Bacterial virulence analysis using brine shrimp as an infection model in relation to the importance of quorum sensing and proteases.
Brine shrimp are aquatic crustaceans belonging to a genus of Artemia. This organism is widely used for testing the toxicity of chemicals. In this study, brine shrimp were evaluated as an infection model organism to study bacterial virulence. Artemia nauplii were infected with various pathogenic bacteria, such as Vibrio vulnificus, Pseudomonas aeruginosa, Burkholderia vietnamiensis, Staphylococcus aureus, and Escherichia coli, and the susceptibility to these bacteria was investigated by counting the survival of the infected nauplii. While all of the tested bacteria have significant virulence to brine shrimp, killing the nauplii in a few days, V. vulnificus showed the strongest virulence. P. aeruginosa also showed a dose-dependent virulence to brine shrimp, but the virulence was weaker than that of V. vulnificus. The virulence tests using the virulence-attenuated mutants of V. vulnificus and P. aeruginosa, such as quorum sensing (QS) mutants or protease-deficient mutants showed a significant attenuation of virulence, demonstrating that the QS mechanism is important in the virulence of these bacteria to brine shrimp. B. vietnamiensis, S. aureus, and E. coli were also virulent to brine shrimp and the virulence was correlated with dosage within 24 h under our conditions. Salmonella enterica Typhimurium and Bacillus subtilis were also virulent to brine shrimp, but the virulence was weak and slowly exerted compared with that of other bacteria. Taken together, we suggest that brine shrimp are a good infection model to assay bacterial virulence, especially for V. vulnificus and P. aeruginosa, and QS is important in the bacterial virulence to brine shrimp.


Mating disruption of citrus leafminer mediated by a noncompetitive mechanism at a remarkably low pheromone release rate.
The citrus leafminer, Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), is a worldwide pest of citrus. A season-long investigation was conducted that evaluated mating disruption for this pest. Effective disruption of the male P. citrella orientation to pheromone traps (98%) and reduced flush infestation by larvae was achieved for 221 d with two deployments of a 3:1 blend of (Z,Z,E)-7,11,13-hexadecatrienal/(Z,Z)-7,11-hexadecadienal at a remarkably low rate of 1.5 g active ingredient (AI)/ha per deployment. To gain insight into the mechanism that mediates the disruption of P. citrella, male moth catch was quantified in replicated plots of citrus treated with varying densities of pheromone dispensers. The densities of septum dispensers compared were: 0 (0/ha, 0.0 g AI/ha), 0.2 (one every fifth tree or 35/ha, 0.05 g AI/ha), 1 (215/ha, 0.29 g AI/ha), and 5 per tree (1,100/ha, 1.5 g AI/ha). Profile analysis by previously published mathematical methods matched predictions of noncompetitive mating disruption. Behavioral observations of male P. citrella in the field revealed that males did not approach mating disruption dispensers in any of the dispenser density treatments. The current report presents the first set of profile analyses combined with direct behavioral observations consistent with previously published theoretical predictions for a noncompetitive mechanism of mating disruption. The results suggest that disruption of P. citrella should be effective even at high population densities given the density-independent nature of disruption for this species and the remarkably low rate of pheromone per hectare required for efficacy.


Molecular cloning, characterization, and mRNA expression of two Cryptochrome genes in Helicoverpa armigera (Lepidoptera: Noctuidae).
Light is a major environmental signal for insect circadian. In this study, we isolated two cryptochrome (cry) genes from Helicoverpa armigera (Hübner) by reverse transcription polymerase chain reaction and RACE-PCR strategies, designated as Ha-cryl (GenBank accession GQ896502) and Ha-cry2 (GenBank accession GQ896503). Ha-CRY1 encoded a fly-like protein of 548 amino acids, while Ha-CRY2 encoded a mammal-like protein of 657 amino acids. Both of these proteins had two conserved domains: a DNA photolyase domain and a flavin adenine dinucleotide (FAD) binding seven domain, and alignment of the amino acid sequence indicated that there was a high degree of homology between the CRYs of H. armigera and other insects. Real-time polymerase chain reaction revealed that: 1) Ha-cry1 and Ha-cry2 mRNA expressions were neither organ-specific nor developmental-stage-specific. 2) Under the light-dark cycle (16:8 L:D), Ha-cry1 abundance tended to increase during the day, then decrease in the night, whereas the expression pattern of Ha-cry2 was opposite. 3) The cyclings of Ha-cry1 and Ha-cry2 expression were disturbed by constant light and darkness. Our study has significant importance for the further study of the functions of the Ha-cry genes and potential control of the cotton bollworm.


Combretastatin A-4-phosphate effectively increases tumor retention of the therapeutic antibody, 131I-A5B7, even at doses that are sub-optimal for vascular shut-down.
Radioimmunotherapy using 131I-A5B7, an anti-CEA antibody, in combination with the vascular disrupting agent, combretastatin A4-phosphate (CA-4-P, 200 mg/kg), has produced tumor cures in SW1222 colorectal xenografts. CA-4-P causes acute tumor blood vessel shutdown, which can be monitored in clinical trials using dynamic contrast enhanced magnetic resonance imaging (DCE-MRI). The purpose of this study was to determine the magnitude of the anti-vascular effect of CA-4-P in the SW1222 tumor, at 200 mg/kg and at lower, more clinically relevant doses, using conventional assays; relate effects to changes in DCE-MRI parameters and determine the corresponding effects on tumor retention of 131I-A5B7. The tumor vascular effects of 30, 100 and 200 mg/kg CA-4-P were determined, at 4- and 24-h post-treatment, using DCE-MRI, uptake of Hoechst 33342 for tumor vascular volume and conventional histology for necrosis. The effect of CA-4-P on tumor and normal tissue 131I-A5B7 retention was also determined. A significant reduction in tumor DCE-MRI kinetic parameters, the initial area under the contrast agent concentration time curve (IAUGC) and the transfer constant (Ktrans), was demonstrated at 4 h after CA-4-P, for all dose levels. These effects persisted for at least 24 h for the 200 mg/kg group but not for lower doses. A similar pattern was seen for vascular volume and necrosis. Despite this dose response, all three dose levels increased tumor retention of radio labeled antibody to a similar degree. These results demonstrate that moderate tumor blood flow reduction following antibody administration is sufficient to improve tumor antibody retention. This is encouraging for the combination of CA-4-P and 131I-A5B7 in clinical trials.


Collagenofibrotic glomerulonephropathy in a cynomolgus macaque (Macaca fascicularis).
Collagenofibrotic glomerulonephropathy (CFGN) is characterized by the deposition of type III collagen within the mesangial matrix and the absence of mesangial cell proliferation. A case of CFGN in a 2.7-year-old female cynomolgus macaque was investigated in the present study. Clinically, the animal was shown to have severe systemic edema along with hypoproteinemia. At necropsy, the kidneys were swollen and pale. The glomerular lesions were characterized by massive diffuse and global accumulation of fibrous materials in the mesangial areas. Neither mesangial cell proliferation nor changes in other organs were found. The fibrous materials were confirmed by the results of immunohistochemical and electron microscopic findings to consist mainly of randomly arranged, curve-shaped, twisted, and entwined type III collagen. This is the first case report of CFGN in nonhuman primates to date.


Secondary signet-ring cell tumour of the prostate derived from a primary gastric malignancy.
Primary signet-ring cell carcinoma of the prostate is infrequent and even more so as secondary spread of this pathologic sub-type to the prostate. We describe the sixth reported case with a diagnosis of a secondary signet-ring cell tumour of the prostate secondary to a gastric cancer. Five years post-gastrectomy to resect signet-ring cell carcinoma, we detected a secondary intra-prostatic spread with urinary tract obstruction. The physical appearance of the tumour cells was similar to that of the previously-resected signet-cell carcinoma of the stomach. There were no metastases in other sites and the patient was treated with radiotherapy. When confronted with intra-prostatic signet-ring cell adenocarcinoma it is necessary to distinguish between primary and secondary aetiology since this would reflect in the choice of treatment and prognosis.


Use of research electronic data capture (REDCap) in a COVID-19 randomized controlled trial: a practical example.
Randomized controlled trials (RCT) are considered the ideal design for evaluating the efficacy of interventions. However, conducting a successful RCT has technological and logistical challenges. Defects in randomization processes (e.g., allocation sequence concealment) and flawed masking could bias an RCT's findings. Moreover, investigators need to address other logistics common to all study designs, such as study invitations, eligibility screening, consenting procedure, and data confidentiality protocols. Research Electronic Data Capture (REDCap) is a secure, browser-based web application widely used by researchers for survey data collection. REDCap offers unique features that can be used to conduct rigorous RCTs. In September and November 2020, we conducted a parallel group RCT among Indiana University Bloomington (IUB) undergraduate students to understand if receiving the results of a SARS-CoV-2 antibody test changed the students' self-reported protective behavior against coronavirus disease 2019 (COVID-19). In the current report, we discuss how we used REDCap to conduct the different components of this RCT. We further share our REDCap project XML file and instructional videos that investigators can use when designing and conducting their RCTs. We reported on the different features that REDCap offers to complete various parts of a large RCT, including sending study invitations and recruitment, eligibility screening, consenting procedures, lab visit appointment and reminders, data collection and confidentiality, randomization, blinding of treatment arm assignment, returning test results, and follow-up surveys. REDCap offers powerful tools for longitudinal data collection and conduct of rigorous and successful RCTs. Investigators can make use of this electronic data capturing system to successfully complete their RCTs. The RCT was prospectively (before completing data collection) registered at ClinicalTrials.gov; registration number: NCT04620798 , date of registration: November 9, 2020.


GaN nanowires grown by halide chemical vapour deposition as photoanodes for photo-electrochemical water oxidation reactions.
Manifold morphologies of GaN nanowires (NWs) were fabricated using halide chemical vapour deposition (HCVD) on an n-Si (111) substrate and demonstrated to be a promising photoelectrode for photo-electrochemical (PEC) water splitting applications. We report a substantial enhancement in the photocurrent for vertically-grown GaN NWs on a buffer layer as compared to other counterparts such as GaN whiskers, tapered nanostructures and thin films. GaN NWs grown on Si have advantages due to the absorption of photons in a wide spectral range from ultraviolet to infrared and thus are directly involved in PEC reactions. A GaN NW photoanode was demonstrated with a saturation photocurrent density of 0.55 mA cm-2 under 1 sun of illumination, which is much greater than its counterparts. The role of the buffer layer and the carrier density on the PEC performance of vertically-grown GaN NW photoanodes is further elucidated. Photo-electrochemical impedance spectroscopy and Mott-Schottky characterizations were employed to further explain the PEC performance of GaN NW embedded photoanodes. Here, photoanodes based on diverse GaN nanostructures were examined for a better PEC evaluation in order to support the conclusion. The results may pave the way for the fabrication of efficient photoelectrodes and GaN as a protective layer against corrosion for improved photo-stability in an NaOH electrolyte for enhancing the efficiency of water splitting.


