Transient receptor potential A1 (TRPA1) activity in the human urethra--evidence for a functional role for TRPA1 in the outflow region.
A role for the transient receptor potential (TRP) A1 ion channel in rat lower urinary tract (LUT) sensation and disease has been proposed, but in the human LUT no information on TRPA1 activity is available. To investigate the distribution of TRPA1 in the human urethra and to study the effect of TRPA1 agonists on isolated urethral strip preparations. Urethral specimens were obtained preoperatively from 10 patients and were freshly prepared for Western blot, immunohistochemistry, and functional in vitro investigations. The expression patterns of TRPA1 were studied with Western blot and immunohistochemistry. The effects of allyl isothiocyanate (AI), cinnamaldehyde (CA), and NaHS (donor of H(2)S) on tension of urethral strips were investigated in tissue baths. TRPA1 immunoreactivity (-IR) was found in nerve fibres in the suburothelial space and was also located to nerve fibres of the muscle layer. Single TRPA1-IR nerves extended into the urothelium. A majority, but not all TRPA1-IR nerves also expressed immunoreactivity for CGRP or TRPV1. In the urothelium, TRPV1 was located to the outer layers whereas TRPA1 was observed in basal urothelial cells. Interspersed between strands of smooth muscle cells of the urethral wall, TRPA1- and vimentin-IR cells containing central nuclei and slender cytoplasmatic extensions were observed. In functional experiments, TRPA1-agonists had no contractile effect in urethral preparations. After precontraction with phenylephrine, AI, CA, and NaHS caused concentration-dependent relaxations of urethral strip preparations. The localization of TRPA1 to nerves that also express TRPV1 and CGRP, and in urothelial cells and interstitial cells, as well as the findings that TRPA1 agonists can modify tone of urethral preparations, propose a role for TRPA1 in afferent and efferent sensory signaling of the human outflow region.


Retrotransposon activation during Drosophila metamorphosis conditions adult antiviral responses.
Retrotransposons are one type of mobile genetic element that abundantly reside in the genomes of nearly all animals. Their uncontrolled activation is linked to sterility, cancer and other pathologies, thereby being largely considered detrimental. Here we report that, within a specific time window of development, retrotransposon activation can license the host's immune system for future antiviral responses. We found that the mdg4 (also known as Gypsy) retrotransposon selectively becomes active during metamorphosis at the Drosophila pupal stage. At this stage, mdg4 activation educates the host's innate immune system by inducing the systemic antiviral function of the nuclear factor-κB protein Relish in a dSTING-dependent manner. Consequently, adult flies with mdg4, Relish or dSTING silenced at the pupal stage are unable to clear exogenous viruses and succumb to viral infection. Altogether, our data reveal that hosts can establish a protective antiviral response that endows a long-term benefit in pathogen warfare due to the developmental activation of mobile genetic elements.


Group IVA phospholipase A2-mediated production of fibronectin by oxidized LDL in mesangial cells.
The deposition of atherogenic lipoproteins such as oxidized low-density lipoprotein (oxLDL) within the mesangium is involved in the overproduction of extracellular matrix proteins, a key event in the progression of glomerular diseases including glomerulosclerosis. To clarify the mechanisms underlying the oxLDL-induced production of extracellular matrix proteins, we examined the possible involvement of group IVA phospholipase A(2) (PLA(2)) using human mesangial cells and group IVA PLA(2)-deficient mouse mesangial cells. oxLDL accelerated the production of fibronectin and collagen (type IV), components of extracellular matrix proteins, with the preceding release of arachidonic acid. Methyl arachidonyl fluorophosphonate (MAFP), known as an inhibitor of group IVA PLA(2), markedly suppressed the oxLDL-induced production of fibronectin as well as the release of arachidonic acid, whereas it did not inhibit the production of collagen. The inhibitory effect of MAFP on the production of fibronectin was reversed by adding arachidonic acid and 12-hydroxyeicosatetraenoic acid. Furthermore, we found that in group IVA PLA(2)-deficient mouse mesangial cells, the production of fibronectin in response to oxLDL was weak as compared with that in wild-type cells. However, the production by oxLDL of collagen was not suppressed in the group IVA PLA(2)-deficient cells. These findings suggest that group IVA PLA(2) is involved in the production of fibronectin in oxLDL-stimulated mesangial cells.


Effectiveness of Paint Psoralen and Ultraviolet-A in Alopecia Areata - Our Experience in the National Skin Center.
Alopecia areata (AA) is usually a benign cause of patchy hair loss that often resolves within a few weeks to months. Most treatment modalities are ineffective in the treatment of severe AA. The use of paint psoralen and ultraviolet-A (PUVA) in the treatment of patients with severe forms of AA has been reported in the literature. The aim of this study was to evaluate the effectiveness of paint PUVA therapy in the treatment of AA in Singapore. We performed a 10-year retrospective analysis of patients who underwent paint PUVA for AA. We evaluated patient demographics and treatment outcomes in the form of percentage change in baseline severity of alopecia tool score and final amount of hair regrowth and relapse rate. Ten patients were included in this study. With paint PUVA therapy, significant hair regrowth was seen in six patients. Paint PUVA therapy in our study showed minimal side effects. PUVA gives fair response in AA in a reasonable time as per our center's experience in Singapore.


Ultrasound stimulation attenuates resorption of tooth root induced by experimental force application.
Root resorption is an adverse outcome of orthodontic tooth movement. However, there have been no available approaches for the protection and repair of root resorption. The aim of this study was to evaluate the effects of low-intensity pulsed ultrasound (LIPUS) on root resorption during experimental tooth movement and the effects of LIPUS in the RANKL/OPG mechanism in osteoblasts and cementoblasts in vitro. Twenty four Wistar strain male rats of 12-week-old were used in this study. The upper first molars were subjected to experimental movement in the mesial direction for 1-3weeks. Through the experimental periods, the right upper first maxillary molar was exposed to LIPUS (LIPUS group) every day for 1, 2 or 3weeks. The nature of root resorption was observed and then quantified by histomorphometric analysis. In the 2weeks period, significantly greater amount of tooth movement was observed in the LIPUS group (p<0.05). In addition, LIPUS group showed less root resorption lacunae and lower number of odontoclasts. In the period of 3weeks, LIPUS group presented significantly shorter length of root resorption lacunae and smaller amount of root resorption area (p<0.01). The number of odontoclasts and osteoclasts was also significantly lower in the LIPUS group (p<0.01 and p<0.05, respectively). However, no significant differences could be found regarding the amount of tooth movement. It is shown that LIPUS exposure significantly reduced the degree of root resorption during tooth movement without interrupting tooth movement. In vitro experiments showed that MC3T3-1 constitutively expressed higher levels of RANKL and RANTES mRNA comparing to OCCM-30. However, OPG mRNA expression was much higher in OCCM-30. LIPUS stimulation significantly increased the mRNA expression of RANKL in MC3T3-E1 at 4 (p<0.01) and 12h (p<0.05), although OPG mRNA expression was not affected by LIPUS. In contrast, the expression of RANKL and OPG mRNAs were both significantly increased by LIPUS in OCCM-30 at 12h (p<0.01). Moreover, LIPUS application suppressed the up-regulation of RANKL mRNA induced by compression force in OCCM-30, but no similar effect could be observed in MC3T3-E1. In conclusion, it is suggested that LIPUS exposure significantly reduces root resorption by the suppression of cementoclastogenesis by altering OPG/RANKL ratio during orthodontic tooth movement without interfering tooth movement. LIPUS may be an effective tool to prevent root resorption during tooth movement and is applicable to clinical use in near future.


