Transdermal testosterone pretreatment in poor responders undergoing ICSI: a randomized clinical trial.
Does pretreatment with transdermal testosterone increase the number of cumulus-oocyte complexes (COCs) retrieved by more than 1.5 in poor responders undergoing intracytoplasmic sperm injection (ICSI), using recombinant follicle stimulating hormone (FSH) and gonadotrophin releasing hormone agonists (GnRHa)? Testosterone pretreatment failed to increase the number of COCs by more than 1.5 as compared with no pretreatment in poor responders undergoing ICSI (difference between medians: 0.0, 95% CI: -1.0 to +1.0). Androgens are thought to play an important role in early follicular development by enhancing ovarian sensitivity to FSH. In a recent meta-analysis, testosterone pretreatment resulted in an increase of 1.5 COCs as compared with no pretreatment. However, this effect was based on the analysis of only two randomized controlled trials (RCTs) including 163 patients. Evidently, there is a need for additional RCTs that will allow firmer conclusions to be drawn. The present RCT was designed to detect a difference of 1.5 COCs (sample size required = 48 patients). From 02/2014 until 04/2015, 50 poor responders fulfilling the Bologna criteria have been randomized (using a randomization list) to either testosterone pretreatment for 21 days ( ITALIC! n = 26) or no pretreatment ( ITALIC! n = 24). All patients underwent a long follicular GnRHa protocol. Recombinant FSH stimulation was started on Day 22 following GnRHa initiation. In the testosterone pretreatment group, a daily dose of 10 mg of testosterone gel was applied transdermally for 21 days starting from GnRHa initiation. Results are expressed as median (interquartile range). No differences in baseline characteristics were observed between the two groups compared. Testosterone levels [median (interquartile range)] were significantly higher in the testosterone pretreatment on the day of initiation of FSH stimulation [114 (99.5) ng/dl versus 20 (20) ng/dl, respectively, ITALIC! P < 0.001]. Duration of FSH stimulation [median (interquartile range)] was similar between the groups compared [12.5 (3.0) days versus 12 (3.0) days, respectively, ITALIC! P = 0.52]. The number of COCs retrieved [median (interquartile range)] was not different between the testosterone pretreatment and the no pretreatment groups [3.5 (4.0) versus 3.0 (3.0), 95% CI for the median: 2.0-5.0 versus 2.7-4.3, respectively; difference between medians: 0.0, 95% CI: +1.0 to -1.0). Similarly no differences were observed regarding fertilization rates [median (interquartile range)] [66.7% (32.5) versus 66.7% (42.9), respectively, ITALIC! P = 0.97] and live birth rates per randomized patient (7.7% versus 8.3%, respectively, rate difference: -0.6%, 95% CI: -19.0 to +16.9). The study was not powered to detect differences less than 1.5 COCs, although it is doubtful whether these differences would be clinically relevant. Moreover, due to sample size restrictions, no conclusions can be drawn regarding the probability of live birth. The results of this randomized clinical trial, suggesting that pretreatment with 10 mg of transdermal testosterone for 21 days does not improve ovarian response by more than 1.5 oocytes, could be used to more accurately consult patients with poor ovarian response. However, an improvement in IVF outcome using a higher dose of testosterone or a longer pretreatment period cannot be excluded. The study was partially funded by a Scholarship from the Academy of Athens. C.A.V. reports personal fees and non-financial support from Merck, Sharp and Dome, personal fees and non-financial support from Merck Serono, personal fees and non-financial support from IPSEN Hellas S.A., outside the submitted work. B.C.T. reports grants from Merck Serono, grants from Merck Sharp & Dohme, personal fees from Merck Serono, personal fees from Merck Sharp & Dohme, personal fees from IBSA & Ferring, outside the submitted work. NCT01961336. 10 October 2013. 02/2014.


Cysteamine-capped gold-copper nanoclusters for fluorometric determination and imaging of chromium(VI) and dopamine.
Highly emissive cysteamine-capped gold-copper bimetallic nanoclusters (CA-AuCu NCs) with a quantum yield of 18% were synthesized via one-pot anti-galvanic reduction. The CA-AuCu NCs were characterized by HR-TEM, XPS, FTIR, MALDI-TOF mass spectrometry, DLS, and zeta potential analyses. The NCs are shown to be viable fluorescent probes for Cr(VI) ions and dopamine (DA) via quenching of the blue fluorescence, typically measured at excitation/emission wavelengths of 350/436 nm. During DA recognition, a dark brown color appears, which is distinguishable from that of Cr(VI) detection. The aggregation induced quenching due to electron transfer was demonstrated by photoluminescence, HR-TEM, FTIR, DLS, and zeta potential interrogations. In buffer of pH 7, response is linear in the 0.2 ~ 100 μM for Cr(VI) and from 0.4 ~ 250 μM for DA. The respective detection limits are 80 and 135 nM. The method was applied to the determination of both Cr(VI) and DA in (spiked) tap, lake and sea water, and in human urine samples. The low toxicity of CA-AuCu NCs was validated by the MTT assay, and their responses to Cr(VI) ions and DA was also proven by Raw 264.7 cell imaging. Graphical abstractCysteamine capped Au-Cu nanoclusters (CA-AuCu NCs) were synthesized via one-pot anti-galvanic reduction and utilized in sensing of Cr(VI) ions and dopamine (DA) with demonstrated real/urine and cell imaging applications.


[Protein functional groups of the ova in the roach and carp-bream of varying degrees of maturity].
The Black Sea roach and carp-bream eggs of different degree of maturity differ from each other by the content of SH-, S--S-groups of proteins and non-protein low molecular weight thiol compounds. Definite differences were found in the optical density of soluble proteins and degree of ionization of their side groups.


Laryngocele--an anaesthetic hazard.
The authors report a case in which a previously unrecognized laryngocele in an eight-year-old child was accidentally intubated and inflated after induction of anaesthesia for tonsillectomy. The nature and diagnosis of laryngocele are discussed.


Acute pancreatitis occurring in gastric aberrant pancreatic tissue.
The occurrence of acute pancreatitis in gastric aberrant pancreas is described. The patient presented initially with acute abdominal pain and a palpable epigastric mass. The symptoms were severe and recurrent, and laparotomy with antrectomy was required.


Inhibition of phospholipase D1 ameliorates hepatocyte steatosis and non-alcoholic fatty liver disease.
Phospholipase D1 (PLD1), a phosphatidylcholine-hydrolysing enzyme, is involved in cellular lipid metabolism. However, its involvement in hepatocyte lipid metabolism and consequently non-alcoholic fatty liver disease (NAFLD) has not been explicitly explored. NAFLD was induced in hepatocyte-specific Pld1 knockout (Pld1(H)-KO) and littermate Pld1 flox/flox (Pld1-Flox) control mice feeding a high-fat diet (HFD) for 20 wk. Changes of the lipid composition in the liver were compared. Alpha mouse liver 12 (AML12) cells and mouse primary hepatocytes were incubated with oleic acid or sodium palmitate in vitro to explore the role of PLD1 in the development of hepatic steatosis. Hepatic PLD1 expression was evaluated in liver biopsy samples in patients with NAFLD. PLD1 expression levels were increased in the hepatocytes of patients with NAFLD and HFD-fed mice. Compared with Pld1-Flox mice, Pld1(H)-KO mice exhibited decreased plasma glucose and lipid levels as well as lipid accumulation in liver tissues after HFD feeding. Transcriptomic analysis showed that hepatocyte-specific deficiency of PLD1 decreased Cd36 expression in steatosis liver tissues, which was confirmed at the protein and gene levels. In vitro, specific inhibition of PLD1 with VU0155069 or VU0359595 decreased CD36 expression and lipid accumulation in oleic acid- or sodium palmitate-treated AML12 cells or primary hepatocytes. Inhibition of hepatocyte PLD1 significantly altered lipid composition, especially phosphatidic acid and lysophosphatidic acid levels in liver tissues with hepatic steatosis. Furthermore, phosphatidic acid, the downstream product of PLD1, increased the expression levels of CD36 in AML12 cells, which was reversed by a PPARγ antagonist. Hepatocyte-specific Pld1 deficiency ameliorates lipid accumulation and NAFLD development by inhibiting the PPARγ/CD36 pathway. PLD1 may be a new target for the treatment of NAFLD. The involvement of PLD1 in hepatocyte lipid metabolism and NAFLD has not been explicitly explored. In this study, we found that the inhibition of hepatocyte PLD1 exerted potent protective effects against HFD-induced NAFLD, which were attributable to a reduction in PPARγ/CD36 pathway-mediated lipid accumulation in hepatocytes. Targeting hepatocyte PLD1 may be a new target for the treatment of NAFLD.


Comparison of Long-term Survival of Patients With Early-Stage Non-Small Cell Lung Cancer After Surgery vs Stereotactic Body Radiotherapy.
Previous comparisons of surgery and stereotactic body radiotherapy (SBRT) for early-stage (ES) non-small cell lung cancer (NSCLC) did not account for the extent of regional lymph node examination (LNE) during surgery. To compare long-term overall survival (OS) of patients with ES NSCLC after surgery vs SBRT when the extent of regional LNE in patients undergoing surgery is thoroughly considered. Cohort study with survival comparisons using the multivariable Cox proportional hazards model and after propensity score matching. Data from the National Cancer Database were analyzed from October 28, 2018, through April 18, 2019. Patients with ES NSCLC diagnosed between January 1, 2004, and December 31, 2015, who underwent any curative-intent surgery or SBRT were included. Long-term OS. Of 104 709 total patients, 91 330 underwent surgery (42 508 [46.5%] male; median [interquartile range] age, 68 [61-75] years) and 13 379 received SBRT (6065 [45.3%] male; median [interquartile range] age, 75 [68-81] years). Surgery, especially lobectomy (hazard ratio [HR], 0.53; 95% CI, 0.50-0.56), and regional LNE, especially when more than 10 lymph nodes were examined (HR, 0.73; 95% CI, 0.69-0.77), were associated with better long-term OS (P < .001). Pneumonectomy was not associated with reduced mortality risk when 0 nodes were examined (HR for stage T1, 1.43; 95% CI, 0.67-3.06; P = .35; HR for stage T2-T3, 0.62; 95% CI, 0.34-1.13; P = .12) or when more than 15 nodes were examined for stage T1 disease in patients younger than 80 years (HR, 0.77; 95% CI, 0.54-1.09; P = .14) or when patients aged 80 years or older received regional LNE of any extent (>15 nodes examined: HR for stage T1, 0.65; 95% CI, 0.16-2.64; P = .54; HR for stage T2-T3, 0.90; 95% CI, 0.50-1.60; P = .71). Less extensive surgery was not associated with improved OS when 0 nodes were examined in patients aged 80 years or older with stage T2 to T3 tumors (HR for lobectomy, 0.90; 95% CI, 0.65-1.25; P = .53) and in selected operable patients older than 75 years with stage T1 tumors (HR for lobectomy, 1.07; 95% CI, 0.57-2.00; P = .84). This study found that, overall, surgery coupled with regional LNE of appropriate extent was associated with the best long-term OS in patients with ES NSCLC.


