Targeted, homology-driven gene insertion in stem cells by ZFN-loaded 'all-in-one' lentiviral vectors.
Biased integration remains a key challenge for gene therapy based on lentiviral vector technologies. Engineering of next-generation lentiviral vectors targeting safe genomic harbors for insertion is therefore of high relevance. In a previous paper (Cai et al., 2014a), we showed the use of integrase-defective lentiviral vectors (IDLVs) as carriers of complete gene repair kits consisting of zinc-finger nuclease (ZFN) proteins and repair sequences, allowing gene correction by homologous recombination (HR). Here, we follow this strategy to engineer ZFN-loaded IDLVs that insert transgenes by a homology-driven mechanism into safe loci. This insertion mechanism is driven by time-restricted exposure of treated cells to ZFNs. We show targeted gene integration in human stem cells, including CD34(+) hematopoietic progenitors and induced pluripotent stem cells (iPSCs). Notably, targeted insertions are identified in 89% of transduced iPSCs. Our findings demonstrate the applicability of nuclease-loaded 'all-in-one' IDLVs for site-directed gene insertion in stem cell-based gene therapies.


The effect of subcutaneous nerve stimulation (SCNS) on pain associated with osteoarthritis of the hip.
A novel form of subcutaneous nerve stimulation (SCNS) was recently introduced for the relief of chronic pain. We present a study using this form of SCNS applied over the radial, median and saphenous nerves in patients with clinically diagnosed osteoarthritis of the hip. Acceptable pain relief was obtained in 60% of patients receiving stimulation, however, comparable analgesia was achieved in a control group, who received no electrical stimulation through similarly placed needles. We suggest that these results may be explained by the ability of SCNS to evoke a placebo response. The efficacy of the placebo effect and the ethical implications of its use in clinical practice are discussed.


Ocular genetics: current understanding.
Over the past decade, there has been an exponential increase in our knowledge of heritable eye conditions. Coincidentally, our ability to provide accurate genetic diagnoses has allowed appropriate counseling to patients and families. A summary of our current understanding of ocular genetics will prove useful to clinicians, researchers, and students as an introduction to the subject.


Comparison between the EUCAST procedure and the Etest for determination of the susceptibility of Candida species isolates to micafungin.
We compared the ability of the EUCAST EDef 7.2 and the Etest to detect the susceptibility to micafungin of 160 Candida and non-Candida clinical isolates. Agreement was higher when Etest MICs were obtained after 24 h of incubation; essential agreement was 90%, and categorical agreement was >90%. False susceptibility was seen only for Candida krusei (10%), and false resistance was observed in 6% of the isolates, ranging from 2.6% (C. glabrata) to 13% (C. albicans).


Crystal structure of the bacterial ribosomal decoding site complexed with a synthetic doubly functionalized paromomycin derivative: a new specific binding mode to an a-minor motif enhances in vitro antibacterial activity.
The crystal structure of the complex between oligonucleotide containing the bacterial ribosomal decoding site (A site) and the synthetic paromomycin analogue 1, which contains the gamma-amino-alpha-hydroxybutyryl (L-haba) group at position N1 of ring II (2-DOS ring), and an ether chain with an O-phenethylaminoethyl group at position C2'' of ring III, is reported. Interestingly, next to the paromomycin analogue 1 specifically bound to the A site, a second molecule of 1 with a different conformation is observed at the crystal packing interface which mimics the A-minor interaction between two bulged-out adenines from the A site and the codon-anticodon stem of the mRNA-tRNA complex. Improved antibacterial activity supports the conclusion that analogue 1 might affect protein synthesis on the ribosome in two different ways: 1) specific binding to the A site forces maintenance of the "on" state with two bulged out adenines, and 2) a new binding mode of 1 to an A-minor motif which stabilizes complex formation between the ribosome and the mRNA-tRNA complex regardless of whether the codon-anticodon stem is of the cognate or near-cognate type.


Early results with restorative proctocolectomy.
The first 25 restorative proctocolectomies (RP) performed by one surgeon since 1986 are reviewed. Ulcerative colitis (23) and idiopathic megarectum (2) were the indications for surgery. The initial seven patients had a submucosal proctectomy and transanal hand sewn pouch-anal anastomosis. Eighteen patients had a totally stapled RP. One patient with malignancy died from factors unrelated to surgery. Complications developed in eight patients after pouch-anal anastomosis and in four patients after ileostomy closure. There was one pouch failure due to pelvic sepsis. No pelvic sepsis has occurred following a stapled ileal pouch-anal anastomosis with defunctioning ileostomy. After a mean follow up period of 20.5 months all patients are continent (mean stool frequency = 4/day and 0.5/night). Soiling occurred in three patients who had a transanal hand sewn pouch-anal anastomosis. All patients in the stapled group have satisfactory control. Restorative proctocolectomy produces satisfactory operative and functional results which have improved as our experience has increased.


Genomic risk of hepatitis C-related hepatocellular carcinoma.
To identify the genetic susceptibility factor(s) for hepatitis C virus-induced hepatocellular carcinoma (HCV-induced HCC), we conducted a genome-wide association study using 432,703 autosomal SNPs in 721 individuals with HCV-induced HCC (cases) and 2890 HCV-negative controls of Japanese origin. Eight SNPs that showed possible association (P <1 10(⁻5)) in the genome-wide association study were further genotyped in 673 cases and 2596 controls. We found a previously unidentified locus in the 50 flanking region of MICA on 6p21.33 (rs2596542, P(combined) = 4.21 10(⁻13), odds ratio = 1.39) to be strongly associated with HCV induced HCC. Subsequent analyses using individuals with chronic hepatitis C (CHC) indicated that this SNP is not associated with CHC susceptibility (P = 0.61) but is significantly associated with progression from CHC to HCC (P = 3.13 10(⁻8)). We also found that the risk allele of rs2596542 was associated with lower soluble MICA protein levels in individuals with HCV-induced HCC (P = 1.38 10(⁻13)).


Parasitic myoma after transabdominal hysterectomy for fibroids: a case report.
Parasitic myomas typically occur after a pedunculated subserosal fibroid loses its uterine blood supply and parasitizes other organs or after a surgery involving morcellation techniques. Parasitic myomas that occur after transabdominal surgery are extremely rare and may not be sufficiently documented. Here, we present a case of parasitic myoma in the anterior abdominal wall following a transabdominal hysterectomy for fibroids. The patient was a 46-year-old Chinese woman who had undergone surgery for uterine myomas at our hospital 1 year prior. The patient later revisited our department with a palpable mass in her abdomen, and imaging revealed a mass in the iliac fossa. The possibility of a broad ligament myoma or solid ovarian tumor was considered before surgery, and laparoscopic exploration was performed under general anesthesia. A tumor measuring approximately 4.5 × 4.0 cm was found in the right anterior abdominal wall, and a parasitic myoma was considered. The tumor was completely resected. Pathological analysis of the surgical specimens suggested leiomyoma. The patient recovered well and was discharged on postoperative day 3. This case suggests that parasitic myoma should be considered in the differential diagnosis of patients presenting with abdominal or pelvic solid tumors with a history of surgery for uterine leiomyomas, even without a history of laparoscopic surgery using a power morcellator. Thorough inspection and washing of the abdominopelvic cavity at the end of surgery is vital.


Ultrasonographic aspects of the Leontopithecus gestation (Lesson, 1840-Callitrichidae, Primates).
There is a concern about conservation of endangered species today. Among this species, the Leontopithecus (Lesson, 1840) is outstanding. Its population has been whirling reduced. So far the reproductive physiology of Leontopithecus has few studies, it is fundamental requisite to preserve this species. Obstetric sonography has become an essential method in reproductive management of primates. This method is very helpful to detect early pregnancy and evaluate some deficiency of fetal growth. In this study, 14 pregnancies were monitored using real-time abdominal sonography. During each evaluation, the number of fetus was recorded, gestational sac and heart beats were observed, and biparietal diameter was measured. The results showed that abdominal sonography is a reliable method for observation of gross morphological changes during pre-natal development and to estimate gestational age. No statistically differences were observed between twins and singletons. This study is the first investigation of pre-natal growth in Leontopithecus.


Molecular epidemiology, antimicrobial susceptibility, and characterization of fluoroquinolone non-susceptible Streptococcus pyogenes in Japan.
Streptococcus pyogenes (Group A streptococci: GAS) are known to cause a wide variety of human illnesses, some of which can be life-threatening. Usually, penicillin is the first-choice agent for the treatment of GAS infections. For patients with penicillin or beta-lactam antibiotics allergies, macrolide drugs are recommended as an alternative therapy. However, an increased prevalence of macrolide-resistant GAS (MRGAS) has been reported in many countries. Furthermore, fluoroquinolone non-susceptible GAS has been reported. The present study was focused on determining the features of fluoroquinolone non-susceptible strains collected from children with pharyngotonsillitis in the southwestern areas of Japan. To reveal the characteristics of fluoroquinolone non-susceptible GAS, we investigated the MIC, T-serotype, emm typing, and PFGE of 298 GAS strains isolated in the Fukuoka southwest area of Japan between 2011 and 2013. We determined that fluoroquinolone non-susceptibility shows a MIC to tosufloxacin of ≧1 μg/ml. We identified 33 (11.1%) fluoroquinolone non-susceptible GAS strains. In these strains, 6 T-serotypes and 9 emm/MLST patterns were detected. The predominant combinations were emm6/ST382 (14 strains, 42.4%) and emm89/ST101 (5 strains, 15.2%). PFGE classified 10 pulsotypes, and each was quite different. These results showed that fluoroquinolone non-susceptible GAS strains have a variety of origins. The usage of fluoroquinolone drugs could have a negative effect on the antimicrobial drug sensitivity of GAS in Japan. Considering such a situation, continuous monitoring of quinolone non-susceptible GAS is necessary.


The risk stratification of adverse neonatal outcomes in women with gestational diabetes (STRONG) study.
To assess the risk of adverse neonatal outcomes in women with gestational diabetes (GDM) by identifying subgroups of women at higher risk to recognize the characteristics most associated with an excess of risk. Observational, retrospective, multicenter study involving consecutive women with GDM. To identify distinct and homogeneous subgroups of women at a higher risk, the RECursive Partitioning and AMalgamation (RECPAM) method was used. Overall, 2736 pregnancies complicated by GDM were analyzed. The main outcome measure was the occurrence of adverse neonatal outcomes in pregnancies complicated by GDM. Among study participants (median age 36.8 years, pre-gestational BMI 24.8 kg/m2), six miscarriages, one neonatal death, but no maternal death was recorded. The occurrence of the cumulative adverse outcome (OR 2.48, 95% CI 1.59-3.87), large for gestational age (OR 3.99, 95% CI 2.40-6.63), fetal malformation (OR 2.66, 95% CI 1.00-7.18), and respiratory distress (OR 4.33, 95% CI 1.33-14.12) was associated with previous macrosomia. Large for gestational age was also associated with obesity (OR 1.46, 95% CI 1.00-2.15). Small for gestational age was associated with first trimester glucose levels (OR 1.96, 95% CI 1.04-3.69). Neonatal hypoglycemia was associated with overweight (OR 1.52, 95% CI 1.02-2.27) and obesity (OR 1.62, 95% CI 1.04-2.51). The RECPAM analysis identified high-risk subgroups mainly characterized by high pre-pregnancy BMI (OR 1.68, 95% CI 1.21-2.33 for obese; OR 1.38 95% CI 1.03-1.87 for overweight). A deep investigation on the factors associated with adverse neonatal outcomes requires a risk stratification. In particular, great attention must be paid to the prevention and treatment of obesity.