Differential gene expression associated with euryhalinity in sea bream (Sparus sarba).
Certain fish have the remarkable capability of euryhalinity, being able to withstand large variations in salinity for indefinite periods. Using the highly euryhaline species, silver sea bream (Sparus sarba), as an experimental model, some of the molecular processes involved during ion regulation (Na+-K+-ATPase), cytoprotection [heat shock protein (hsp) 70], and growth (somatotropic axis) were studied. To perform these studies, seven key genes involved in these processes were cloned, and the tissue-specific expression profiles in fish adapted to salinities of 6 parts per thousand (ppt; hypoosmotic), 12 ppt (isoosmotic), 33 ppt (seawater), and 50 ppt (hypersaline) were studied. In gills, the transcriptional and translational expression profiles of Na+-K+-ATPase alpha- and beta-subunit genes were lowest in isoosmotic-adapted fish, whereas in kidneys the expression of the beta-subunit increased in seawater- and hypersaline-adapted groups. The hsp70 multigene family, comprising genes coding for heat shock cognate (hsc70), inducible heat shock protein (hsp70), and a heat shock transcription factor (hsf1), was found to be highly upregulated in gills of seawater- and hypersaline-adapted fish. In liver, hsc70 expression was lowest in isoosmotic groups, and in kidneys the hsp70 multigene family remained unchanged over the salinity range tested. The regulation of the somatotropic axis was studied by measuring pituitary growth hormone expression and liver IGF-I expression in salinity-adapted fish. The expression amounts of both genes involved in the somatotropic axis were highest in fish maintained at an isoosmotic salinity. The results of this study provide new information on key molecular processes involved in euryhalinity of fish.


An iterative and regenerative method for DNA sequencing.
This paper presents, to our knowledge, the first iterative DNA sequencing method that regenerates the product of interest during each iterative cycle, allowing it to overcome the critical obstacles that impede alternative iterative approaches to DNA sequencing: loss of product and the accumulation of background signal due to incomplete reactions. It can sequence numerous double-stranded (ds) DNA segments in parallel without gel resolution of DNA fragments and can sequence DNA that is almost entirely double-stranded, preventing the secondary structures that impede sequencing by hybridization. This method uses ligation of an adaptor containing the recognition domain for a class-IIS restriction endonuclease and digestion with a class-IIS restriction endonuclease that recognizes the adaptor's recognition domain. This generates a set of DNA templates that are each composed of a short overhang positioned at a fixed interval with respect to one end of the original dsDNA fragment. Adaptor ligation also appends a unique sequence during each iterative cycle, so that the polymerase chain reaction can be used to regenerate the desired template-precursor before class-IIS restriction endonuclease digestion. Following class-IIS restriction endonuclease digestion, sequencing of a nucleotide in each overhang occurs by template-directed ligation during adaptor ligation or through a separate template-directed polymerization step with labeled ddNTPs. DNA sequencing occurs in strides determined by the number of nucleotides separating the recognition and cleavage domains for the class-IIS restriction endonuclease encoded in the ligated adaptor, maximizing the span of DNA sequenced for a given number of iterative cycles. This method allows the concurrent sequencing of numerous dsDNA segments in a microplate format, and in the future it can be adapted to biochip format.


Reliability and relative validity of a diet index score for adults derived from a self-reported short food survey.
Assess the reliability and relative validity of a diet index score for adults derived using a 38-item online survey. The short food survey (SFS) measured 'usual' intake of seven food groups, three food choice indicators and variety; and was completed by 61 adults aged 19-50 years from Adelaide, Australia. A score was applied to assess compliance with the 2013 Australian Dietary Guidelines. Reliability of the survey was measured between two administrations one week apart; and validity by comparing the first administration to the average of three 24-hour dietary recalls. Statistical analyses included paired samples t-tests, intra-class correlation coefficient (ICC), percentage agreement, Cohen's kappa coefficients and Bland-Altman plots. Estimates of daily food group servings were reliable (within 0.3 servings, P > 0.05) but not valid for all foods groups. The mean total index score was approximately 70 points (out of 100) on both SFS administrations (ICC = 0.71 (95% CI 0.56:0.81). Relative validity analysis showed moderate correlation between SFS#1 and 24-hour recalls (ICC = 0.43 (0.21:0.62, P < 0.001)), with 51% agreement in allocation to tertiles of diet quality between methods (k = 0.262, P = 0.004). The survey overestimated the diet index score by an average of 12.7 points out of 100 [-20.11:42.94] in comparison to recalls. The survey overestimated compliance with guidelines relative to dietary recalls. It demonstrated good reliability; however, the validity of estimating intake of some food groups needs improvement. Future refinement will provide a valuable online tool to assess compliance with the Australian dietary recommendations.


Meeting report of the 2017 KidGen Renal Genetics Symposium.
The 2017 KidGen Renal Genetics Symposium was held at the Royal Children's Hospital and Murdoch Children's Research Institute, Melbourne, from 6 to 8 December 2017. This meeting addressed clinical, diagnostic, and research aspects of inherited kidney disease. More than 100 clinicians, researchers, and patient representatives attended the conference. The overall goal was to improve the understanding and direction of genomics in renal medicine in Australia and discuss barriers to the use of genomic testing within this area. It also aimed to strengthen collaborations between local, state, and global research and diagnostic and clinical groups.


Endogenous Na+ transport inhibitor in human hypertension: further biochemical and chemical studies.
Endogenous digitalis-like compound(s) (endalin) has(ve) been reported to be involved in some diseases. Endalin activity is increased in plasma and urine of some essential hypertensives, and in Na+-dependent experimental hypertension. The aims of this study are to compare the biological properties of one endalin extracted from urine of hypertensive patients and of normotensive offspring of hypertensive subjects to those of ouabain and to determine the chemical nature of such an urine-derived endalin. The donors were selected on the basis of the highest Na+,K+-ATPase inhibition produced by extract from their 24-h urine. They consisted of 8 hypertensive patients, 21 normotensive subjects with family history of hypertension and 6 normotensive subjects with no known family history of hypertension. Endalin was semi-purified from 500 liters of pooled urine by flash chromatography on RP 18 packing (40 microns) followed by anion exchange chromatography and two HPLCs on RP 18 reversed phase. Endalin was traced by its capability of inhibiting dog kidney Na+,K+-ATPase activity and 3H-ouabain binding to the enzyme, by its cross-reaction with anti-digoxin antibodies and by its natriuretic effect in rat bioassay. The mechanism of Na+,K+-ATPase inhibition by a semi-purified urine-derived endalin and its consequences on Na-transport were studied and compared to those of ouabain. Semi-purified urine-derived endalin was similar to ouabain in that: it reversibly and specifically inhibited Na+,K+-ATPase activity; it inhibited Na+,K+-ATPase non-competitively with ATP; its inhibitory effect was facilitated by Na+; K+ decreased its inhibitory effect on Na+,K+-ATPase.(ABSTRACT TRUNCATED AT 250 WORDS)


Ursodeoxycholic acid attenuates experimental autoimmune arthritis by targeting Th17 and inducing pAMPK and transcriptional corepressor SMILE.
Ursodeoxycholic acid (UDCA) has been known that UDCA has prominent effects on liver, however, there is little known about its influence on autoimmune disease. Here, the benefit of UDCA on arthritis rheumatoid (RA) in vivo was tested. RA mouse were induced using collagen II (CIA, collagen induced arthritis) where the disease severity or UDCA-related signaling pathway such as AMP-activated protein kinase (AMPK) or small heterodimer partner interacting leucine zipper protein (SMILE) was evaluated by westerblot and immunohistochemical staining. Gene expression was measured by realtime-polymerase chain reaction (PCR). The administration of UDCA effectively alleviated the arthritic score and incidence with decreased cartilage damage and lipid metabolic parameters. UDCA also suppressed the secretion of pro-inflammatory cytokines. It was confirmed that UDCA upregulated the expression of SMILE and transcriptional activity of PPARγ via controlling AMPK or p38 activity. In the present study, the therapeutic effect of UDCA inducing SMILE through AMPK activation in rheumatoid arthritis mouse as well as other autoimmune disease was proposed.


[The investigation the combined effect of SNP rs9939609 (gene FTO) and rs4994 (gene ADRB3) polymorphisms on risk of obesity].
The genetic factor plays a significant role in the development of obesity, by present time the association of hundreds genetic polymorphisms with the risk of this disease is established. However, the combined influence of genetic polymorphisms remains practically unstudied. We aimed to investigate the combined effect SNP rs9939609 (gene FTO) and rs4994 (gene ADRB3) polymorphisms on risk of obesity. A case-control study was conducted, including255 obese case (BMI>30 kg/m2) and 427 non obese controls (BMI<30 kg/m2). Genotyping was performed using allele-specific amplification, detection results in real time using TaqMan-probes complementary DNA polymorphic sites. It has been shown, that presence of one mutant allele of rs9939609 (gene FTO) and rs4994 (gene ADRB3) leads to statistically significant association with obesity. Presence of two mutant alleles in different polymorphic variants increases risk of obesity by 15%, presence of three mutant alleles - by 2.63 fold. The quantity surveyed, suffering obesity, increased depending on the number of mutant alleles in studied genetic polymorphisms. Presence of one or two mutant alleles in one polymorphic variant increased the number of patients with obesity by 13.4%, presence of two or three mutant alleles in different polymorphic variants -by 18-19%.


The nuclear receptor REV-ERBα integrates circadian clock and energy metabolism.
The physiological processes of mammals show rhythmic changes in a 24-h cycle. Circadian rhythms are under the subtle control of the autonomous circadian clock, and dysregulation of the circadian system can lead to health problems such as metabolic disorders. REV-ERBα, a member of the nuclear receptor superfamily, is an important component of the mammalian circadian clock. REV-ERBα regulates various physiological processes, including the regulation of metabolism, inflammation and immunity as well as the circadian rhythm, making it a potential therapeutic target for metabolic syndrome, inflammatory diseases and cancers. In recent years, an array of new REV-ERBα ligands have been discovered, most of which have potential applications in the treatment of diseases. In this review, we focus on the regulatory role of nuclear receptor REV-ERBα in energy metabolism and inflammation, in order to provide new strategies for the therapy of metabolic syndrome and its related diseases.


