimination and mental health across diverse ethnic minority immigrant youth from different cultural backgrounds. We examined the association between perceived discrimination and past week symptoms of depression and anxiety and whether patterns of problem and emotion-focused coping moderate these relationships among Somali and Hispanic immigrant youth (N = 353) in an urban midwestern setting (mean age = 15; 53% male, 39% first generation, 75% low income). Path analysis models examined the main effects of perceived discrimination for depression and anxiety and whether problem and emotion-focused coping moderated these associations. Path analysis models suggest that perceived discrimination was positively associated with past week symptoms of depression (β = .37, standard error = .06) and anxiety (β = .16, standard error = .06) across ethnicity. However, adolescents who reported high levels of discrimination and who used predominantly problem-focused coping strategies experienced fewer internalizing problems than youth who relied predominantly on emotion-focused coping strategies. Our findings suggest that strengthening youths' problem-focused coping strategies in the face of discriminatory stress is a promising health promotion and risk prevention approach.


Voluntary exercise during extinction of auditory fear conditioning reduces the relapse of fear associated with potentiated activity of striatal direct pathway neurons.
Relapse of previously extinguished fear presents a significant, pervasive obstacle to the successful long-term treatment of anxiety and trauma-related disorders. Thus, identification of a novel means to enhance fear extinction to stand the passage of time and generalize across contexts is of the utmost importance. Acute bouts of exercise can be used as inexpensive, noninvasive treatment strategies to reduce anxiety, and have been shown to enhance memory for extinction when performed in close temporal proximity to the extinction session. However, it is unclear whether acute exercise can be used to prevent relapse of fear, and the neural mechanisms underlying this potential effect are unknown. The current study therefore examined whether acute exercise during extinction of auditory fear can protect against the later relapse of fear. Male F344 rats lacking an extended history of wheel running were conditioned to fear a tone CS and subsequently extinguished within either a freely mobile running wheel, a locked wheel, or a control context lacking a wheel. Rats exposed to fear extinction within a freely mobile wheel ran during fear extinction, and demonstrated reduced fear as well as attenuated corticosterone levels during re-exposure to the extinguished CS during the relapse test in a novel context 1week later. Examination of cfos mRNA patterns elicited by re-exposure to the extinguished CS during the relapse test revealed that acute exercise during extinction decreased activation of brain circuits classically involved in driving fear expression and interestingly, increased activity within neurons of the direct striatal pathway involved in reward signaling. These data suggest that exercise during extinction reduces relapse through a mechanism involving the direct pathway of the striatum. It is suggested that a positive affective state could become associated with the CS during exercise during extinction, thus resulting in a relapse-resistant extinction memory.


Percutaneous pancreatography: case report and presentation of technique.
A case of traumatic pancreatitis with a radiopaque calculus producing pain by obstructing the distal pancreatic duct is presented. Preoperative ductal anatomy was defined by a percutaneous pancreatogram that established the presence of mechanical duct obstruction as the cause of pain, and the potential for operative relief of duct obstruction. A remission from pain resulted from pancreatic duct decompression by a lateral pancreaticojejunostomy. The potential value of percutaneous pancreatography is discussed.


Potential Cardiovascular Applications of Total-body PET Imaging.
Cardiovascular conditions can exist as part of a systemic disorder (eg, sarcoidosis, amyloidosis, or vasculitis) or have systemic consequences as a result of the cardiovascular insult (eg, myocardial infarction). In other circumstances, multisystem evaluation of metabolism and blood flow might be key for evaluation of multisystemic syndromes or conditions. Long axial field-of-view PET/computed tomography systems hold the promise of transforming the investigation of such systemic disorders. This article aims at reviewing some of the potential cardiovascular applications of this novel instrumentation device.