Central nervous system dysfunction after anesthesia in the geriatric patient.
Cerebral infarction, bleeding, long-lasting hypofusion, and profound hypoxia are well-known factors behind central nervous system dysfunction after anesthesia. Other explanations may be the metabolic-endocrine stress response and psychologic factors related to changes in the environment. The clinical presentations can be obvious as in cerebral death or stroke but delirium also may be readily recognized. A more subtle and long-lasting deterioration in cognitive function is called postoperative cognitive dysfunction. This condition can only be detected with the use of neuropsychologic testing and recently, postoperative cognitive dysfunction has been detected as the most common cerebral complication after noncardiac surgery in elderly patients.


Transmission of the quantum interaction of erythrocytes.
The previously demonstrated long-range quantum mechanical interaction between human erythrocytes suspended in plasma also occurs with artificial media. Certain macromolecules dissolved in phosphate-buffered saline will transmit the interaction provided their concentration is above a minimum. Extended macromolecules transmit the interaction, whereas a compact macromolecule (albumin) does not.


Macrocyclic lactone resistance in Dirofilaria immitis: risks for prevention of heartworm disease.
Heartworm disease, caused by Dirofilaria immitis, can be lethal in dogs and cats. It is transmitted by mosquitoes, and occurs in many parts of the world. Prevention relies on macrocyclic lactones. Macrocyclic lactones used are ivermectin, selamectin, abamectin, eprinomectin, milbemycin oxime and moxidectin, administered at 30-day intervals during the transmission season. Some moxidectin formulations are long-acting injectables. In the USA, preventives are recommended throughout the year. Loss of efficacy of macrocyclic lactone preventives was reported in 2005 and proof of resistance in the USA was published a decade later. Understanding factors which promote resistance is important to maintain control. Factors important for resistance development are discussed. Better, inexpensive tests to confirm resistance are needed. Infection in animals under chemoprophylaxis per se does not imply resistance because lack of compliance in preventive use could be the reason. In vivo confirmation of resistance is expensive, slow and ethically questionable. A microfilariae suppression test can be a surrogate test, but requires a high dose of a macrocyclic lactone and repeated blood microfilaria counts 2-4 weeks later. DNA single nucleotide polymorphism markers have been successfully used. However, the specific genetic changes which cause resistance are unknown. Surveys to map and follow the extent of resistance are needed. Long acting mosquito repellants and insecticides can play a useful role. High dose rate formulations of moxidectin, coupled with mosquito biting mitigation may reduce transmission of resistant genotypes. Doxycycline, daily for 28 days, as anti-Wolbachia treatment, can reduce transmission and remove adult parasites. However, new classes of heartworm preventives are needed. While any preventive strategy must be highly effective, registration requirements for 100% efficacy may hinder development of useful new classes of preventives. Continued reliance on macrocyclic lactone preventives, when they do not work against resistant genotypes, will spread resistance, and allow for more disease.


Craniofacial characteristics and velopharyngeal function in cleft lip/palate children with and without adenoidectomy.
The association between velopharyngeal function, craniofacial morphology and adenoidectomy was investigated using 27 craniofacial and nasopharyngeal variables taken from lateral cephalograms. The sample consisted of 96 boys with cleft palates with or without cleft lips. They were examined at 6 years of age when cephalograms were obtained and perceptual speech assessments were performed. The subjects were divided into three groups: (1) velopharyngeal competence (VPC, n = 45); (2) mild incompetence not requiring velopharyngoplasty (VPI, n = 36); and (3) previous incompetence operated on with velopharyngoplasty ad modum Hoenig (VPP, n = 15) before the 6-year examination. The groups were further divided into two subgroups according to previous adenoidectomy (Ad+, Ad-). The cranial base, size and interrelationship of the maxilla and mandible and their relationship to the cranial base or the bony nasopharynx did not differ among the VPC, VPI and VPP groups. The sagittal depth of the nasopharyngeal airway (Pm-ad1, Pm-ad2, Pm-ad3) was significantly wider in the VPP group than in the the VPC and VPI groups. The previous adenoidectomy decreased the thickness of the posterior pharyngeal wall (ad1-Ba, ad2-so) and thus increased airway size. The length of the velum did not differ between the three groups or their subgroups with and without adenoidectomy. The results showed that adenoidectomy is a risk to velopharyngeal function by widening the nasal airway, but velopharyngeal incompetence cannot definitely be attributed to adenoidectomy.


Psychological measures of patient's decision-making for prenatal genetic testing.
To use a questionnaire to determine the levels of maternal decision-related distress, clarity of the pros and cons, and certainty when considering prenatal genetic diagnostic testing; and to assess the relationship between these constructs and patient characteristics. Cross-sectional study. Voluntary, anonymous questionnaires distributed 2017-2019 to women referred for invasive prenatal genetic testing. Excluded: English or Spanish illiterate. Maternal characteristics were collected. Questions evaluated distress, decisional certainty, and decisional clarity on a 5-point Likert scale (range: 0 = low/uncertain/unclear to 4 = high/certain/clear). Analysis: non-parametric Kruskal-Wallis, correlation statistics, and ANOVA. Forty-four female patients completed it. Most were married, white, Catholic, and multiparous. 58% had already made a testing decision. Patients expressed low distress levels (mean 1.18 ± 0.80) and expressed high decisional certainty (mean 3.28 ± 0.76) and clarity (mean 3.30 ± 0.99). Decisional certainty and clarity were positively correlated (r = 0.47, p < .01), whereas distress was negatively correlated with decisional certainty (r = -0.8136, p < .0005) and decisional clarity (r = -0.49, p = .007). No significant differences by religion or parity. Greater distress (p < .05) and less decisional clarity (p = .07) occurred between those still debating testing vs those who had decided. Higher maternal distress scores were associated with lower decisional certainty and decisional clarity in women considering prenatal genetic testing.


Subendocardial viability ratio estimated by arterial tonometry: a critical evaluation in elderly hypertensive patients with increased aortic stiffness.
1. Increased aortic stiffness predisposes to myocardial ischaemia by increasing the systolic tension-time index and by decreasing aortic pressure throughout diastole. The tonometric subendocardial viability ratio (SEVR) is a non-invasive estimate of myocardial perfusion relative to cardiac workload. The hypothesis that SEVR is impaired in elderly hypertensives with high aortic pulse pressure (PP) was tested in the present study. 2. The SEVR was calculated by radial applanation tonometry in 203 subjects. In addition, diastolic time (DT), systolic time (ST) and mean diastolic and systolic aortic pressures (Pd and Ps, respectively) were calculated. First, 60 subjects matched for age and gender were analysed (20 controls, 20 hypertensives with pulse pressure (PP) < or = 60 mmHg, 20 hypertensives with PP > 60 mmHg; mean (+/-SD) age 64 +/- 9 years; 24 women, 36 men). The remaining 143 subjects, aged 53 +/- 10 years, were analysed subsequently. 3. The SEVR was similar in the three elderly groups (1.39 +/- 0.34, 1.39 +/- 0.28 and 1.35 +/- 0.25, in controls and hypertensive patients with PP < or = 60 and > 60 mmHg, respectively). The SEVR was positively related to DT/ST (r(2) = 0.89) and to DT (r(2) = 0.73) and was negatively related to heart rate (r(2) = 0.56; P < 0.001 each). However, SEVR was not related to ST, PP, mean Pd or mean Ps. At a given DT/ST, SEVR tended to be lower in hypertensives with PP > 60 mmHg than in hypertensives with normal PP. The positive linear relationship between SEVR and DT/ST was confirmed in the remaining 143 subjects (r(2) = 0.90), with no influence of aortic pressure. 4. The tonometric SEVR was not impaired in elderly hypertensive patients with increased aortic stiffness. In resting elderly and middle-aged individuals, the tonometric SEVR was mainly related to DT/ST ratio, not to aortic pressure.


Prevalence of serogroups and virulence genes in Escherichia coli associated with postweaning diarrhoea and edema disease in pigs and a comparison of diagnostic approaches.
Identification of Escherichia coli causing porcine postweaning diarrhoea (PWD) or edema disease (ED) requires knowledge regarding the prevalent pathotypes within a given region. This study was undertaken to determine the present distribution of serogroups, hemolytic activity and virulence factor gene profiles among porcine pathogenic E. coli isolates in Denmark and to compare detection of these characteristics as diagnostic approaches. Five hundred and sixty-three E. coli were serogrouped using E. coli O-antisera and investigated for hemolytic activity. Of these, 219 isolates were further characterized using a 5'-nuclease PCR assay detecting genes for adhesion factors, enterotoxins and verocytotoxin 2e (VT2e). Forty-two different serogroups were found. The most prevalent serogroup was O149 accounting for 49.9% of all isolates, followed by O138 (14.9%), O139 (6.9%), O141 (4.1%) and O8 (3.7%). Hemolytic activity was detected in 87.7% of all isolates. Virulence factor genes detected were F4 (44.7%), F18 (39.3%), intimin (1.4%), F6 (0.9%), STb (77.6%), EAST1 (65.8%), LT (61.6%), STa (26.5%) and VT2e (16.4%). Six pathotypes accounted for 65.7% of all isolates investigated. Using possession of virulence factor genes as reference, O-serogrouping employing a selection of antisera representing common pig pathogenic serogroups and detection of hemolysis were evaluated as epidemiological markers for pathogenicity. Both criteria were associated with pathogenicity (P<0.001, for both), however, both methods also resulted in false classifications regarding pathogenicity for 11.9 and 13.2% of isolates, respectively. Detection of adhesion factor genes F4, F18 and intimin is suggested as an operational alternative when diagnosing PWD and ED.


Malignant transformation of human skin fibroblasts by two alternative pathways.
We developed a telomerase-positive, infinite life span human fibroblast cell strain (MSU-1.0) by transfection of a v-MYC oncogene and spontaneous over-expression of transcription factors SP1/SP3. Loss of expression of p14(ALT) and enhanced expression of SPRY2 gave rise to the MSU-1.1 cell strain. Unlike MSU-1.0 cells, the MSU-1.1 cells can be malignantly transformed by expression of N-RAS(LYS61) or H-Ras(v12) oncoproteins (driven by their original promoters) and expression of a SRC-family protein, v-FES. MSU-1.1 cells can also be malignantly transformed by high expression of these RAS oncogenes or the v-K-RAS oncogene. PDGF-B transformed MSU-1.1 cells give rise to benign tumors (fibromas) in athymic mice. A second route to malignant transformation of the MSU-1.1 cells involves loss of functional TP53 protein by carcinogen treatment and loss of expression of wild type p16(INK). These studies indicate 6-8 "hits" are required to activate the oncogenes and inactivate the suppressor genes we identified.