Placental transfer and fetal urinary excretion of gentamicin during constant rate maternal infusion.
Placental transfer and fetal urinary excretion of gentamicin was studied in midtrimester goat and human previable fetuses during constant rate maternal infusion with the drug. Gentamicin was not detected in the serum of any of the goat fetuses, even when maternal serum concentrations ranged from 15.2 mug/ml to 20.9 mug/ml. However, gentamicin was present in the amniotic fluid of four animals. Gentamicin was also present in fetal urine collected from three animals. In contrast human fetal central venous serum concentrations of gentamicin were 21-37% of those in maternal serum after constant rate infusion of the mother. In addition, gentamicin was present in human fetal urine in concentrations 2-- 3 times those in fetal serum. The observed difference in fetal serum concentration of gentamicin between the two species represents a difference in placental permeability to gentamicin and/ or a difference in fetal renal clearance of the drug.


RSM process optimization of biodiesel production from rapeseed oil and waste corn oil in the presence of green and novel catalyst.
In the scenario of global warming and pollution, the green synthesis and use of biodiesel has acquired utmost priority. Due to several limitations of homogeneous catalysis, organobase immobilized heterogeneous catalyzed production of biodiesel has come out as a favored route. The present report demonstrates the design and synthesis of Peganum harmala spice seed extract modified GO-CuFe2O4 (SSE@GO-CuFe2O4) nanocomposite as an organobase functionalized high surface area magnetic nanocatalyst. Pistachio leaves were used in the green reduction of precursor salts to synthesize CuFe2O4 NPs. The as-synthesized nanomaterial was characterized physicochemically by Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), energy dispersive X-Ray analysis (EDX), elemental mapping, transmission electron microscopy (TEM), X-Ray diffraction (XRD), thermogravimetric analysis (TGA) and vibrating sample magnetometer techniques (VSM). Subsequently, the catalyst was explored in the efficient synthesis of biodiesels by trans-esterification of two substrates, the rapeseed oil and waste corn oil. The optimum conditions for biodiesel production were determined through response surface methodology based on Box-Behnken design including the study of calibration curves and 3D contour plots. Easy separation and workup, use of green medium, excellent reused for several times and short reaction time are outstanding benefits of this study.


A Critical Appraisal of the New Competency-Based Medical Undergraduate Curriculum in Biochemistry.
The new competency-based medical education undergraduate curriculum (CBMC) was launched for the 2019 admission batch of MBBS students. The programme is designed to create an "Indian Medical Graduate" (IMG) possessing the requisite knowledge, skills, attitudes, values and responsiveness, so that the graduate may function appropriately and effectively as a physician of first contact with the community while being globally relevant. Given that implementation of this curriculum is still in its infancy across the country, we stand to gain from a unified approach to its implementation. Phase I of the curriculum includes anatomy, physiology, and biochemistry along with professional and personal development modules. Biochemistry enjoys an enviable position in the medical curriculum as it explains the molecular basis of diseases. We present an appraisal of the curriculum in Biochemistry by reviewing the components against Harden's six themes which are considered when planning or developing a curriculum. Further, five core components of CBME are selected on the basis of three research papers to characterize underlying assumptions of CBME to suggest ways of logical implementation for achieving the competencies expected of the Indian Medical Graduate. The insight gained shall help students to be equipped with competencies which they shall be able to use in their day- to- day work, which shall ultimately help benefit patient care and the society at large. The online version contains supplementary material available at 10.1007/s12291-022-01088-y.


Derivation and validation of an equation to determine the optimal ventilator setting in children undergoing intracranial revascularization surgery: A single-center retrospective study.
It can be difficult to determine the appropriate ventilator settings to maintain normocapnia in children undergoing general anesthesia for surgery for moyamoya disease, especially immediately following anesthesia induction. We conducted this study to attempt to derive an equation to predict the appropriate ventilator settings and subsequently validated the accuracy of the equation. A retrospective study of 91 pediatric patients less than 18 years of age who underwent cerebral revascularization for moyamoya disease at our institution. Fifty-eight patients were used to derive the equation, and the subsequent 33 patients were used to validate the equation. We calculated the required respiratory rate to attain normocapnia based on the median of all values of the minute volume during normocapnia (estimated partial pressure of arterial carbon dioxide of 38-42 mm Hg) and the assumption that the tidal volume was 8 mL/kg body weight. We derived the regression equation from the derivation data set where the required respiratory rate to attain normocapnia was represented by age. We simplified the equation by rounding coefficients to the nearest integer. The level of agreement between the respiratory rate predicted from the equation and the actual required respiratory rate was assessed in the validation group using Bland-Altman analysis. The derived equation is tidal volume = 8 mL/kg body weight, respiratory rate = 24-age/min. Bland-Altman analysis in the validation group revealed that the mean bias between the predicted and actual respiratory rate was 0.29 (standard deviation, 3.67). The percentage of cases where the predicted rate was within ± 10% and ± 20% of the actual rate was 42.4% and 66.7%, respectively. We derived and validated a simple and easily applicable equation to predict the ventilator settings required to attain normocapnia during general anesthesia in children with moyamoya disease.


The quantitation of human ventricular myosin light chain 1 in serum after myocardial necrosis and infarction.
To determine the clinical utility of human ventricular myosin light chain 1 (HVLC1) in the diagnosis of myocardial necrosis, we established a radioimmunoassay for serum HVLC1 using polyclonal antibodies raised against the purified human protein. HVLC1 levels were measured in sera from 110 control patients and 38 patients immediately after cardiovascular surgery and in serial specimens from 10 patients with uncomplicated myocardial infarctions. The HVLC1 assay was found to be equal in sensitivity for the first 48 hours to the CK-MB for myocardial necrosis after cardiovascular surgery and for myocardial infarction. In virtually all myocardial infarct patients, HVLC1 levels in sera rose within hours of the onset of chest pain. From thereon, two sub-trends were discerned. For many, the HVLC1 levels remain elevated for 9-12 days beyond. In some, the HVLC1 levels returned to normal levels 1-2 days after the initial rise, but became elevated again for the next 5-9 days. In either instance, the diagnosis of myocardial infarction was permitted as late as 10-12 days after the onset of chest pain which is in stark contrast to all conventionally used biochemical markers.


Analysis of Adeno-Associated Virus Serotype 8 (AAV8)-antibody complexes using epitope mapping by molecular imprinting leads to the identification of Fab peptides that potentially evade AAV8 neutralisation.
Gene therapy is a promising approach for treating genetic disorders by delivering therapeutic genes to replace or correct malfunctioning genes. However, the introduced gene therapy vector can trigger an immune response, leading to reduced efficacy and potential harm to the patient. To improve the efficiency and safety of gene therapy, preventing the immune response to the vector is crucial. This can be achieved through the use of immunosuppressive drugs, vector engineering to evade the immune system, or delivery methods that bypass the immune system altogether. By reducing the immune response, gene therapy can deliver therapeutic genes more effectively and potentially cure genetic diseases. In this study, a novel molecular imprinting technique, combined with mass-spectrometry and bioinformatics, was used to identify four antigen-binding fragments (Fab) sequences of Adeno-Associated Virus (AAV) - neutralising antibodies capable of binding to AAV. The identified Fab peptides were shown to prevent AAV8's binding to antibodies, demonstrating their potential to improve gene therapy efficiency by preventing the immune response.


Post-mortem imaging in traffic fatalities: from autopsy to reconstruction of the scene using freely available software.
CT scan coupled with autopsy is the gold standard for the forensic investigation of fatal road traffic accidents. The objective of the present paper is to demonstrate that from this, it is possible to reconstruct elements of an accident with minimal human and material resources using basic knowledge of three-dimensional imaging software. This is illustrated by a case implicating a pedestrian and a motor vehicle in which the impact areas were matched using freely available computer-aided design software. Such an approach aims to improve the visualisation of forensic elements, which is crucial for the understanding of all parties involved in the legal implications of such accidents and which could become the standard practice in many institutes.


Synthesis of human U1 RNA. II. Identification of two regions of the promoter essential for transcription initiation at position +1.
We have analyzed the requirements for human U1 RNA transcription catalyzed by RNA polymerase II. In Xenopus laevis oocytes, a human U1 RNA gene with only 231 and 35 nucleotides of the 5' and 3' flanking regions, respectively (Lund, E. and Dahlberg, J. E. (1984) J. Biol. Chem. 259, 2013-2021), is able to support accumulation of human U1 RNA. We show that the point in the template corresponding to the 5' end of U1 RNA is a site of transcription initiation. That result rules out the possibility that the 5' end of U1 RNA is generated by cleavage and capping of a precursor RNA. The accumulation of correctly initiated human U1 RNA transcripts requires at least two essential upstream elements. The region between positions -231 and -203 is indispensable for transcription both in oocytes and in vitro. The other region, between positions -105 and -6, fixes the location of the 5' ends of the U1 RNA transcripts in oocytes while not altering the overall level of transcription. This latter region contains a sequence located around position -50, which we propose serves as the analog of the T-A-T-A sequence in U1 and U2 RNA genes.


Expression level and methylation status of three tumor suppressor genes, DLEC1, ITGA9 and MLH1, in non-small cell lung cancer.
Despite therapeutic advances, lung cancer remains one of the most common causes of cancer-related death in the world. There is a need to develop biomarkers of diagnostic and/or prognostic value and to translate findings in basic science research to clinical application. Tumor suppressor genes (TSGs) represent potential useful markers for disease detection, progression and treatment target. We tried to elucidate the role of three 3p21.3 TSGs: DLEC1, ITGA9 and MLH1, in non-small cell lung cancer (NSCLC). We assessed their expression pattern by qPCR in 59 NSCLC tissues and in the matched macroscopically unchanged lung tissues. Additionally, we analyzed gene promoter methylation status by methylation-specific PCR in NSCLC samples. We did not find significant correlations between gene expression and methylation. In case of DLEC1 and ITGA9, expression levels were decreased in 71-78 % of tumor samples and significantly different between tumor and normal tissues (P = 0.0001). It could point to their diagnostic value. ITGA9 could also be regarded as a diagnostic marker differentiating NSCLC subtypes, as its expression level was significantly lower in squamous cell carcinoma (P = 0.001). The simultaneous down-regulation of DLEC1 and ITGA9 was observed in 52.5 % of NSCLCs. MSPs revealed high frequencies of gene promoter methylation in NSCLCs: 84 % for DLEC1 and MLH1 and 57 % for ITGA9. Methylation indexes reflected moderate gene methylation levels: 34 % for ITGA9, 27 % for MLH1 and 26 % for DLEC1. However, frequent simultaneous methylation of the studied genes in more than 50 % of NSCLCs suggests the possibility of consider them as a panel of epigenetic markers.