Interaction of replication protein A with photoreactive DNA structures.
A new photoreactive oligonucleotide derivative was synthesized with a perfluoroarylazido group attached to the 2'-position of the ribose fragment of the 5'-terminal nucleotide. Using this conjugate, photoreactive DNA duplexes were produced which contained single-stranded regions of different length, single-stranded breaks (nicks), and also ds duplex with a photoreactive group inside one of the chains. These structures imitate DNA intermediates generated at different stages of DNA replication and repair. The interaction of replication protein A (RPA) with the resulting DNA structures was studied using photoaffinity modification and gel retardation assay. Independently of the DNA structure, only the large subunit of RPA (p70) was crosslinked to photoreactive DNAs, and the intensity of its labeling increased with decrease in the size of the single-stranded region and was maximal in the case of the nick-containing DNA structure. By gel retardation, the most effective binding of RPA to this structure was shown, whereas the complexing of RPA with DNA containing the unmodified nick and also with the full duplex containing the photoreactive group inside the chain was significantly less effective. The data suggest that RPA should be sensitive to such damages in the double-stranded DNA structure.


Occupational exposure to cadmium and kidney dysfunction.
Investigations were carried out in an alkaline battery factory. The study group consisted of 102 persons and the control group of 85 persons. Cadmium in blood (Cd-B) and cadmium in urine (Cd-U), as well as beta 2-microglobulin (B2-M), retinol binding protein (RBP), amino acids in urine were determined. Exposure to cadmium was high; Cd-B and Cd-U concentrations were higher than recommended, 10 micrograms/l and 10 micrograms/g creat. In 65% and 56% of workers, respectively. Excretion of B2-M and RBP in urine was higher than the accepted upper limits of 380 and 130 micrograms/g creat. in about 20% of the workers. A significant correlation was observed between: log Cd-U.log Cd-B (r = 0.85), log B2-M.log RBP (r = 0.66), log Cd-U.log B2-M (r = 0.52), and log Cd-U.log RBP (r = 0.55). To evaluate the admissible period of occupational exposure to cadmium, an integrated exposure index (Cd-B x years of exposure) is proposed. According to the dose-response relationship, an increase of low molecular protein excretion in urine can be expected in 10% of the cases at Cd-U amounting to 10 to 15 micrograms/g creat. and Cd-B x years of about 300 to 400.


Morphological study of CT image of posterior pilon variant fracture and its possible clinical significance.
The incidence of posterior pilon variant fractures has been underestimated. The purpose was to study the characteristics of posteromedial (PM) and posterolateral (PL) fragments in CT imaging of posterior pilon variant fractures, and to provide help for clinical diagnosis and treatment. CT imaging data of 109 cases of posterior pilon variant fractures in our hospital from January 2013 to December 2020 were retrospectively analyzed. According to Mason and Molloy classification, PM fragments were further divided into pilon subtypes and avulsed subtypes. The largest actual area of fragments in axial and sagittal were selected as the study plane, and the maximum axial lengths of X, Y and Z, α angle, β angle, fragment area (S1-7) and fragment area ratio (FAR1-4), interfragmentary (IF) angle, and back of tibia (BT) angle were measured. A total of 109 cases were included in this study, 61 of whom were pilon subtypes [90.16% were supination-external rotation (SER) injuries]. 48 cases were avulsed subtypes [81.25% were pronation-external rotation (PER) injuries]. Pilon subtypes were larger than avulsed subtypes in X, Y, Z, α2 Angle, β2 Angle, fragment area and ratio, and IF and BT angle (P < 0.05). There was no difference between α1 and β1 angle (P > 0.05). The morphology of pilon subtype was larger than that of avulsion subtype. According to fragment size, morphology, and injury mechanism, two fragments of pilon subtype should be anatomic reduction and fixation. However, the PL fragment of avulsion subtype should to be fixed, while PM fragment may only need conservative treatment.