Is the Bone-Conduction HeadBand test useful for predicting the functional outcome of a round window active middle ear implant?
To estimate the reliability of the Bone Conduction-HeadBand (BC-HB) test for predicting the postoperative functional outcome of a round-window (RW) vibroplasty. Within-subject comparison of the functional results of the BC-HB test, which is routinely used for the preoperative evaluation of a bone-conduction transducer, with an active middle ear implant (AMEI) placed onto the round window. Tertiary referral university hospital center. Seven patients with similar anatomic (absent stapes superstructure) and functional (moderate, mixed hearing loss) sequelae from open tympanoplasty technique. All subjects underwent preoperative audiologic assessment with the BC-HB. Subsequently, all subjects underwent surgical placement of an AMEI onto the round window. Pure tone and speech audiometry in quiet and noise were assessed. Additionally, evaluation of specific satisfactory targets was performed using the Client Oriented Scale of Improvement. Pure tone and speech audiometry in quiet established that both devices had very similar performance and provided remarkable improvement compared with the unaided condition. However, high-frequency gain and speech audiometry in noise demonstrated better performance with RW-AMEI. In patients presenting with mixed hearing loss as a sequela from middle ear surgery, the preoperative BC-HB test may be helpful in predicting the final functional outcome and patient satisfaction with RW-AMEI.


Ortner's Syndrome Due to Giant Thoracic Aortic Aneurysm.
Ortner's syndrome is hoarseness of the voice caused by enlarged cardiac structures compressing the left recurrent laryngeal nerve (LRLN). The LRLN is vulnerable for impingement by cardiac structures due to its anatomical course. LRLN is longer and it hooks below the arch of the aorta, posterior to the ligamentum arteriosum before ascending toward the tracheo- esophageal groove. Ortner's syndrome has been commonly described with left atrial enlargement due to mitral stenosis; however, other causes like aortic aneurysm, pulmonary artery aneurysm, and aortic dissection have been described. Although rare, hoarseness of the voice could be the only presenting complaint in aortic aneurysms as in the case described herein.


Carcinogenic glutamic acid pyrolysis product in the dialysate of uremic patients treated by continuous ambulatory peritoneal dialysis.
By using a high-performance liquid chromatography (HPLC) method, we determined the contents of 2-amino-6-methyldipyrido[1,2-a:3', 2'-d] imidazole (Glu-P-1) and 2-aminodipyrido[1,2-a:3', 2'-d]imidazole (Glu-P-2) in the dialysate of patients with uremia. The total dialysate (approximately 6 liters) per day of the patients who had received continuous ambulatory peritoneal dialysis (CAPD) was examined. The amounts of Glu-P-1 and Glu-P-2 in the total dialysate per day were 552.2 +/- 266.9 pmole and 386.3 +/- 146.0 pmole (mean +/- SD, n = 10), respectively. The recoveries of Glu-P-1 and Glu-P-2 in the dialysate on our assay method were 68.2% and 54.3%, respectively. Based on the recoveries, the average amounts of Glu-P-1 and Glu-P-2 in the dialysate of uremic patients were assumed to be 809.7 pmole and 711.4 pmole, respectively.


Intratracheal adenoviral-mediated delivery of iNOS decreases pulmonary vasoconstrictor responses in rats.
We hypothesized that adenovirus-mediated inducible nitric oxide synthase (iNOS) gene transduction of the lung would result in time-dependent iNOS overexpression and attenuate the vascular constrictor responses to a thromboxane mimetic, U-46619. Rats were treated via the trachea with surfactant alone (sham), surfactant containing an adenoviral construct with a cytomegalovirus promoter-regulated human iNOS gene (Adeno-iNOS), or an adenoviral construct without a gene insert (Adeno-Control). Adeno-iNOS-transduced rats demonstrated human iNOS mRNA and increased iNOS protein levels only in the lungs. Immunohistochemistry of lungs from Adeno-iNOS-treated animals demonstrated transgene expression in alveolar wall cells. In the lungs from Adeno-iNOS-transduced rats, the expression of iNOS protein and exhaled nitric oxide concentrations were increased on days 1-4 and 7 but returned to baseline values by day 14. The administration of the selective iNOS inhibitor L-N6-(1-iminoethyl)lysine dihydrochloride (L-NIL) decreased exhaled nitric oxide concentrations to levels found in Adeno-Control-transduced lungs. In a second group of rats, the segmental vasoconstrictor responses to U-46619 were determined in isolated, perfused lungs 3 days after transduction. Lungs from rats transduced with Adeno-iNOS had reduced total, arterial, and venous vasoconstrictor responses to U-46619 compared with sham, Adeno-Control, and control groups. In a third set of experiments, the response to 400 nM U-46619 in the presence of 10 microM L-NIL was not different in the isolated lungs from Adeno-Control- and Adeno-iNOS-transduced rats. We conclude that adenovirus-mediated iNOS gene transduction of the lung results in time-dependent iNOS overexpression, which attenuates the vascular constrictor responses to the thromboxane mimetic U-46619.


Gating of reafference in the external cuneate nucleus during self-generated movements in wake but not sleep.
Nervous systems distinguish between self- and other-generated movements by monitoring discrepancies between planned and performed actions. To do so, corollary discharges are conveyed to sensory areas and gate expected reafference. Such gating is observed in neonatal rats during wake-related movements. In contrast, twitches, which are self-generated movements produced during active (or REM) sleep, differ from wake movements in that they reliably trigger robust neural activity. Accordingly, we hypothesized that the gating actions of corollary discharge are absent during twitching. Here, we identify the external cuneate nucleus (ECN), which processes sensory input from the forelimbs, as a site of movement-dependent sensory gating during wake. Whereas pharmacological disinhibition of the ECN unmasked wake-related reafference, twitch-related reafference was unaffected. This is the first demonstration of a neural comparator that is differentially engaged depending on the kind of movement produced. This mechanism explains how twitches, although self-generated, trigger abundant reafferent activation of sensorimotor circuits in the developing brain.


Processing and storage methods affect oral and gut microbiome composition.
In microbiome studies, fecal and oral samples are stored and processed in different ways, which could affect the observed microbiome composition. In this study, we compared storage and processing methods applied to samples prior to DNA extraction to determine how each affected microbial community diversity as assessed by 16S rRNA gene sequencing. We collected dental swabs, saliva, and fecal samples from 10 individuals, with three technical replicates per condition. We assessed four methods of storing and processing fecal samples prior to DNA extraction. We also compared different fractions of thawed saliva and dental samples to fresh samples. We found that lyophilized fecal samples, fresh whole saliva samples, and the supernatant fraction of thawed dental samples had the highest levels of alpha diversity. The supernatant fraction of thawed saliva samples had the second highest evenness compared to fresh saliva samples. Then, we investigated the differences in observed community composition at the domain and phylum levels and identified the amplicon sequence variants (ASVs) that significantly differed in relative abundance between the conditions. Lyophilized fecal samples had a greater prevalence of Archaea as well as a greater ratio of Firmicutes to Bacteroidetes compared to the other conditions. Our results provide practical considerations not only for the selection of storage and processing methods but also for comparing results across studies. Differences in processing and storage methods could be a confounding factor influencing the presence, absence, or differential abundance of microbes reported in conflicting studies.


Karyotype differentiation in tellin shells (Bivalvia: Tellinidae).
Although Tellinidae is one of the largest and most diverse families of bivalves, its taxonomy is utterly chaotic. This is mainly due to the morphological diversity and homoplasy displayed by their shells and to the scarcity of the molecular phylogenetic studies performed on them. A molecular cytogenetic analysis of four tellin shell species, Bosemprella incarnata, Macomangulus tenuis, Moerella donacina and Serratina serrata, was performed. To molecularly characterize the analyzed specimens, the sequence of a fragment of the mitochondrial cytochrome c oxidase subunit I (COI) was also studied. The karyotypes of the four species were composed of different amounts of bi-armed and telocentric chromosomes. The chromosomal mapping of 45S and 5S rDNA and H3 histone gene clusters by fluorescent in situ hybridization also revealed conspicuous differences on the distribution of these DNA sequences on their karyotypes. Vertebrate type telomeric sequences were located solely on both ends of each chromosome in all four tellin shells. We present clear evidence of the valuable information provided by FISH signals in both analyzing chromosome evolution in Tellinidae and as a further tool in identifying tellin shell specimens for molecular phylogenies.


Golgi pH, Ion and Redox Homeostasis: How Much Do They Really Matter?
Exocytic and endocytic compartments each have their own unique luminal ion and pH environment that is important for their normal functioning. A failure to maintain this environment - the loss of homeostasis - is not uncommon. In the worst case, all the main Golgi functions, including glycosylation, membrane trafficking and protein sorting, can be perturbed. Several factors contribute to Golgi homeostasis. These include not only ions such as H+, Ca2+, Mg2+, Mn2+, but also Golgi redox state and nitric oxide (NO) levels, both of which are dependent on the oxygen levels in the cells. Changes to any one of these factors have consequences on Golgi functions, the nature of which can be dissimilar or similar depending upon the defects themselves. For example, altered Golgi pH homeostasis gives rise to Cutis laxa disease, in which glycosylation and membrane trafficking are both affected, while altered Ca2+ homeostasis due to the mutated SCPA1 gene in Hailey-Hailey disease, perturbs various protein sorting, proteolytic cleavage and membrane trafficking events in the Golgi. This review gives an overview of the molecular machineries involved in the maintenance of Golgi ion, pH and redox homeostasis, followed by a discussion of the organelle dysfunction and disease that frequently result from their breakdown. Congenital disorders of glycosylation (CDGs) are discussed only when they contribute directly to Golgi pH, ion or redox homeostasis. Current evidence emphasizes that, rather than being mere supporting factors, Golgi pH, ion and redox homeostasis are in fact key players that orchestrate and maintain all Golgi functions.


Extension of oxygen tolerance by treatment with endotoxin: means to improve its potential therapeutic safety in man.
Treatment of adult rats with low doses of bacterial lipopolysaccharide (endotoxin) consistently results in a marked protective effect against O2-induced lung damage and lethality. We report here two means to improve the therapeutic ratio of endotoxin (ratio of dose producing desired beneficial effect/dose producing undesired toxic effects), which could make it a more acceptable pharmacologic agent for possible use in patients who require prolonged hyperoxic therapy. (a) Rats made "tolerant" to the lethal/toxic effects of high doses of endotoxin (25 mg/kg) by pretreatment with very low doses of endotoxin (10 ng----10 micrograms/kg) were found to still respond to a standard protective dose of endotoxin (500 micrograms/kg) with marked resistance to O2 toxicity. (Survival in greater than 95% O2 X 72 h = 19/20 (95%), vs. 4/17 (24%) for controls.) (b) Two chemically modified native endotoxin preparations ("endotoxoids"), with approximately 100 X decreased toxic potential, were found to have retained their ability to protect adult rats from prolonged hyperoxic exposure (90%-100% survival rates). These two experimental manipulations (use of the "endotoxin tolerance" phenomenon and treatment with partially detoxified "endotoxoids") were associated with increased lung antioxidant enzyme activities during O2 exposure in the treated animals. Continued research may eventuate in the possible clinical application of a safe form of endotoxin treatment for the prevention of O2 toxicity in humans.