The effect of various grading scales on student grade point averages.
To investigate changes in and the impact of grading scales from 2005 to 2010 and explore pharmacy faculty and student perceptions of whole-letter and plus/minus grading scales on cumulative grade point averages (GPAs) in required courses. Grading scales used in 2010 at the University of Cincinnati College of Pharmacy were retrospectively identified and compared to those used in 2005. Mean GPA was calculated using a whole-letter grading scale and a plus/minus grading scale to determine the impact of scales on GPA. Faculty members and students were surveyed regarding their perceptions of plus/minus grading. Nine unique grading scales were used throughout the curriculum, including plus/minus (64%) and whole-letter (21%) grading scales. From 2005 to 2010 there was transition from use of predominantly whole-letter scales to plus/minus grading scales. The type of grading scale used did not affect the mean cumulative GPA. Students preferred use of a plus-only grading scale while faculty members preferred use of a plus/minus grading scale. The transition from whole-letter grading to plus/minus grading in courses from 2005 to 2010 reflects pharmacy faculty members' perception that plus/minus grading allows for better differentiation between students' performances.


Uric acid and gamma-glutamyl-transferase in children and adolescents with obesity: Association to anthropometric measures and cardiometabolic risk markers depending on pubertal stage, sex, degree of weight loss and type of patient care: Evaluation of the adiposity patient follow-up registry.
Associations between body mass index (BMI)- standard deviation score (SDS)/waist-to-height ratio (WHtR) were studied with (i) serum uric acid (sUA)/gamma-glutamyl-transferase (GGT) and (ii) cardiometabolic risk markers in children with obesity, considering sex, pubertal development, and degree of weight loss/type of patient care. 102 936 children from the Adiposity-Follow-up registry (APV; 47% boys) were included. Associations were analysed between sUA/GGT and anthropometrics, transaminases, lipids, fasting insulin (FI), homeostasis model assessment of insulin resistance (HOMA-IR), triglycerides to HDL-cholesterol (TG/HDL)-ratio. Follow-up analyses (3-24 months after baseline) considered a BMI-SDS reduction ≥0.2 (n = 11 096) or ≥0.5 (n = 3728). Partialized correlation analyses for sex and BMI-SDS were performed, taking pubertal development into consideration. At baseline, BMI-SDS showed the strongest correlations to sUA (r = 0.35; n = 26 529), HOMA-IR/FI (r = 0.30; n = 5513 /n = 5880), TG/HDL-ratio (r = 0.23; n = 24 501), and WHtR to sUA (r = 0.32; n = 10 805), GGT (r = 0.34; n = 11 862) and Alanine-aminotransferase (ALAT) (r = 0.33; n = 11 821), with stronger correlations in boys (WHtR and GGT: r = 0.36, n = 5793) and prepubertal children (r = 0.36; n = 2216). GGT and sUA (after partializing effects of age, sex, BMI-SDS) showed a correlation to TG/HDL-ratio (r = 0.27; n = 24 501). Following a BMI-SDS reduction ≥0.2 or ≥0.5, GGT was most strongly related to Aspartate-aminotransferase (ASAT)/ ALAT, most evident in prepuberty and with increasing weight loss, and also to TG/HDL-ratio (r = 0.22; n = 1528). Prepubertal children showed strongest correlations between BMI-SDS/WHtR and GGT. ΔBMI-SDS was strongly correlated to ΔsUA (r = 0.30; n = 4160) and ΔGGT (r = 0.28; n = 3562), and ΔWHtR to ΔGGT (r = 0.28; n = 3562) (all p < 0.0001). Abdominal obesity may trigger hyperuricemia and hepatic involvement already in prepuberty. This may be stronger in infancy than anticipated to date. Even moderate weight loss has favourable effects on cardiometabolic risk profile and glucose homeostasis.


Arterial blood gases in undisturbed resting birds: measurements in chicken and duck.
Arterial blood was sampled in resting, unrestrained and undisturbed birds using a remote-controlled sampling device. With this technique a blood sample of 0.45 ml was obtained for analysis of PO2, PCO2 and pH at the body temperature of the animal. In 5 domestic hens the following mean values were obtained: PA02-82 torr; PACO2=33 torr; pHA=7.52. The average body temperature was TB=41.0 degrees C. In 6 domestic ducks the analysis yielded values of PAO2=82 torr; PACO2=38 torr; pH2=7.49; Tb=41.0 degrees C. Although these values are close to those reported in the literature some discrepancies may be recognized; they can be attributed to respiratory and metabolic changes of blood composition possibly induced by classical sampling procedures.


Physiological and thermoregulatory effects of oral taurine supplementation on exercise tolerance during forced convective cooling.
AbstractWe investigated the effects of taurine supplementation on cycling time to exhaustion in cold conditions. Eleven males cycled to exhaustion at a power output equivalent to the mid-point between ventilatory threshold and maximum aerobic power following 15-min rest in the cold (apparent temperature of ∼ 4°C; air flow of 4.17 m s-1). Two hours before, participants ingested taurine (50 mg·kg-1) or placebo beverage. Pulmonary gases, carbohydrate (CHO) and fat oxidation, body temperatures, mean local sweat rate, heart rate, rate of perceived exertion (RPE) and thermal comfort were recorded. Time to exhaustion was not different between trials (taurine = 14.6 ± 4.7 min; placebo = 13.4 ± 5.6 min, P = 0.061, d = 0.27). There were no effects (P > 0.05) of taurine on core temperature, mean skin temperature or local sweat rates. However, the placebo condition showed greater (P < 0.05) reductions in arm-to-finger temperature gradient (i.e. vasodilation) across pre-exercise passive cold exposure and increased CHO oxidation (P < 0.05). Participants also reached a thermally 'comfortable' level quicker in the taurine condition (P < 0.05). A 50 mg·kg-1 dose of taurine did not statistically benefit endurance exercise after moderate cold exposure but conferred some potential vascular and metabolic effects.


CAMP-dependent protein kinase: prototype for a family of enzymes.
Protein kinases represent a diverse family of enzymes that play critical roles in regulation. The simplest and best-understood biochemically is the catalytic (C) subunit of cAMP-dependent protein kinase, which can serve as a framework for the entire family. The amino-terminal portion of the C subunit constitutes a nucleotide binding site based on affinity labeling, labeling of lysines, and a conserved triad of glycines. The region beyond this nucleotide fold also contains essential residues. Modification of Asp 184 with a hydrophobic carbodiimide leads to inactivation, and this residue may function as a general base in catalysis. Despite the diversity of the kinase family, all share a homologous catalytic core, and the residues essential for nucleotide binding or catalysis in the C subunit are invariant in every protein kinase. Affinity labeling and intersubunit cross-linking have localized a portion of the peptide binding site, and this region is variable in the kinase family. The crystal structure of the C subunit also is being solved. The C subunit is maintained in its inactive state by forming a holoenzyme complex with an inhibitory regulatory (R) subunit. This R subunit has a well-defined domain structure that includes two tandem cAMP binding domains at the carboxy-terminus, each of which is homologous to the catabolite gene activator protein in Escherichia coli. Affinity labeling with 8N3 cAMP has identified residues that are in close proximity to the cAMP binding sites and is consistent with models of the cAMP binding sites based on the coordinates of the CAP crystal structure. An expression vector was constructed for the RI subunit and several mutations have been introduced. These mutations address 1) the major site of photoaffinity labeling, 2) a conserved arginine in the cAMP binding site, and 3) the consequences of deleting the entire second cAMP binding domain.


12-0-tetradecanoylphorbol-13-acetate and concanavalin A enhance glucose uptake in thymocytes by different mechanisms.
The effects of the tumor promoter 12-0-tetradecanoylphorbol-13-acetate (TPA) and the plant lectin concanavalin A (Con A) on glucose uptake in murine thymocytes were studied. TPA induces a rapid dose-dependent increase in the uptake of 2-deoxyglucose and in the transport of 3-0-methylglucose. Con A also elicits a time- and dose-dependent enhancement of 2-deoxyglucose uptake. The effect of Con A, however, is less pronounced. The effect of combined treatment of thymocytes with Con A and TPA is not additive. Cytochalasin B completely inhibits the basal, as well as TPA- and Con A-enhanced, 2-deoxyglucose uptake. Dexamethasone markedly inhibits basal 2-deoxyglucose uptake, but is less inhibitory to enhanced 2-deoxyglucose uptake induced by TPA and Con A. The effect of TPA on 2-deoxyglucose uptake and 3-0-methylglucose transport is refractory to inhibition by isobutyl methyl xanthine, dibutyryl cyclic AMP, and ethyleneglycol tetraacetic acid. These agents markedly inhibit the enhancement of 2-deoxyglucose (2-DOG) uptake by Con A. p-Bromophenacyl bromide, an inhibitor of phospholipase A2, also selectively inhibits Con A enhancement of 2-DOG uptake. Taken together, the results suggest that Con A and TPA exert their stimulatory effect on glucose uptake by different activating mechanisms, but they may share a final common transport pathway.


Association of outer retinal layer morphology with visual acuity in patients with retinal vein occlusion: SCORE Study Report 13.
To assess associations between visual acuity (VA) and the status of the photoreceptor inner segment-outer segment (IS-OS) junction in a subset of patients in the Standard Care vs COrticosteroid for REtinal Vein Occlusion (SCORE) Study. High-resolution time domain optical coherence tomography (OCT) scans of study eyes from a single site participating in the SCORE Study were evaluated. Integrity of the IS-OS junction in the central subfield was evaluated using a three-step scale: absent, abnormal or normal. Associations of the IS-OS status with ETDRS VA letter score and center point thickness (CPT) were investigated. Baseline OCTs of 42 eyes were evaluated. The IS-OS junction was absent in 30 (71%) and abnormal in 12 (29%). At month 12, the IS-OS junction was absent in 18 (43%), abnormal in 12 (28%), and normal in 12 (28%) eyes. At baseline, IS-OS status was significantly associated with CPT, but not with VA. At month 12, IS-OS status was significantly associated with CPT and VA, that is, absent or abnormal IS-OS was associated with increased CPT and worse VA. Change in IS-OS status was not associated with change in CPT (P=0.8). Worsening of IS-OS status was associated with loss of VA and improvement in IS-OS status to normal was associated with gain in VA (P=0.03). In this data set with long-term follow-up of OCTs as part of the SCORE Study, there is a correlation between change in IS-OS status and VA. This supports further evaluation of outer retinal morphology in larger data sets.