Emergence of daptomycin non-susceptibility in colonizing vancomycin-resistant Enterococcus faecium isolates during daptomycin therapy.
Infections due to vancomycin-resistant enterococci (VRE) are of significant importance in high-risk populations, and daptomycin is a bactericidal antibiotic to treat multidrug-resistant VRE in these patients. The emergence of daptomycin non-susceptibility invasive VRE during daptomycin therapy is a major clinical issue. Here the hypothesis was tested that systemic daptomycin therapy also induces the emergence of daptomycin non-susceptible (DNS-) isolates in colonizing VRE populations. 11 vancomycin-resistant Enterococcus faecium strain pairs recovered from rectal swabs were available for analysis. All initial isolates exhibited daptomycin MICs within the wild type MIC distribution of E. faecium (MIC≤4 mg/L). In follow-up isolates from five patients a 4-16-fold daptomycin MIC increase was detected. All patients carrying DNS-VRE received daptomycin (14-28 days) at 4 mg/kg body weight, while two patients in whom no DNS-VRE emerged were only treated with daptomycin for 1 and 4 days, respectively. Comparative whole genome sequencing identified DNS-VRE-specific single nucleotide polymorphisms (SNP), including mutations in cardiolipin synthase (Cls), and additional SNPs in independent genes potentially relevant for the DNS phenotype. Mutations within cls were also identified in three additional, colonizing DNS-VRE. Of these, at least one strain was transmitted within the hospital. In none of the VRE isolates tested, pre-existing or de novo mutations in the liaFSR operon were detected. This is the first report documenting the emergence of DNS-VRE in colonizing strains during daptomycin treatment, putting the patient at risk for subsequent DNS-VRE infections and priming the spread of DNS-VRE within the hospital environment.


Photoperiod effects in a freshwater community: Amphibian larvae develop faster and zooplankton abundance increases under an early-season photoperiod.
Organisms that shift their phenologies in response to global warming will experience novel photic environments, as photoperiod (daylength) continues to follow the same annual cycle. How different organisms respond to novel photoperiods could result in phenological mismatches and altered interspecific interactions. We conducted an outdoor mesocosm experiment exposing green frog (Rana clamitans) larvae, gray treefrog (Hyla versicolor) larvae, phytoplankton, periphyton, and zooplankton to a three-month shift in photoperiod: an early-season photoperiod (simulating April) and a late-season photoperiod (simulating July). We manipulated photoperiod by covering and uncovering tanks with clear or light-blocking lids to mimic realistic changes in daylength. We assessed amphibian life history traits and measured phytoplankton, periphyton, and zooplankton abundances. Green frog larvae and gray treefrog metamorphs were more developed under the early-season photoperiod. Gray treefrog total length was also reduced, but photoperiod did not affect green frog total length. Although phytoplankton and periphyton abundances were not affected by photoperiod, copepod nauplii were in greater abundance under the early-season photoperiod. Overall, this simplified aquatic community did not exhibit significant changes to structure when exposed to a three-month shift in photoperiod. Temperate amphibians that breed earlier in the year may develop faster, which may have long-term costs to post-metamorphic growth and performance. Asynchronous shifts in zooplankton abundances in response to altered photoperiods could subsequently affect freshwater community structure. While photoperiod has been shown to individually affect freshwater organisms, our study using replicated outdoor wetland communities shows that the comprehensive effects of photoperiod may be less important than other cues such as temperature and precipitation.


Surgical treatment of traumatic carotid cavernous fistulas.
Six cases of traumatic corotid cavernous fistulae treated by a combination of trapping and embolization are reported. The advantages and disadvantages of various operative procedures in the treatment of carotid cavernous fistulas are discussed. In our opinion a combination of trapping and embolization has advantages over other methods in the treatment of traumatic carotid cavernous fistulae.