Detection and perception of generic host volatiles by mosquitoes: responses to CO2 constrains host-seeking behaviour.
Natural selection has favoured specialization in anthropophilic mosquito host choice, yet in the absence of human hosts, females feed on a selected range of vertebrates. For host recognition, we hypothesize that mosquitoes primarily rely on generic host volatiles. Detection and perception of such compounds would provide the mosquito with a flexible, yet constrained, odour coding system that could delineate host preference. In this study, we show that the quintessential generic volatile for host-seeking, carbon dioxide, activates and attracts the malaria mosquito, Anopheles coluzzii, and the arbovirus vectors, Aedes aegypti and Culex quinquefasciatus, within boundaries set by the dynamic range and coding capacity of the CO2-sensitive olfactory receptor neurons. These boundaries are sufficiently broad to elicit behavioural responses to various hosts within their preferred host range. This study highlights the significance of the sensitivity of the carbon dioxide detection system and its regulation of host seeking and recognition.


Indoleamine 2,3-dioxygenase and iron are required for Mycobacterium leprae survival.
Our previous study has demonstrated that IL-10 may modulate both indoleamine 2,3-dioxygenase (IDO) and CD163 expression in lepromatous leprosy (LL) cells, favoring Mycobacterium leprae persistence through induction of regulatory pathways and iron storage. Here, we observed that in LL lesion cells there is an increase in the expression of proteins involved in iron metabolism such as hemoglobin (Hb), haptoglobin, heme oxygenase 1 and transferrin receptor 1 (TfR1) when compared to tuberculoid leprosy (BT) cells. We also found increased iron deposits and diminished expression of the iron exporter ferroportin 1 in LL lesion cells. Hemin, but not FeSO4 stimulation, was able to enhance M. leprae viability by a mechanism that involves IDO. Analysis of cell phenotype in lesions demonstrated a predominance of M2 markers in LL when compared with BT lesion cells. A positive correlation between CD163 and PPARG with the bacillary index (BI) was observed. In contrast, TNF, STAT1 and CSF2 presented a negative correlation with the BI. In summary, this study demonstrates that iron may regulate IDO expression by a mechanism that involves IL-10, which may contribute for the predominance of M2-like phenotype in LL lesions that favors the phagocytosis and maintenance of M. leprae in host cells.


[24-Hour blood pressure profile in patients with mild and moderate arterial hypertension and sleep apnea/hypopnea].
To evaluate 24-hour blood pressure (BP) profile in arterial hypertension (AH) patients (pts) with desaturation signs of sleep apnea/hypopnea syndrome (SAHS). We investigated 61 pts (44 males and 17 females) aged between 23-70 (52 +/- 2) years with mild to moderate AH. BP monitoring was performed with multisensor system TM-2425 (A&D, Japan). We assessed the following parameters: mean 24-h, awake, sleep systolic (S), diastolic (D) and pulse (P) BPs, systolic and diastolic BP loads ("normalized area under the curve"--NAUC). A normal circadian rhythm of BP was defined when nocturnal fall of SBP was > 10% and < 20%. The morning rise of BP we assessed by speed of increase of mean BP from 4 a.m. to 12 a.m. The nocturnal monitoring of arterial oxygen saturation(SaO2) was performed with pulseoximeter "NONIN 8500M" (USA). The analysis of the results was performed with the original program ARM-SaO2". The presence of SAHS was confirmed when the number of 4% desaturations were greater than 15 per hour or in the presence of group episodes of 4% desaturation below 90%. In 19 pts we revealed desaturation signs of SAHS. The comparison group included pts without SAHS (n = 42). We compared the groups regarding 24-h BP profile parameters. SAHS group had the following parameters significantly higher: mean 24-h (151.7 +/- 4.5 vs 142.9 +/- 2.4 mm Hg, p < 0.07) and sleep SBPs (142.8 +/- 5.1 vs 132.7 +/- 2.6 mm Hg, p < 0.05); mean 24-h (65.2 +/- 2.6 vs 55.9 +/- 1.9 mm Hg, p < 0.008), daytime (65.6 +/- 2.7 vs 56.6 +/- 2.0 mm Hg, p < 0.01) and sleep PBPs (64.1 +/- 2.7 vs 53.1 +/- 1.9 mm Hg, p < 0.002); 24-h (20.1 +/- 3.8 vs 12.6 +/- 1.8 mm Hg, p < 0.05) and sleep NAUC of SBP (24.6 +/- 4.4 vs 15.3 +/- 2.2 mm Hg, p < 0.03). In the group with SAHS were significantly higher the frequency of abnormal circadian rhythm of SBP (84 vs 57%, p < 0.05) and the speed of morning rise of mean BP (23.3 +/- 5.9 vs 8.5 +/- 2.8 mm Hg/h, p < 0.01). Our results suggest that pts with desaturation signs of SAHS are characterized by unfavourable changes in 24-h BP profile parameters, first of all owning to sleep systolic and pulse blood pressures with alteration of circadian rhythm and high speed of morning rise of BP.


Data augmentation based on multiple oversampling fusion for medical image segmentation.
A high-performance medical image segmentation model based on deep learning depends on the availability of large amounts of annotated training data. However, it is not trivial to obtain sufficient annotated medical images. Generally, the small size of most tissue lesions, e.g., pulmonary nodules and liver tumours, could worsen the class imbalance problem in medical image segmentation. In this study, we propose a multidimensional data augmentation method combining affine transform and random oversampling. The training data is first expanded by affine transformation combined with random oversampling to improve the prior data distribution of small objects and the diversity of samples. Secondly, class weight balancing is used to avoid having biased networks since the number of background pixels is much higher than the lesion pixels. The class imbalance problem is solved by utilizing weighted cross-entropy loss function during the training of the CNN model. The LUNA16 and LiTS17 datasets were introduced to evaluate the performance of our works, where four deep neural network models, Mask-RCNN, U-Net, SegNet and DeepLabv3+, were adopted for small tissue lesion segmentation in CT images. In addition, the small tissue segmentation performance of the four different deep learning architectures on both datasets could be greatly improved by incorporating the data augmentation strategy. The best pixelwise segmentation performance for both pulmonary nodules and liver tumours was obtained by the Mask-RCNN model, with DSC values of 0.829 and 0.879, respectively, which were similar to those of state-of-the-art methods.


DTCO optimizes critical path nets to improve chip performance with timing-aware OPC in deep ultraviolet lithography.
Design technology co-optimization (DTCO) is a potential approach to tackle the escalating expenses and complexities associated with pitch scaling. This strategy offers a promising solution by minimizing the required design dimensions and mitigating the pitch scaling trend. It is worth noting that lithography has played a significant role in dimensional scaling over time. This paper proposes a DTCO flow to reduce the impact of the process variation (PV) band and edge placement error (EPE). First, we performed the digital back-end design of the high-performance processor and got the test layout; second, we executed timing analysis on the test layout to get the critical path net that affects the chip performance; third, we proposed the timing-aware optimized optical proximity correction (OPC) method to optimize the PV band and EPE by adjusting the weights of critical path net merit points, optimizing the generation of the sub-resolution assistant feature, giving tighter EPE specs for merit points on the critical path net, and placing denser merit points as well as denser breakpoints for the critical path net to obtain greater freedom in the OPC process. Finally, it is verified that our proposed DTCO process can significantly reduce the EPE and lead to a slight decrease in the PV band of the chip while maintaining the same process windows.


The Effects of Long-Term 40-Hz Physioacoustic Vibrations on Motor Impairments in Parkinson's Disease: A Double-Blinded Randomized Control Trial.
Recent studies have suggested that vibration therapy may have a positive influence in treating motor symptoms of Parkinson's disease (PD). However, quantitative evidence of the benefits of vibration utilized inconsistent methods of vibration delivery, and to date there have been no studies showing a long-term benefit of 40 Hz vibration in the PD population. The objective of this study was to demonstrate the efficacy of vibration administered via a physioacoustic therapy method (PAT) on motor symptoms of PD over a longer term, completed as a randomized placebo-controlled trial. Overall motor symptom severity measured by the Unified Parkinson's Disease Rating Scale III showed significant improvements in the treatment group over 12 weeks. Specifically, all aspects of PD, including tremor, rigidity, bradykinesia, and posture and gait measures improved. To our knowledge, this is the first study to quantitatively assess 40-Hz vibration applied using the PAT method for potential long-term therapeutic effects on motor symptoms of PD.


Trilateral overlap of tuberculosis, diabetes and HIV-1 in a high-burden African setting: implications for TB control.
The diabetes mellitus burden is growing in countries where tuberculosis (TB) and HIV-1 remain major challenges, threatening TB control efforts. This study determined the association between TB and diabetes/impaired glucose regulation in the context of HIV-1.A cross-sectional study was conducted at a TB clinic in Cape Town (South Africa). Participants were screened for diabetes and impaired glucose regulation using fasting plasma glucose, oral glucose tolerance test and glycated haemoglobin (HbA1c).414 TB and 438 non-TB participants were enrolled. In multivariable analysis, diabetes was associated with TB (OR 2.4, 95% CI 1.3-4.3; p=0.005), with 14% population-attributable risk fraction; however, this association varied by diagnostic test (driven by HbA1c). The association remained significant in HIV-1-infected individuals (OR 2.4, 95% CI 1.1-5.2; p=0.030). A high prevalence of impaired glucose regulation (65.2% among TB cases) and a significant association with TB (OR 2.3, 95% CI 1.6-3.3; p<0.001) was also found.Diabetes and impaired glucose regulation prevalence was high and associated with TB, particularly in HIV-1-infected individuals, highlighting the importance of diabetes screening. The variation in findings by diagnostic test highlights the need for better glycaemia markers to inform screening in the context of TB and HIV-1.


[Food toxicology].
The complex problems of food toxicology and especially of mutagenesis and carcinogenesis require continuing efforts for a better understanding of the mechanisms, risk evaluation and prevention. Essential progress was the recognition of mutation. In vitro tests now provide reproducible results within a short time. Risk evaluation remains a difficult problem, since is has not been possible yet to establish thresholds values for genotoxic substances. However, threshold levels for carcinogen promoters gain increasing importance as demonstrated for 3 representative classes of substances: mycotoxines , nitrosamines and pesticides.


Future concerns and recent life events of elderly community residents.
Fifty-eight elderly community residents completed a 37-item inventory that measured their level of concern about potentially stressful events or emotions. Afterwards, they indicated whether selected life events had actually occurred in the last six months. All data were intercorrelated to determine if there were relationships between total scores, personal characteristics, and recent life events. Results showed elderly persons in this sample to be most worried about immediate economic survival and disability. Those in poorer health who had less education, were living with their children, and had less tenure in the neighborhood evidenced greater total concern.