Confining donor conformation distributions for efficient thermally activated delayed fluorescence with fast spin-flipping.
Fast spin-flipping is the key to exploit the triplet excitons in thermally activated delayed fluorescence based organic light-emitting diodes toward high efficiency, low efficiency roll-off and long operating lifetime. In common donor-acceptor type thermally activated delayed fluorescence molecules, the distribution of dihedral angles in the film state would have significant influence on the photo-physical properties, which are usually neglected by researches. Herein, we find that the excited state lifetimes of thermally activated delayed fluorescence emitters are subjected to conformation distributions in the host-guest system. Acridine-type flexible donors have a broad conformation distribution or bimodal distribution, in which some conformers feature large singlet-triplet energy gap, leading to long excited state lifetime. Utilization of rigid donors with steric hindrance can restrict the conformation distributions in the film to achieve degenerate singlet and triplet states, which is beneficial to efficient reverse intersystem crossing. Based on this principle, three prototype thermally activated delayed fluorescence emitters with confined conformation distributions are developed, achieving high reverse intersystem crossing rate constants greater than 106 s-1, which enable highly efficient solution-processed organic light-emitting diodes with suppressed efficiency roll-off.


Immunotherapy of melanoma.
The rationale for immunotherapy of human melanoma is based on the knowledge acquired in the molecular characterization of T cell defined antigens which are recognized in vitro by patients' lymphocytes. Based on this information, active immunotherapy (vaccination) and adoptive immunotherapy trials were conducted in metastatic melanoma patients. This review highlights the most important clinical studies and discuss their limits, in terms of both immune and clinical response considering the formulation of the vaccine (cellular, peptide/protein; DNA, etc.) or the features of immune cells used in adoptive immunotherapy. This new knowledge, along with that of escape mechanisms, should allow to improve significantly the clinical response rate in the immunotherapy of melanoma.


Dose-rate dependence of heat radiosensitization.
The dose rate dependence of heat radiosensitization was studied using rat astrocytoma cells in culture and a clinically relevant protocol of heat dose and heat radiation sequence. Cells were treated with a minimally toxic heat dose of 43 degrees C for 30 minutes, after which they were irradiated with varying doses of radiation at dose rates ranging from 0.567 to 300 cGy/min. This heat dose substantially reduced the extrapolation number (n), but had little effect on Do of the radiation survival curve at dose rates of 50 cGy/min or greater. At dose rates less than 10 cGy/min, 43 degrees C for 30 min had little effect on n and only for the lowest dose rate studied (0.567 cGy/min) was there a significant reduction in Do (60%). The thermal enhancement ratio did not vary inversely with radiation dose rate over the dose rate range studied but, instead, was maximal at the two dose rate extremes (0.567 and 300 cGy/min). These data demonstrate that a clinically relevant heat dose enhances very low dose rate, as well as high dose rate, ionizing radiation, but suggest that little benefit is to be gained from using dose rates intermediate between conventional radiotherapeutic high dose rates or dose rates representative of interstitial implants.


Associations Between Children's Telomere Length, Parental Intrusiveness, and the Development of Early Externalizing Behaviors.
Shorter telomeres mark cellular aging and are linked to chronic stress exposure as well as negative physical and psychological outcomes. However, it is unclear whether telomere length mediates associations between early stress exposure and later externalizing problems, or whether boys and girls differ in pathways to these concerns. We therefore examined associations between telomere length, early stress via negative caregiving, and children's externalizing symptom development over time in 409 three-year-old children and their parents. Telomere length mediated the association between early parental intrusiveness and later rule-breaking behavior; however, this association was moderated by children's biological sex such that parent intrusiveness was related only to boys' rule-breaking. Findings support the notion that children's telomere length may mark individual differences in responses to negative early caregiving, and highlight a potential mechanism contributing to the development of rule-breaking problems in boys.


Pharmacokinetics of midazolam in neonates undergoing extracorporeal membrane oxygenation.
Although the pharmacokinetics of midazolam in critically ill children has been described, there are no such reports in extracorporeal membrane oxygenation. The pharmacokinetics of midazolam and 1-hydroxy midazolam after continuous infusion (50-250 microg. kg(-1). h(-1)) were determined in 20 neonates undergoing extracorporeal membrane oxygenation. Patients were randomized into two groups: group 1 (n = 10) received midazolam extracorporeally (into the circuit), and group 2 received drug via central or peripheral access. Blood samples for determination of plasma concentrations were taken at baseline, 2, 4, 6, 12, 18, and 24 h, then every 12 h. Population pharmacokinetic analysis and model building was conducted using WinNonMix (Pharsight Corporation, Mountain View, CA). The 1-hydroxy midazolam/midazolam metabolic ratio was determined as a surrogate marker of cytochrome P450 3A activity. The parameter estimates (n = 19) were based on a one-compartment model with time-dependent change in volume of distribution. Volume (mean +/- standard error) expanded monoexponentially from the onset of extracorporeal membrane oxygenation to a maximum value, 0.8 l +/- 0.5 and 4.1 +/- 0.5 l/kg, respectively. Consequently, plasma half-life was substantially prolonged (median [range]) from onset to steady-state: 6.8 (2.2-39.8) and 33.3 (7.4-178) h, respectively. Total body clearance was determined as (mean +/- standard error) 1.4 +/- 0.15 ml. kg-1. min-1. The median metabolic ratio was 0.17 (0.03-0.9). No significant differences were observed between the two groups with respect to parameter estimates. Simulations of plasma concentration profiles revealed excess levels at conventional doses. These results reveal significantly increased volume of distribution and plasma half-life in neonates receiving extracorporeal membrane oxygenation. Altered kinetics may reflect sequestration of midazolam by components of the extracorporeal membrane oxygenation circuit.


Regulation of gene expression in adult rat hepatocytes cultured on a basement membrane matrix.
Freshly isolated adult rat hepatocytes, when cultured on type I collagen (commercially available as Vitrogen), assume a polygonal shape, form a stable monolayer within 24 hours, but lose the capacity to express some liver-specific functions over time in culture. We incubated hepatocytes in a serum-free medium on a reconstituted basement membrane gel, "matrigel" (prepared from an extract of extracellular matrix of the murine Engelbreth-Holm-Swarm sarcoma), and observed that the cells adhered firmly, remained rounded as single cells or clusters, and maintained liver-specific gene expression for more than 1 week in vitro. Hepatocytes on matrigel secreted substantially higher amounts of albumin, transferrin, haptoglobin, and hemopexin, Northern blot analyses of extracted cellular RNA, expressed increased amounts of mRNA for the liver-specific protein albumin (as compared with cells on vitrogen). In cultures treated with phenobarbital, cytochrome P-450b, and cytochrome P-450e, mRNAs and proteins were barely detectable in cells on Vitrogen but were induced to levels similar to those in the liver in vivo in matrigel cultures. Likewise, the use of matrigel greatly enhanced the induction of mRNA and protein for P-450c by 3-methylcholanthrene and for P-450p by steroidal and nonsteroidal inducers. However, neither substratum permitted induction of P-450d by 3-methylcholanthrene, suggesting that the effects of matrigel are selective even for expression in liver of members of the superfamily of cytochrome P-450 genes. Within 5 days in cultures on Vitrogen, hepatocytes expressed detectable amounts of fetal liver aldolase activity and also mRNA for vimentin and type I collagen, each considered a phenotypic change reflecting hepatocyte "dedifferentiation." None of these was present in cells on matrigel. Responsiveness to mitogenic stimuli, as judged by incorporation of 3H-thymidine into DNA, was also decreased in hepatocytes cultured on matrigel. Finally, there was a remarkable increase in the levels of both matrices during the first 2 days in culture. However, the continuously cytoskeleton mRNA over time in culture than did the rounded cells on matrigel. We conclude that hepatocytes cultured on matrigel, as opposed to the standard collagen, exhibit remarkably enhanced expression of many liver-specific functions.