Down-modulation of B cell signal transduction by ligation of mucins to CD22.
Epithelial cancer cells secrete mucins carrying carbohydrate antigens such as a sialyl-Tn antigen into cancer tissues and/or the bloodstream, in which mucins may interact with CD22 (Siglec-2). Mucins isolated from colon cancer cells and bovine submaxillary mucins bound to CD22 cDNA transfectants and a human B cell line, Daudi cell, and the binding of soluble recombinant CD22 to the mucins was confirmed by means of a plate assay. The binding specificity was demonstrated by the fact that the mucins bound to the recombinant CD22 with an intact ectodomain but not to that with a mutated ectodomain. Daudi cells were stimulated with anti-IgM F(ab')(2) in the presence or absence of mucins. Ligation of mucins to CD22 decreased the phosphorylation of CD22 and SHP-1 recruitment, and the phosphorylation of ERK-1/2 prominently. The in vivo effect of mucins on splenic B cells in the tumor-bearing state was investigated using mucin-producing (TA3-Ha) and non-producing (TA3-St) mammary adenocarcinoma-bearing mice. When fluorescence-labeled epiglycanins were administered to normal mice, a portion of them was taken up by the spleen and became associated with splenic B cells. We found that splenic B cells were reduced in TA3-Ha-bearing mice but not in TA3-St-bearing ones. These results suggest that in the tumor-bearing state a portion of the mucins in the bloodstream was taken up by the spleen and ligated to CD22 expressed on splenic B cells, which may have led to down-regulation of signal transduction.


Digital screen time and suicidality during high school: How important is cyberbullying? A mediation analysis using the youth risk behavioral surveillance survey, 2011-2019.
Elevated digital screen time (i.e., 2+ hours per day) is associated with suicidal ideations, planning, and attempts during adolescence. Recent studies suggest quality, rather than duration, of digital screen time is most impactful on adolescent mental health. We investigate the role of cyberbullying victimization on the relationship between elevated digital screen time and risk factors for completed suicide. We pooled five years of biennial Youth Risk Behavioral Surveillance (YRBS) survey data (2011-2019). Participants were n = 73,011 high school students in the United States (US). Elevated digital screen time reflected spending 3 h (or more) per day on leisure, digital media. Outcome variables were: (1) feeling sad/hopeless; (2) suicidal ideation; (3) suicidal planning; and (4) suicide attempt. Structural equation models (SEMs) tested the mediating effects of self-reported online bullying victimization (i.e., cyberbullying) on the effect of elevated digital screen time on suicidality. We controlled for age, sex, race/ethnicity, and survey year. Subsample analyses stratified by sex were also conducted. Overall, 40.5% of high school students reported elevated digital screen time and 15.4% reported cyberbullying. Cyberbullying mediated a substantial proportion of the relationship between digital screen time and feeling sad/hopeless (16%), suicidal ideation (18%), planned suicide attempt (18%), and past suicide attempt (26%), among high school students, controlling for covariates. Similar mediating effects were observed in models stratified by sex. Findings reinforce prior research demonstrating that the quality of leisure, digital media strongly influences the relationship between digital screen time and mental health during adolescence. Findings need replication via longitudinal designs.


Combined antihypertensive effect of unripe Rubus coreanus Miq. and Dendropanax morbiferus H. Lév. Extracts in 1 kidney-1 clip hypertensive rats and spontaneously hypertensive rats.
We previously showed that enzymatically hydrolyzed Dendropanax morbiferus H. Lév. leaf (Hy-DP) and unripe Rubus coreanus Miq. (5-uRCK) extracts exhibit potent vasodilator effects on isolated aortic rings from rats partly through endothelium-dependent and endothelium-independent mechanisms. These two extracts have different mechanisms of action; however, their combined effect on antihypertensive activity has not been explored. The present study aims to investigate the effect of a chronic optimized mixture (HDR-2, composed of Hy-DP and 5-uRCK in a 2:1 mass ratio) on vascular tension and blood pressure in two different hypertensive rat models. The results showed that HDR-2 concentration-dependently relaxed endothelium-intact and endothelium-denuded aortic rings precontracted with phenylephrine. Antihypertensive effects were assessed in vivo on a 1 kidney-1 clip (1 K-1C) rat model of hypertension and spontaneously hypertensive rats (SHRs). Acute HDR-2 treatment significantly decreased systolic blood pressure (SBP) 3 h posttreatment in both models. Chronic HDR-2 administration also significantly decreased SBP in the hypertensive rat models. Moreover, HDR-2 increased eNOS protein expression and phosphorylation levels in the aorta. Chronic HDR-2 administration may effectively improve vascular function by decreasing plasma angiotensin-converting enzyme (ACE) activity and AngII levels. HDR-2 significantly improved acetylcholine (ACh)-induced aortic endothelium-dependent relaxation and affected sodium nitroprusside (SNP)-induced endothelium-independent relaxation in SHRs.