Devosia subaequoris sp. nov., isolated from beach sediment.
A novel marine bacterium was isolated from a sediment sample from Hwasun Beach in Jeju, Republic of Korea. The cells were found to be Gram-negative, aerobic, oxidase-positive, catalase-positive, motile rods. The organism required natural seawater or artificial sea salts for growth. The temperature and pH ranges for growth were 20-42 degrees C and pH 5.1-12.1, respectively. Phylogenetic analyses based on 16S rRNA gene sequences showed that the organism belonged to the order Rhizobiales and formed a robust cluster with members of the genus Devosia. Its phylogenetic neighbours were the type strains of Devosia riboflavina (96.8 % 16S rRNA gene sequence similarity), Devosia neptuniae (96.7 %), Devosia soli (96.5 %) and Devosia limi (96.2 %), 'Devosia terrae' DCY11 (96.2 %) and 'Candidatus Devosia euplotis' (96.2 %). The predominant ubiquinone was Q-10, the major fatty acids were C(18 : 1), C(18 : 0) and C(16 : 0) and the G+C content of the DNA was 59.1 mol%. On the basis of phenotypic data and the results of phylogenetic analyses, strain HST3-14(T) represents a novel species of the genus Devosia, for which the name Devosia subaequoris sp. nov. is proposed. The type strain is strain HST3-14(T) (=KCTC 12772(T) =JCM 14206(T)).


Ion transport in Hydrodictyon africanum.
The concentrations of K, Na, and Cl in the cytoplasm and vacuole, the tracer fluxes of these ions into and out of the cenocyte, and the electrical potential difference between bathing solution and vacuole and cytoplasm, have been measured in Hydrodictyon africanum. If the ions were acted on solely by passive electrochemical forces, a net efflux of K and Cl and a net influx of Na would be expected. Tracer fluxes indicate a net influx of K and Cl and efflux of Na in the light; these net fluxes are consequently active, with an obligate link to metabolism. The effects of darkness and low temperature indicate that most of the tracer K and Cl influx and Na efflux are linked to metabolism, while the corresponding tracer fluxes in the direction of the free energy gradient are not. Ouabain specifically inhibits the metabolically linked portions of tracer K influx and Na efflux. Alterations in the external K concentration have similar effects on metabolically mediated K influx and Na efflux. It would appear that K influx and Na efflux are linked, at least in the light.


Global and Multi-National Prevalence of Fungal Diseases-Estimate Precision.
Fungal diseases kill more than 1.5 million and affect over a billion people. However, they are still a neglected topic by public health authorities even though most deaths from fungal diseases are avoidable. Serious fungal infections occur as a consequence of other health problems including asthma, AIDS, cancer, organ transplantation and corticosteroid therapies. Early accurate diagnosis allows prompt antifungal therapy; however this is often delayed or unavailable leading to death, serious chronic illness or blindness. Recent global estimates have found 3,000,000 cases of chronic pulmonary aspergillosis, ~223,100 cases of cryptococcal meningitis complicating HIV/AIDS, ~700,000 cases of invasive candidiasis, ~500,000 cases of Pneumocystis jirovecii pneumonia, ~250,000 cases of invasive aspergillosis, ~100,000 cases of disseminated histoplasmosis, over 10,000,000 cases of fungal asthma and ~1,000,000 cases of fungal keratitis occur annually. Since 2013, the Leading International Fungal Education (LIFE) portal has facilitated the estimation of the burden of serious fungal infections country by country for over 5.7 billion people (>80% of the world's population). These studies have shown differences in the global burden between countries, within regions of the same country and between at risk populations. Here we interrogate the accuracy of these fungal infection burden estimates in the 43 published papers within the LIFE initiative.


Astragaloside IV inhibits doxorubicin-induced cardiomyocyte apoptosis mediated by mitochondrial apoptotic pathway via activating the PI3K/Akt pathway.
Doxorubicin (DOX) is a widely used antitumor drug whose application is seriously limited by its cardiotoxicity. Mitochondria-mediated cardiomyocyte apoptosis plays a critical role in DOX-induced cardiotoxicity (DIC). The aim of the present study was to investigate the protective effect of astragaloside IV (3-O-beta-D-xylopyranosyl-6-O-beta-D-glucopyranosyl-cycloastragenol, AS-IV), a pure saponin isolated from Astragalus membranaceus, against DOX-induced cardiomyocyte apoptosis in primary cultured neonatal rat cardiomyocytes. Immunocytochemistry and Microculture Tetrazolium (MTT) assays showed that AS-IV significantly reduced DOX-induced cardiomyocyte loss. Additionally, AS-IV markedly ameliorated DOX-caused cardiomyocyte dysfunction via restoring the beating cell ratio and beating rate in cardiomyocytes. Furthermore, AS-IV substantially reduced the mitochondrial reactive oxygen species (ROS) production and lactate dehydrogenase (LDH), creatine kinase-MB isoenzyme (CK-MB) and cytochrome c (CytC) release, and restored the reduced ATP level, succinate dehydrogenase (SDH) and ATP synthase activities induced by DOX, suggesting that AS-IV significantly attenuated DOX-induced mitochondrial damage and dysfunction. It was further observed that DOX-induced cardiomyocyte apoptosis, as qualitatively evaluated by Hoechst 33258 staining and accurately quantified by flow cytometry, was markedly inhibited by AS-IV. Western blot analysis manifested that AS-IV significantly inhibited the activation of mitochondrial apoptotic pathway (MAP) via inducing the phosphorylation of Akt and Bad. Furthermore, phosphatidylinositol 3-kinase (PI3K) inhibitor 2-(4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride (LY294002) remarkably inhibited the anti-apoptotic effect of AS-IV. Moreover, AS-IV didn't compromise the antitumor activity of DOX. Taken together, our findings indicate that AS-IV ameliorates DIC, and this beneficial effect appears to be dependent on the activation of the PI3K/Akt pathway. Thus, AS-IV may hold promise as an efficient cardioprotective agent against DIC.


Cochlear implantation in the world's largest medical device market: utilization and awareness of cochlear implants in the United States.
Provision of cochlear implants (CIs) for those within the criteria for implantation remains lower in the United States than in some other developed nations. When adults and children are grouped together, the rate of utilization/provision remains low at around 6%. For children, the provision rate is about 50% of those who could benefit from an implant, compared with figures of about 90% for the Flanders part of Belgium, the United Kingdom and other European countries. The probable reasons for this underprovision include: low awareness of the benefits of CIs among the population; low awareness among health-care professionals; the lack of specific referral pathways; some political issues relating to the Deaf Community; and financial issues related to health provision. Such financial issues result in situations which either fail to provide for access to implants or provide too low a level of the necessary funding, especially for low-income individuals covered by public health-care programs such as Medicaid. These issues might be mitigated by adoption and publication of standards for best clinical practices for CI provision, availability of current cost-effectiveness data, and the existence of an organization dedicated to cochlear implantation. Such an organization, the American Cochlear Implant Alliance (ACI Alliance), was recently organized and is described in the paper by Niparko et al. in this Supplement.


Controlled Nano-Bio Interface of Functional Nanoprobes for in Vivo Monitoring Enzyme Activity in Tumors.
Engineering inorganic nanoparticles with a biocompatible shell to improve their physicochemical properties is a vital step in taking advantage of their superior magnetic, optical, and photothermal properties as multifunctional molecular imaging probes for disease diagnosis and treatment. The grafting/peeling-off strategy we developed for nanoparticle surface coating can fully control the targeting capability of functional nanoprobes by changing their colloidal behaviors such as diffusion and sedimentation rates at the desired sites. We demonstrated that a cleavable coating layer initially immobilized on the surface of magnetic resonance imaging probes not only makes the nanoparticles water-soluble but also can be selectively removed by specific enzymes, thereby resulting in a significant decrease of their water solubility in an enzyme-rich environment. Upon removal of surface coating, the changes in hydrodynamic size and surface charges of nanoprobes as a result of interacting with biomolecules and proteins lead to dramatic changes in their in vivo colloidal behaviors ( i. e., slow diffusion rates, tendency to aggregate and precipitate), which were quantitatively evaluated by examining changes in their hydrodynamic sizes, magnetic properties, and count rates during the size measurement. Because the retention time of nanoprobes within the tumor tissues depends on the uptake and excretion rate of the nanoprobes through the tumors, selective activation of nanoprobes by a specific enzyme resulted in much higher tumor accumulation and longer retention time within the tumors than that of the inactive nanoprobes, which passively passed through the tumors. The imaging contrast effect of tumors using activatable nanoprobes was significantly improved over using inactive probes. Therefore, the grafting/peeling-off strategy, as a general design approach for surface modification of nanoprobes, offers a promising and highly efficient way to render the nanoparticles suitable for targeted imaging of tumors.


Diversity and host range of Shiga toxin-encoding phage.
Shiga toxin 2 (Stx2) from the foodborne pathogen Escherichia coli O157:H7 is encoded on a temperate bacteriophage. Toxin-encoding phages from C600::933W and from six clinical E. coli O157:H7 isolates were characterized for PCR polymorphisms, phage morphology, toxin production, and lytic and lysogenic infection profiles on O157 and non-O157 serotype E. coli. The phages were found to be highly variable, and even phages isolated from strains with identical pulsed-field gel electrophoresis profiles differed. Examination of cross-plaquing and lysogeny profiles further substantiated that each phage is distinct; reciprocal patterns of susceptibility and resistance were not observed and it was not possible to define immunity groups. The interaction between Shiga toxin-encoding phage and intestinal E. coli was examined. Lytic infection was assessed by examining Shiga toxin production following overnight incubation with phage. While not common, lytic infection was observed, with a more-than-1,000-fold increase in Stx2 seen in one case, demonstrating that commensal E. coli cells can amplify Shiga toxin if they are susceptible to infection by the Shiga toxin-encoding phages. Antibiotic-resistant derivatives of the Stx2-encoding phages were used to examine lysogeny. Different phages were found to lysogenize different strains of intestinal E. coli. Lysogeny was found to occur more commonly than lytic infection. The presence of a diverse population of Shiga toxin-encoding phages may increase the pathogenic fitness of E. coli O157:H7.


A Japanese Family Meeting the Clinical Diagnostic Criteria for MEN1 with a MEN1 Variant of Uncertain Significance.
Multiple gastroenteric, pancreatic, and pituitary neuroendocrine neoplasms (NENs) were diagnosed in a 74-year-old man with a history of primary hyperparathyroidism (PHPT). Germline testing demonstrated a variant of MEN1 (c.1694T>A, p.L565Q), whose pathogenicity was classified as a variant of uncertain significance (VUS) according to the ACMG/AMP guidelines. The same germline variant was detected in the patient's son and daughter, who also showed PHPT or hypercalcemia and met the clinical diagnostic criteria for multiple endocrine neoplasia Type 1 (MEN1). During surveillance of the son, multiple pancreatic tumors suggestive of NENs were detected. The pathogenicity of the current MEN1 variant was re-evaluated as likely pathogenic, based on additional family data.