Management of osteoradionecrosis of the jaws with pentoxifylline-tocopherol: a systematic review of the literature and meta-analysis.
Osteoradionecrosis (ORN) of the jaws remains among the most commonly encountered and challenging complications of radiotherapy to the head and neck. The purpose of this study was to provide a review of the medical management for ORN and evaluate the reported outcomes with the use of pentoxifylline and tocopherol (PENTO), by means of a systematic review and meta-analysis. The predictor variable was the use of PENTO in the treatment of ORN. The outcome variable was the proportion of full recovery or significant improvement not requiring further intervention. The likelihood function was used to combine the studies and estimate the proportion and standard deviation of each outcome by the maximum likelihood estimation. Seven studies met the inclusion criteria. A total 211 patients were treated. One hundred twenty-six patients recovered fully or improved significantly not requiring further intervention. Sixty patients remained the same, 10 were lost to follow-up, and the disease progressed in 15. The current literature supports the use of PENTO in the treatment of ORN of the jaws. Additional well-designed prospective studies are needed in order to further validate the regimen that can then be employed in the treatment of ORN.


24-hour esophageal pH monitoring before and after medical therapy for reflux esophagitis.
Medical treatment of gastroesophageal reflux disease often results in improvement of symptoms. The purpose of this study was to determine if improvement in symptoms and endoscopic appearance after treatment was associated with a reduction in reflux, as measured with 24-hr pH recordings. Twenty patients with severe chronic reflux esophagitis participated in an eight-week double-blind trial of medical therapy with metoclopramide and cimetidine versus placebo and cimetidine. Significant symptom score improvement was noted in 11 patients. Eleven patients also had improvement in the endoscopic appearance of the esophageal mucosa, and eight of these patients had significant symptom improvement. Initial 24-hr pH recordings were abnormal in all patients, evidenced by an esophageal pH less than 4 during 20% of the study period. Improvement in 24-hr results was noted in only five patients--three with clinical and endoscopic improvement, and two with no improvement. In conclusion, there was no relationship between clinical improvement and the results of 24-hr pH recordings. Successful symptom relief and endoscopic healing of esophagitis during medical treatment may occur despite persistent reflux of gastric contents.


Major histocompatibility complex (MHC) antigens polymorphism and alloimmunization study in thalassemia patients with febrile non-hemolytic transfusion reaction (FNHTR).
HLA alloimmunization is one of the most troublesome consequence of regular transfusion which is itself a mainstay measure to provide longevity to the thalassemia patients. Febrile non-hemolytic transfusion reaction (FNHTR) is one of the most common complication which might be related to the HLA alloimmunization. Here, we studied the HLA antigenic system and alloimmunization rate in the Iranian β-thalassemia patients who suffered from FNHTR compare to the β-thalassemia patients without FNHTR. Total of 60 β-thalassemia patients with FNHTR (case group) and 20 β-thalassemia patients without FNHTR (control group) randomly have been selected and enrolled in the study. All were tested for HLA-A and -B loci by PCR-SSP method and also for the presence of anti-lymphocyte antibodies by LIFT method. Comparisons between two groups were performed by Pearson's χ2 test. Totally, a significant predominance was noted for two HLA alleles, HLA-A*24 (P = 0.029) and B*55 (P = 0.034) which have higher prevalence in control group. Although no significant association was found between the presence of anti-leukocyte antibodies and the development of FNHTR, the HLA-A*32 (P = 0.047) allele was considered as possible genetic markers in the susceptibility to the development of anti-leukocyte antibodies. Here some evidences about the possible role of HLA polymorphism in susceptibility to FNHTR are provided. Those results indicated that HLA-A*24 and HLA-B*55 might play protective role on inducing FNHTR in β-thalassemia patients. Further studies which investigate the allele level of HLA-I alongside with specific reactivity of HLA-I antibodies might reveal more deep data about these phenomena.


Does sevoflurane inhibit serum cholinesterase in children?
Serum cholinesterase activity was measured at induction, and following anaesthesia in 41 children aged between 4 and 30 months. The median exposure to sevoflurane was 273%.min. The results did not demonstrate any significant difference in cholinesterase activity, when expressed by gram of serum proteins following inhalation anaesthesia using sevoflurane or intravenous anaesthesia using propofol. The values (SD) obtained were 175 (42) UI.g(-1) before anaesthesia and 177 (43) UI.g(-1) following anaesthesia. The only change in cholinesterase activity detected was related to heamodilution. We conclude that plasma fluoride concentration following sevoflurane administration [13.8 (4.2) microm x l(-1)] is too low to exert an inhibiting effect on in vivo cholinesterase activity and that the previously reported decrease in mivacurium requirements during sevoflurane anaesthesia is unlikely to be due to inhibition by fluoride ions.


Implementation of an intraleukocitary cystine quantification method for diagnosis of cystinosis.
Cystinosis is an inborn error of metabolism, clinically characterised by severe renal involvement and development of corneal cystine deposits, especially in the adult form of the disease. Cystinosis is a treatable condition. Therefore, an early diagnosis is necessary to start therapy. For biochemical confirmation of the condition it is necessary to quantify intracellular cystine concentrations. For this, different methods have been described with variations in cell isolation strategies and the amino acid quantification techniques used. In order to improve confirmatory biochemical diagnosis in our setting, a protocol for intraleukocitary cystine quantification was established. A high performance liquid chromatography based method for cystine quantification in polymorphonuclear cells was implemented. Evaluation of the best anticoagulant to use and temperature stability of the sample at 4̊C were performed. In addition, we established reference values for our population. It was determined that intraleukocitary cystine quantification must be performed in blood samples containing acid-citrate-dextrose as anticoagulant. Samples must be processed immediately due to their poor stability even when refrigerated. Based on the results from 50 healthy individuals, the cut-off point established for our population was 0.34nmol 1/2 cystine/mg. The adaptation performed to the cystine quantification method here presented the highest control population that has been reported in the literature so far. Our results highlight the need for making available a cystine quantification method locally and confirm the convenience for each laboratory to establish its own reference values to provide greater reliability for interpreting results.


Effects of cholesterol oxidation derivatives on cholesterol esterifying and cholesteryl ester hydrolytic enzyme activity of cultured rabbit aortic smooth muscle cells.
The effects of 5 micrograms/ml of 25-hydroxycholesterol; cholestane-3 beta,5 alpha,6 beta-triol; and cholesterol on acyl CoA cholesterol acyltransferase, acid cholesteryl ester hydrolase and neutral cholesteryl ester hydrolase was studied in cultured rabbit aortic smooth muscle cells. After 1 hour incubation, 25-hydroxycholesterol resulted in a fourfold stimulation of acyl CoA cholesterol acyltransferase activity. No stimulation by 25-hydroxycholesterol was noted before 15 minutes or after 5 hours of incubation. Neither cholestane-3 beta,5 alpha,6 beta-triol nor cholesterol influenced acyl CoA cholesterol acyltransferase activity at any time interval. No significant effects of any of the sterols were noted on acid cholesteryl ester hydrolase or neutral cholesteryl ester hydrolase activity. The imbalance between acyl CoA cholesterol acyl transferase and hydrolase activities induced by 25-hydroxycholesterol could result in cholesteryl ester accumulation by arterial smooth muscle cells, which may be associated with atherosclerosis.


Peripheral migration of pacemaker electrodes.
A 55-year-old male was admitted with right ileo-femoral phlebitis caused by an unused pacemaker electrode fragment which had migrated from the right subclavian vein to the right iliac vein. Vena cava plication was followed by removal of the electrode fragment and complete resolution of the phlebitis. The exposed metal coil may have contributed to the severe inflammatory reaction.


A histopathological and ultrastructural study of eccrine porocarcinoma with special reference to its subtypes.
Five cases of eccrine porocarcinoma were studied by light and electron microscopy. Histopathologically, these could be classified into two types; the common and the giant cell type. The common type was characterized by almost uniform medium-sized cuboidal tumour cells and a formation of well-developed intracytoplasmic lumina. A broad diversity of histopathological and ultrastructural features was seen in these tumours. The tumours of the giant cell type consisted of mononuclear polygonal cells and bizarre giant cells. This type was considered to be an undifferentiated form of porocarcinoma.


The efficacy of topical gentamycin application on prophylaxis and treatment of wound infection: A systematic review and meta-analysis.
The purpose of this study was to conduct a systematic review and meta-analysis in patients with local wound infection or infective risk, evaluating effects of topical gentamycin application on prophylaxis and treatment of wound infection. Embase, the Cochrane Library, Pubmed, Medline (from Ovid) and three Chinese literature databases (CNKI, VIP and WANFANG) were searched. Randomised controlled studies (RCTs) and observational studies (OSs) that assessed the efficacy of topical gentamycin application on prophylaxis and treatment of local wound infection were included. The primary outcome was clinical efficacy. Secondary outcomes included duration of recovery time and length of hospital stay. Fifteen studies (1781 patients) met inclusion criteria. Twelve studies were RCTs and other three studies were OSs. Compared with non-gentamycin group, topical gentamycin application had significantly higher rates of clinical efficacy (OR = 3.57, 95% CI 2.52-5.07). In terms of duration of wound healing, it's taken shorter time in gentamycin group than non-gentamycin group (OR = -4.94, 95% CI -8.37 to -1.51). However, the length of hospital stay had no significantly difference between the two groups (OR = -3.40, 95% CI -8.42 to 1.63). Subgroup analyses were conducted according to study design (RCTs or OSs), purpose and administration type. And the results showed that there were no significant difference of clinical efficacy in study design (P = 0.21, I2 = 35.4%), purpose (P = 0.32, I2 = 0%) and administration type subgroup (P = 0.74, I2 = 0%). However, topical gentamycin application had significantly shorter duration of wound healing in randomly controlled trials compared with observational studies, but had no difference in terms of administration type(P = 0.20, I2 = 38.6%). Studies to date show that topical gentamycin application significantly increases the rate of clinical efficacy and decreases the duration of wound healing in patients with local wound infection or infective risk.