X-ray crystallographic analysis of 6-aminohexanoate-dimer hydrolase: molecular basis for the birth of a nylon oligomer-degrading enzyme.
6-Aminohexanoate-dimer hydrolase (EII), responsible for the degradation of nylon-6 industry by-products, and its analogous enzyme (EII') that has only approximately 0.5% of the specific activity toward the 6-aminohexanoate-linear dimer, are encoded on plasmid pOAD2 of Arthrobacter sp. (formerly Flavobacterium sp.) KI72. Here, we report the three-dimensional structure of Hyb-24 (a hybrid between the EII and EII' proteins; EII'-level activity) by x-ray crystallography at 1.8 A resolution and refined to an R-factor and R-free of 18.5 and 20.3%, respectively. The fold adopted by the 392-amino acid polypeptide generated a two-domain structure that is similar to the folds of the penicillin-recognizing family of serine-reactive hydrolases, especially to those of d-alanyl-d-alanine-carboxypeptidase from Streptomyces and carboxylesterase from Burkholderia. Enzyme assay using purified enzymes revealed that EII and Hyb-24 possess hydrolytic activity for carboxyl esters with short acyl chains but no detectable activity for d-alanyl-d-alanine. In addition, on the basis of the spatial location and role of amino acid residues constituting the active sites of the nylon oligomer hydrolase, carboxylesterase, d-alanyl-d-alanine-peptidase, and beta-lactamases, we conclude that the nylon oligomer hydrolase utilizes nucleophilic Ser(112) as a common active site both for nylon oligomer-hydrolytic and esterolytic activities. However, it requires at least two additional amino acid residues (Asp(181) and Asn(266)) specific for nylon oligomer-hydrolytic activity. Here, we propose that amino acid replacements in the catalytic cleft of a preexisting esterase with the beta-lactamase fold resulted in the evolution of the nylon oligomer hydrolase.


Evaluation of the clinical cardiac safety of pemigatinib, a fibroblast growth factor receptor inhibitor, in participants with advanced malignancies.
Pemigatinib is a potent inhibitor of the fibroblast growth factor receptor (FGFR) family of receptors that is approved for the treatment of cholangiocarcinoma with FGFR2 fusion or other rearrangements. Data from a first-in-human clinical study were used to assess the potential for pemigatinib to produce clinically significant effects on heart rate (HR) and cardiac repolarization (QTc). A central tendency analysis for electrocardiogram (ECG) outliers and a plasma concentration-QTc analysis were conducted to assess cardiac safety in the first-in-human pemigatinib study (FIGHT-101; NCT02393248). The study included 113 participants who received at least one dose of pemigatinib as monotherapy and had at least one pair of plasma pharmacokinetic (PK) and ECG data points collected. Timed 12-lead ECGs were performed within 15 min of PK blood draws. The ECG parameters for each dose group in the study varied within expectations for patients with advanced malignancies. Categorical analysis of QT interval corrected for HR by Fridericia's method did not reveal dose dependence in the incidence of outliers, and the results of the central tendency and concentration-QTc analyses did not suggest a dose- or concentration-dependent drug effect. Least squares mean change from baseline in HR was small and did not indicate a clinically relevant effect on HR, and no effect was observed on cardiac conduction as assessed by PR and QRS intervals. In conclusion, pemigatinib does not exhibit any clinically significant prolongation of QTc or dose-dependent changes in HR. Clinical trial registration: ClinicalTrials.gov NCT02393248.