Myocardial protection with blood cardioplegia in ischemically injured hearts: reduction of reoxygenation injury with allopurinol.
Myocellular injury mediated by oxygen radicals potentially limits myocardial protection in ischemically damaged hearts. This damage may be greater with oxygen-carrying blood cardioplegic solutions. A major mechanism of oxygen radical production is the conversion of hypoxanthine to uric acid by xanthine oxidase. In 16 anesthetized dogs, we studied whether adding allopurinol, a xanthine oxidase inhibitor, to blood cardioplegia would improve recovery of left ventricular (LV) performance and oxygen consumption. Millar transducer-tipped catheters and minor axis ultrasonic crystals were placed to assess LV performance by the slope of the end-systolic pressure-minor axis diameter relationships (Emax). Following total vented bypass, the hearts underwent 30 minutes of normothermic ischemia and then hypothermic blood cardioplegia with 1 mM allopurinol (N = 8) or without allopurinol (N = 8). Postischemic LV performance was significantly better with allopurinol than without (49.5 +/- 8.0 versus 17.4 +/- 4.1% of preischemic Emax; p less than 0.004). Postischemic LV oxygen consumption in the beating working state, calculated from LV blood flow (15 microm microspheres) and oxygen extraction, was comparable to preischemic values with and without allopurinol (10.2 +/- 1.2 versus 8.6 +/- 1.2 ml O2/100 gm/min). We conclude that allopurinol enhancement of blood cardioplegia increases myocardial protection in severely ischemic ventricles.


Structure and Functional Characterization of Human Aspartate Transcarbamoylase, the Target of the Anti-tumoral Drug PALA.
CAD, the multienzymatic protein that initiates and controls de novo synthesis of pyrimidines in animals, associates through its aspartate transcarbamoylase (ATCase) domain into particles of 1.5 MDa. Despite numerous structures of prokaryotic ATCases, we lack structural information on the ATCase domain of CAD. Here, we report the structure and functional characterization of human ATCase, confirming the overall similarity with bacterial homologs. Unexpectedly, human ATCase exhibits cooperativity effects that reduce the affinity for the anti-tumoral drug PALA. Combining structural, mutagenic, and biochemical analysis, we identified key elements for the necessary regulation and transmission of conformational changes leading to cooperativity between subunits. Mutation of one of these elements, R2024, was recently found to cause the first non-lethal CAD deficit. We reproduced this mutation in human ATCase and measured its effect, demonstrating that this arginine is part of a molecular switch that regulates the equilibrium between low- and high-affinity states for the ligands.


[Amplification, rearrangement and expression of the c-myc and c-Ha-ras1 proto-oncogenes in patients with breast cancer].
The restriction analysis of breast cancer DNA revealed amplification of c-myc and c-Ha-ras oncogenes in 3 of 19 and 4 of 22 carcinomas, respectively. In two tumours containing amplified c-Ha-ras1-fragment an additional ras-band absent in unchanged lymphatic nodule and blood leukocytes of the same patients was revealed. Amplification of myc gene was registered in poorly differentiated carcinomas with high metastatic potential. c-Ha-ras1 activation (amplification and elevated expression) was identified in nonaggressive tumours with prolonged progression.


Prostatic Artery Embolization in Nonindex Benign Prostatic Hyperplasia Patients: Single-center Outcomes for Urinary Retention and Gross Prostatic Hematuria.
To present outcomes for prostatic artery embolization (PAE) to treat urinary retention and gross prostatic hematuria in nonindex benign prostatic hyperplasia patients. Seventy-five patients undergoing PAE from December 2013 to August 2018 (age = 77.5 ± 8.6, age-adjusted Charlson comorbidity index = 4.6 ± 2.0, prostate volume = 224 mL ± 135 mL) for retention (n = 46) and/or gross prostatic hematuria (n = 55) were retrospectively reviewed. Twenty-six patients had both problems. Urinary retention patients (UR, n = 46, catheterization = 162.4 ± 148.1 days) underwent voiding trials 1-2 months post-PAE, with International Prostate Symptom Score (IPSS), Quality of Life (QoL), and postvoid residual (PVR) recorded at 3, 6, 12, 24, and 36 months. Pre- and post-PAE hematuria-related visits were compared for gross hematuria patients (GH, n = 39), as were transfusion rates for severe hematuria patients requiring bladder irrigation (SH, n = 16). Ninety-day adverse event tabulation used Clavien-Dindo classification. Three months post-PAE, 33/38(87%) UR patients were catheter-free (IPSS = 8.9 ± 5.3, QoL = 1.6 ± 1.7, PVR = 158 mL ± 207 mL). Results were similar at 6 months (catheter-free = 26/28(93%), IPSS = 6.5 ± 4.4, QoL = 1.1 ± 0.9, PVR = 149 mL ± 139 mL), 12 months (catheter-free = 19/20(95%), IPSS = 4.7 ± 4.8, QoL = 0.6 ± 0.9, PVR = 125 mL ± 176 mL), 24 months (catheter-free = 11/12(92%), IPSS = 4.4 ± 3.0, QoL = 0.9 ± 0.8, PVR = 66 mL ± 68 mL), and 36 months (catheter-free = 5/6(83%), IPSS = 5.8 ± 3.8, QoL = 0.8 ± 1.0, PVR =99 mL ± 71 mL). Out of 37, 34(92%) GH patients remained hematuria-free at 483 ± 137 days, with 22 hematuria-related visits pre-PAE vs none post-PAE. Hematuria resolved <48 hours post-PAE in 14/16(87.5%) SH patients, with 36 blood units transfused pre-PAE, 4 units transfused <48 hours post-PAE, and none thereafter. Subsequently, 13/16(81%) remained hematuria-free at 500 ± 501 days; 2/16(13%) required fulguration; 1/16(6%) developed bladder tumor. There were 2 deaths <30 days post-PAE, and 8(11%) Grade-II urinary infections. PAE provided safe, effective, and durable treatment for retention and gross hematuria in nonindex benign prostatic hyperplasia patients.


Dynamics of metastasis suppressor gene inactivation.
For most cancer cell types, the acquisition of metastatic ability leads to clinically incurable disease. Twelve metastasis suppressor genes (MSGs) have been identified that reduce the metastatic propensity of cancer cells. If these genes are inactivated in both alleles, metastatic ability is promoted. Here, we develop a mathematical model of the dynamics of MSG inactivation and calculate the expected number of metastases formed by a tumor. We analyse the effects of increased mutation rates and different fitness values of cells with one or two inactivated alleles on the ability of a tumor to form metastases. We find that mutations that are negatively selected in the main tumor are unlikely to be responsible for the majority of metastases produced by a tumor. Most metastases-causing mutations will be present in all (or most) cells in the main tumor.


Factors contributing to relapse in rigidly fixed mandibular setbacks.
The incidence of, and factors accounting for, relapse in 25 subjects who underwent mandibular setbacks via a bilateral sagittal split osteotomy with rigid fixation were studied. Fourteen had single-jaw operations, and the remaining 11 had concomitant maxillary procedures. Cephalometric radiographs were reviewed preoperatively, immediately postoperatively, and 6 months to 3 years after surgery. Relapse was defined as forward movement of pogonion during the postoperative period. No difference in the movement of the mandible in one- or two-jaw cases was noted. Even with excellent occlusal results, there was a tendency for the mandible (chin point) to rotate forward. In the one-jaw cases 43.7% relapse was noted, whereas 53.4% was seen in the two-jaw cases. A regression analysis showed that the magnitude of setback was the single factor that significantly predicted relapse in one-jaw cases, whereas alteration of the proximal segment accounted for relapse in two-jaw procedures. These results seem interrelated when considering alterations in the spatial arrangement of the muscular tissues and their attachments.


Primary spinal intradural extramedullary cysticercosis.
Spinal cysticercosis represents an uncommon localization of a common parasitic disease and, in most cases, is associated with intracranial involvement. Once confirmed, the entire neuraxis should be evaluated. We present an unusual case of a 60-year-old male patient with isolated localization of spinal intradural extramedullary cysticercosis. The patient was treated successfully with the combination of surgical excision and orally administered albendanzole. We discuss the treatment options, the diagnostic screening, and the possible route of dissemination. Because this is a rare but potentially life-threatening disease, clinicians should always take it into consideration in the differential diagnosis of spinal space-occupying lesions.