A circadian susceptibility/resistance rhythm for potassium cyanide in male BALB/cCr mice.
Circadian rhythms in mortality and/or survival time following a single intraperitoneal injection of a LD50 of potassium cyanide were studied. In two investigations, different but comparable subgroups of inbred male BALB/cCr mice were treated at 4-h intervals (under conditions standardized for chronobiologic study) during 24-h spans. Mice were observed for exact time-to-death during the first hour after treatment as well as overall mortality during the entire 24-h post-injection span following each KCN treatment timepoint. In both studies, mortality from KCN exhibited a 24-h rhythm. Highest mortality occurred in mice injected at 1600 (80% mortality) in Experiment 1 and 2000 (100% mortality) in Experiment II. Lowest mortality occurred at 0400 (40% mortality) in Experiment I and 0800 (30% mortality) in Experiment II. The need to consider the circadian organization of physiologic function when bioassaying toxicity is discussed.


Tracer uptake by photoreceptor synaptic terminals. I. Dark-mediated effects.
This electron microscope study examines the sites of uptake and subsequent redistribution of the extracellularly applied tracer, horseradish peroxidase (HRP), in photoreceptor synaptic terminals during dark adaptation. First, chicks are sacrificed by perfusion fixation at various intervals after the addition of tracer to in situ eyecups. It is demonstrated that when terminals contain only one tracer-labeled organelle, this may be either a synaptic or a coated vesicle. At later times vacuoles in terminals become labeled. Some of these are seen as involutions of terminal membrane and some are observed in the cell body. Therefore, it seems likely that some involutions of terminal membrane pinch off and are transported retrogradely. Second, chicks are given intraocular injections of HRP at various intervals in a 12-hr dark period and sacrificed 1 hr later. Terminals of chicks sacrificed at 1 hr into the dark period contain predominantly tracer-labeled synaptic vesicles and vacuoles, whereas terminals of chicks sacrificed at 3 hr into the dark period contain tracer-labeled coated vesicles also. Diverticular membrane, thought to be the result of synaptic vesicle fusion and coalescence with the presynaptic membrane, waxes during the first hour of darkness and then wanes over a period of several hours (N. Cooper and B. McLaughlin, 1982, J. Ultrastruc. Res. 79, 58-73). These observations, together with those presented here, suggest that not all synaptic vesicles coalesce to form the diverticular membrane and that during the waxing stage, some synaptic vesicles are recycled rapidly without passing through a coated vesicle intermediate stage. This is discussed in the context of current models of localized synaptic vesicle membrane recycling.


LifeSoaks: a tool for analyzing solvent channels in protein crystals and obstacles for soaking experiments.
Due to the structural complexity of proteins, their corresponding crystal arrangements generally contain a significant amount of solvent-occupied space. These areas allow a certain degree of intracrystalline protein flexibility and mobility of solutes. Therefore, knowledge of the geometry of solvent-filled channels and cavities is essential whenever the dynamics inside a crystal are of interest. Especially in soaking experiments for structure-based drug design, ligands must be able to traverse the crystal solvent channels and reach the corresponding binding pockets. Unsuccessful screenings are sometimes attributed to the geometry of the crystal packing, but the underlying causes are often difficult to understand. This work presents LifeSoaks, a novel tool for analyzing and visualizing solvent channels in protein crystals. LifeSoaks uses a Voronoi diagram-based periodic channel representation which can be efficiently computed. The size and location of channel bottlenecks, which might hinder molecular diffusion, can be directly derived from this representation. This work presents the calculated bottleneck radii for all crystal structures in the PDB and the analysis of a new, hand-curated data set of structures obtained by soaking experiments. The results indicate that the consideration of bottleneck radii and the visual inspection of channels are beneficial for planning soaking experiments.


Analytical applications of a carbon nanotubes composite modified with copper microparticles as detector in flow systems.
In this work we report on the successful use of a composite prepared by dispersion of multi-wall carbon nanotubes (1-5 microm length, 20-50 nm diameter) and copper microparticles within mineral oil as detector for amino acids quantification in flow injection analysis and capillary electrophoresis. The resulting electrode displays a highly sensitive amperometric detection of amino acids, based on the copper dissolution facilitated by the strong activity of amino acids as ligands of Cu(II). The sensor makes possible the detection of amino acids, electroactive or not, at very low potentials (0.000 V) and physiological pH. A correlation between the sensitivity for the amino acids and the amount of copper within the composite is observed, demonstrating the importance of the metal in the sensor response. The best analytical performance is obtained for the electrode containing 12.0% (w/w) copper. The excellent results obtained with the carbon nanotube paste electrodes containing copper (CNTPE-Cu) as detector in flow systems makes them an interesting alternative for further analytical applications involving different bioanalytes.


Cone-beam computed tomography study of root and canal morphology of maxillary first and second molars in an Indian population.
The aim of this study was to investigate the root and canal morphology of maxillary first and second molars in an Indian population by using cone-beam computed tomography (CBCT). Maxillary first (n = 220) and second (n = 205) molars were collected from an indigenous Indian population and scanned by using a CBCT scanner at a constant slice thickness of 125 μm/slice. Volume rendering and multiplanar volume reconstruction were performed. The number of root canals was examined, and root canal system configurations were classified by using historical and contemporary classifications. Single-rooted first and second molars commonly showed types I, IV (0.5%) and type III (1%) canal systems, respectively. Buccal roots of two-rooted first molars showed 2 canal systems, type I and type IV, whereas second molars with 2 roots showed wide variations in canal anatomy. The most common canal morphology in the mesiobuccal roots of three-rooted first and second molars was type I (51.8% and 62%, respectively), followed by type IV (38.6% and 50%, respectively). The distobuccal and palatal roots of first and second molars showed predominantly type I canal morphology. Additional canal types were identified in 2.2% and 9.3% of the first and second molars, respectively. The root number, morphology, and canal morphology of Indian maxillary molars showed features that were different from both Caucasian and Mongoloid traits. CBCT is an exciting and clinically useful tool in studying root canal morphology.


A case report of 131I therapy for Graves' disease patient with hemiagenesis.
Thyroid hemiagenesis is a rare congenital dysplasia, whereas a variety of pathological changes may occur in residual thyroid lobe. The most frequently described pathology in residual thyroid lobe is Graves' hyperthyroidism. Although I therapy has been generally recommended as the preferred treatment for Graves' disease (GD), subjects relating to hemiagenesis are very limited, especially in China. A 43-year-old female patient presented to our hospital on November 2014, with a 1-year history of palpitation, fatigue, and hand tremor. Her situation was getting worse within 2 months. The thyroid function tests were suggestive of thyrotoxicosis. The technetium thyroid scintigraphy only showed an enlarged right lobe with increased tracer uptake. Then, the agenesis of left lobe and isthmus was confirmed by ultrasound and magnetic resonance imaging (MRI). Thus, a diagnosis of GD with hemiageneis of the left lobe and isthmus of thyroid was made. Thiamazole was discontinued because of drug-induced hepatic injury. According to our procedures, the patient was treated by I. Hypothyroidism was observed 3 months after I therapy. After replacement therapy with L-thyroxine (LT4), the state of euthyroid maintained. Once hypothyroidism occurs, regular application of LT4 and review of thyroid function is very important. Thus, patients' compliance needs to be strengthened. Besides, we could not convince the family members of our patient to undergo ultrasonographic examination. The genetic factor of the agenesis could not be proved in this case.


Severity of DSS-induced colitis is reduced in Ido1-deficient mice with down-regulation of TLR-MyD88-NF-kB transcriptional networks.
Indoleamine 2,3 -dioxygenase 1 (IDO1) catalyzes L-tryptophan to kynurenine in the first and rate-limiting step of tryptophan metabolism. IDO1 is expressed widely throughout the body, with especially high expression in colonic intestinal tissues. To examine the role of IDO1 in the colon, transcriptome analysis was performed in both Ido1(-/-) and Ido1(+/+) mice. Gene set enrichment analysis identified the Inflammatory Response as the most significant category modulated by the absence of IDO1. This observation prompted us to further investigate the function of IDO1 in the development of tissue inflammation. By using DSS-induced experimental colitis mice models, we found that the disease in Ido1(-/-) mice was less severe than in Ido1(+/+) mice. Pharmacological inhibition of IDO1 by L-1MT attenuated the severity of DSS-colitis as well. Transcriptome analyses revealed that pathways involving TLR and NF-kB signaling were significantly down-regulated by the absence of IDO1. Furthermore, dramatic changes in TLR and NF-kB signaling resulted in substantial changes in the expression of many inflammatory cytokines and chemokines. Numbers of inflammatory cells in colon and peripheral blood were reduced in IDO1 deficiency. These findings suggest that IDO1 plays important roles in producing inflammatory responses and modulating transcriptional networks during the development of colitis.


Synthesis of 1,4,5-trisubstituted 1,2,3-triazoles amicable for automation.
A three-component 3+2 cycloaddition reaction followed by Suzuki coupling reaction was carried out to synthesize a library of compounds using automation (parallel synthesizer). Scaffolds that are unexplored in literature were used for the synthesis of library. The iodo-triazoles formed by 3+2 cycloaddition reaction were coupled with boronic acids to get tri-substituted triazoles.


Detection of conformational changes in human chorionic gonadotropin upon binding to rat gonadal receptors.
After binding to rat testicular or ovarian luteinizing hormone (LH) receptors, human chorionic gonadotropin (hCG) and mammalian LH can be detected with monoclonal antibodies directed against a conserved epitope on the beta subunit of the hormones. Two such anti-hCG/anti-LH monoclonal antibodies, known as B105 and B110, compete with one another for binding to this epitope region on free and receptor-bound hormone. By comparing the affinities of B105 and B110 for these two forms of hCG, we have detected apparent changes in the structure of the hormone which develop subsequent to receptor binding. Whereas the affinity of B105 for receptor-bound hCG is approximately 10-fold lower than that for free hCG, the affinity of B110 for receptor-bound hCG is nearly 20-fold greater than that for free hCG. Both B105.hCG and B110.hCG complexes bind to the receptor; however, they have approximately 25 and 50% lower affinity than hCG. Thus, although B110 binds better to the form of hCG which is bound to receptors, binding of B110 to hCG does not appear to induce a conformational change in the hormone which facilitates hormone-receptor binding. Consequently, both B105 and B110 partially inhibit binding of hCG to its receptors. Fab fragments of B105 and B110 are as effective as intact B105 and B110 in inhibiting the binding of labeled B105 and B110 to hCG-receptor complexes, suggesting that circular complexes which might be formed by the interaction of divalent antibody, two molecules of hCG, and two membrane-bound receptors or one divalent receptor are not contributing to the affinity of the antibodies for receptor-bound hCG. Alternatively, formation of circular complexes can explain an increase in apparent affinity of B105 for ovine or bovine LH-receptor complexes. Data obtained with B105 suggest either that the structure of the epitope is altered following binding or that a portion of the epitope is partially obscured when hCG binds to the receptor. In contrast, the data obtained using B110 are not explained by models in which steric factors reduce the affinity of the antibody for the hormone-receptor complex. Therefore, as a minimal explanation for these observations, we postulate that the conformation of the B105/B110 epitope region is altered following binding of the hormone to receptors. The nature of the conformational change and its relationship to LH/hCG action is unknown.