Value Based Care and Patient-Centered Care: Divergent or Complementary?
Two distinct but overlapping care philosophies have emerged in cancer care: patient-centered care (PCC) and value-based care (VBC). Value in healthcare has been defined as the quality of care (measured typically by healthcare outcomes) modified by cost. In this conception of value, patient-centeredness is one important but not necessarily dominant quality measure. In contrast, PCC includes multiple domains of patient-centeredness and places the patient and family central to all decisions and evaluations of quality. The alignment of PCC and VBC is complicated by several tensions, including a relative lack of patient experience and preference measures, and conceptions of cost that are payer-focused instead of patient-focused. Several strategies may help to align these two philosophies, including the use of patient-reported outcomes in clinical trials and value determinations, and the purposeful integration of patient preference in clinical decisions and guidelines. Innovative models of care, including accountable care organizations and oncology patient-centered medical homes, may also facilitate alignment through improved care coordination and quality-based payment incentives. Ultimately, VBC and PCC will only be aligned if patient-centered outcomes, perspectives, and preferences are explicitly incorporated into the definitions and metrics of quality, cost, and value that will increasingly influence the delivery of cancer care.


Univariate near infrared methods for determination of pesticides in agrochemicals.
It has been developed a general strategy for the determination of pesticides in agrochemicals by Fourier transform near infrared (FT-NIR) spectroscopy. The methodology is based on previous extraction of the active principles through sonication with acetonitrile and direct determination by transmission measurements, using glass vials as measurement cells and univariate calibration at selected wavenumbers as a function of the pesticide to be analyzed. Chlorsulfuron, metamitron, iprodione, pirimicarb, procymidone and tricyclazole were used as test molecules and data obtained by FT-NIR for 20 samples containing one of these active principles compare well with those found by reference liquid chromatography (HPLC) procedures. In short, FT-NIR provides relative standard deviations between 0.04 and 0.7% and limit of detection values ranging from 0.004 to 0.17 mg g(-1) with recovery values between 96.4 and 100.5%. On the other hand, FT-NIR offers a 10 times faster methodology than chromatography ones and an environmentally friendly alternative which reduces the reagent consumption to 1 mL acetonitrile per sample.


Lithium-ion batteries: runaway risk of forming toxic compounds.
Lithium-ion batteries are stabilized by an ultrathin protective film that is 10-50 nanometers thick and coats both electrodes. Here we artificially simulate the 'thermal-runaway' conditions that would arise should this coating be destroyed, which could happen in a battery large enough to overheat beyond 80 degrees C. We find that under these conditions the reaction of the battery electrolyte with the material of the unprotected positive electrode results in the formation of toxic fluoro-organic compounds. Although not a concern for the small units used in today's portable devices, this unexpected chemical hazard should be taken into account as larger and larger lithium-ion batteries are developed, for example for incorporation into electric-powered vehicles.


On the tertiary structure of the extracellular domains of the epidermal growth factor and insulin receptors.
Alignment of the sequences, the identification of conserved residue patterns and secondary structure predictions indicate that the extra-cellular regions of the human and Drosophila epidermal growth factor (EGF), c-erb-B2 and human insulin receptors each contain two large, homologous domains (L) which are probably comprised of at least four short alpha-helices followed by turns of conserved length and beta-strands. In the human and Drosophila EGF and c-erb-B2 receptors these homologous domains are each followed by a series of smaller cystine-rich domains (S) to give a gene-duplicated structure of L1S11S12S13L2S21S22S23. In the human insulin receptor, the second series of cystine domains is replaced by a different sequence. These duplicated structures are probably organised as a pseudo-symmetrical dimer. There are two 'hyper-variable' regions, one at the end of the large domains and one in the cystine-rich sequences, which are candidates for hormone or growth-factor binding.


[RAPD-based analysis of introgression of barley genetic material into the genome of alloplasmic wheat lines (Hordeum geniculatum All./ Triticum aestivum L.)].
Genomes of three alloplasmic wheat lines obtained on the basis of barley--wheat hybrid Horderum geniculatum All. (2n = 28) x Triticum aestivum L. (2n = 42)(Pyrotrix 28) were examined using random amplified polymorphic DNA (RAPD) analysis. Line L-29 was obtained after first backcross of the initial hybrid with the wheat variety Pyrotrix 28 and ten subsequent self-pollinating generations. This line was represented by euploid plants with typical to the common wheat chromosome number (2n = 42), as well as by aneuploids, which contained an additional telocentric chromosome in the main karyotype (2n = 42 + t). Lines L-26 and L-27 were obtained by two backcrosses of one BC1 plant with the wheat variety Novosibirskaya 67 and one subsequent self-polination of one BC3 plant. Chromosome number in all these plants corresponded to 2n = 40 + 4t. RAPD analysis was carried out using seven primers, which were previously proved to be effective for identification of the barley genome fragments within hybrid genomes of alloplasmic lines. The presence of barley genome fragments in line L-29 was revealed by use of five primers, while in lines L-26 and L-27 these fragments were detected by use of one primer. The significant difference in the number of barley RAPD fragments in the genomes of alloplasmic lines obtained at different backcrossing stages suggests more intense displacement of barley genome during backcrossing compared to self-pollination in BC1 plants.


A simple and economical in-house phage technique for the rapid detection of rifampin, isoniazid, ethambutol, streptomycin, and ciprofloxacin drug resistance in Mycobacterium tuberculosis, directly on decontaminated sputum samples.
The early detection of drug resistance would be a boon for TB control programs. The aim of the present study was to set up a rapid phage assay for the testing of drug susceptibility of Mycobacterium tuberculosis to rifampin, isoniazid, ethambutol, streptomycin, and ciprofloxacin, directly on decontaminated sputum samples. Mueller-Hinton broth was used instead of 7H9 broth to make the method more economical. Vancomycin and polymyxin B were added to the concentrated sputum samples to reduce the bacterial contamination. The phage assay on decontaminated sputum samples was compared with the proportion method using M. tuberculosis isolates from the same sputum samples. Phage assay results were available within 48h for rifampin and streptomycin and within 72h for all the other drugs. In contrast the proportion method required 4-6 weeks from the primary cultures. The sensitivity of the phage assay was in the range of 93% to 100% and specificity in the range of 96% to 100% for all the drugs tested. The interpretation of results was possible for 334 of the 370 (90.3%) acid-fast bacillus (AFB) smear-positive sputum samples by the phage assay. The phage assay for the detection of drug resistance on direct decontaminated sputum samples is economical, easy to perform, and rapid.


Hydrogen metabolism in aerobic hydrogen-oxidizing bacteria.
A survey on organisms able to use molecular hydrogen as electron donor in the energy-yielding process is presented. In the group of the aerobic hydrogen-oxidizing bacteria so far two types of hydrogenases have been encountered, a NAD-reducing, soluble enzyme (H2 : NAD oxidoreductase) and a membrane-bound enzyme unable to reduce pyridine nucleotides. With respect to the distribution of both types of hydrogenases three groups of hydrogen-oxidizing bacteria can be diffentiated containing (i) both types (Alcaligenes eutrophus), (ii) a soluble enzyme only (Nocardia opaca lb), and (iii) a membrane-bound hydrogenase only (majority of genera and species). The results of studies on the NAD-specific hydrogenase of A. eutrophus are summarized. Results on the solubilization and purification of the membrane-bound hydrogenase of A. eutrophus are presented in detail. The enzyme was solubilized from purified membranes by Triton X-100 and sodium desoxycholate or phospholipase D. The crude membrane extract was fractionated by ammonium sulfate precipitation and chromatography on carboxymethylcellulose at pH 5.5. The enzyme was stable in potassium phosphate buffer; it resembles the soluble enzyme with respect to stability under oxidizing conditions. Further biochemical and immunological data indicate, however, that both enzymes are different with respect to their native structure.


Single-centre prospective comparison between contrast-enhanced ultrasound and computed tomography angiography after EVAR.
To evaluate contrast-enhanced ultrasound (CEUS) as an effective alternative to CT-angiography (CTA) for endoleak detection and aneurismal sac diameter measurement in the follow-up after endovascular abdominal aortic aneurysm repair (EVAR). From January 2006 to December 2010, 395 patients underwent EVAR follow-up with both CTA and CEUS. The diameter of the aneurismal sac and the presence of endoleaks were evaluated in all the 395 paired examinations. Bland-Altman plots showed a good agreement in aneurismal sac diameter evaluation between the two imaging modalities. The mean diameter was 54.93 mm (standard deviation (SD) ±12.57) with CEUS and 56.01 mm (SD ± 13.23) with CTA. The mean difference in aneurismal sac diameter was -1.08 mm ± 3.3543 (95% confidence interval (CI), -0.75 to -1.41), in favour of CTA. The number of observed agreement in endoleak detection was 359/395 (90.89%). The two modalities detected the same type I and type III endoleaks. McNemar's χ(2) test confirmed that CTA and CEUS are equivalent in endoleak detection. CEUS demonstrated to be as accurate as CTA in endoleak detection and abdominal aortic aneurysm diameter measurements during EVAR follow-up, without carrying the risks of radiation exposure or nephrotoxicity. Even if it cannot be proposed as the sole imaging modality during follow-up, our analysis suggests that it should have a major role.


Plant blindness and diversity in AI language models.
Large language models (LLMs) will benefit science by accelerating task performance. We explored whether answers generated by ChatGPT (generative pretrained transformer) to questions of biology are sufficiently diverse. 'Plant awareness' in ChatGPT answers was found to be highly variable, illustrating the importance of scientists being involved in validating the data and methods used to train artificial intelligence (AI) models.