Dihydropyridine BAY-K-8644 activates chromaffin cell calcium channels.
Douglas and Rubin suggested that "the role of acetylcholine as a transmitter at the adrenal medulla is to cause some brief change in medullary cells which allows Ca ions to penetrate them and trigger the catecholamine ejection process". The Ca2+-channel blocking agents, verapamil, nifedipine and nitrendipine, have been used widely to investigate the properties of slow Ca2+ channels in a variety of tissues, including the adrenomedullary chromaffin cell. Recently, small modifications to the nifedipine molecule produced a derivative, BAY-K-8644 (methyl-1,4-dihydro-2, 6-dimethyl-3-nitro-4-(2-trifluoromethylphenyl)-pyridine-5-carboxylate), that in contrast to the Ca2+-channel blocking agents, stimulated cardiac and vascular smooth muscle contractility. We have tested whether this compound behaves as a Ca2+-channel activator at the chromaffin cell membrane as shown by Schramm et al. in smooth muscle cells. The experiments described here strongly suggest that it does so.


Efficient production of an ezetimibe intermediate using carbonyl reductase coupled with glucose dehydrogenase.
Ezetimibe is a top-selling hypolipidemic drug for the treatment of cardiovascular diseases. Biosynthesis of (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one ((S)-ET-5) using carbonyl reductase has shown advantages including high catalytic efficiency, excellent stereoselectivity, mild reaction conditions, and environmental friendness, and was considered as the key step for ezetimibe production. The regeneration efficiency of the cofactor, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) is one of the main restricted factor. Recombinant Escherichia coli strain (smCR125) coexpressing carbonyl reductase (CR125) and glucose dehydrogenase were successfully constructed and applied for the production of (S)-ET-5 for the first time. Without extra addition of the coenzyme NADPH, the yield of 99.8% and the enantiomeric excess (e.e.) of 99.9% were achieved under ET-4 concentration of 200 g/L. Using a substrate fed-batch strategy, under the optimal conditions, the substrate ET-4 concentration was increased to 250 g/L with the yield of 98.9% and the e.e. of 99.9% after 12 hr reaction. The space-time yield of 494.5 g L-1 d-1 and the space-time yield per gram biocatalyst of 24.7 g L-1 d-1 g-1 DCW were achieved, which were higher than ever reported for the biosynthesis of the ezetimibe intermediate.


Early and Accurate Detection of Radiation-induced Heart Damage by Cardiodynamicsgram.
Cardiodynamicsgram (CDG) has emerged recently as a noninvasive spatiotemporal electrocardiographic method for subtle cardiac dynamics information analysis within electrocardiogram (ECG). This study explored the feasibility of CDG for detecting radiation-induced heart damage (RIHD) in a rat model. A single radiation dose of 40 Gy was delivered to the cardiac apex of female Wistar rats. First, CDG was generated through dynamic modeling of ECG signals using the deterministic learning algorithm. Furthermore, CDG indexes were calculated using the wavelet transform and entropy. In this model, CDG entropy indexes decreased significantly after radiotherapy. The shape of CDG changed significantly after radiotherapy (irregular shape) compared with controls (regular shape). Macrophage and fibrosis in myocardium of rats increased significantly after radiotherapy. CDG changes after radiotherapy were significantly correlated with histopathological changes and occurred significantly earlier than histopathological changes. This study provides an experimental basis for the clinical application of CDG for the early detection of RIHD.


An automated microscopy workflow to study Shigella-neutrophil interactions and antibiotic efficacy in vivo.
Shigella are Gram-negative bacterial pathogens responsible for bacillary dysentery (also called shigellosis). The absence of a licensed vaccine and widespread emergence of antibiotic resistance has led the World Health Organisation (WHO) to highlight Shigella as a priority pathogen requiring urgent attention. Several infection models have been useful to explore the Shigella infection process; yet, we still lack information regarding events taking place in vivo. Here, using a Shigella-zebrafish infection model and high-content microscopy, we developed an automated microscopy workflow to non-invasively study fluorescently labelled bacteria and neutrophils in vivo. We applied our workflow to antibiotic-treated zebrafish, and demonstrate that antibiotics reduce bacterial burden and not neutrophil recruitment to the hindbrain ventricle. We discovered that nalidixic acid (a bactericidal antibiotic) can work with leukocytes in an additive manner to control Shigella flexneri infection and can also restrict dissemination of Shigella sonnei from the hindbrain ventricle. We envision that our automated microscopy workflow, applied here to study the interactions between Shigella and neutrophils as well as antibiotic efficacy in zebrafish, can be useful to innovate treatments for infection control in humans.


[Breast cancer: postoperative care (author's transl)].
Sixty percent of breast cancer patients show postoperative morbidity due to local recurrence or metastases. For these women and those who suffer from therapy-dependent somatic or psychologic disorders, organized postoperative care is necessary. Mode of action and organization of the Heidelberg postoperative care unit are described. An evaluation of postoperative care programs in Germany, Austria and Switzerland reveals the necessity of increasing our activities. An interdisciplinary approach in this field is of great importance.


Enterovirus infection in a semi-closed community.
Infection with enteroviruses was studied over a 61-week period (during 1960-61) in a semi-closed child community in the Detention Home of the Allegheny County Juvenile Court in Pittsburgh, Pennsylvania. While most of the viruses isolated were known adeno- and enterovirus types, three apparently represent new enterovirus types or 'prime strains'. Viruses were isolated in all but 6 weeks of the 61-week study period from the gastro-intestinal tracts of 110 children out of a total population of 514 (21%); of these 110, 24 children (22%) excreted virus at the time of admission. The population averaged 37 children (more than half of whom were under 5 years of age), with a turnover of about nine per week. Spread of infection in this community on introduction of a new virus was demonstrated, with virus shedding of variable duration after infection. Thirteen of the 110 positive children (12%) showed mixed virus infections. During the entire study period, no clinical diarrheal illness was found associated with the viral infections detected.


Chronic kidney disease prevalence and awareness in middle age and young old: Regional comparative study in Japan.
Patients with chronic kidney disease are not aware of the illness because of its asymptomatic nature, but the association of disease progression and awareness in general population has not been sufficiently analysed on a large scale. We analysed the nationwide annual specific health checkup covering more than a half of the overall population at aged 40-74 in Japan, approximately 29.4 million people as of 2018, in combination with parameters to represent regional characteristics. The rate of the examinees with kidney dysfunction, an estimated glomerular filtration rate of <45 mL/min/1.73 m2 , was 1.0%, while that of examinees with of dipstick proteinuria ≥ (+) was 3.7%. Next, we conducted a regional comparative study on 335 medical administrative areas divided in the country. The regional rate of examinees aged 65-74 over the total examinees was positively correlated with the prevalence of kidney dysfunction (r = 0.72, p < .0001). Additionally, the mean rate of examinees aware of their 'chronic kidney failure' was 0.6%, and the awareness rate was correlated with the prevalence of both kidney dysfunction (r = 0.36, p < .001) and positive dipstick proteinuria (r = 0.31, p < .001) in those aged 65-74 at the regional level. Association of nephrology care resources with the prevalence or awareness was unclear at the regional level. We found a regional association of chronic kidney disease prevalence and awareness in a recent young old population in Japan. Further studies are needed to evaluate the patient screen and referral at the individual level.


Comparative study of intracranial aneurysms in advanced age.
A correlative angiographic and clinical analysis of 108 intracranial subdural aneurysms in patients from 60 to 84 years of age is presented. Angiographically defined spasm was present in about 54% of the cases. Spasm grades I and II had no apparent influence on clinical course, and grades III and IV were in some instances associated with arterial occlusion and infarction. There was no clear evidence that the spasm was related to hypertension, and no relationship could be established between preoperative and postoperative spasm. Atherosclerotic alterations in some cases were combined with a persistent narrowing of the arterial lumen.


Antigen-based immunotherapy of melanoma: Canvaxin therapeutic polyvalent cancer vaccine.
As yet there are no FDA-approved cancer vaccines for malignant melanoma, but encouraging response rates and low toxicities reported in phase I/II trials suggest that antigen-based active immunotherapy may complement current treatment modalities. The cumulative data for Canvaxin therapeutic polyvalent cancer vaccine represent the largest phase II clinical trial of any cancer vaccine. Univariate and multivariate analyses of these data have demonstrated the prognostic significance of this allogeneic whole-cell preparation as a postoperative adjuvant treatment for patients with stage III and IV melanoma. The vaccine has also been shown promising results after resection of stage II melanoma and in patients with regional in-transit disease. The consistent correlation between immune and clinical responses to the vaccine suggests that immune parameters may be used to monitor a patient's response to vaccine therapy.