Covalency-reinforced oxygen evolution reaction catalyst.
The oxygen evolution reaction that occurs during water oxidation is of considerable importance as an essential energy conversion reaction for rechargeable metal-air batteries and direct solar water splitting. Cost-efficient ABO3 perovskites have been studied extensively because of their high activity for the oxygen evolution reaction; however, they lack stability, and an effective solution to this problem has not yet been demonstrated. Here we report that the Fe(4+)-based quadruple perovskite CaCu3Fe4O12 has high activity, which is comparable to or exceeding those of state-of-the-art catalysts such as Ba(0.5)Sr(0.5)Co(0.8)Fe(0.2)O(3-δ) and the gold standard RuO2. The covalent bonding network incorporating multiple Cu(2+) and Fe(4+) transition metal ions significantly enhances the structural stability of CaCu3Fe4O12, which is key to achieving highly active long-life catalysts.


Biogas stripping of ammonia from fresh digestate from a food waste digester.
The efficiency of ammonia removal from fresh source-segregated domestic food waste digestate using biogas as a stripping agent was studied in batch experiments at 35, 55 and 70°C, at gas flow rates of 0.125 and 0.250Lbiogasmin(-1)L(-1)digestate with and without pH adjustment. Higher temperatures and alkaline conditions were required for effective ammonia removal, and at 35°C with or without pH adjustment or 55°C with unadjusted pH there was little or no removal. Results were compared to those from earlier studies with digestate that had been stored prior to stripping and showed that ammonia removal from fresh digestate was more difficult, with time constants 1.6-5.7 times higher than those previously reported. This has implications for the design of large-scale systems where continuous stripping of fresh digestate is likely to be the normal operating mode. A mass balance approach showed that thermal-alkaline stripping improved hydrolysis.


Proposing Melasma Severity Index: A New, More Practical, Office-based Scoring System for Assessing the Severity of Melasma.
Melasma Area and Severity Index (MASI), the scoring system in melasma, needs to be refined. To propose a more practical scoring system, named as Melasma Severity Index (MSI), for assessing the disease severity and treatment response in melasma. Four dermatologists were trained to calculate MASI and also the proposed MSI scores. For MSI, the formula used was 0.4 (a × p(2)) l + 0.4 (a × p(2)) r + 0.2 (a × p(2)) n where "a" stands for area, "p" for pigmentation, "l" for left face, "r" for right face, and "n" for nose. On a single day, 30 enrolled patients were randomly examined by each trained dermatologist and their MASI and MSI scores were calculated. Next, each rater re-examined every 6(th) patient for repeat MASI and MSI scoring to assess intra- and inter-rater reliability of MASI and MSI scores. Validity was assessed by comparing the individual scores of each rater with objective data from mexameter and ImageJ software. Inter-rater reliability, as assessed by intraclass correlation coefficient, was significantly higher for MSI (0.955) as compared to MASI (0.816). Correlation of scores with objective data by Spearman's correlation revealed higher rho values for MSI than for MASI for all raters. Sample population belonged to a single ethnic group. MSI is simpler and more practical scoring system for melasma.


Cost-Effectiveness Analysis of Temocillin Treatment in Patients with Febrile UTI Accounting for the Emergence of Antibiotic Resistance.
Antibiotic resistance has been listed as one of the biggest threats to global health today. A recent study has shown that treating febrile urinary tract infections with temocillin instead of cefotaxime leads to a reduced selection of antibiotic-resistant bacteria. However, a potential challenge with prioritizing temocillin over cefotaxime is the cost consequences. This study aimed to assess the cost effectiveness of using temocillin compared to cefotaxime in treating febrile urinary tract infections in a model that takes the emergence of antibiotic resistance into account. We used a Markov cohort model to estimate the costs and health effects of temocillin and cefotaxime treatment in febrile urinary tract infections in a Swedish setting. Health effects were assessed in terms of quality-adjusted life-years, and the primary outcome was the cost per quality-adjusted life-year gained with temocillin compared to cefotaxime. We used a 5-year time horizon. The model results showed that temocillin treatment led to better health outcomes at a higher total cost. The cost per quality-adjusted life-year gained was approximately 38,400 EUR. Results from the sensitivity analysis suggested a 63% probability of temocillin being cost effective at a threshold of 50,000 EUR. Furthermore, results showed that the cost effectiveness of temocillin in febrile urinary tract infections is highly dependent on the drug cost. As antibiotic consumption is a driving force of resistance, it is essential to consider the development of resistance when studying the health economic consequences of antibiotic treatments. In doing so, this study found temocillin to be cost effective for febrile urinary tract infections.


Antepartum aminophylline treatment for prevention of the respiratory distress syndrome in premature infants.
The frequency of idiopathic respiratory distress (IRD) among premature offspring born of women who were given aminophylline before the thirty-fourth week of pregnancy was evaluated. Sixty-seven premature deliveries were included in the aminophylline group and 75 in the control group. The perinatal death rate was 7.1% in the aminophylline group and 17.9% in the control group (p less than 0.05). A statistically significant difference was noted between the aminophylline and control groups in the frequency of IRD which was three times lower (10%) in the aminophylline group than in the control group (29.5%) for the total of premature infants. If the time of rupture of membranes is taken into consideration, a significant decrease in the frequency of IRD following aminophylline administration is noted in the infants, whose mothers had ruptured membranes for more than 24 hours. No complications or side effects of aminophylline administration were noted in the mothers or their infants.


The WHO/START study in New Caledonia: A psychological autopsy case series.
Limited information is available about suicidal behavior in the Pacific Islands. Forty percent of the New Caledonian population is indigenous Kanak; insights into the characteristics of suicide deaths in this population compared to other ethnic groups would be valuable. The aim of this paper is to deepen our understanding of the cultural underpinnings of suicide in New Caledonia by presenting the results of the WHO/START psychological autopsy study. A case-series psychological autopsy study was conducted based on medical, police files, and interviews with relatives of 52 individuals who died by suicide in 2014 and 2015 in New Caledonia. Kanak indigenous individuals represented more than half of suicides. Prevalence of mental disorders was identified in 62% of suicide decedents; a previous suicide attempt was frequent (37% of cases). A serious argument with a partner was the most prevalent life event (60%). Few warning signs were present among young people and Kanak in general. Comparison with information on the general population suggests indigenous Kanak are more vulnerable and that having a mental health disorder plays an important role in suicide. Violent arguments with a partner could also be a major risk factor. Traditional protective factors (being employed, living with their family or partner, religion) appeared to have limited effect on suicide. Mental health promotion, prevention, and care should be prioritized. Prevention strategies, including domestic violence prevention are recommended in New Caledonia. Further research is needed to better identify young and Kanak subjects at risk of suicide.


The Subglottic Stenosis 6 Questionnaire: A Novel Quality-of-Life Survey Validated to Predict Need for Intervention in Patients With Idiopathic Subglottic Stenosis.
Idiopathic subglottic stenosis (iSGS) is a recurrent, chronic disease defined by fibroinflammatory narrowing of the subglottic airway. A key challenge in treatment is monitoring disease progression, which may be debilitating and unpredictable in its timing. Can the Subglottic Stenosis 6 (SGS-6) questionnaire be validated as a novel quality-of-life (QOL) instrument to monitor breathing, disease progression, and disease severity proactively in patients with iSGS? Panel data from 51 patients with iSGS were collected from January 2012 through June 2022, representing 1,684 patient encounters including routine office visits and treatment encounters. Subjective QOL scores (including the novel SGS-6 and established RAND-36 and EuroQol Five Dimensions [EQ-5D] Visual Analog Scale) and objective pulmonary function test (PFT) results were collected at each visit. Subjective SGS-6 QOL scores were repeated within 1 week of initial reporting. Panel regression analyses were performed to assess the relationship between SGS-6 scores, PFT results, and a patient's need for intervention. Minimal clinically important differences (MCIDs) for SGS-6 and peak expiratory flow percentage (PEF%) were assessed using receiver operating characteristic (ROC) curve analysis and a patient's need for intervention as the external anchor. Each one-point increase in SGS-6 score (of a maximum of 27) was associated with a 3.26% decrease in PEF%, a 1.93-point decrease in RAND-36 Physical Health composite score, a 1.27-point decrease in RAND-36 Mental Health composite score, and a 0.88-point decrease in EQ-5D Visual Analog Scale score. The intracorrelation coefficient for the SGS-6 composite score is 0.838 (95% CI, 0.770-0.888). Compared with patient baselines, SGS-6 scores were 4.66 points greater at the time of intervention with an MCID of 2.25 from a patient's baseline. The area under the ROC curve for SGS-6 and a patient's intervention point was 0.81. iSGS disease severity can be modeled using the SGS-6 questionnaire, offering physicians and patients a potentially new method of tracking disease progression and need for intervention remotely.


Circulating thrombospondin 2 levels reflect fibrosis severity and disease activity in HCV-infected patients.
Among several secreted glycoproteins belonging to the thrombospondin family, thrombospondin 2 (TSP2) is involved in various functions, including collagen/fibrin formation. Liver/serum TSP2 levels have been correlated to liver fibrosis stage and disease activity in nonalcoholic fatty liver disease. This study investigated whether serum TSP2 was associated with clinicopathological features in hepatitis C virus (HCV)-infected patients as well. A total of 350 patients with HCV who had undergone liver biopsy were retrospectively enrolled and divided into a discovery cohort (n = 270) and a validation cohort (n = 80). In the discovery cohort, serum TSP2 levels were moderately correlated with both liver fibrosis stage (r = 0.426, P < 0.0001) and activity grade (r = 0.435, P < 0.0001). The area under the receiver operating characteristic curve of TSP2 for predicting severe fibrosis (≥ F3) was 0.78 and comparable to or better than those of autotaxin (0.78), FIB-4 index (0.78), and APRI (0.76). The discovery cohort findings were closely replicated in the validation cohort. Moreover, comprehensive liver genetic analysis of HCV-infected patients confirmed that the expression of the THBS2 gene encoding TSP2 was significantly higher in severely fibrotic F4 than in F1 patients. Circulating TSP2 levels may reflect the severity of hepatic fibrosis/inflammation in HCV-infected patients.