Crude Extracts of Marine-derived and Soil Fungi of the Genus Neosartorya Exhibit Selective Anticancer Activity by Inducing Cell Death in Colon, Breast and Skin Cancer Cell Lines.
The crude ethyl acetate extracts of marine-derived fungi Neosartorya tsunodae KUFC 9213 (E1) and N. laciniosa KUFC 7896 (E2), and soil fungus N. fischeri KUFC 6344 (E3) were evaluated for their in vitro anticancer activities on a panel of seven human cancer cell lines. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed, after 48 h treatments with different concentrations of extracts, to determine their concentration of the extract or Dox that inhibits cell viability by 50% for each cell line. The effects of the crude extracts on DNA damage, clonogenic potential and their ability to induce cell death were also assessed. E1 was found to the void of anti-proliferative effects. E2 was shown to decrease the clonogenic potential in human colorectal carcinoma cell line (HCT116), human malignant melanoma cell line (A375), human breast adenocarcinoma cell line (MCF7), and human caucasian colon adenocarcinoma Grade II cell line (HT29) cells, whereas E3 showed such effect only in HCT116 and MCF7 cells. Both extracts were found to increase DNA damage in some cell lines. E2 was found to induce cell death in HT29, HCT116, MCF7, and A375 cells while extract E3 increased cell death in MCF7 and HCT116 cell lines. The results reveal that E2 and E3 possess anticancer activities in human colon carcinoma, breast adenocarcinoma, and melanoma cells, validating the interest for an identification of molecular targets involved in the anticancer activity. The crude ethyl acetate extract of N. tsunodae (E1) did not decrease cell viability in any of the tested cell linesThe crude ethyl acetate extracts of N. laciniosa (E2) and N. fischeri (E3) decreased cell proliferation in some human cancer cell lines tested at both short- and long-termN. laciniosa (E2) induced a significant increase in the number of cell death, in part, due to the induction of DNA damageN. fischeri (E3) induce cell death but in some cell lines without induction of DNA damage detected by comet assayCrude ethyl extracts of N. laciniosa (E2) and N. fischeri (E3) exert an anticancer activity in human colon carcinoma, breast adenocarcinoma, and malignant melanoma cells. Abbreviations Used: A375: Human malignant melanoma cell line; A549: Human non-small lung cancer cell line; DAPI: 4,6-diamidino-2-phenylindole; DMEM: Dulbecco's Modified Eagle Medium; DMSO: Dimethylsulfoxide; Dox: Doxorubicin; E1: Neosartorya tsunodae KUFC 9213; E2: Neosartorya laciniosa KUFC 7896; E3: Neosartorya fischeri KUFC 6344; FBS: Fetal bovine serum; HCT116: Human colorectal carcinoma cell line; HEPES: (N-[2-hydroxyethyl] piperazine-N'-[2-ethane-sulfonic acid]); HepG2: Human hepatocellular carcinoma cell line; HT29: Human caucasian colon adenocarcinoma Grade II cell line; IC50: Concentration of the extract or Dox that inhibits cell viability by 50%; MCF7: Human breast adenocarcinoma cell line; MEM: Minimum Essential Medium Eagle; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NCI-H460: Human non-small lung cancer cell line; PBS: Phosphate buffered saline; PE: Plating efficiency; RPMI: Roswell park memorial institute medium; SF: Surviving fraction; U-251: Human malignant glioblastoma cell line.


N6-methyladenosine profiling reveals that Xuefu Zhuyu decoction upregulates METTL14 and BDNF in a rat model of traumatic brain injury.
The traditional Chinese herbal formula Xuefu Zhuyu decoction (XFZYD) is a classic formula in the category of invigorating blood circulation and resolving blood stasis. It has been proven to improve the neurological and ethological prognosis of traumatic brain injury. XFZYD promotes synaptic and axonal regeneration after traumatic brain injury, which is functionally modulated by the N6-methyladenosine (m6A) modification of RNA. However, the epigenetic effects of XFZYD on m6A modification remain unknown. To explore how XFZYD protects against traumatic brain injury induced by controlled cortical impact (CCI) injury by altering RNA m6A modification. The modified neurological severity scoring and Morris water maze were performed to evaluate the neuroprotective effects of XFZYD for 14 days and screen the dose. Then, dot blot, western blotting, and methylated RNA immunoprecipitation sequencing (MeRIP-Seq) were used to explore changes in RNA m6A modification in the perilesional cortex. The Metascape platform was used to analyze the Gene Ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Reactome pathway of the differential m6A-tagged genes. Furthermore, MeRIP-qPCR was conducted to quantify differences in the hub differential m6A modification gene brain-derived neurotrophic factor (Bdnf). XFZYD significantly ameliorated the neurological deficits, spatial learning, and memory impairments in rats post-CCI on day 14. XFZYD enhanced the m6A level, and the expression of METTL14 and YTHDC2 in the perilesional cortex of CCI rats. In all three groups, the 3'-untranslated regions and coding sequence were primarily enriched for m6A peaks. XFZYD reversed the increased proportion of 3'-untranslated regions, and the decreased proportion of coding sequence and 5'-untranslated regions post-CCI. Moreover, XFZYD markedly downregulated 41 elevated m6A-tagged transcripts and upregulated 119 decreased m6A-tagged transcripts following CCI. Gene ontology and KEGG pathway analysis revealed that XFZYD-regulated m6A-tagged transcripts were predominantly enriched in synapse assembly, synaptic plasticity, learning or memory, and MAPK signaling pathway. Then, the hub-regulated m6A-tagged gene BDNF was identified. Both the m6A methylation level and the protein level of BDNF were ascended by XFZYD treatment. XFZYD improves neurological deficits, spatial learning and memory impairments in rats post-TBI probably through increasing the expression of METTL14 and BDNF in the cortex. Our study highlights a novel post-transcriptional regulation mechanism mediated by herbal medicine for traumatic brain injury treatment.


The facile fabrication of few-layer graphene and graphite nanosheets by high pressure homogenization.
We demonstrated a facile and efficient approach for the fabrication of few-layer graphene in DMF and graphite nanosheets in water with the assistance of high pressure homogenization. This work provides a physical perspective for understanding the exfoliation of other layered materials in liquid phase exfoliation.


Surgery of the skull base.
Tumors involving the base of the skull are often occult and may become quite large before detection is possible. Symptomology varies depending upon the nature of the tumor and its placement. These neoplasms have frequently been considered inoperable simply because of their location, but in recent years microsurgical technique and high speed air drills have allowed the modern temporal bone surgeon to gain improved access to the skull base. A wide variety of surgical approaches to tumors in this area have been described in the past 20 years: translabyrinthine, middle fossa, transcochlear, retrolabyrinthine, retrosigmoid, transpalatal-transclival, and through the jugular bulb. The purpose of this paper is to review the indications, complications, and results of each of these procedures in relation to specific tumors involving the skull base. Case reports illustrate the diagnosis and surgical management of a variety of unusual neoplasms, including an extradural meningioma of the temporal bone and clivus, a low grade squamous cell carcinoma on the tegmen in a radical cavity, a large primary cholesteatoma, an osteoblastoma of the temporal and occipital bones, an XIth nerve neuroma in the jugular bulb area, and an osseous hemangioma involving the facial nerve at the geniculate ganglion.


New Insights into the Mycolate-Containing Compound Biosynthesis and Transport in Mycobacteria.
Mycolic acids are extremely-long-chain fatty acids that compose a large family of mycolate-containing compounds, major envelope lipid components and critical pathogenicity factors of Mycobacterium tuberculosis. In recent years there have been major advances in understanding their metabolic pathway. Unknown enzymes of the fatty acid synthase type II elongation system and the condensation system that builds the mycolic acid scaffold were identified. Missing links with the mycolate-containing compound biosynthesis-such as the mechanisms of transfer onto trehalose and of translocation through the inner membrane-were deciphered, while recycling processes have emerged. Beyond the more accurate picture of the biosynthesis and translocation pathways dedicated to these unique molecules, major issues that should be addressed in the future are also discussed.


Neural and Hormonal Correlates of Sexual Arousal in Transgender Persons.
Whereas mounting work has begun to document the neural correlates underlying sexual arousal (SA) in humans, the associations between gender identity and the brain correlates of SA as well as their hormonal contributions remain unknown. This study investigated neural activation to sexual arousal in transgender and cisgender persons. 20 transgender men and 19 transgender women (TW) already living in their identified gender were compared to 21 cisgender men (CM) and 19 cisgender women. Participants viewed erotic and neutral video clips while undergoing 3 Tesla magnetic resonance imaging. Group-specific brain activation, brain functional connectivity, and brain-hormone associations within the neurophenomenological model of sexual arousal (Stoleru et al, 2012). Consistent with the model, participants activated most of its components. However, between-group differences were mostly showing larger activation for CM relative to any of the other 3 groups. Moreover, functional connectivity analyses (psychophysiological interactions) indicated unique patterns for CM, cisgender women, and TW in how different components of SA communicated with one another. Finally, androgens in transgender men and estrogens in TW correlated negatively with parietal cortex and primary (sensori-) motor regions, respectively, while CM showed positive correlations of androgens with parietal cortex, somatosensory regions, and the insula. Data provide information on neurobiological changes in sexual arousal during treatment with gender-affirming hormone therapy. Although a limitation is the lack of pretreatment data, the present study provides comprehensive information including brain activation, functional connectivity, and hormonal associations in a large sample. The results highlight a complex picture of the neural correlates of SA in gender identity and sex assigned at birth. Mueller SC, Wierckx K, T'Sjoen G. Neural and Hormonal Correlates of Sexual Arousal in Transgender Persons. J Sex Med 2020;17:2495-2507.


Electrophysiological analysis of the neural circuitry underlying opiate effects in the nucleus accumbens septi.
Systemically administered opiates inhibit or excite spontaneously active single units of the nucleus accumbens septi (NAS) but do not affect fimbria-evoked NAS responses. However, microinjections of morphine to the dopamine cell bodies of the ventral tegmental area (VTA-M) do inhibit these evoked responses. Since systemic heroin can reverse the inhibitory effect of VTA-M on the fimbria-evoked NAS responses, systemic opiates might exert actions on other brain regions that oppose the inhibitory effect of VTA-M on the NAS. Evidence is provided indicating that this opposing action can arise from opiate actions at the ventral subiculum of the hippocampus.


Binary Classification for Failure Risk Assessment.
Survival analysis is tremendously powerful, and is a popular methodology for analyzing time to event models in bioinformatics. Furthermore, several of its extensions can simultaneously perform variable selection in conjunction with model estimation. While this flexibility is extremely desirable, under certain scenarios, binary class variable selection and classification methods might provide more reliable risk estimates. Synthetic simulations and real data case studies suggest that when (1) randomly censored points comprise only a small portion of data, (2) biological markers are weak, (3) it is desired to compute risk across predetermined time intervals, and (4) the assumptions of the competing time to event models are violated, binary class models tend to perform superior. In practice, it might be prudent to test both model families to guarantee adequate analysis. Here we describe the pipeline of binary class feature selection and classification for time to event risk assessment.


A practical approach to method validation in pharmaceutical analysis.
Guidelines issued by Regulatory Authorities make it clear that validation of analytical methodology is now widely required in support of registration dossiers. Although some attempts are made at defining terms and some vague indications are sometimes provided within these guidelines, no clear advice is provided on how validations should be conducted and what results should be expected. In this paper it is attempted to suggest some practical approaches to conducting validation and in particular to the determination of accuracy, linearity and limit of detection/quantitation.


Sedimentation and polar order of active bottom-heavy particles.
Self-propelled particles in an external gravitational field have been shown to display both an increased sedimentation length and polar order even without particle interactions. Here, we investigate self-propelled particles which additionally are bottom-heavy, that is they feel a torque aligning them to swim against the gravitational field. For bottom-heavy particles the gravitational field has the two opposite effects of i) sedimentation and ii) upward alignment of the particles' swimming direction. We perform a multipole expansion of the one-particle distribution of non-interacting particles with respect to orientation and derive expressions for sedimentation length and mean particle orientation which we check against Brownian Dynamics simulations. For large strength of gravity or small particle speeds and aligning torque, we observe sedimentation with increased sedimentation length compared with passive colloids but also active colloids without bottom-heaviness. Increasing, for example, swimming speed the sedimentation profile is inverted and the particles swim towards the top wall of the enclosing box. We find maximal orientational order at intermediate swimming speeds for both cases of particles with bottom-heaviness and those without. Ordering unsurprisingly is increased for the bottom-heavy particles, but this difference disappears at higher levels of activity and for very high activities ordering goes to zero in both cases.