Application of theophylline metabolite assays to the exploration of liver microsome oxidative function in man.
The effects on theophylline oxidative metabolism of 3 inhibitors of liver microsome activity--cimetidine, troleandomycin and ketoconazole--were investigated in 6 healthy volunteers. The 3 compounds increased plasma theophylline half-life by 73.6 +/- 15.6% (P less than 0.01), 107.8 +/- 9.7% (P less than 0.001) and 21.7 +/- 6.8% (P less than 0.02), respectively, and reduced plasma theophylline clearance by 38.3 +/- 4.8% (P less than 0.001), 51.4 +/- 2.4% (P less than 0.001), and 8.9 +/- 7.8% (NS), respectively. Troleandomycin inhibited to the same extent the 2 theophylline metabolism pathways: N-demethylation resulting in the formation of 1-methyluric acid (1-MU) and 3-methylxanthine (3-MX), and 8-hydroxylation resulting in the formation of 1,3-dimethyluric acid (1,3-DMU). The production clearances of these metabolites were almost equally depressed by 60.2 +/- 3.9%, 60.2 +/- 2.1%, and 51.7 +/- 4.5%, respectively. Cimetidine predominantly inhibited the N-demethylation pathway; the production clearances of 1-MU and 3-MX were depressed by 58.5 +/- 4.0% and 57.5 +/- 4.1% (P less than 0.001), respectively, whereas the production clearance of 1,3-DMU was depressed by 38.3 +/- 6.1% (P less than 0.001). Ketoconazole had no significant effect on the produciton clearances of theophylline metabolites. Measurement of theophylline metabolite formation clearances might be a useful test to explore liver microsome oxidative function.


Reactivity of Paul-Bunnell type heterophile antibody in sera from infectious mononucleosis patients with the surface of lymphoid cells carrying Epstein-Barr virus genomes.
The possible presence of Paul-Bunnell (PB) antigen on Epstein-Barr virus (EBV)-transformed lymphocytes were investigated. Of 23 EBV genome-positive lymphoblastoid cell lines tested all but one absorbed PB type antibody from the serum of an infectious mononucleosis patient. The one EBV-negative B cell line tested did not absorb the heterophile antibody. PB antibody, purified by an immunoadsorbent procedure using beef cell antigen, reacted with the EBV producer P3HR-1 cell line in an indirect membrane immunofluorescence test and was shown to be IgM antibody. Titers of heterophile agglutinin and reactivity with the cell surface were reduced to the same degree by absorption with beef cell antigen but not with guinea pig kidney antigen. PB antibody was distinct from IgM antibody against the EBV-determined membrane antigen, since the latter was not absorbed by beef cell antigen. PB antibody was also reactive with other EBV-positive B cell lines (QIMR-WIL, NC-37, and Raji) which were free of surface IgM. No reaction occurred with the nonproducer P3HR-1 line, a null cell line, and two T cell lines. The results suggest the presence of PB antigen on most EBV-transformed B lymphocytes, and its appearance in each of the transformed lymphocytes of patients with acute infectious mononucleosis.