Enhanced effect of DNA immunization plus in vivo electroporation with a combination of hepatitis B virus core-PreS1 and S-PreS1 plasmids.
To develop a novel, effective HBV therapeutic vaccine, we constructed two HBV DNA immunogens that contained PreS1, HBSS1, and HBCS1. Several delivery methods, such as intramuscular (i.m.) injection, intramuscular injection plus electroporation (i.m.-EP), and intradermal injection plus electroporation (i.d.-EP) were used in a murine model to analyze and compare the immune responses that were induced by the DNA immunogens. We found that i.d.-EP accelerated specific antibody seroconversion and produced high antibody (anti-PreS1, anti-S, and anti-C antibody) titers after HBSS1 and HBCS1 immunization. Combining the HBSS1 and HBCS1 DNA immunogens with i.d.-EP produced the strongest multiantigen (PreS1, S, and C)-specific cellular immune response and the highest specific PreS1 antibody levels. The results indicated that DNA immunization using HBSS1 and HBCS1 might be an ideal candidate, with its ability to elicit robust B and T cell immune responses against multiantigen when combined with optimized delivery technology. The present study provides a basis for the design and rational application of a novel HBV DNA vaccine.


Production of bacterial cellulose from alternate feedstocks.
Production of bacterial cellulose by Acetobacter xylinum ATCC 10821 and 23770 in static cultures was tested from unamended food process effluents. Effluents included low-solids (LS) and high-solids (HS) potato effluents, cheese whey permeate (CW), or sugar beet raffinate (CSB). Strain 23770 produced 10% less cellulose from glucose than did strain 10821 and diverted more glucose to gluconate. Unamended HS, CW, and CSB were unsuitable for cellulose production by either strain, and LS was unsuitable for production by strain 10821. However, strain 23770 produced 17% more cellulose from LS than from glucose, indicating that unamended LS could serve as a feedstock for bacterial cellulose.


[Tuberculosis in the developing countries and the work of the international Union for the control of tuberculosis (author's transl)].
In the developing countries tuberculosis is one of the most common diseases. According to estimates the number of cases of infectious tuberculosis is 3-4 millions. The guidelines for the effective control of the disease in the developing countries were laid down in a report by a Spezial Committee of the W.H.O. in 1974. The emphasis is 1) on protecting young people as far as possible by BCG vaccination, 2) on the early detection and treatment of as many cases of infectious tuberculosis as possible. The diagnosis does not depend on (expensive) radiographic equipment, as it can be made simply by microscopic examination of the sputum. For treating the infection various efficacious and not too expensive drugs are available which succeed within a few weeks in reducing the process to a non-infectious stage. Sanatorium or hospital treatment is generally not necessary.


Labeled bleomycin as a tumor localizing agent III. Selectivity of tumor tissue uptake of the different forms of [57Co] bleomycin A2 and B2.
The cell uptake of the different forms (I and II) of [57Co]bleomycin A2 and B2 was studied in a Rhabdomyosarcoma cell culture. The results show that the uptake of form I appears to be significantly higher than the uptake of form II. The evidence presented indicates that form I is formed in vivo as well as in vitro from form II by biotransformation. Transferrin stimulates the uptake of Co-bleomycin B2 form I only. As a result of trypsin treatment, it is suggested that form II binds only on the outer cell membrane and is not able to pass this membrane.


Evaluation of Four Methods for the Assessment of Sarcopenia in Older Adults in Nursing Homes.
We aimed to evaluate the comparative effectiveness and applicability of using calf circumference (CC), strength, need for assistance with walking, rising from a sitting position, climbing stairs, and the incidence of falls (SARC-F), as well as SARC-F plus CC (SARC-CalF) and the Ishii test, for assessing sarcopenia in older adult nursing home occupants. In this cross-sectional study, the diagnostic criteria of the AWGS2019 were used as the standard, and the accuracy of the four screening methods determined by indicators, including sensitivity, specificity, receiver operating characteristic (ROC) curve, positive predictive values (PPV), and negative predictive values (NPV). One hundred and ninety-nine older adults, 97 male and 102 female, were included. The prevalence of sarcopenia was 48.7%. Over all the participants, the sensitivity and specificity of CC were 74.22% and 51.96%, respectively, and 40.21% and 83.33%, respectively, for SARC-F. The use of SARC-CalF raised the SARC-F sensitivity (71.14%) while reducing the specificity (60.78%). The Ishii test had a sensitivity of 89.7% and a specificity of 74.51%. The PPV and NPV of CC, SARC-F, SARC-CalF, Ishii test were 0.6/0.68, 0.7/0.59, 0.55/0.63, and 0.77/0.88, respectively. The areas under the curve for CC, SARC-F, SARC-CalF, and the Ishii test were 0.67(95%CI, 0.59-0.74), 0.71(95%CI, 0.64-0.79), 0.71(95%CI, 0.64-0.79), and 0.86 (95% CI,0.81-0.92), respectively. The sensitivity and specificity of CC, SARC-F, SARC-CalF, and the Ishii test for sarcopenia screening in males were 71.69%/56.41%, 29.31%/79.49%, 67.24%/64.10%, and 94.83%/56.41%, respectively, and in females were 79.49%/49.21%, 56.41%/85.71%, 76.92%/58.73%, and 82.05%/85.71%, respectively. The PPV and NPV of CC, SARC-F, SARC-CalF, and the Ishii test in males were 0.71/0.56, 0.68/0.43, 0.74/0.57, and 0.76/0.88, respectively, while in females the values were 0.49/0.79, 0.71/0.76, 0.54/0.8, and 0.78/0.89, respectively.The areas under the curve for CC, SARC-F, SARC-CalF, and the Ishii test in males were 0.7(95%CI, 0.59-0.8), 0.63(95%CI, 0.52-0.75), 0.68(95%CI, 0.57-0.8), and 0.86(95% CI, 0.78-0.94), respectively, and in females 0.69(95%CI, 0.58-0.8), 0.81(95%CI, 0.72-0.89), 0.76(95%CI, 0.67-0.86), and 0.85 (95%CI, 0.77-0.94), respectively. The overall screening ability of the Ishii test for sarcopenia was superior to that of the CC, SARC-F, and SARC-CalF in older adults in nursing homes.


Longitudinal sleep phenotypes among offspring of bipolar parents and community controls.
Sleep disturbances are a prominent feature of bipolar disorder (BP). However, it remains unclear how sleep phenotypes may evolve among at-risk youth, and their relevance to BP onset. Pittsburgh Bipolar Offspring Study (BIOS) offspring (ages 10-18) and their parents completed assessments approximately every two years pertaining to current psychopathology and offspring sleep habits. A latent transition analysis (LTA) identified latent sleep groups within offspring based on their ratings of six sleep domains using the School Sleep Habits Survey. Demographic and clinical characteristics were compared between sleep groups. Logistic regression tested links between sleep group and BP onset at the subsequent assessment. The LTA model identified latent groups of good, poor, and variable sleepers. We observed an overall trend of good sleep becoming variable, and then poor, as youth age. Offspring in the poor sleep group were more likely to have psychopathology. Adjusting for age and depression, poor sleepers had nearly twice the odds of developing BP relative to good (OR=1.99, CI=0.45-8.91) or variable (OR=2.03, CI=0.72-5.72) sleepers. Limitations include the use of proximal sleep phenotypes to predict BP onset, and a self-report measure of sleep CONCLUSIONS: We found three non-overlapping sleep phenotype groups in a large sample of offspring of bipolar probands and offspring of demographically-matched community control parents. Clinicians should consider that youth will likely experience variable and/or poor sleep as they age, and that at-risk youth with poor sleep may be at increased risk of developing MDD and BP at their next assessment.