Patient preferences for in-center intense hemodialysis.
There is a lack of data on patient preferences for intense hemodialysis (IHD). In this study, we conducted a cross-sectional survey to identify patient preferences and patient-centered barriers for IHD. A questionnaire on preferences and anticipated barriers, anticipated benefits, and quality of life for three in-center IHD schedules (daytime 2 hr six times/week [DHD], nocturnal 8 hr three times/week [ND3], and nocturnal 8 hr six times/week [ND6]) was administered to 100 chronic hemodialysis patients. A majority of patients (68%) were willing to undergo DHD for symptomatic benefits or increase in survival. An increase in energy level (94%) and improvement in sleep (57%) were the most common potential benefits that would justify DHD, but only 19% would undergo DHD for an increase in survival of < or =3 years. Only 20% and 7% would consider ND3 and ND6, respectively. The most common reported barriers were inadequate time for self (50%) and family (53%), followed by transportation difficulties (53%). Most patients would undergo DHD for symptomatic or survival benefits, but not ND3 or ND6. Disruption of personal time, however, is an important consideration. Success of DHD program would depend on arrangements for transportation to dialysis unit.


Variable patterns of actin filaments in mammalian skeletal muscle.
The ultrastructural organization of myofilaments in skeletal muscle was studied in four mammalian species (mouse, rat, hamster, goat). In all these species, myofibrils showing irregularly distributed arrays of a variable number of actin filaments (from 6 to 11) were observed. The proportion of such myofibrils and the predominant patterns of actin filaments varied from one species to another. These results are in agreement with those previously reported for human skeletal muscle.


Phytochemical and Pharmacological Investigations on Nymphoides indica Leaf Extracts.
Nymphoides indica (L.) Kuntze (Menyanthaceae) is traditionally used in the Indian subcontinent. However, scientific data reporting its constituents are poor. This study aimed at evaluating its phytochemical constituents and various biological activities. Phytochemical investigations of the extracts and fractions resulted in the isolation of 5 lipophilic compounds, i.e. azelaic (nonanedioic) acid (1) and 4-methyl-heptanedioic acid (3), hexadecanoic (2) and stearic acid (5) and the fatty alcohol hexadecanol (4); 3 seco-iridoids, i.e. 7-epiexaltoside (6), 6″,7″-dihydro-7-epiexaltoside (7) and menthiafolin (8); 3 flavonoids, i.e. 3,7-di-O-methylquercetin-4'-O-β-glucoside (9), 3-O-methylquercetin-7-O-β-glucoside (10) and 3,7-di-O-methylquercetin (11); scopoletin (12) and ferulic acid (13); and the monoterpenoids foliamenthoic acid (14) and 6,7-dihydrofoliamenthoic acid methyl ester (15). Compounds 1-5 showed moderate antimicrobial activities, whereas compound 9 presented mild antiprotozoal activities against Trypanosoma brucei (IC50 8 μM), Leishmania infantum (IC50 32 μM) and Trypanosoma cruzi (IC50 30 μM). Antiglycation activity was shown by compounds 7 (IC50 0.36 mM), 10 (IC50 0.42 mM) and 15 (IC50 0.61 mM). Finally α-glucosidase inhibition was shown by compounds 7, 9, 11 and 13-15. It could be concluded that N. indica leaf extracts possess mild to moderate antimicrobial, antiprotozoal, antioxidant and antidiabetic activities. Copyright © 2016 John Wiley & Sons, Ltd.


Microradiographic aspect on iliac bone tissue in postmenopausal women with and without vertebral crush fractures.
Bone mass, microstructure and microconnectivity of cancellous bone tissue, microporosity, and microdensity of cortical bone have been evaluated with an image analysing computer on microradiographs in 34 iliac crest bone biopsies from two individual, paired age-matched groups of postmenopausal women. Group 1 comprised 17 cases without vertebral crush fractures (mean age 66 years, range 45-80 years) and time since menopause 20.5 +/- 11.5 years. The second group were 17 cases with vertebral crush fractures (mean age 67 years, range 48-86 years) and time since menopause 16.5 +/- 7.6 years. No differences were observed either in bone mass or in Parfitt's indices of the microarchitecture of their cancellous bone tissue. However, both groups 1 and 2 were different by the number of the structure elements per 30 mm2 of tissue volume and the trabecular bone pattern factor (TBPF) being higher in group 1 than in group 2 (p < 0.005)--showing a better connectivity of bone network in group 2 with crush vertebral fractures than in group 1. The number of terminus to terminus struts of the trabecular bone pattern is increased in group 1 (p < 0.001). On 7-microns stained sections the active cancellous eroded perimeter (Cn.Oc.Pm/B.Pm) is greater in group 1 than in group 2 (p < 0.02). Therefore, in these two populations with a similar degree of osteopenia, a decrease of trabecular connectivity does not provide an explanation of bone fragility and the occurrence of vertebral crushed fractures.


Evolutionary dynamics of Ceratonova species (Cnidaria: Myxozoa) reveal different host adaptation strategies.
The myxozoan parasite Ceratonova shasta is an important pathogen that infects multiple species of Pacific salmonids. Ongoing genetic surveillance has revealed stable host-parasite relationships throughout the parasite's endemic range. We applied Bayesian phylogenetics to test specific hypotheses about the evolution of these host-parasite relationships within the well-studied Klamath River watershed in Oregon and California, USA. The results provide statistical support that different genotypes of C. shasta are distinct lineages of one species, which is related to two other Ceratonova species in the same ecosystems; Ceratonova X in speckled dace and C. gasterostea in threespine stickleback. Furthermore, we found strong support for the hypothesis that C. shasta type 0 in native steelhead trout and type I in Chinook salmon each evolved with a specialist host adaptation strategy, while C. shasta type II in coho salmon resulted from a generalist host adaptation strategy. Inferred date and host species of the most recent common ancestor of extant Klamath basin types indicate that it occurred between 14,000 and 21,000 years ago, and most likely infected a native steelhead or rainbow trout host.


Biotransformation studies of textile dye Remazol Orange 3R.
In the present study, biotransformation of Remazol Orange 3R (RO3R) was studied using well-known bacterial isolate Pseudomonas aeruginosa strain BCH. The dye was decolorized up to 98 % within 15 min. The induction in the level of various oxidoreductive enzymes viz. laccase, tyrosinase, veratryl alcohol oxidase and DCIP reductase were observed in the cells obtained after decolorization of RO3R, which supports their role in decolorization. The metabolites of RO3R obtained after biodegradation were identified and characterized by various analytical techniques viz, HPLC, FTIR, and GC-MS. The RO3R was transformed to the N-(7 amino 8 hydroxy-napthalen-2yl) actamide (m/z, 198), Acetamide (m/z, 59) and Napthalen-1-ol (m/z, 144).


Inverse birth cohort effects in ovarian cancer: Increasing risk in BRCA1/2 mutation carriers and decreasing risk in the general population.
BRCA1/2 carriers are at increased risk of ovarian cancer, and some reports suggest an increasing risk in more recent birth cohorts. In contrast, decreasing incidences have been observed in the general population. The aim was to assess the birth cohort effect on ovarian cancer risk in BRCA1/2 carriers relative to their background general population. Data on ovarian cancer incidence was collected for a cohort of 1050 BRCA1/2 mutation carriers ascertained by our regional clinic and retrieved from the general Dutch population cancer registry. Birth cohorts were categorized as pre-1935, 1935-1953, post-1953. Birth cohort effects on the ovarian cancer risk were estimated using hazard ratios (HRs) in BRCA1/2 carriers and Poisson rate ratios in the general population. Standardized incidence ratios (SIRs) were calculated to compare populations. HRs were adjusted for mutation position and family history. Compared to the pre-1935 cohort, BRCA1 carriers in the 1935-1953 and post-1953 cohorts had an increased ovarian cancer risk of HRadjusted 1.54 (95% CI 1.11-2.14) and 2.40 (95% CI 1.56-3.69), respectively. BRCA2 carriers in the 1935-1953 cohort had an HRadjusted of 3.01 (95% CI 1.47-6.13). The SIRs for the 1935-1953 and post-1953 cohorts were 1.7 and 2.7, respectively, for the BRCA1 carriers and 1.6 times and 2.4 times, respectively, for BRCA2 carriers. Mutation carriers, particularly BRCA1 carriers, born in the most recent cohorts, have the highest additional ovarian cancer risk as compared to the general population.


Persistent hiccups and Horner's syndrome in a case of primary CNS lymphoma with diffuse cerebral, hypothalamic and lateral brainstem involvement - An exercise in clinical neuroanatomy.
We present a case of a 42-year-old male presenting with persistent hiccups and a Horner's syndrome, among other symptoms and signs of hypothalamic and brainstem dysfunction. He had a biopsy-proven diffuse infiltrative large primary CNS B-cell lymphoma involving the left fronto-temporal hemisphere, diencephalon and brainstem. The aim of this case report is to highlight key clinical and neuro-anatomical correlations that bring light to the art of the clinical examination.


Heterologous expression, purification and characterization of three novel esterases secreted by the lignocellulolytic fungus Penicillium purpurogenum when grown on sugar beet pulp.
The lignocellulolytic fungus, Penicillium purpurogenum, grows on a variety of natural carbon sources, among them sugar beet pulp. Culture supernatants of P. purpurogenum grown on sugar beet pulp were partially purified and the fractions obtained analyzed for esterase activity by zymograms. The bands with activity on methyl umbelliferyl acetate were subjected to mass spectrometry to identify peptides. The peptides obtained were probed against the proteins deduced from the genome sequence of P. purpurogenum. Eight putative esterases thus identified were chosen for future work. Their cDNAs were expressed in Pichia pastoris. The supernatants of the recombinant clones were assayed for esterase activity, and five of the proteins were active against one or more substrates: methyl umbelliferyl acetate, indoxyl acetate, methyl esterified pectin and fluorescein diacetate. Three of those enzymes were purified, further characterized and subjected to a BLAST search. Based on their amino acid sequence and properties, they were identified as follows: RAE1, pectin acetyl esterase (CAZy family CE 12); FAEA, feruloyl esterase (could not be assigned to a CAZy family) and EAN, acetyl esterase (former CAZy family CE 10).