Laparoscopic sleeve gastrectomy as an alternative to gastric bypass in patients with multiple intraabdominal adhesions.
Laparoscopic sleeve gastrectomy and its indications are currently being evaluated. The objective of this study was to show the preliminary results obtained with this technique indicated as an alternative to gastric bypass in patients with multiple intraabdominal adhesions, therefore preserving the benefits of the laparoscopic approach. An analysis of all patients who underwent a laparoscopic sleeve gastrectomy for the above indication was done. Data included demographics, number of previous surgeries, operative time, morbidity, mortality, and %EWL at 3 and 6 months. Fifteen patients underwent laparoscopic sleeve gastrectomy as an alternative to gastric bypass because of multiple intraabdominal adhesions. No patient required conversion to an open procedure; morbidity was 6% with no mortality. %EWL at 3 months was 41% and at 6 months was 44%. Mean follow-up was 6 months. In our initial experience, laparoscopic sleeve gastrectomy proved to be a safe and effective alternative to gastric bypass for patients with multiple intraabdominal adhesions.


Prostaglandin-mediated hyperemia and renin-mediated hypertension during acute ureteral obstruction.
Acute elevation of ureteral pressure to 100 mm Hg in anesthetized dogs (n=7) resulted in an increase (P less than 0.005) in systemic blood pressure form 151 +/- 7 to 163 +/- 7 mm Hg, a transient (approximately 15 min) increase (P less than 0.05) in renal blood flow from 413 +/- 27 to 465 +/- 27 ml/min and a rise (P less than 0.05) in plasma renin activity from 6.0 +/- 1.6 to 10.3 +/- 2.1 ng/ml/hr. Pretreatment with a competitive inhibitor of angiotensin II, i.e. sar1gly8AII, abolished the hypertensive response to acute ureteral obstruction, and pretreatment with 2 mg/kg of either indomethacin (n=6) or meclofenamate (n=3), 15 min before obstruction, prevented the hyperemic response. These results suggest that acute ureteral obstruction leads to hypertension via activation of the renin-angiotensin system and hyperemia via a prostaglandin-initiated mechanism.


Oxytocin-induced facilitation of lordosis behaviour in rats is progesterone-dependent.
Oxytocin was administered intracerebroventricularly to ovariectomized female rats and its effect on lordosis was examined. In Experiment 1a, oxytocin (0.17 mU in 4 microliters saline) failed to facilitate lordosis behaviour in animals primed acutely with 10 micrograms estradiol benzoate (EB) and 300 micrograms progesterone (P). In Experiment 1b, however, oxytocin significantly facilitated lordosis in animals primed acutely with 10 micrograms EB and 250 or 200 micrograms P. In Experiments 2a and 2b, oxytocin failed to facilitate lordosis in animals treated acutely with 10 micrograms EB, or chronically with 0.8 micrograms EB daily for eight days prior to testing. These results support the hypothesis that the facilitation of lordosis in ovariectomized rats by centrally-administered oxytocin is progesterone-dependent.


Tempol prevents chronic sleep-deprivation induced memory impairment.
Sleep deprivation is associated with oxidative stress that causes learning and memory impairment. Tempol is a nitroxide compound that promotes the metabolism of many reactive oxygen species (ROS) and has antioxidant and neuroprotective effect. The current study investigated whether chronic administration of tempol can overcome oxidative stress and prevent learning and memory impairment induced by sleep deprivation. Sleep deprivation was induced in rats using multiple platform model. Tempol was administered to rats via oral gavages. Behavioral studies were conducted to test the spatial learning and memory using radial arm water maze. The hippocampus was dissected; antioxidant biomarkers (GSH, GSSG, GSH/GSSG ratio, GPx, SOD, and catalase) were assessed. The result of this project revealed that chronic sleep deprivation impaired both short and long term memory (P<0.05), while tempol treatment prevented such effect. Furthermore, tempol normalized chronic sleep deprivation induced reduction in the hippocampus activity of catalase, GPx, and SOD (P<0.05). Tempol also enhanced the ratio of GSH/GSSG in chronically sleep deprived rats treated with tempol as compared with only sleep deprived rats (P<0.05). In conclusion chronic sleep deprivation induced memory impairment, and treatment with tempol prevented this impairment probably through normalizing antioxidant mechanisms in the hippocampus.