Machine learning, the kidney, and genotype-phenotype analysis.
With biomedical research transitioning into data-rich science, machine learning provides a powerful toolkit for extracting knowledge from large-scale biological data sets. The increasing availability of comprehensive kidney omics compendia (transcriptomics, proteomics, metabolomics, and genome sequencing), as well as other data modalities such as electronic health records, digital nephropathology repositories, and radiology renal images, makes machine learning approaches increasingly essential for analyzing human kidney data sets. Here, we discuss how machine learning approaches can be applied to the study of kidney disease, with a particular focus on how they can be used for understanding the relationship between genotype and phenotype.


[Morphological and clinical aspects of the so-called sinus tumours of the ovary (author's transl)].
Data from literature as well as our own studies are used in the discussion of differential diagnosis of the so-called endodermal sinus tumours. Clinical and paraclinical aspects of this once lethal tumour species are reported. New therapeutic strategies are presented consisting of the introduction of new, effective combinations with adjuvant chemotherapy. Thus at early stages a conservative surgical approach seems to allow the preservation of fertility in these generally young women. But even in more advanced cases the administration of a modern adjuvant polychemotherapy is likely to result in higher survival rates.


Megakaryocyte volume modulates bone marrow niche properties and cell migration dynamics.
All hematopoietic cells that develop in the bone marrow must cross the endothelial barrier to enter the blood circulation. Blood platelets, however, are released by bigger protrusions of huge progenitor cells, named megakaryocytes, and enter the blood stream as so-called proplatelets before fragmenting into mature platelets. Recently, a second function of megakaryocytes has been identified, as they modulate the quiescence of hematopoietic stem cells, mostly via different soluble factors. We know from light sheet fluorescence microscopy images that megakaryocytes are distributed throughout the bone marrow facing a dense vascular network. Here, we used such three-dimensional images to provide a realistic simulation template reflecting the in vivo cell-vessel distributions resulting in reliable whole-bone analysis in silico Combining this approach with an automated image analysis pipeline, we found that megakaryocytes influence migration of neutrophils and hematopoietic stem cells, and thus act as biomechanical restrainers modulating cell mobility and extravasation. Indeed, as a consequence of increased megakaryocyte volumes in platelet-depleted mice neutrophil mobility was reduced in these animals.


Neonatal thyroid function born to mothers living with long-term excessive iodine intake from drinking water.
The effects of long-term excessive maternal iodine intake on neonatal thyroid function are less known. This study aimed to assess the effects of maternal excessive iodine intake from drinking water on thyroid functions of both mothers and their neonates. This observational study was performed in high iodine (HI) areas and adequate iodine (AI) intake areas, including 384 healthy pregnant women in late gestation (mean week 39·3 ± 1·6 weeks) and their newborns. Blood and urine samples were obtained from pregnant women, while cord blood samples were obtained from neonates. Urinary iodine concentration (UIC) and thyroid function were evaluated. The median maternal UIC was 1241 and 217 μg/l in HI and AI areas, respectively (P < 0·001). The concentrations of serum thyroid-stimulating hormone (TSH), free triiodothyronine (FT3) and free thyroxine (FT4) in neonates in HI areas were 7·33 mIU/l (range 5·47, 11·06 mIU/l), 2·93 ± 0·59 and 15·03 ± 1·92 pmol/l, respectively, while that were 4·71 mIU/l (range 3·96, 6·04 mIU/l), 2·31 ± 0·28 and 16·50 ± 1·35 pmol/l in AI neonates (P < 0·05). Similar changes were also observed in neonates in HI areas when excluding the effect of maternal thyroid autoimmunity. Cord blood TSH concentration (r = 0·31, P = 0·001) and FT3 concentration (r = 0·43, P = 0·001) were positively correlated with maternal UIC. Cord blood FT4 concentration was negatively correlated with maternal UIC (r = -0·25, P = 0·001). Mothers living in HI areas (β = 0·296, 95% CI: 0·163, 0·255) and with subclinical hypothyroidism (β = 0·360, 95% CI: 0·034, 0·175) contributed to elevated cord blood TSH concentration in neonates, while male neonates were more likely to present with higher TSH concentration compared with female infants (β = -0·760, 95% CI: -0·119, -0·033). Excessive iodine intake during pregnancy was associated with an increased rate of hyperthyrotropinaemia in neonates and their mothers, especially in male neonates.


Virulence-Associated Gene Profiles of Escherichia coli Isolated from Chickens with Colibacillosis in Japan and Their Correlation with Pathogenicity in Chicken Embryos.
Colibacillosis, an infectious disease of chickens, is caused by avian pathogenic Escherichia coli (APEC); however, in addition to APEC, other pathogens are also frequently isolated from chickens affected with colibacillosis. Therefore, experimental infections in chickens are necessary to evaluate the pathogenicity of APEC isolates. Recent studies have shown that embryo lethality assays can be used as an alternative method to evaluate the pathogenicity of E. coli. In this study, to determine the important virulence genes associated with the pathogenicity of E. coli, 67 strains of E. coli that possessed different combinations of eight representative virulence genes (cva/cvi, vat, tsh, iucD, papC, irp2, iss, and astA) were isolated from broilers with colibacillosis in Japan, and the chicken embryo lethal assay was conducted. The genes vat, papC, and irp2 showed strong correlation with the level of virulence in E. coli. Our study provides useful information about the important virulence-associated genes in relation to the pathogenicity of E. coli in Japanese chickens.


Effect of a new fungicide (PNDP) on the minor nucleotide content of Fusarium oxysporum.
The minor nucleotide constituents of rRNA from the plant pathogenic fungus Fusarium oxysporum were investigated. When the fungus was grown for 8 days in aerated liquid culture the following minor nucleotides were identified from 18S and 25S rRNA: m6A, m2A , m2(6)A, m1G, m2G , m2(2)G, m3C , Cm, m5C, m3U , Um, cm5U , rT and psi. PNDP (+/- threo-1-phenyl-2-nitro-1,3 diacetoxy-propane), a new fungicide of unknown biochemical mode of action, caused a great decrease in the minor nucleotide content of the fungus. Only seven minor nucleotides were present in the rRNA of the fungus grown in PNDP -containing medium. A hypermodified nucleotide, mcm5S2U , was found in treated but not in untreated fusarium. On comparing the effect of PNDP with that of known protein synthesis inhibitors such as cycloheximide and chloramphenicol, it was concluded that PNDP acted similarly to cycloheximide, a cytoplasmic protein synthesis inhibitor. It is suggested that the mode of action of PNDP may be the inhibition of the synthesis of RNA modifying enzymes of F. oxysporum.


[Analysis of interventions designed to improve clinical supervision of student nurses in Benin].
The absence of an explicit and coherent conception of the articulation between theory and practice in the reform of nursing training in Benin has resulted in poor quality clinical supervision of student nurses. The objective of this article is to analyze two interventions designed to improve the quality of supervision. A student welcome booklet developed by means of a consultative and provocative participatory approach was tested with twelve student nurses versus a control group. Content analysis of the data collected by individual semi-directed interviews and during two focus groups demonstrated the value of this tool. Student nurses were also taught to use to training diaries inspired by the ?experiential learning? Training diaries were analysed using a grid based on the descriptive elements of the five types of Scheepers training diaries (2008). According to the student nurses, the welcome booklet provided them with structured information to be used as a reference during their training and a better understanding of their teachers, and allowed them to situate the resources of the training course with a lower level of stress. Fifty-eight per cent of the training diaries were are mosaics, reflecting the reflective practice and self-regulated learning of student nurses. This activity also promoted metacognitive dialogue with their supervisors. The student welcome booklet appeared to facilitate integration of student nurses into the clinical setting and promoted professional and organizational socialization. The training diary improved the quality of clinical learning by repeated reflective observation of student nurses and helped to maintain permanent communication with the supervisors.


Ulcer healing effect of autologous mixed sheets consisting of fibroblasts and peripheral blood mononuclear cells in rabbit ischemic hind limb.
We developed mixed cell sheets consisting of fibroblasts and peripheral blood mononuclear cells that had high potency for secreting vascular endothelial growth factor. The purpose of this study was to confirm the therapeutic effects of mixed sheets in rabbits suffering from ulcers at the ischemic hind limbs. We used the ulcer model, which was constructed by implantation and sandwiching the skin between two magnets to be a representative of human refractory cutaneous ulcer. The ulcer healing rate of mixed cell sheets was higher than that of the control at an early stage of healing. The calf blood pressure and angiographic score, which were considered to reflect rough collateral blood flow, did not vary amongmixed cell sheets. However, through laser Doppler perfusion image, implantation of mixed cell sheets revealed a significant improvement in microvascular blood flow in the healed skin of the ischemic limb compared to trafermin, a recombinant human basic fibroblast growth factor, and the control. These results suggest that mixed cell may operate predominantly on the surface of the ischemic tissue by their angiogenic potency, thereby promoting healing of the ischemic ulcer. Mixed cell sheets could become a promising therapeutic material for refractory cutaneous ulcers.


Unusual reactivity of methylene group adjacent to pyridine-2-carboxamido moiety in iron(III) and cobalt(III) complexes.
The Fe(III) and Co(III) complexes of the ligand N-(2-picolyl)picolinamide (pmpH; H represents the dissociable amide hydrogen), namely, [Fe(pmp)(2)]BF(4) (1) and [Co(pmp)(2)]ClO(4) (2), have been synthesized and structurally characterized. The [bond]CH(2)[bond] moiety of pmp(-) in [M(pmp)(2)](+) (M = Fe, Co) is very reactive and is readily converted to carbonyl (C[double bond]O) group upon exposure to dioxygen. Such conversion results in [M(bpca)(2)]ClO(4) complexes (M = Fe (3), Co (5); bpcaH = bis(2-pyridylcarbonyl)amine) which have been characterized by spectroscopy and X-ray diffraction. The structure of 5 is reported here for the first time. The reactivity of the [bond]CH(2)[bond] moiety of pmp(-) has so far precluded the isolation of 1 although other metal complexes of pmp(-) have been reported years ago. The CH(2) --> C[double bond]O transformation arises from the tendency of the coordinated pmp(-) ligand to achieve further conjugation in the ligand framework and provides a better way to synthesize the metal complexes of bpcaH ligand. Reaction of 3 with NaH affords Fe(II) complex [Fe(bpca)(2)] (4) without any reduction of the ligand frame.