Production of extracellular polymeric substances (EPS) by Serratia sp.1 using wastewater sludge as raw material and flocculation activity of the EPS produced.
Growth profile and extracellular polymeric substances (EPS) production of Serratia sp.1 was studied in shake flask fermentation for 72 h using wastewater sludge as raw material. Maximum cell concentration of 6.7 × 10(9) cfu/mL was obtained at 48 h fermentation time. EPS dry weight, flocculation activity and dewaterability of different EPS (tightly bound or TB-EPS, loosely bound or LB-EPS and broth-EPS or B-EPS) were also measured. The highest concentration of LB-EPS (2.45 g/L) and TB-EPS (0.99 g/L) were attained at 48 h of fermentation. Maximum flocculation activity and dewaterability (ΔCST) of TB-EPS (76.4%, 14.5s and 76.5%, 15.5s), LB-EPS (67.8%, 8.1s and 64.7%, 7.6s) and broth EPS (61%, 6.1s and 70.4%, 6.8s) were obtained at 36 and 48 h of growth. Higher flocculation activity and dewaterability were achieved with TB-EPS than with the two other EPS. Characterization of TB-EPS and LB-EPS was done in terms of their protein and carbohydrate content. Protein content was much higher in TB-EPS where as carbohydrate content was only slightly higher in TB-EPS than LB-EPS. Morphology of the Serratia strain after fermentation in sludge and TSB was observed under a scanning electron microscope and the cell size was found to be bigger in the sludge medium than the TSB medium.


Allelic association analysis of the 5-HT2C receptor gene in bipolar affective disorder.
We have examined a structural variant of the 5-HT2C receptor (Cys23Ser) for allelic association with bipolar affective disorder in 88 cases and 113 controls. Overall, there was no significant difference in allele frequencies between the two groups, indicating that the 5-HT2C gene is not a major risk factor for bipolar affective disorder. However, when the subjects were analysed according to sex, there was a small excess of the serine ser23 allele in female cases (P = 0.04) and this effect was also seen if the ser23 allele was considered recessive (P = 0.03). A small increase in significance was found if only female cases with a known family history were included (P = 0.01). These results suggest that the ser23 allele may increase susceptibility to bipolar affective disorder in women.


Evaluation of pharmacochaperone-mediated rescue of mutant V2 receptor proteins.
Nephrogenic Diabetes Insipidus is a rare disorder which is characterized by severe water imbalance in the body. The disease can be acquired or inherited. AVPR2 (arginine vasopressin type 2 receptor) mutations are responsible for genetical type of the disorder. Mutations in the AVPR2 gene may cause loss-of-function due to conformational defects. According to the mutation type, the three-dimensional structure of AVPR2 may be affected even if it is functional and therefore it may not reach the plasma membrane where it is functional. Consequently, it is generally trapped in the Endoplasmic reticulum or Golgi apparatus, which are the quality control systems of the cell. Pharmacological chaperones have been used to retrieve these mutant AVPR2s from these quality control systems of the cell and take them to the plasma membrane for therapeutic purposes. In this study, in order to analyze the effects of two pharmacological chaperones, SR121463B and SR49059, we performed total ELISA and surface ELISA studies and cAMP accumulation assays on mutant receptors (G12E, R68W, V88M, ΔR67_G69/G107W, R106C, V162A and T273M). We observed that pharmacological chaperones may act differently on mutated AVPR2s. Cell surface expression of the mutant receptors and cAMP accumulation response, after stimulation with AVP, were mostly improved by these pharmacological chaperones. We believe that, this study presents important results with respect to the process of the variable type of mutated proteins in the cell and may help in developing a process of new types of chaperones.


Clinicopathologic Analyses of Chronic Vascular Rejection After Kidney Transplantation.
We discuss the clinicopathologic analyses of cases of biopsy specimens (BS) after renal transplantation and clarify the mechanisms underlying the development and prognostic significance of chronic vascular rejection (CVR). CVR was diagnosed in 30 renal allograft BS obtained from 23 renal transplant patients being followed up at the Department of Urology and Transplant Surgery, Toda Chuo General Hospital, between January 2010 and August 2017. CVR was diagnosed at a median of 33.1 months post-transplantation. Among the 23 patients, 14 had a history of rejection. Among the 30 BS showing evidence of CVR, the CVR was mild (cv1 on Banff's classification) in 19, moderate (cv2) in 6, and severe (cv3) in 5. We then classified the 30 BS showing evidence of CVR by their overall histopathologic features as follows: cv alone was seen in 9 (30%), cv + antibody-mediated rejection (AMR) in 11 (37%), and cv + T-cell-mediated rejection (TCMR) in 8 (27%). Loss of the renal allograft occurred during the observation period in 2 patients (9%). Of the remaining patients with functioning grafts, deterioration of the renal allograft function after the biopsies occurred in 6 (26%). Our study results suggest that AMR contributes to CVR in 30% to 40% of cases, TCMR in 20% to 30% of cases, isolated v lesions in 10% of cases, and cv lesions alone in 30%. The prognosis of the graft exhibiting CVR was not too poor even under the present immunosuppressive protocol.


The management of pain during pediatric hematopoietic stem cell transplantation: A qualitative study of contextual factors that influenced pain management practices.
Children hospitalized following hematopoietic stem cell transplantation (HSCT) experience complex and prolonged pain in response to the intensity of this treatment. To describe how pain was managed for children during HSCT therapy and how contextual factors related to the clinical environment influenced healthcare providers' and parents' pain management practices. A qualitative case study was conducted and involved semi-structured interviews at two time points following transplantation (30 and 90 days) with parents (n = 10) and naturalistic observations of pain-related care provided to children (n = 29) during HSCT therapy by their healthcare providers (n = 10). Semi-structured interviews were also conducted with healthcare providers (n = 14). The effectiveness of pain management interventions was hindered by the multifactorial nature of pain children experienced, a gap in the provision of psychosocial interventions for pain and a lack of evidence-based guidelines for the sustained, and often long-term, administration of opioids and adjuvant medications. Misconceptions were demonstrated by healthcare providers about escalating pain management according to pain severity and differentiating between opioid tolerance and addiction. Parents were active in the management of pain for children, especially the provision of nonpharmacological interventions. Collaboration with external pain services and the impact of caring for children in protective isolation delayed timely management of pain. There is a pressing need to create evidence-based supportive care guidelines for managing pain post transplantation to optimize children's relief from pain. If parents and children are to be involved in managing pain, greater efforts must be directed toward building their capacity to make informed decisions.


The association of cognitive phenotypes with postoperative outcomes after epilepsy surgery in patients with temporal lobe epilepsy.
The concept of cognitive phenotypes has been developed to categorize the heterogeneity of neuropsychological profiles in patients with temporal lobe epilepsy (TLE). This study examines the utility of cognitive phenotypes derived from clinical criteria in the prediction of postoperative outcomes. Scores from 9 standardized neuropsychological tests were used to sample preoperative performance in 4 core domains (intellectual, memory, language, & executive function) in 445 patients with TLE (206 right: 236 left). Patients were grouped into 3 clinical phenotypes using clinical criteria: 1. intact cognition, 2. isolated memory and/or language impairment, and 3. widespread impairment. Patients who did not meet the criteria for these phenotypes were characterized as having a mixed profile phenotype. Approximately half of the sample had intact cognitive function, with one-quarter demonstrating isolated impairments in language and memory function. The remainder demonstrated widespread impairment or a mixed pattern of cognitive impairments. The clinically derived cognitive phenotypes were associated with demographic and clinical characteristics. Patients with widespread cognitive impairments had an earlier onset of seizures than those with other cognitive phenotypes. They also reported higher levels of depression. Higher levels of anxiety were reported in those with isolated memory/language impairments. Phenotypes were not associated with postoperative seizure outcome or postoperative declines in verbal memory or language function, but an intact phenotype was associated with a greater risk of decline in visual learning than right-sided surgery. Distinct cognitive phenotypes in TLE can be identified using clinical criteria and may reflect neurodevelopmental influences and mood in addition to progression of the disease. Phenotype may be a more powerful predictor of postoperative decline in visual memory than laterality of surgery.


Increased expression of microRNA-221 inhibits PAK1 in endothelial progenitor cells and impairs its function via c-Raf/MEK/ERK pathway.
Coronary artery disease (CAD) is associated with high mortality and occurs via endothelial injury. Endothelial progenitor cells (EPCs) restore the integrity of the endothelium and protect it from atherosclerosis. In this study, we compared the expression of microRNAs (miRNAs) in EPCs in atherosclerosis patients and normal controls. We found that miR-221 expression was significantly up-regulated in patients compared with controls. We predicted and identified p21/Cdc42/Rac1-activated kinase 1 (PAK1) as a novel target of miR-221 in EPCs. We also demonstrated that miR-221 targeted a putative binding site in the 3'UTR of PAK1, and absence of this site was inversely associated with miR-221 expression in EPCs. We confirmed this relationship using a luciferase reporter assay. Furthermore, overexpression of miR-221 in EPCs significantly decreased EPC proliferation, in accordance with the inhibitory effects induced by decreased PAK1. Overall, these findings demonstrate that miR-221 affects the MEK/ERK pathway by targeting PAK1 to inhibit the proliferation of EPCs.


Stressing out over survival: glutamine as an apoptotic modulator.
The amino acid glutamine (GLN) has received considerable attention as a potential therapeutic adjuvant in critical illness and in improving postoperative clinical outcomes. Most studies on the role of GLN in cellular physiology have historically focused on its anabolic roles in specific cell types and its contribution to growth in cancer cells. However, an emerging body of work that examines the consequences of GLN deprivation on cellular survival and gene expression has constructed a new paradigm for this amino acid, namely, that limited extracellular GLN supplies modulate stress and apoptotic responses. A survey of the scientific literature was conducted on GLN in cell survival signaling and apoptosis. Work from our laboratory in liver cancer cells also was included in this review. Most studies on this topic have used mammalian cell lines derived from the gut, immune system (including hybridomas), and various cancers. GLN limitation, even in the presence of an adequate glucose supply, impacts stress-related gene expression, differentially modulates receptor-mediated apoptosis, and directly elicits apoptosis through signaling mechanisms and caspase cascades that are specific to cell type. To date, GLN transporters, cellular hydration, glutaminyl-tRNA synthetase, ATP levels, mRNA stability, and glutathione economy have been variably implicated in GLN-dependent survival signaling. The cell type-specific mechanisms underlying the regulatory role of GLN in cell survival continue to unfold at a steady pace through in vitro studies. These results have collectively provided testable hypotheses for further in vivo studies into their physiological relevance during GLN "nutritional pharmacology."


Raloxifene nanomicelles reduce the growth of castrate-resistant prostate cancer.
Castrate-resistant prostate cancer (CRPC) patients are characterised by a 5-year relative survival rate of ∼25-33%. Recently, our laboratory encapsulated a selective oestrogen receptor modulator, raloxifene, into poly(styrene-co-maleic acid) (SMA-raloxifene), which demonstrated superior in vitro cytotoxicity compared with free drug against several CRPC cell lines. To validate SMA-raloxifene for the management of CRPC using a mouse xenograft model. The internalisation and retention of micellar and free raloxifene in vitro were measured by HPLC. A PC3-CRPC xenograft model was used to compare the biodistribution of both raloxifene formulations, as well as their effect on tumour progression where mice received free raloxifene (1 or 5 mg/kg, i.v.) or SMA-raloxifene (1 mg/kg, i.v.) weekly for 4 weeks. SMA-raloxifene exhibited 75% higher intracellular content compared to free drug after 48 h in PC3 cells. Biodistribution of raloxifene was 69% higher in tumours following SMA-raloxifene compared with free raloxifene. Weekly administration of 1 mg/kg free raloxifene reduced tumour progression by 20% after 4 weeks, whereas 1 mg/kg SMA-raloxifene and 5 mg/kg free raloxifene reduced progression by 40%. Encapsulation of raloxifene increased its therapeutic potential for the management of CRPC.