Keratin polypeptide composition as a biochemical tool for the discrimination of benign and malignant epithelial lesions in man.
An investigation was undertaken of the keratin polypeptides of various benign and malignant tumors of the human skin and vaginal epithelium by one and two dimensional gel electrophoresis. The keratin patterns of benign tumors were found to be similar to the patterns of normal epithelium or stratum corneum. The relative proportion of stratum-corneum associated keratin polypeptides to those polypeptides characteristic for the living layers corresponds to morphological features (e.g., hyperkeratosis, acanthosis). In contrast to benign tumors, epithelial carcinomas totally lack the group of high-molecular-weight keratins. This finding may be helpful in the diagnostic discrimination between benign and malignant epithelial lesions.


The effects of hypothermia on myocardial oxygen consumption and transmural coronary blood flow in the potassium-arrested heart.
Hypothermia remains the primary adjunct employed to lower cellular metabolism during various cardiac procedures. In these experiments, left ventricular myocardial oxygen consumption (MVO2) and transmural blood flow (TBF) were measured during cardiopulmonary bypass with the range of temperatures used clinically. Determinations were made in empty beating normothermic hearts and after potassium cardioplegia at 37, 32, 28, 22, 18, and 15 degrees (K+ = 15--37 meq/L: Hct 25 volumes %). Oxygen content of the total coronary sinus collection was compared with a large volume arterial sample using a Lex-O2-Con-TL analyzer (vs Van Slyke, R = 0.98). Transmural blood flow was measured at each temperature using microspheres (8 microns), and perfusion was maintained at 80 mmHg. Asystole (37 degrees) alone decreased MVO2 from 5.18 +/- 0.55 to 1.85 +/- 0.20 ml O2/min/100 g of left ventricle or approximately 65% (p less than 0.001). With progressive cooling to 15 degrees an additional 82% decrement in oxygen uptake occurred during asystole (p less than 0.001). During asystole at 37 degrees the decrease in MVO2 was reflected mainly by a large decrement (p less than 0.01) in TBF (1.27 +/- 0.19 to 0.74 +/- 0.17 ml/min/g of mean left ventricular flow). However, with cooling below 32 degrees, the arteriovenous oxygen difference narrowed progressively (p less than 0.001) while TBF paradoxically returned to control levels. Endocardial/epicardial flow ratios were not altered by cooling. These data not only confirm earlier reports describing a sequential drop in MVO2 with incremental myocardial cooling, but also establish MVO2 levels for perfused hearts arrested by potassium at lower temperatures (18--15 degrees). Moreover, as transmural blood flow becomes independent of metabolic necessity during hypothermia, coronary autoregulation appears to be impaired, possibly affecting detrimental tissue over perfusion.


Preeclampsia activates circulating immune cells with engagement of the NF-kappaB pathway.
Compelling evidence implicates peripheral immune activation in the pathophysiology of preeclampsia. Polymorphonuclear neutrophils appear to be the cells most strongly affected, with changes in expression of surface markers and release of granule enzymes. Here, we investigated activation in additional leukocyte populations among women with preeclampsia. We used flow cytometry to evaluate changes in leukocyte markers in preeclampsia compared with uncomplicated pregnancy. To gain insights into intracellular pathways involved in leukocyte activation, we monitored the NF-kappaB signal transduction pathway. Plasma levels of interleukin-6 (IL-6) were also studied as an additional indication of cellular activation. Preeclampsia is associated with changes in L-selectin (CD62L) on neutrophils (P = 0.004), monocytes (P = 0.013), and T cells (P = 0.048) when compared with normal pregnancy. These changes include an increase in nuclear translocation of NF-kappaB and increased levels of IL-6 (P = 0.005). These findings are consistent with the presence of a generalized phenomenon of immune activation in preeclampsia.


Presentation of a flashing light following training produces amnesia.
Long-Evans rats given exposure to a flashing-light stimulus following one-trial fear conditioning showed amnesia relative to controls not given the flashing-light experience. Together with previous results obtained in this laboratory, it is apparent that the flashing light modulates memory processing.


AST/ALT ratio as a significant predictor of the incidence risk of prostate cancer.
To investigate the effect of serum aspartate transaminase/alanine transaminase (AST/ALT) on the risk of prostate cancer. A total of 404 patients undergoing prostate biopsy from April 2016 to July 2019 were enrolled. One hundred and ninety-four patients with prostatic cancer (PCa) were diagnosed by pathology. Two hundred and ten patients were diagnosed with benign prostatic hyperplasia (BPH). Multivariate logistic regression was used to analyze the effect of AST/ALT ratio and other factors on the incidence of PCa. AST/ALT ratio was significantly higher in PCa than in BPH patients (OR 2.313, 95%CI 1.337-4.003, P = .002). ROC curve indicated that the best cutoff was 1.155 in predicting the incidence risk of PCa. The age of PCa patients is higher than BPH patients (OR 1.054, 95%CI 1.027-1.082, P < .001). We also found that platelets were lower in PCa than in BPH patients. Multivariate analysis showed that AST/ALT ratio could be used as an independent predictor to assess the incident risk of PCa(OR 1.043, 95%CI 1.014-1.072, P = .003). However, AST/ALT ratio did not predict the incidence in high-risk or low-risk PCa. AST/ALT ratio was an independent factor in predicting the incidence of PCa. When the level of AST/ALT ratio in serum raised, the incidence risk of PCa was significantly increased, which was helpful for the clinical diagnosis of PCa. We still needed a multicenter study to verify the role of AST/ALT ratio in the development of PCa.


Portal vein thrombosis as the first sign of nephrotic syndrome.
A 52-year-old man presented to hospital having experienced abdominal pain, abdominal distention and oliguria for 1 week. An abdominal CT scan revealed thrombosis in the superior mesenteric vein and in the right branch and the trunk of the portal vein. Physical examination, blood and urine analyses, color Doppler ultrasonography and abdominal CT scan. Nephrotic syndrome complicated by portal vein thrombosis. Treatment with batroxobin, low-molecular-weight heparin, prostaglandin E, dipyridamole and methylprednisolone.


The purification of nuclease-free T4-RNA ligase.
RNA ligase has been highly purified in good yields from bacteriophage T4-infected Escherichia coli by a rapid and reproducible procedure. The enzyme is free of phosphomonoesterase and ribonuclease activities and is therefore suitable for the synthesis of oligoribonucleotides and for the labeling of the 3'-terminus of RNA. Greater than 90% of the protein in the enzyme preparation migrates as a single band on gradient polyacrylamide gels containing sodium dodecyl sulfate during electrophoresis. For use as a DNA synthesis reagent the enzyme may be reliably freed of deoxyribonuclease activity by an additional chromatographic procedure using a commercially avialable resin.


Using spike train distances to identify the most discriminative neuronal subpopulation.
Spike trains of multiple neurons can be analyzed following the summed population (SP) or the labeled line (LL) hypothesis. Responses to external stimuli are generated by a neuronal population as a whole or the individual neurons have encoding capacities of their own. The SPIKE-distance estimated either for a single, pooled spike train over a population or for each neuron separately can serve to quantify these responses. For the SP case we compare three algorithms that search for the most discriminative subpopulation over all stimulus pairs. For the LL case we introduce a new algorithm that combines neurons that individually separate different pairs of stimuli best. The best approach for SP is a brute force search over all possible subpopulations. However, it is only feasible for small populations. For more realistic settings, simulated annealing clearly outperforms gradient algorithms with only a limited increase in computational load. Our novel LL approach can handle very involved coding scenarios despite its computational ease. Spike train distances have been extended to the analysis of neural populations interpolating between SP and LL coding. This includes parametrizing the importance of distinguishing spikes being fired in different neurons. Yet, these approaches only consider the population as a whole. The explicit focus on subpopulations render our algorithms complimentary. The spectrum of encoding possibilities in neural populations is broad. The SP and LL cases are two extremes for which our algorithms provide correct identification results.


Proactive staff help avert patient safety crisis in OR.
Quality improvement program encourages employees to proactively report situations. Greasy feel, wrong color in liquid spurs employees into action. When staff have concerns, key phrases will automatically stop a process.


CEST imaging of the APT and ssMT predict the overall survival of patients with glioma at the first follow-up after completion of radiotherapy at 3T.
Outcome prediction of patients with glioma early after the completion of radiotherapy represents a major clinical challenge. Previously, the prognostic value of chemical exchange saturation transfer (CEST) imaging has been demonstrated in patients with newly diagnosed glioma. The objective of this study was to assess the potential of amide proton transfer (APT)-, relayed nuclear Overhauser effect (rNOE)- and semi-solid magnetization transfer (ssMT)-imaging according to Zhou et al. (APTwasym), Goerke et al. (MTRRexAPT, MTRRexNOE and MTRRexMT) and Mehrabian et al. (PeakAreaAPT, PeakAreaNOE and MTconst) for the prognostication of the overall survival (OS) of patients with glioma at the first follow-up after the completion of radiotherapy. 49 of 72 participants with diffuse glioma, who underwent CEST MRI at 3T between July 2018 and December 2021 4 to 6 weeks after the completion of radiotherapy, were analyzed. Contrast-enhancing tumor (CE) and whole tumor (WT) volumes were segmented on T2w-FLAIR and contrast-enhanced T1w images. Kaplan-Meier analysis and logrank-test were used for statistical analyses. APTw imaging demonstrated the strongest association with OS (HR = 4.66, p < 0.001). The MTconst (HR = 2.54, p = 0.044) was associated with the OS of participants with residual contrast-enhancing glioma tissue, whilst the MTRRexAPT (HR = 2.44, p = 0.056) showed a trend in this sub-cohort. The MTRRexNOE, MTRRexMT and PeakAreaNOE were not associated with survival. Imaging of the APT and ssMT at the first follow-up 4 to 6 weeks after the completion of radiotherapy at 3T were associated with the overall survival of study participants with glioma.


A new fasciocutaneous flap model identifies a critical role for endothelial Notch signaling in wound healing and flap survival.
Flap surgery is a common treatment for severe wounds and a major determinant of surgical outcome. Flap survival and healing depends on adaptation of the local flap vasculature. Using a novel and defined model of fasciocutaneous flap surgery, we demonstrate that the Notch ligand Delta-like 1 (Dll1), expressed in vascular endothelial cells, regulates flap arteriogenesis, inflammation and flap survival. Utilizing the stereotyped anatomy of dorsal skin arteries, ligation of the major vascular pedicle induced strong collateral vessel development by end-to-end anastomosis in wildtype mice, which supported flap perfusion recovery over time. In mice with heterozygous deletion of Dll1, collateral vessel formation was strongly impaired, resulting in aberrant vascularization and subsequent necrosis of the tissue. Furthermore, Dll1 deficient mice showed severe inflammation in the flap dominated by monocytes and macrophages. This process is controlled by endothelial Dll1 in vivo, since the results were recapitulated in mice with endothelial-specific deletion of Dll1. Thus, our model provides a platform to study vascular adaptation to flap surgery and molecular and cellular regulators influencing flap healing and survival.