Cytogenetic and morphologic typing of 58 papillary renal cell carcinomas: evidence for a cytogenetic evolution of type 2 from type 1 tumors.
We evaluated clinical characteristics, patient outcome (mean follow-up, 47 months), and cytogenetic abnormalities in the largest as yet reported cytogenetic series of 47 primary and 11 secondary papillary renal cell carcinomas for differences between the recently proposed type 1 and type 2 subtypes. Secondary tumors were more often of type 2 morphology (P = 0.02), whereas primary type 2 tumors were associated with higher clinical stage (P = 0.001) and worse patient outcome (P = 0.02). Although both subtypes had at least one of the primary chromosomal gains at 17q, 7, and 16q, type 2 tumors had moderately lower frequencies of primary gains at 17p (61 versus 94%; P = 0.007) and 17q (72 versus 97%; P = 0.02). On the other hand, type 2 tumors overall had more chromosomal alterations than type 1 tumors (P = 0.01), particularly gains of 1q (28 versus 3%; P = 0.02) and losses of 8p (33 versus 0%; P = 0.001), 11 (28 versus 3%; P = 0.02), and 18 (44 versus 9%; P = 0.01). Hierarchical clustering suggested cytogenetic patterns common but not restricted to type 2 morphology, one characterized by multiple additional gains, and another predominantly showing additional losses. These findings provide genetic evidence that type 1 and type 2 tumors arise from common cytogenetic pathways and that type 2 tumors evolve from type 1 tumors. Independently of type, losses of 9p were statistically correlated with advanced disease (P = 0.0008) and may serve as a potential adverse prognostic marker in papillary renal cell carcinomas.


Human alpha-subunit analogs act as partial agonists to the thyroid-stimulating hormone receptor: differential effects of free and yoked subunits.
The alpha-subunit is common to the heterodimeric glycoprotein hormones and has been highly conserved throughout vertebrate evolution. In an effort to determine if wild-type and engineered human alpha analogs can serve as agonists or antagonists to the human thyroid-stimulating hormone (TSH) receptor (TSHR), a potent alpha mutant, obtained by replacing four amino acid residues with lysine (alpha4K), was assayed and compared with the wild-type alpha-subunit. When added to CHO cells expressing TSHR, alpha4K, and to a very limited extent the fused homodimer, alpha4K-alpha4K, but not alpha, exhibited agonist activity as judged by cAMP production. When yoked to TSHR to yield fusion proteins, neither alpha, alpha4K, alpha-alpha, nor alpha4K-alpha4K activated TSHR, although yoked alpha4K and alpha4K-alpha4K were weak inhibitors of TSH binding to TSHR. The yoked subunit-receptor complexes were, however, functional as evidenced by increased cAMP production in cells co-expressing human TSHbeta and alpha-TSHR, alpha4K-TSHR, alpha-alpha-TSHR, and alpha4K-alpha4K-TSHR. These results demonstrate that agonists to TSHR can be obtained with alpha-subunit analogs and suggest that rational protein engineering may lead to more potent alpha-based derivatives. The differences found between the experimental paradigms of adding free alpha analogs to TSHR and covalent attachment are attributed to con-formational constraints imposed by fusion of the alpha-subunit analog and receptor, and may suggest an important role for a free (C-terminal) alpha-carboxyl in the absence of the beta-subunit.


Ergosteryl 2-naphthoate, An Ergosterol Derivative, Exhibits Antidepressant Effects Mediated by the Modification of GABAergic and Glutamatergic Systems.
Phytosterols are a kind of natural component including sitosterol, campesterol, avenasterol, ergosterol (Er) and others. Their main natural sources are vegetable oils and their processed products, followed by grains, by-products of cereals and nuts, and small amounts of fruits, vegetables and mushrooms. In this study, three new Er monoester derivatives were obtained from the reflux reaction with Er: organic acids (furoic acid, salicylic acid and 2-naphthoic acid), 1-Ethylethyl-3-(3-dimethyllaminopropyl) carbodiimide hydrochloride (EDCI) and 4-dimethylaminopyridine (DMAP) in dichloromethane. Their chemical structures were defined by IR and NMR. The present study was also undertaken to investigate the antidepressant-like effects of Er and its derivatives in male adult mice models of depression, and their probable involvement of GABAergic and glutamatergic systems by the forced swim test (FST). The results indicated that Er and its derivatives display antidepressant effects. Moreover, one derivative of Er, ergosteryl 2-naphthoate (ErN), exhibited stronger antidepressant activity in vivo compared to Er. Acute administration of ErN (5 mg/kg, i.p.) and a combination of ErN (0.5 mg/kg, i.p.), reboxetine (2.5 mg/kg, i.p.), and tianeptine (15 mg/kg, i.p.) reduced the immobility time in the FST. Pretreatment with bicuculline (a competitive γ-aminobutyric acid (GABA) antagonist, 4 mg/kg, i.p.) and N-methyl-d-aspartic acid (NMDA, an agonist at the glutamate site, 75 mg/kg, i.p.) effectively reversed the antidepressant-like effect of ErN (5 mg/kg, i.p.). However, prazosin (a α1-adrenoceptor antagonist, 1 mg/kg, i.p.) and haloperidol (a non-selective D2 receptor antagonist, 0.2 mg/kg, i.p.) did not eliminate the reduced immobility time. Altogether, these results indicated that ErN produced antidepressant-like activity, which might be mediated by GABAergic and glutamatergic systems.


The Usefulness of Surgical Treatment in Slow-Flow Vascular Malformation Patients.
Many difficulties exist in establishing a treatment plan for slow-flow vascular malformation (SFVM). In particular, little research has been conducted on the surgical treatment of SFVMs. Thus, we investigated what proportion of SFVM patients were candidates for surgical treatment in clinical practice and how useful surgical treatment was in those patients. This study included 109 SFVM patients who received care at the authors' clinic from 2007 to 2015. We classified the patients as operable or non-operable, and analyzed whether the operability and the extent of the excision varied according to the subtype and location of the SFVM. Additionally, we investigated complications and self-assessed satisfaction scores. Of the 109 SFVM patients, 59 (54%) were operable, while 50 (46%) were nonoperable. Total excision could be performed in 44% of the operable SFVM patients. Lymphatic malformations were frequently non-operable, while capillary malformations were relatively operable (P=0.042). Total excision of venous malformations could generally be performed, while lymphatic malformations and combined vascular malformations generally could only undergo partial excision (P=0.048). Complications occurred in 11% of the SFVM patients who underwent surgery; these were minor complications, except for 1 case. The average overall satisfaction score was 4.19 out of 5. Based on many years of experience, we found that approximately half (54%) of SFVM patients were able to undergo surgery, and around half (44%) of those patients were able to fully recover after a total excision. Among the patients who underwent surgical treatment, high satisfaction was found overall and relatively few complications were reported.


Sleep-potentiated epileptiform activity in early thalamic injuries: Study in a large series (60 cases).
The study aims at a better definition of continuous spike-waves during sleep (CSWS) with an early thalamic lesion, focusing on various grades of sleep-potentiated epileptiform activity (SPEA). Their possible relationship with different clinical features was studied to try to define prognostic factors of the epileptic disorder, especially relating to behavior/cognitive outcome, in order to improve prevention and treatment strategies. Sixty patients with early thalamic injury were followed since the first registration of SPEA with serial neurological, long term EEG monitoring and neuropsychological examinations, as well as neuroimaging and a detailed clinical history. They were classified in three different groups according to the sleep spike-waves (SW) quantification: electrical status epilepticus during sleep (ESES), more than 85% of slow sleep; overactivation between 50% and 85% and simple activation between 10 and 50%). Results were then examined also with a statistical analysis. In our series of CSWS occurring in early brain injured children with unilateral thalamic involvement there is a common neuropathologic origin but with various grades of SPEA severity. Statistical analysis showed that patients evolving toward ESES presented more commonly the involvement of the mediodorsal part of thalamus nuclei and a bilateral cortico-subcortical brain injury, epilepsy was more severe with a delayed onset; moreover, in the acute stage .ESES patients presented the worst behavior/cognitive performances. As to cognitive and behavior outcome, longer SPEA duration as well as bilateral brain injury and cognitive/behavior impairment in acute phase appear linked to a poor outcome; some particular neuropathology (ischemic stroke and haemorrhagic infarction) as well as hydrocephalus shunting are associated with behavior disorders. Discrete features seem to support different underlying mechanisms in ESES patients in comparison with less severe SPEA; they represent negative prognostic factors. Longer SPEA duration as well as bilateral brain injury and cognitive/behavior impairment in acute phase seem predictive of a worse cognitive/behavior outcome.


Allogenic Adipose Tissue-Derived Stromal/Stem Cells and Vitamin D Supplementation in Patients With Recent-Onset Type 1 Diabetes Mellitus: A 3-Month Follow-Up Pilot Study.
Objective: To evaluate the short term safety and potential therapeutic effect of allogenic adipose tissue-derived stromal/stem cells (ASCs) + cholecalciferol in patients with recent-onset T1D. Methods: Prospective, phase II, open trial, pilot study in which patients with recent onset T1D received ASCs (1 × 106 cells/kg) and cholecalciferol 2000 UI/day for 3 months (group 1) and were compared to controls with standard insulin therapy (group 2). Adverse events, C-peptide (CP), insulin dose, HbA1c, time in range (TIR), glucose variability (continuous glucose monitoring) and frequency of CD4+FoxP3+ T-cells (flow cytometry) were evaluated at baseline (T0) and after 3 months (T3). Results: 13 patients were included (8: group 1; 5: group 2). Their mean age and disease duration were 26.7 ± 6.1 years and 2.9 ± 1.05 months. Adverse events were transient headache (n = 8), mild local reactions (n = 7), tachycardia (n = 4), abdominal cramps (n = 1), thrombophlebitis (n = 4), mild floaters (n = 2), central retinal vein occlusion (n = 1, complete resolution). At T3, group 1 had lower insulin requirement (0.22 ± 0.17 vs. 0.61±0.26IU/Kg; p = 0.01) and HbA1c (6.47 ± 0.86 vs. 7.48 ± 0.52%; p = 0.03) than group 2. In group 1, 2 patients became insulin free (for 4 and 8 weeks) and all were in honeymoon at T3 (vs. none in group 2; p = 0.01). CP variations did not differ between groups (-4.6 ± 29.1% vs. +2.3 ± 59.65%; p = 0.83). Conclusions: Allogenic ASCs + cholecalciferol without immunosuppression was associated with stability of CP and unanticipated mild transient adverse events in patients with recent onset T1D. ClinicalTrials.gov registration: NCT03920397.


Changing characteristics of the empathic communication network after empathy-enhancement program for medical students.
The Empathy-Enhancement Program for Medical Students (EEPMS) comprises five consecutive weekly sessions and aims to improve medical students' empathic ability, an essential component of humanistic medical professionalism. Using a graph theory approach for the Ising network (based on l1-regularized logistic regression) comprising emotional regulation, empathic understanding of others' emotion, and emotional expressivity, this study aimed to identify the central components or hubs of empathic communication and the changed profile of integration among these hubs after the EEPMS. Forty medical students participated in the EEPMS and completed the Depression Anxiety Stress Scale-21, the Empathy Quotient-Short Form, the Jefferson Scale of Empathy, and the Emotional Expressiveness Scale at baseline and after the EEPMS. The Ising model-based network of empathic communication was retrieved separately at two time points. Agitation, self-efficacy for predicting others' feelings, emotional concealment, active emotional expression, and emotional leakage ranked in the top 20% in terms of nodal strength and betweenness and closeness centralities, and they became hubs. After the EEPMS, the 'intentional emotional expressivity' component became less locally segregated (P = 0.014) and more directly integrated into those five hubs. This study shows how to quantitatively describe the qualitative item-level effects of the EEPMS. The key role of agitation in the network highlights the importance of stress management in preserving the capacity for empathic communication. The training effect of EEPMS, shown by the reduced local segregation and enhanced integration of 'intentional emotional expressivity' with hubs, suggests that the EEPMS could enable medical students to develop competency in emotional expression, which is an essential component of empathic communication.