Conduction velocity changes along the processes of rat primary sensory neurons.
Conduction velocities of rat L4, L5 and L6 dorsal root ganglion neurons were measured in vitro, from several points on the peripheral nerve and dorsal root. Conduction velocities calculated from a single stimulation point (12-26 mm from the ganglion) proved accurate for fibres conducting up to 17 m/s in the peripheral nerve and up to 14 m/s in the dorsal root, but tended to underestimate the value for faster fibres. C-fibre neurons of the L4 and L5 ganglia had a unimodal distribution of conduction velocities below 1.3 m/s. These were discontinuous with A-fibre conduction velocities, which also had a unimodal distribution and had no clear A delta peak. In contrast, conduction velocities of L6 ganglion neurons showed no discontinuity between C- and A-fibres, but had a clear A delta peak. In A-fibre neurons, dorsal root conduction velocities were on average about 14% slower than, and linearly related to, those in the peripheral nerve. However, in individual neurons the dorsal root conduction velocity could be slower or faster than that in the peripheral nerve. In C-fibre neurons dorsal root conduction velocities were almost always slower (average 28%) but not correlated with those of the peripheral nerve. Slowing of conduction velocity along the sciatic nerve was seen in most fibres conducting at less than 2 m/s, but not in faster fibres. The slowing was substantial (up to 60%), sometimes from the A delta to the C-fibre range, and sudden, occurring at a distance of between 15 and 29 mm from the ganglion.


Outcome of primary tubular tubulopathies diagnosed in pediatric age.
Primary tubulopathies are rare and usually present at pediatric age. Recent advances in genetic diagnosis and treatment have changed its natural history. This study provides the clinical spectrum of a series of primary tubulopathies diagnosed in a Pediatric Nephrology Unit and to offer long-term follow-up data regarding growth, estimated glomerular filtration and intercurrent complications. Observational study in 53 patients with primary tubulopathies and identified genetic defect: Gitelman syndrome (36%), distal renal tubular acidosis (15%), cystinuria (11%), X-linked hypophosphatemic rickets (7%), Dent-syndrome Lowe (7%), cystinosis (6%), and 1-2 cases of other tubulopathies. Demographic, analytical and clinical data were collected at diagnosis, during evolution and at the time of the study. The age (median and interquartile range) at diagnosis was 5.08 years (1.33-8.50). The most frequent presentation manifestations were metabolic decompensations associated with intercurrent processes (40%) and short stature (38%). Height (mean±SD) was -1.39±1.49 at diagnosis and 1.07±1.54 after a follow-up of 18.92 (6.25-24.33) years. Sixteen (32%) developed an estimated glomerular filtration<90ml / min / 1.73m2. Three patients required replacement renal replacement. Eleven patients had metabolic decompensations that required hospitalization, 9 renal colic and / or kidney stones and 10 mental problems. Six of 8 patients with distal renal tubular acidosis developed sensorineural deafness. Primary tubulopathies are a heterogeneous group of diseases that cause growth impairment, largely reversible with treatment, risk of estimated glomerular filtration reduction and significant extrarenal complications derived or associated.


Animal models of neuromuscular diseases: pathophysiology and implications for rehabilitation.
This article explains how animal models can be utilized to develop rehabilitation approaches to minimize the pathophysiology of neuromuscular diseases (NMD). Homologous animal models can be used to understand the pathogenesis of each NMD and the effect of physical interventions. Much of the current work using animals has been conducted to show how gene therapy could be used to ameliorate these diseases. Although we applaud this work, animal models should also be used to determine how the phenotypic expression of the disease can be modified by such physical interventions as exercise, stretching, and immobilization. Methods that can be experimentally applied to animals but not to humans can be used to determine the underlying causes of the pathologic process and to identify potential therapies for NMD.


The role of the IRE1 pathway in PBDE-47-induced toxicity in human neuroblastoma SH-SY5Y cells in vitro.
Polybrominated diphenyl ethers (PBDEs) are widely used as flame retardants. As one of the dominant congeners, 2,2', 4,4'-tetrabromodiphenyl ether (PBDE-47) has been shown to be neurotoxic to neuronal cells although the mechanisms remain unclear. To test whether PBDE-47's toxicity was related to endoplasmic reticulum (ER) stress and the unfolded protein response (UPR), human neuroblastoma cells (SH-SY5Y cells) were treated with different concentrations of PBDE-47. Reactive oxygen species (ROS), apoptosis and the expressions of the inositol-requiring enzyme 1 (IRE1) pathway-related molecules were detected. PBDE-47 exposure increased ROS production and activated the UPR by increasing the expressions of glucose-regulated protein 78 (GRP78), IRE1, X-box-binding protein-1 (XBP1), phosphorylation of c-jun N-terminal kinase (JNK) and GADD153/C/EBP homologous protein (CHOP) genes in SH-SY5Y cells. The apoptotic rate increased with the remarkable up-regulation of the Bax/Bcl-2 ratio after IRE1 knockdown, demonstrating the anti-apoptotic role of IRE1. Furthermore, the expressions of CHOP, XBP1 and JNK were down-regulated indicating that IRE1 may activate these key molecules related to apoptosis. PBDE-47 exposure can increase ROS production and activate the IRE1 pathway of the UPR in SH-SY5Y cells contributing to its toxicity. The IRE1 pathway may have both protective and proapoptotic effects on SH-SY5Y cells.


Electrophysiological evidence for terminal sprouting of locus coeruleus neurons following repeated mild stress.
To see if repeated mild stress causes plastic changes in central noradrenergic terminal axons, the density of terminal axons arising in locus coeruleus (LC) neurons of rats was quantified by antidromic stimulation technique. After the termination of stress treatments (immersion in warm water for 10 min daily) for 1 or 2 weeks, electrophysiological experiments were performed under urethane anesthesia. The frequency of LC neurons activated antidromically from the cerebral cortex increased in rats stressed for 2 weeks but not for 1 week. Since the increased frequency of antidromic responses was not due to a change in terminal excitability, the change observed here is considered to be morphological (terminal sprouting) rather than a physiological consequence. The results suggest that LC neurons dynamically alter their terminal morphology in response to environmental stimuli.


Functional CT imaging in oncology.
The two-compartment pharmacokinetics exhibited by iodinated contrast media makes these agents well suited to the study of tumour angiogenesis in which new vessels are not only produced in greater number but also are abnormally permeable to circulating molecules. The temporal changes in contrast enhancement of tumours on CT have been shown to correlate with histopathological assessments of angiogenesis with the intravascular and extravascular phases of contrast enhancement reflecting microvessel density and vascular permeability, respectively. By quantifying tumour contrast enhancement to capture physiological information about the vascular system, functional CT can provide a useful adjunct to the anatomical information afforded by MDCT in oncology, aiding with tumour diagnosis, risk stratification and therapy monitoring. By simultaneously assessing tumour vascularity and metabolic demand, the broader expansion of integrated MDCT/PET imaging will support highly sophisticated assessments of tumour biology within a single examination.


Large-Scale Multiple Testing of Correlations.
Multiple testing of correlations arises in many applications including gene coexpression network analysis and brain connectivity analysis. In this paper, we consider large scale simultaneous testing for correlations in both the one-sample and two-sample settings. New multiple testing procedures are proposed and a bootstrap method is introduced for estimating the proportion of the nulls falsely rejected among all the true nulls. The properties of the proposed procedures are investigated both theoretically and numerically. It is shown that the procedures asymptotically control the overall false discovery rate and false discovery proportion at the nominal level. Simulation results show that the methods perform well numerically in terms of both the size and power of the test and it significantly outperforms two alternative methods. The two-sample procedure is also illustrated by an analysis of a prostate cancer dataset for the detection of changes in coexpression patterns between gene expression levels.


Antibiotic resistance genes (ARGs) in agricultural soils from the Yangtze River Delta, China.
As an important reservoir of intrinsic antimicrobial resistance, soil is subjected to increasing anthropogenic activities that creates sustained selection pressure for the prevalence of antibiotic resistance genes (ARGs), thus constituting an important environmental dissemination pathway to human exposure. This study investigated the levels and spatial distributions of three classes of ARGs in relation to a range of co-occurring chemical mixtures and soil properties at a regional scale of the Yangtze River Delta (YRD), China. The selected eight ARGs were all detected in 241 agricultural soil samples with relative abundances ranging from 1.01 × 10-7 to 2.31 × 10-1 normalized to the 16S rRNA gene. The sulII and tetG were the dominant ARGs with a mean relative abundance of 6.67 × 10-3 and 5.25 × 10-3, respectively. The ARGs were mainly present in agricultural soils alongside Taihu Lake and Shanghai municipality, the most agriculturally and economically vibrant area of the YRD region. Antibiotics, rather than other co-occurring pollutants and soil properties, remain to be the dominant correlate to the ARGs, suggesting their co-introduction into the soils via irrigation and manure application or the sustained selection pressure of antibiotics from these sources for the proliferation of ARGs in the soils. While the current dataset provided useful information to assess the ARGs pollution for mitigation, future studies are warranted to reveal the complete picture on the potential transfer of antimicrobial resistance from soil to agricultural produces to human consumption and associated health implications.


Antibacterial and cytotoxic prenylated dihydrochalcones from Eriosema montanum.
Two new prenylated dihydrochalcones (1,2) and eighteen known secondary metabolites (3-20) were isolated from the CH2Cl2-MeOH (1:1) extracts of the roots, the stem bark and the leaves of Eriosema montanum Baker f. (Leguminosae). The structures of the isolated compounds were characterized by NMR, IR, and UV spectroscopic and mass spectrometric analyses. The structures of compounds 5, 10, 11 and 13 were confirmed by single crystal X-ray diffraction. The antibacterial activity of the crude extracts and the isolated constituents were established against Gram-positive and Gram-negative bacteria. Among the tested compounds, 1-4 and 10 showed strong activity against the Gram-positive bacterium Bacillus subtilis with minimum inhibitory concentration (MIC) ranging from 3.1 to 8.9 μM, as did the leaf crude extract with an MIC of 3.0 μg/mL. None of the crude extracts nor the isolated compounds were active against Escherichia coli. Compounds 1, 3 and 4 showed higher cytotoxicity, evaluated against the human breast cancer cell line MCF-7, with EC50 of 7.0, 18.0 and 18.0 μM, respectively. These findings contribute to the phytochemical understanding of the genus Eriosema, and highlight the pharmaceutical potential of prenylated dihydrochalcones.