Self-reported history of comorbidities and markers of ovarian reserve among subfertile women.
To investigate whether history of comorbidities is associated with markers of ovarian reserve among subfertile women. This observational study includes 645 women seeking fertility care at the Massachusetts General Hospital who enrolled in the Environment and Reproductive Health (EARTH) study (2005-2019). Women completed a comprehensive questionnaire including medical diagnosis of comorbidities. Ovarian reserve markers including antral follicle count (AFC), assessed by transvaginal ultrasound, and circulating serum levels of day 3 FSH and AMH, are assessed by immunoassays. We fit linear regression models to evaluate the association between history of comorbidities and markers of ovarian reserve while adjusting for confounders. Self-reported history of hypertension, cancer, and neurological disorders was negatively associated with AFC in unadjusted models and in adjusted models for age, smoking, physical activity, comorbidity count, and BMI. Adjusted mean AFC (95% CI) was lower among women with history of hypertension, compared to women with no self-reported history of hypertension (11.5 vs 15.6, p value 0.0001). In contrast, day 3 FSH levels were positively related to history of eating disorders in both unadjusted and adjusted models (10.8 vs. 7.43 IU/L, p value ≤ 0.0001). Self-reported history of other comorbidities was unrelated to AFC, day 3 FSH, and AMH levels. History of hypertension, cancer, and neurological disorders was negatively associated with AFC, and eating disorders were positively related to day 3 FSH levels. The prevention of common comorbidities among women in reproductive age may help increase women's fertility given the declining birth rates and increasing use of assisted reproductive technologies in the past years.


New NO-donors with antithrombotic and vasodilating activities, II: 3-alkyl-N-nitroso-5-sydnone imines.
Fifteen 3-alkyl-, four 3-cycloalkyl-N-nitroso-5-sydnone imines and five 3-alkyl-N-nitro-5-sydnone imines were synthesized and their ability to inhibit platelet aggregation induced by collagen (Born-test) was studied in vitro. Dependent on the chemical structure, the IC50-values for the inhibition of platelet aggregation were in the range of 0.2-140 mumol/L. It is suggested that this scale reflects different binding properties of the nitrosimines with respect to the platelet membrane. Highest activities were observed for the 3-hexyl (2f) and the 3-cyclohexyl (2p) derivative. Three nitrimines (3e, 3f, 3i) also showed IC50 values below 10 mumol/L. For the nitrosimines 2a, 2f, and 2m antithrombotic activity was demonstrated in vivo. They inhibited laser induced arterial thrombosis in anesthetized rats up to 70% two h after oral administration. In conscious renal-hypertensive dogs, the decrease in systolic blood pressure and left ventricular enddiastolic pressure suggests an antianginal activity of the compound 2a similar to that of molsidomine (M). The smoother onset and the longer duration of action of the new compound as compared to M could be a significant advantage of 2a in the therapy of angina pectoris.


Risk factors associated with an unsuccessful pessary fitting trial in women with pelvic organ prolapse.
Our purpose was to estimate the percentage of women with symptomatic pelvic organ prolapse who had a successful pessary fitting trial, to identify pelvic examination parameters that are associated with an unsuccessful pessary fitting trial, and to identify the most commonly used pessary for each stage of prolapse. In a prospective, observational study, 100 consecutive women with symptomatic pelvic organ prolapse were examined, staged, and fitted with either a ring or Gellhorn pessary. A successful pessary fitting trial occurred if a woman continued to use a pessary 1 week after being fitted. Women were then divided into two groups on the basis of whether they had a successful pessary fitting trial: group 1 was successful, and group 2 was unsuccessful. Pelvic examination parameters were evaluated as risk factors for an unsuccessful pessary fitting trial. Group 1 had 73 women, and group 2 had 27 women. A short vaginal length (< or =6 cm) and a wide vaginal introitus (4 fingerbreadths accommodated) were associated with group 2 (11% vs 0%, P=.02, and 22% vs 7%, P=.04, respectively). Ring pessaries were used more with stage II and III prolapse (100% and 71%, respectively), whereas Gellhorn pessaries were used more with stage IV prolapse (64%, P<.001). Seventy-three women (73%) with symptomatic pelvic organ prolapse had a successful pessary fitting trial. A short vaginal length and a wide vaginal introitus were risk factors for an unsuccessful pessary fitting trial. Gellhorn pessaries are more often needed with stage IV prolapse.


Gliomatosis peritonei with endometriosis externa.
A case of gliomatosis peritonei developed 9 years after the removal of an immature ovarian teratoma is reported. An unique association of gliomatosis with endometriosis externa was found without the usual symptoms of endometriosis. The prognosis of gliomatosis is generally excellent and depends on the histological grade of the primary tumor and on the presence of organ metastasis or immature tumor elements in the implants. Pathogenetical aspects of both lesions are discussed briefly.


Size structure of testate amoebae (Arcellinida and Euglyphida) in different habitats from a lake in the upper Paraná River floodplain.
This study aimed to investigate the relationship between the size structure of testate amoebae in distinct habitats, i.e. plankton, aquatic macrophytes and aquatic sediment. The samples were taken from a floodplain lake of the upper Paraná River. The assumptions we strived to scrutinize were that (i) larger mean sizes of testate amoebae would be recorded in the sediment of the lake; and (ii) temporally, smaller individuals would be registered during the high water period in all habitats. The sampling was done monthly, from April 2007 to March 2008, in triplicates for each habitat. Testate amoebae were represented by individuals sized between 20 and 400 μm. The smaller individuals predominated in plankton samples, while in the aquatic sediment the larger ones were chiefly represented. These differences were probably associated with metabolic activities, i.e. the energy needs of these unicellular organisms, in each habitat. Two-way ANOVA yielded significant differences between hydrological periods. During the high water period, the increase in rainfall and consequently in water flow, decreased the stability of the system and increased turbulence and water column circulation. Therefore, environmental stability seems to be one of the main factors driving the temporal variation in the size structure of these specific organisms.


Erratum to "Use of a real-time location system to detect cows in distinct functional areas within a barn" (JDS Commun. 2:217-222).
[This corrects the article DOI: 10.3168/jdsc.2020-0050.].


Evaluation of the Biological Activity of Folic Acid-Modified Paclitaxel-Loaded Gold Nanoparticles.
Gold nanoparticles (AuNPs) with good physical and biological properties are often used in medicine, diagnostics, food, and similar industries. This paper explored an AuNPs drug delivery system that had good target selectivity for folate-receptor overexpressing cells to induce apoptosis. A novel drug delivery system, Au@MPA-PEG-FA-PTX, was developed carrying paclitaxel (PTX) on folic acid (FA) and polyethylene glycol (PEG)-modified AuNPs. The nanomaterial was characterized by transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), and ultraviolet-visible spectroscopy (UV-Vis). Also, the biological activity of the AuNPs drug delivery system was examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in HL-7702, Hela, SMMC-7721, and HCT-116 cells. Furthermore, apoptotic activity using annexin V-FITC, mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) levels was estimated by flow cytometry and fluorescence microscopy. Au@MPA-PEG-FA-PTX exhibited a distinct core-shell structure with a controllable size of 28±1 nm. Also, the AuNPs maintained good dispersion and spherical shape uniformity before and after modification. The MTT assay revealed good antitumor activity of the Au@MPA-PEG-FA-PTX against the Hela, SMMC-7721, and HCT-116 cells, while Au@MPA-PEG-FA-PTX produced better pharmacological effects than PTX in isolation. Further mechanistic investigation revealed that effective internalization of AuNPs by folate-receptor overexpressing cancer cells induced cell apoptosis through excessive production of intracellular ROS. The AuNPs drug delivery system showed good target selectivity for folate-receptor overexpressing cancer cells to induce target cell-specific apoptosis. These AuNPs may have great potential as theranostic agents such as in cancer.