Zika Virus: A Critical Analysis and Pharmaceutical Perspectives.
Zika virus belongs to family of viruses 'Flaviviridae' and spreads mostly by daytime-active Aedes mosquitoes. The resulting infection is known as Zika fever. It is usually asymptomatic or often causes mild symptoms, which are very similar to dengue fever. The Aedes aegypti mosquito is also responsible for dengue and chikungunya viruses. Zika virus can spread by crossing the placental barrier from a pregnant mother to a fetus, which can result in microcephaly, severe brain malformations including Guillain-Barre syndrome (GBS), and other birth defects. Until now, there is no specific treatment of Zika fever disease, and Zika virus illness cannot be prevented by medications or vaccines. According to WHO, no vaccine is likely to be available until 2020. The only way of preventing this disease is to prevent the mosquito bites. This article presents the history of Zika virus, its reported cases including microcephaly and GBS, and a comparison of its symptoms with those of dengue and chikungunya diseases, as well as preventive measures. With advances in research and technology, knowledge about the Zika virus has grown, yet some questions remain unanswered regarding Zika virus's genetic diversity, pathophysiology, transmission vectors and reservoirs, potential synergetic of coinfection with other related arboviruses, and treatment. These problems highlight the need for further research to achieve adequate the surveillance, infection management, optimized treatment, and public health mediations in Zika virus outbreaks. This article contributes to our understanding of the disease mechanism, genome structure, diagnosis, transmission, and preventive strategies to combat Zika virus infection.


Management of early vocal cord carcinoma.
Endoscopic stripping or laser therapy are the primary modalities for patients with premalignant vocal cord lesions and patients with T1 carcinomas of the mid-portion of the vocal cord. Partial laryngectomy is advocated for patients whose resection margins are positive following laser cordectomy, or where tumor extends to the arytenoid, anterior commissure, opposite vocal cord or subglottis. Radiotherapy is reserved for patients with tumors involving the interarytenoid region, professional voice patients, or patients unfit for general anesthesia.


Growth kinetics of Listeria monocytogenes in broth and beef frankfurters--determination of lag phase duration and exponential growth rate under isothermal conditions.
The objective of this study was to develop a new kinetic model to describe the isothermal growth of microorganisms. The new model was tested with Listeria monocytogenes in tryptic soy broth and frankfurters, and compared with 2 commonly used models-Baranyi and modified Gompertz models. Bias factor (BF), accuracy factor (AF), and root mean square errors (RMSE) were used to evaluate the 3 models. Either in broth or in frankfurter samples, there were no significant differences in BF (approximately 1.0) and AF (1.02 to 1.04) among the 3 models. In broth, the mean RMSE of the new model was very close to that of the Baranyi model, but significantly lower than that of the modified Gompertz model. However, in frankfurters, there were no significant differences in the mean RMSE values among the 3 models. These results suggest that these models are equally capable of describing isothermal bacterial growth curves. Almost identical to the Baranyi model in the exponential and stationary phases, the new model has a more identifiable lag phase and also suggests that the bacteria population would increase exponentially until the population approaches to within 1 to 2 logs from the stationary phase. In general, there is no significant difference in the means of the lag phase duration and specific growth rate between the new and Baranyi models, but both are significantly lower than those determined from the modified Gompertz models. The model developed in this study is directly derived from the isothermal growth characteristics and is more accurate in describing the kinetics of bacterial growth in foods.


[Expression and Significance of Notch1, P21WAF1 and involucrin in nasopharyngeal carcinoma].
Abnormal expression of Notch1 protein was often found in many kinds of primary tumors, but its correlation with nasopharyngeal carcinoma (NPC) is still unclear. This study was designed to investigate the expression of Notch1 and its downstream proteins P21(WAF1) and Involucrin in NPC, and analyze their correlations with the differentiation of NPC cells. The expression of Notch1, P21(WAF1), and Involucrin in 101 specimens of NPC and 20 specimens of chronic inflammatory nasopharyngeal mucosa were detected by SP immunohistochemistry. The positive rates of Notch1, P21(WAF1), and Involucrin were 100%, 90.0%, and 100% in chronic inflammatory nasopharyngeal mucosa, and were 77.2%, 89.1%, and 80.1% in NPC, respectively. The expression of Notch1, P21(WAF1), and Involucrin were significantly suppressed along with descending differentiation of NPC (P=0.000, P=0.026, and P=0.000). The positive rates of Notch1, P21(WAF1), and Involucrin were significantly higher in keratinizing squamous cell carcinoma (KSCC) than in differentiated nonkeratinizing carcinoma (DNKC) and undifferentiated carcinoma (UDC) (P<0.05), and were significantly higher in DNKC than in UDC (P<0.05). The expression of Notch1 in NPC was positively correlated with the expression of P21(WAF1) (r=0.306, P=0.002) and Involucrin (r=0.325, P=0.001). No significant correlation was found between the expression of P21(WAF1) and Involucin. The expression of Notch1, P21(WAF1), and Involucrin are closely correlated to the differentiation of NPC cells.


Bone Grafts, Substitutes, and Augments in Benign Orthopaedic Conditions Current Concepts.
Musculoskeletal tumors are relatively rare diagnoses made by orthopaedic surgeons. While approximately 2,500 primary bone sarcomas are diagnosed annually in the USA, the number of benign orthopaedic tumors encountered annually is far more difficult to quantify. Some studies have documented between 3% and 10% of the general population having benign bony lesions. Many of these conditions can be simply observed, while others will require surgical intervention. Surgical treatments for benign conditions range from a one-step curettage to extensive resection and reconstruction. With treatment of larger lesions, significant bony defects may need to be addressed surgically. Treatment options have evolved over time with the use of various bone graft and bone void fillers, including methyl methacrylate cement, autograft, allograft bone chips, struts and osteoarticular segments, synthetic bone graft substitutes, and metal augments. This review provides an overview of the present status of bone graft, substitutes, and augment options for the orthopaedic surgeon treating benign musculoskeletal conditions.


Regenerative growth of corticospinal tract axons via the ventral column after spinal cord injury in mice.
Studies that have assessed regeneration of corticospinal tract (CST) axons in mice after genetic modifications or other treatments have tacitly assumed that there is little if any regeneration of CST axons in normal mice in the absence of some intervention. Here, we document a previously unrecognized capability for regenerative growth of CST axons in normal mice that involves growth past the lesion via the ventral column. Mice received dorsal hemisection injuries at thoracic level 6-7, which completely transect descending CST axons in the dorsal and dorsolateral column. Corticospinal projections were traced by injecting biotinylated dextran amine (BDA) into the sensorimotor cortex of one hemisphere either at the time of the injury or 4 weeks after injury, and mice were killed at 20-23 or 46 d after injury. At 20-23 d after injury, BDA-labeled CST axons did not extend past the lesion except in one animal. By 46 d after injury, however, a novel population of BDA-labeled CST axons could be seen extending from the gray matter rostral to the injury into the ventral column, past the lesion, and then back into the gray matter caudal to the injury in which they formed elaborate terminal arbors. The number of axons with this highly unusual trajectory was small ( approximately 1% of the total number of labeled CST axons rostral to the injury). The BDA-labeled axons in the ventral column were on the same side as the main tract and thus are not spared ventral CST axons (which would be contralateral to the main tract). These results indicate that normal mice have a capacity for CST regeneration that has not been appreciated previously, which has important implications in studying the effect of genetic or pharmacological manipulations on CST regeneration in mice.


Oral Spindle Cell Sarcoma: A Rare Case Report and Review of Literature.
Spindle cell lesions of the head and neck region are diverse in nature by means of clinical and biological heterogeneity. Though few lesions are found to be malignant, several others are benign or merely reactive in nature. Although these lesions are fairly common occurring in other parts of the body, they are very rare in the oral cavity, accounting for less than 1% of all tumours in the oral region. Herein, a case of 48-year-old man who presented with a polypoid lesion of the maxilla has been reported. Histopathological examination and immunohistochemistry revealed spindle cell sarcoma of the left maxillary sinus. We present this rare tumour to contribute to the better understanding and awareness of this rare malignancy where diagnosis can be very challenging.


Effect of different eye speculums on intra ocular pressure.
To measure the difference in intraocular pressure with use of two different types of eye speculums. The study was conducted at the Department of Ophthalmology, Abbasi Shaheed Hospital, Karachi, from November 2011 to May 2012. Through probability convenience sampling, hundred patients were selected and divided into two equal groups. Complete history and ocular examination was done. Intraocular pressure was measured using Goldmann Applanation tonometer on slit lamp. It was again measured with Barraquer wire speculum inserted in the eyes of 50 patients, and Universal eye speculum in the other 50 patients with the help of Goldmann Applanation. SPSS 16 was used for statistical analysis. There were 63 (65%) males and 34 (35%) females with an overall mean age of 45 +/- 14.09 years. The mean age baseline intraocular pressure of the right eye without speculum was 15.04 +/- 3.83 mmHg and that of the left eye was 15.48 +/- 4.00 mmHg. With Barraquer wire speculum, the value for the right eye was 22.21+/- 4.99, and that of the left eye was 23.08 +/- 5.41. With Universal eye speculum, the value for the right eye was 25.51 +/- 3.02, and for the left eye it was 25.78 +/- 3.64.There was a statistically significant difference in baseline and post-speculum measurements (p < 0.0001). Paired sample t-test was also conducted for values of the two speculums and the result was statistically significant (p < 0.0001). Barraquer wire speculum caused less increase in intraocular pressure compared to the Universal eye speculum. It should be preferred for intraocular surgeries to avoid increase in pressure during surgery and prolapse of ocular contents. Not only the effect of speculum, but also the type of speculum should be kept in mind when measuring intraocular pressure during examination under anaesthesia.


The limits of parental authority to accept or refuse medical treatment.
The legal and ethical right of parents to refuse medical treatment for their children differs from the authority possessed by competent adults with decisional capacity. Parents have a duty to act in the best interests of their children from the children's perspective and not to inflict harm. Best interests are determined by weighing benefits and burdens, which includes using evidence-based outcomes and value judgments. The result is placed along a risk/benefit spectrum. If the result is close to low risk/high benefit, the parents have a strong obligation to accept a health care team recommendation. Otherwise, parents may choose between reasonable medical options without threat or coercion.