Management of subdural effusion and hydrocephalus following decompressive craniectomy for posttraumatic cerebral infarction in a patient with traumatic brain injury: a case report.
Subdural effusion with hydrocephalus (SDEH) is a rare complication of traumatic brain injury, especially following decompressive craniectomy (DC) for posttraumatic cerebral infarction. The diagnosis and treatment are still difficult and controversial for neurosurgeons. A 45-year-old man developed traumatic cerebral infarction after traumatic brain injury and underwent DC because of the mass effect of cerebral infarction. Unfortunately, the complications of traumatic subdural effusion (SDE) and hydrocephalus occurred in succession following DC. Burr-hole drainage and subdural peritoneal shunt were performed in sequence because of the mass effect of SDE, which only temporarily improved the symptoms of the patient. Cranioplasty and ventriculoperitoneal shunt were performed ultimately, after which SDE disappeared completely. However, the patient remains severely disabled, with a Glasgow Outcome Scale of 3. It is important for neurosurgeons to consider the presence of accompanying hydrocephalus when treating patients with SDE. Once the diagnosis of SDEH is established and the SDE has no mass effect, timely ventriculoperitoneal shunt may be needed to avoid multiple surgical procedures, which is a safe and effective surgical method to treat SDEH.


A preliminary pharmacokinetic study of 111In-labeled 260F9 anti-(breast cancer) antibody in patients.
The pharmacokinetics of 111In-labeled 260F9, a murine monoclonal antibody directed against a breast-cancer-associated antigen, was determined in seven patients with advanced breast cancer. Six patients were administered 1 mg antibody containing 1 mCi 111In. The seventh patient was administered 20 mg unlabeled antibody followed by 1 mg 111In-labeled antibody all via a peripheral vein. Immunoprecipitation, HPLC and SDS-PAGE gels demonstrated the stability of radiolabel on the antibody. The serum clearance of the radiolabel closely fits (r2 greater than 0.95) a two-compartment model for the first six patients. The apparent volume of distribution of the radiolabel approximated to the plasma volume (31) and its mean residence time was 23.7 h. The radiolabel had an average t 1/2 beta of 22.9 +/- 12.21 h at the 1-mg dose. At the 20-mg dose one-compartment elimination kinetics were observed with the radiolabel and antibody showing similar mean residence times (36-41 h) and a t 1/2 beta of 26-28 h. Whole-body imaging showed that the blood-pool: liver ratio of radioactivity increased fourfold (at 48 h postinfusion) at the higher dose and the percentage of the injected dose of radioactivity in the liver decreased from 25% to 8% (24 h postinfusion). In one patient 7-14 times more radioactivity was localized in a breast tumor than in fat (normal breast). Over the first 25 h an average (cumulative) 7.5% of the total dose was excreted in urine. A study of 260F9 in CDF-1 mice demonstrated that the radiolabel remained associated with the antibody in serum. The antibody, however, cleared 60-fold slower in mice than in patients and showed an increased mean residence time of 191 h. The disparity in the pharmacokinetics of the antibody seen in the mouse and in the clinic, points to the different behavior shown by murine monoclonal antibodies in humans. This points to the need for preliminary studies of antibodies in patients for preclinical evaluations of their effectiveness as drug-targeting agents.


Biological equivalence of natural bovine and recombinant human alpha-endothelial cell growth factors.
The cDNA encoding human alpha-endothelial cell growth factor (alpha-ECGF) has been engineered for high-level expression in Escherichia coli. Induction of bacterial cultures harboring the recombinant plasmid pMJ26 results in the appearance of a prominent 16-kDa polypeptide. This protein has been purified from bacterial lysates using a rapid, 2-step procedure employing heparin-Sepharose affinity based chromatography and reversed-phase high pressure liquid chromatography. Recombinant human alpha-ECGF was compared to bovine brain-derived alpha-ECGF in three biological assays: receptor binding on murine lung capillary endothelial cells (LE-II cells), stimulation of [3H]thymidine incorporation in LE-II cells, and stimulation of human umbilical vein endothelial cell proliferation. The results demonstrate that the recombinant human mitogen has the same biological potency as the bovine brain-derived material. Fluorescence spectroscopy was used to study the interaction between recombinant ECGF and heparin. Heparin-binding resulted in a 40% reduction in the intrinsic fluorescence of ECGF, consistent with a heparin-induced conformational change. The intrinsic fluorescence of ECGF also varied as a function of pH.


Evaluation of an Electronic Platform for Problem Based Learning for Subspecialty Fellows.
Increasing demands on the time of trainees may warrant new self-directed, concise methods of problem based learning. To address these issues in urological oncology CBULP was designed to provide a concise electronic format that could be readily accessed when the fellow was rested and ready to learn. We evaluated the perceived usefulness of this program. Subspecialists from 2 academic urology programs and an educational professional wrote 42 clinical scenarios about various renal and adrenal malignancies, and generated concise learning points. These cases were mailed to various urological oncology fellowships in the United States and Canada. An 18-question survey was delivered electronically 8 weeks later. Responses were recorded anonymously via survey software. Of 36 fellows 30 (83%) responded. Of the respondents 74% completed at least 5 cases and the majority completed more than 10. Of the respondents 93% thought that the cases had the appropriate amount of detail and covered core concepts related to renal/adrenal tumors. No respondent required more than 20 minutes to finish any case. Of the respondents 93% and 100% indicated that the cases effectively illustrated the basic principles of the disease process, and the fundamentals of evaluation and management, respectively. Overall 97% of respondents thought that CBULP could be an effective learning resource for fellows. An electronic case based method of learning appears to be a useful tool for subspecialty fellows. It may be a worthwhile self-directed supplement to traditional educational resources.


Dermatomyositis and Polymyositis.
Dermatomyositis (DM) and polymyositis (PM) are idiopathic inflammatory myopathies characterized by proximal greater than distal muscle weakness, elevated serum creatine kinase levels, electrophysiologic abnormalities, and inflammation on muscle biopsy. Clinically and electrophysiologically, DM and PM appear very similar, and muscle biopsy is the gold standard for diagnosis. Much of the PM literature based the diagnosis on Bohan and Peter's criteria, which is now obsolete given the advances of immunopathology. As diagnostic criteria for the inflammatory myopathies have been refined, it has become apparent that PM is much less common than previously thought, and, in fact, is probably quite rare. More recent literature, using strict histopathologic criteria for diagnosis of PM, has brought into question previously reported associations. Because of this, the clinical entity of PM is poorly defined. The exact incidence of each is unknown because previous epidemiologic studies often grouped them together, but overall the annual incidence of the inflammatory myopathies is approximately one in 100,000. DM and PM respond to immunomodulating therapies. High-dose oral prednisone is generally accepted first-line therapy. In patients who do not respond adequately to prednisone alone, or in whom prednisone cannot be weaned, methotrexate or azathioprine can be added. In the authors' experience, methotrexate works faster and is more effective than azathioprine. However, because of the increased risk of interstitial lung disease with methotrexate, the authors avoid this in patients with anti-Jo-1 antibodies and, obviously, in patients who already have pulmonary disease. If patients do not respond adequately to the combination of prednisone and methotrexate or azathioprine, a trial of intravenous immunoglobulin is administered.


Charbeckia macrophylla gen. et sp. nov. from the Lower Mississippian Price (Pocono) Formation of southeastern West Virginia.
A fossil plant discovery from the Price (Pocono) Formation of southeastern West Virginia provides new information on the poorly known compression floras of the Lower Mississippian of North America. The new plant described herein consists of long tapering bipinnate fronds with imbricate basal pinnae, planate apical pinnae, and unlobed, elliptical to obovate pinnules with open dichotomous venation. Veins concentrated in the medial region of the pinnule curve toward the lateral margin, suggestive of an early stage in the evolution of a midrib. Pinnules exhibit a distinctly revolute or otherwise reinforced margin. The exceptional length of some fronds (over 1m) and pinnule size (some over 3cm by 2cm) strongly contrast with the generally diffusely branched fronds and small or highly dissected pinnule morphology that typify Early Mississippian leaf taxa. A new generic assignment, Charbeckia macrophylla, is thereby justified. The rigid appearance of the tapering rachis, the imbricate pinnae that appear to have resisted compression, and the reinforced pinnule margins imply thick evergreen leaves, perhaps adapted for drought tolerance. A possible Calamopityalean affinity is indicated by the size of the fronds and the stout petioles, which fall within the expected size range of the Kalymma-type petiole bases described from the nearby New Albany Shale of Tournaisian age.


Medieval anesthesia--the spongia somnifera.
The author reviews in detail the literature concerning medieval general anesthesia in general and the so-called spongia somnifera in particular. In spite of claims to the contrary, he concludes that there is no evidence that medieval physicians had at their disposal general anesthesia for their patients undergoing surgery. On the contrary, there is evidence that the spongia somnifera was ineffective and that what kept the tradition of its effectiveness in the literature may have been the everlasting hope for surgical analgesia.


Beneficial Effects of Receiving Johrei on General Health or Hypothermia Tendency.
Johrei is a type of biofield therapy that is said to bring physical and mental well-being to the recipient. This study sought to measure changes in body temperature and circulation resulting from Johrei treatment, for generally healthy subjects and for individuals with a tendency toward hypothermia. A total of 199 qualified Johrei practitioners and 144 non-qualified operators provided Johrei and placebo treatments, respectively. Volunteer subjects -186 in general health and 39 with a hypothermia tendency - participated in this study to receive either or both of these treatments. Each subject was given a 10 min treatment daily by either a qualified practitioner or a non-qualified operator. The effects on subjects of receiving each treatment were compared by observing quantitative changes in blood flow and surface body temperature after a course of treatment. A total of 107 healthy subjects were randomly assigned to the qualified-practitioner group or the non-qualified operator group. Treatment by qualified practitioners significantly enhanced blood flow and surface body temperature in the subjects' designated neck area compared to that in treatment by non-qualified operators. This finding was further corroborated by a comparative experiment in which each healthy subject was treated by both a qualified practitioner and a non-qualified operator. These results indicate that only the qualified-practitioner treatment increased the subject's-blood flow and surface body temperature. Similarly, in a comparative study of qualified-practitioner treatment against non-qualified-operator treatment, subjects tending toward hypothermia showed increased blood flow and elevated body temperature with only the authentic Johrei treatment.


Wnt signaling in testis development: Unnecessary or essential?
Testis development is a fundamental process in sexual development and reproduction. It is under the regulation of multiple factors. Wnt signaling pathway is a classical pathway, which plays an essential role during early development. From a traditional view, Wnt signaling serves as a key regulator of female reproductive system. However, its role in testis development is relatively controversial. This paper reviews Wnt signaling's part in the major events during testis development, including primordial germ cell specification, proliferation and migration, testis determination, spermatogenesis and somatic cell regulation, and summarizes Wnt signaling's impact on testis-related disorders. We evaluate the outcomes of current studies in the field and suggest future research directions.