Incorporation of doxorubicin in different polymer nanoparticles and their anticancer activity.
Background: Nanoparticles are under investigation as carrier systems for anticancer drugs. The expression of efflux transporters such as the ATP-binding cassette (ABC) transporter ABCB1 is an important resistance mechanism in therapy-refractory cancer cells. Drug encapsulation into nanoparticles has been shown to bypass efflux-mediated drug resistance, but there are also conflicting results. To investigate whether easy-to-prepare nanoparticles made of well-tolerated polymers may circumvent transporter-mediated drug efflux, we prepared poly(lactic-co-glycolic acid) (PLGA), polylactic acid (PLA), and PEGylated PLGA (PLGA-PEG) nanoparticles loaded with the ABCB1 substrate doxorubicin by solvent displacement and emulsion diffusion approaches and assessed their anticancer efficiency in neuroblastoma cells, including ABCB1-expressing cell lines, in comparison to doxorubicin solution. Results: The resulting nanoparticles covered a size range between 73 and 246 nm. PLGA-PEG nanoparticle preparation by solvent displacement led to the smallest nanoparticles. In PLGA nanoparticles, the drug load could be optimised using solvent displacement at pH 7 reaching 53 µg doxorubicin/mg nanoparticle. These PLGA nanoparticles displayed sustained doxorubicin release kinetics compared to the more burst-like kinetics of the other preparations. In neuroblastoma cells, doxorubicin-loaded PLGA-PEG nanoparticles (presumably due to their small size) and PLGA nanoparticles prepared by solvent displacement at pH 7 (presumably due to their high drug load and superior drug release kinetics) exerted the strongest anticancer effects. However, nanoparticle-encapsulated doxorubicin did not display increased efficacy in ABCB1-expressing cells relative to doxorubicin solution. Conclusion: Doxorubicin-loaded nanoparticles made by different methods from different materials displayed substantial discrepancies in their anticancer activity at the cellular level. Optimised preparation methods resulted in PLGA nanoparticles characterised by increased drug load, controlled drug release, and high anticancer efficacy. The design of drug-loaded nanoparticles with optimised anticancer activity at the cellular level is an important step in the development of improved nanoparticle preparations for anticancer therapy. Further research is required to understand under which circumstances nanoparticles can be used to overcome efflux-mediated resistance in cancer cells.


A novel cholic acid-based contrast enhancement agent for targeted MRI.
The novel Gd(III) complexes of heptadentate ligands NE3TA and NE3TA-Bn were prepared, and their relaxivities were measured and favorably compared to the commercially available MRI contrast enhancement agent Gd(DOTA). NE3TA was conjugated with cholic acid (CA) to produce CA-NE3TA. TEM images of Gd(CA-NE3TA) indicate that the complex self-assembles forming nano-sized micelles and displays an over threefold increased relaxivity compared to Gd(DOTA). The new cholic acid-conjugated nanoparticle MR contrast enhancement agent, Gd(CA-NE3TA) possesses great promise for use in targeted MRI.


[An adrenal pseudocyst as the cause of chronic back pain].
We present the case of a 36 year old female patient with a 3 month history of severe lower back pain without improvement after physiotherapy and analgesic drug therapy. X-rays of the spine were normal except for a calcified paravertebral lesion on the right side. A cystic tumor projecting on the posterior area of the right liver lobe was seen using ultrasound. MRI-scan showed an extrahepatic localization of the tumor. Intraoperatively, the tumor was located in the right adrenal gland which was then removed. Histopathology revealed an adrenal pseudocyst, and the pain was gone immediately after surgery.


Oxidative DNA damage and plasma antioxidant capacity in type 2 diabetic patients with good and poor glycaemic control.
Diabetes mellitus is a complex metabolic disorder characterized by a disturbance in glucose metabolism. Recent evidence suggests that increased oxidative damage as well as reduction in antioxidant capacity could be related to the complications in patients with type 2 diabetes. The aim of this study was to measure plasma antioxidant status in type 2 diabetic patients with good and poor glycaemic control and its relationship with oxidative DNA damage. Thirty-nine type 2 diabetic patients and eighteen healthy subjects were recruited for this study. We found that diabetic patients had slightly, but not significantly lower antioxidant capacity, measured with the "ferric reducing ability of plasma" (FRAP) assay, than healthy subjects. On the contrary, oxidative DNA damage (measured by the Comet assay) in leukocytes obtained from diabetic patients was significantly higher compared to healthy subjects. Taking into account glucose control, we found that the FRAP level was significantly (p<0.05) lower in diabetic subjects with poor glycaemic control than healthy subjects, while patients with good glycaemic control had FRAP values similar to controls. We also observed an unexpected positive correlation between FRAP values and oxidative DNA damage in diabetic patients; moreover, a positive correlation was found between FRAP and glucose level or HbA(1c) in patients with poor glycaemic control. In conclusion, our results confirm that patients with type 2 diabetes have a higher oxidative DNA damage than healthy subjects and that plasma antioxidant capacity is significantly lower only in patients with poor glycaemic control, moreover, in these patients FRAP values are positively correlated with glycaemic levels and HbA(1c). These observations indicate that a compensatory increase of the antioxidant status is induced as a response to free radical overproduction in type 2 diabetes. Therefore, the addition of antioxidant supplements to the current pharmacological treatment could have potentially beneficial effects in diabetic patients with poor glycaemic control.


World Hepatitis day 2021 -screening and vaccination against Hepatitis B virus in Accra, Ghana.
In Ghana, Hepatitis B virus (HBV) infection remains a major public health threat as in many parts of the world. Even with an effective vaccine, there are shortfalls with low vaccine coverage among adults. To create awareness and encourage vaccination, community engagement and public-private partnerships are needed in endemic settings to help fund campaigns and offer screening and vaccinations at no cost to under privileged people. An awareness and screening exercise was scheduled by University of Ghana-based Hepatitis-Malaria (HEPMAL) project team to coincide with the World Hepatitis Day (WHD) 2021. It was to engage the community in creating awareness of the menace and offer diagnostic services to ascertain prevalence levels and provide needed clinical support. Participants from the University of Ghana community and its immediate environs were registered, taken through pre-counselling sessions where they were educated on hepatitis transmission and prevention before consenting. Eligible participants were screened for HBV markers (HBsAg, HBeAg, HBsAb, HBcAb,HbcAg) with a rapid test kit. All HBsAb-negative participants were recommended for initial vaccination at the event, whilst the subsequent shots were administered at the University Hospital Public Health Department. Hepatitis B surface Antigen-positive participants were counselled and referred for appropriate care. / Outcomes: A total of 297 people, comprising of 126 (42%) males and 171 (58%) females aged between 17 and 67 years were screened during the exercise. Amongst these, 246 (82.8%) showed no detectable protective antibodies against HBV and all of them agreed to and were given the first dose HBV vaccine. Additionally, 19 (6.4%) individuals tested positive for HBsAg and were counselled and referred to specialists from the University Hospital for further assessment and management. We found that 59 (19.9%) of our participants had previously initiated HBV vaccination and had taken at least one dose of the vaccine more than 6 months prior to this screening, 3 of whom tested positive for HBsAg. For the three-dose HBV vaccines deployed, a little over 20% (50/246) and a further 17% (33/196) did not return for the second and the third doses respectively, resulting in an overall 66% (163/246) of persons who completed all three vaccinations. / Lessons learnt: Our medical campaign exercise established an active case prevalence rate of 6.4% and achieved a full vaccination success rate of 66% which is critical in the induction of long-term immunity in the participants. Aside these achievements, we would like to reiterate the importance of the use of different approaches including educational events and WHD activities to target groups and communities to raise awareness. Additionally, home and school vaccination programmes may be adopted to enhance vaccine uptake and adherence to the vaccination schedule. We plan to extend this screening exercise to deprived and/or rural communities where HBV incidence may be higher than in urban communities.


Bradycardia in anorexia nervosa.
We present a case of a 24-year-old woman initially referred for a permanent pacemaker for symptomatic sinus bradycardia. Further consultations revealed significant weight loss and subsequent psychiatric review confirmed a diagnosis of anorexia nervosa.


Evaluation of clinical activity and functional impairment in smokers with ankylosing spondylitis.
Ankylosing spondylitis (AS), a rheumatic disease, incapacitates human beings during the most productive stage of their lives, causes functional disturbances, and thus affects both patients and their relatives. Disease activity, functional impairment, and efficacy of treatment are affected by various environmental and genetic factors, one of which is smoking. We investigated smoking's effect on clinical activity and functional impairment in AS. Forty-eight patients with back pain and limited movements, diagnosed with AS based on clinical, radiological, and laboratory examinations and referred to the Trakya University School of Medicine Physical Medicine and Rehabilitation Department in Turkey were included in this study. Disease activity was evaluated with the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and functional impairment with the Bath Ankylosing Spondylitis Functional Index (BASFI). The Mann-Whitney U test was used to determine relationships between smoking and physical findings, laboratory results, and BASDAI and BASFI scores. Twenty-four patients were smokers. They had significantly higher BASDAI and BASFI scores and significantly lower forced vital capacity values than nonsmokers (P=0.000, P=0.002, and P=0.001, respectively) (Mann-Whitney U test). Smoking is associated with poor clinical aspect and functional impairment in AS. The underlying mechanism can be the physical incapacity caused by smoking.


Resting-state fMRI study on drug-naïve early-stage patients with Parkinson's disease and with fatigue.
Fatigue is one of the most common and debilitating non-motor symptoms in patients with Parkinson's disease (PD), which could manifest during the early stage of the disease and persist through the disease course. However, the treatment options for fatigue remain limited for patients with PD. Using seed-based resting-state functional magnetic resonance imaging, we explored the fatigue-related functional deficiencies in the anterior caudate nucleus, anterior putamen, and posterior putamen in a cohort of early-stage drug-naïve patients with PD. Thirty-eight patients with PD, 19 with and 19 without fatigue, and 31 matched healthy controls were selected. The fatigue status was defined based on the score obtained from the fatigue severity scale (FSS). Patients with PD with fatigue exhibited a decreased connectivity in the cerebellar-striatal, cortico-striatal, and mesolimbic-striatal loops. No increased functional connectivity was observed. The abnormal connections of the dorsal striatum subdivisions overlapped to extensive brain regions, including the cerebellum, inferior frontal gyrus, inferior temporal gyrus, lingual gyrus, rolandic operculum, insular, and hippocampus. Our findings revealed that the widespread functional deficiency in the striatal-cerebellar-cerebral cortical network may be critical to the pathology underlying fatigue in the early-stage PD. The key feature of fatigue-related connectivity was observed between the caudate nucleus and the cerebellum, which could serve as a potential biomarker or treatment target for fatigue in early-stage patients with PD in future studies.