COVID-19: An Indian perspective.
Lockdown, quarantine, self-isolation, personal protection equipment, and social distancing have become words of daily usage ever since the world health organization declared coronavirus disease 2019 (COVID-19) as a pandemic. The impact of COVID-19 extends over the medical field, economy, education, and politics. Though the knowledge of the virus is evolving, we are yet to find a solution. India, country with the second largest population, went into a phase of lockdown from 25th March 2020 to 31st May 2020. There was phased measure to "Unlock" starting from 1st June 2020. This has affected the clinical practice and training of the resident. The challenges faced during this unprecedented time are multifaceted which includes overcrowding, healthcare system, and educational background. Indian Association of Cardiovascular-Thoracic Surgeons kept continuing the educational program through a series of "Masterclass."


Localization of CRF-immunoreactive neurons in the cat medulla oblongata: their presence in the inferior olive.
Corticotropin releasing factor (CRF)-immunoreactive (IR) perikarya, visualized by the indirect immunoperoxidase method in colchicine-pretreated cats, were localized in many discrete regions of the medulla oblongata. They were found mainly in the dorsal aspect and midline of the medulla oblongata, and more rostrally in the ventrolateral portion. Our results also demonstrated CRF-IR neurons in the rostrocaudal extent of the inferior olive, probably projecting to the cerebellar cortex via thick axons visualized along the lateral edge of the medulla. CRF-IR olivary cells were also found in the pontine cat from which the forebrain was removed, but neither in hypophysectomized nor adrenalectomized cats.


Pesticide exposure and the microbiota-gut-brain axis.
The gut microbiota exist within a dynamic ecosystem shaped by various factors that includes exposure to xenobiotics such as pesticides. It is widely regarded that the gut microbiota plays an essential role in maintaining host health, including a major influence on the brain and behaviour. Given the widespread use of pesticides in modern agriculture practices, it is important to assess the long-term collateral effects these xenobiotic exposures have on gut microbiota composition and function. Indeed, exposure studies using animal models have shown that pesticides can induce negative impacts on the host gut microbiota, physiology and health. In tandem, there is a growing body of literature showing that the effects of pesticide exposure can be extended to the manifestation of behavioural impairments in the host. With the increasing appreciation of the microbiota-gut-brain axis, in this review we assess whether pesticide-induced changes in gut microbiota composition profiles and functions could be driving these behavioural alterations. Currently, the diversity of pesticide type, exposure dose and variation in experimental designs hinders direct comparisons of studies presented. Although many insights presented, the mechanistic connection between the gut microbiota and behavioural changes remains insufficiently explored. Future experiments should therefore focus on causal mechanisms to examine the gut microbiota as the mediator of the behavioural impairments observed in the host following pesticide exposure.


Shape effects on reflexive spatial attention are driven by the dorsal stream.
In a modified reflexive spatial attention paradigm, when the cue and the target are at the same spatial location, processing of the target is faster when the cue and the target have different shapes compared to same (shape effect). Recent physiological findings suggest distinct population level encoding of shape in ventral versus dorsal cortical visual streams in monkeys. In human observers, we tested whether the effect of shape on reflexive spatial attention could be attributed to ventral and/or dorsal stream encoding of shape. In the modified reflexive spatial attention paradigm, we varied the shapes of the cue and target. Based on data from monkey physiology (Lehky & Sereno, 2007), we selected four pairs of cue and target shapes. In some pairs, cue and target were similarly encoded (similar encoding distance) by a population of cells in the lateral intraparietal cortex, a dorsal stream area, but more dissimilarly encoded (having a greater encoding distance) by a population of cells in the anterior inferotemporal cortex (AIT), a ventral stream area. In other pairs, cue and target were similarly encoded in AIT and had greater dissimilarity in LIP encoding. We found that pairs of cue and target with greater dissimilarity in LIP encoding produced larger and more consistent shape effects up to a cue to target onset asynchrony (CTOA) of 450 ms. The shape effects for cue and target pairs with greater dissimilarity in AIT encoding were smaller and inconsistent, suggesting that shape effects in reflexive spatial attention are largely driven by the dorsal stream.


Comparative proteomic analysis of compartmentalised Ras signalling.
Ras proteins are membrane bound signalling hubs that operate from both the cell surface and endomembrane compartments. However, the extent to which intracellular pools of Ras can contribute to cell signalling is debated. To address this, we have performed a global screen of compartmentalised Ras signalling. We find that whilst ER/Golgi- and endosomal-Ras only generate weak outputs, Ras localised to the mitochondria or Golgi significantly and distinctly influence both the abundance and phosphorylation of a wide range of proteins analysed. Our data reveal that ~80% of phosphosites exhibiting large (≥1.5-fold) changes compared to control can be modulated by organellar Ras signalling. The majority of compartmentalised Ras-specific responses are predicted to influence gene expression, RNA splicing and cell proliferation. Our analysis reinforces the concept that compartmentalisation influences Ras signalling and provides detailed insight into the widespread modulation of responses downstream of endomembranous Ras signalling.


Idiopathic dilatation of the right atrium. Case report and literature review.
Idiopathic dilatation of the right atrium is a rare malformation characterized by disproportionate growth of the right atrium in the absence of a cause that justifies it. We present the case of a patient with idiopathic dilatation of the right atrium diagnosed since fetal life.


Differential, temporal and spatial expression of genes involved in storage oil and oleosin accumulation in developing rapeseed embryos: implications for the role of oleosins and the mechanisms of oil-body formation.
The temporal and spatial expression of oleosin and delta 9-stearoyl-ACP desaturase genes and their products has been examined in developing embryos of rapeseed, Brassica napus L. var. Topas. Expression of oleosin and stearate desaturase genes was measured by in situ hybridisation at five different stages of development ranging from the torpedo stage to a mature-desiccating embryo. The temporal pattern of gene expression varied dramatically between the two classes of gene. Stearate desaturase gene expression was relatively high, even at the torpedo stage, whereas oleosin gene expression was barely detectable at this stage. By the stage of maximum embryo fresh weight, stearate desaturase gene expression had declined considerably while oleosin gene expression was at its height. In contrast to their differential temporal expression, the in situ labelling of both classes of embryo-specific gene showed similar, relatively uniform patterns of spatial expression throughout the embryo sections. Immunogold labelling of ultra-thin sections from radicle tissue with anti-oleosin antibodies showed similar patterns to sections from cotyledon tissue. However, whereas at least three oleosin isoforms were detectable on western blots of homogenates from cotyledons, only one isoform was found in radicles. This suggests that some of the oleosin isoforms may be expressed differentially in the various types of embryo tissue. The differential timing of stearate desaturase and oleosin gene expression was mirrored by similar differences in the timing of the accumulation of their ultimate products, i.e. storage oil and oleosin proteins.(ABSTRACT TRUNCATED AT 250 WORDS)


Women's experience of the health information process involving a digital information tool before commencing radiation therapy for breast cancer: a deductive interview study.
Individuals undergoing radiation therapy for breast cancer frequently request information before, throughout and after the treatment as a means to reduce distress. Nevertheless, the provision of information to meet individuals needs from their level of health literacy is often overlooked. Thus, individuals information needs are often unmet, leading to reports of discontent. Internet and digital information technology has significantly augmented the available information and changed the way in which persons accesses and comprehends information. As health information is no longer explicitly obtained from healthcare professionals, it is essential to examine the sequences of the health information process in general, and in relation to health literacy. This paper reports on qualitative interviews, targeting women diagnosed with breast cancer who were given access to a health information technology tool, Digi-Do, before commencing radiation therapy, during, and after treatment. A qualitative research design, inspired by the integrated health literacy model, was chosen to enable critical reflection by the participating women. Semi-structured interviews were conducted with 15 women with access to a digital information tool, named Digi-Do, in addition to receiving standard information (oral and written) before commencing radiation therapy, during, and after treatment. A deductive thematic analysis process was conducted. The results demonstrate how knowledge, competence, and motivation influence women's experience of the health information process. Three main themes were found: Meeting interactive and personal needs by engaging with health information; Critical recognition of sources of information; and Capability to communicate comprehended health information. The findings reflect the women's experience of the four competencies: to access, understand, appraise, and apply, essential elements of the health information process. We can conclude that there is a need for tailored digital information tools, such as the Digi-Do, to enable iterative access and use of reliable health information before, during and after the radiation therapy process. The Digi-Do can be seen as a valuable complement to the interpersonal communication with health care professionals, facilitating a better understanding, and enabling iterative access and use of reliable health information before, during and after the radiotherapy treatment. This enhances a sense of preparedness before treatment starts.


Aortic thrombosis diagnosed by ultrasound.
A neonate developed aortic thrombosis following catheterization of the umbilical artery. The thrombus was identified and followed up by ultrasonography. Fibrinolytic therapy led to the almost complete disappearance of the aortic thrombus but the child died of cerebral haemorrhage. Physicians should be alerted to the possibility of aortic thrombosis in neonates following umbilical arterial catheterization. Ultrasonography is an appropriate method for diagnosis and follow-up in these cases.


Enhanced Protein Profiling Arrays for High-Throughput Quantitative Measurement of Cytokine Expression.
The antibody array has become a powerful technology in recent years and is widely used to detect the expression levels of various proteins such as cytokines, growth factors, chemokines, and angiogenic factors, some of which are involved in cancer progression. In this chapter, we describe a protein array technology called enhanced protein profiling array, which can simultaneously and quantitatively measure the expression levels of a few proteins in hundreds or thousands of samples, and an example of its use is presented.


Effect of the Otago Exercise Programme on the frailty of people between 65 and 80 years old.
To evaluate the effect of the Otago Exercise Programme on the frailty of non-institutionalized people between 65 and 80 years of age and study factors associated with frailty. Pre-post test study (baseline and after 12 months) without control group in people aged 65-80 years (living in the community and with independent ambulation) after being trained in the Otago Exercise Programme in 5 sessions: weeks 1, 2, 4 and 8, and a reinforcement session at 6 months. Patients were recruited from 3 health areas (n = 200). Frailty was assessed with the FRAIL scale. There were 198 people who started the study and 161 completed the 12-month follow-up. The average age of the sample was 72.28 years; predominance of female sex 64.65%; low educational level 76.50%; people living alone 24.50%; overweight in 42%, and grade I obesity in 32.32%. Significant differences were observed with a P = .023 in the level of frailty between baseline and 12 months. The exploratory analysis identified significant differences before and after the intervention by sex (being a woman) (P = .018) and condition of living alone (P = .0468). The Otago Exercise Programme prevents positive results in frailty in people 65 to 80 years of age and can help maintain function and prevent deterioration.


Retrospective estimation of diabetes incidence from information in a prevalent population and historical mortality.
If the distribution of onset age of disease in a well-defined prevalent (cross-sectional) population of patients is known, disease incidence rates specific for age and calendar time period may be estimated, assuming known mortality rates and a closed population. This paper develops a method of estimation, illustrates this method on Danish diabetes data, and discusses its general applicability. The prevalent population of diabetic subjects in Fyn County on July 1, 1973 was ascertained from prescriptions, and information on disease onset was obtained from the patients' medical records. In this study only patients with onset of disease before or at age 30 years were studied. The mortality of diabetic subjects in Denmark was estimated from retrospective hospital data covering the period since 1933, and historical age-specific population sizes of Fyn County were obtained from census data. The incidence of diabetes increases with calendar time and with age for most cohorts. The variation with age for a fixed calendar year is more complicated, however, usually displaying a local maximum at about the age of puberty and a higher incidence at the upper end of the studied age range.


Assessing temporal differences in the predictive power of baseline TyG-related parameters for future diabetes: an analysis using time-dependent receiver operating characteristics.
It is known that measuring the triglyceride glucose (TyG) index and TyG-related parameters [triglyceride glucose-body mass index (TyG-BMI), triglyceride glucose-waist circumference (TyG-WC), and triglyceride glucose-waist to height ratio (TyG-WHtR)] can predict diabetes; this study aimed to compare the predictive value of the baseline TyG index and TyG-related parameters for the onset of diabetes at different future periods. We conducted a longitudinal cohort study involving 15,464 Japanese people who had undergone health physical examinations. The subject's TyG index and TyG-related parameters were measured at the first physical examination, and diabetes was defined according to the American Diabetes Association criteria. Multivariate Cox regression models and time-dependent receiver operating characteristic (ROC) curves were constructed to examine and compare the risk assessment/predictive value of the TyG index and TyG-related parameters for the onset of diabetes in different future periods. The mean follow-up period of the current study cohort was 6.13 years, with a maximum of 13 years, and the incidence density of diabetes was 39.88/10,000 person-years. In multivariate Cox regression models with standardized hazard ratios (HRs), we found that both the TyG index and TyG-related parameters were significantly and positively associated with diabetes risk and that the TyG-related parameters were stronger in assessing diabetes risk than the TyG index, with TyG-WC being the best parameter (HR per SD increase: 1.70, 95% CI 1.46, 1.97). In addition, TyG-WC also showed the highest predictive accuracy in time-dependent ROC analysis for diabetes occurring in the short-term (2-6 years), while TyG-WHtR had the highest predictive accuracy and the most stable predictive threshold for predicting the onset of diabetes in the medium- to long-term (6-12 years). These results suggest that the TyG index combined with BMI, WC, and WHtR can further improve its ability to assess/predict the risk of diabetes in different future periods, where TyG-WC was not only the best parameter for assessing diabetes risk but also the best risk marker for predicting future diabetes in the short-term, while TyG-WHtR may be more suitable for predicting future diabetes in the medium- to long-term.


The importance of self-regulatory and goal-conflicting processes in the avoidance of drunk driving among Greek young drivers.
The present study examined self-regulatory and goal-conflicting processes in the avoidance of drunk driving among Greek young drivers. A total of 361 university students in Greece completed a questionnaire, using a retrospective cross-sectional survey design. One-third reported to have driven under the influence of alcohol. Although prior intentions were clearly related to actual avoidance of drunk driving, one out of five respondents had not complied with their intention. An examination of post-intentional correlates of avoidance of drunk driving among positive intenders showed that avoidance of drunk driving was positively related to alcohol limitation plans and alcohol limitation self-efficacy, whereas negative relations were found for goal conflict and behavioural willingness. The present study suggests that people should not only be motivated but also be equipped with self-regulatory strategies aiming at the avoidance of drinking. Finally, goal commitment should be enhanced by increasing the salience of the avoidance goal.


Synergistic osteogenic and angiogenic effects of KP and QK peptides incorporated with an injectable and self-healing hydrogel for efficient bone regeneration.
Irregular defects generated by trauma or surgery in orthopaedics practice were usually difficult to be fitted by the preformed traditional bone graft substitute. Therefore, the injectable hydrogels have attracted an increasing interest for bone repair because of their fittability and mini-invasivity. However, the uncontrollable spreading or mechanical failures during its manipulation remain a problem to be solved. Moreover, in order to achieve vascularized bone regeneration, alternatives of osteogenic and angiogenic growth factors should be adopted to avoid the problem of immunogenicity and high cost. In this study, a novel injectable self-healing hydrogel system (GMO hydrogel) loaded with KP and QK peptides had been developed for enhancing vascularized regeneration of small irregular bone defect. The dynamic imine bonds between gelatin methacryloyl and oxidized dextran provided the GMO hydrogel with self-healing and shear-thinning abilities, which led to an excellent injectability and fittability. By photopolymerization of the enclosed GelMA, GMO hydrogel was further strengthened and thus more suitable for bone regeneration. Besides, the osteogenic peptide KP and angiogenic peptide QK were tethered to GMO hydrogel by Schiff base reaction, leading to desired releasing profiles. In vitro, this composite hydrogel could significantly improve the osteogenic differentiation of BMSCs and angiogenesis ability of HUVECs. In vivo, KP and QK in the GMO hydrogel demonstrated a significant synergistic effect in promoting new bone formation in rat calvaria. Overall, the KP and QK loaded GMO hydrogel was injectable and self-healing, which can be served as an efficient approach for vascularized bone regeneration via a minimally invasive approach.


Social risk factors as predictors for parental report of deviant behaviour in 3-year-old children.
The objective of this study was to examine whether social risk factors adjusted for gender and some well-known perinatal risk factors were related to parental descriptions of deviant behaviour in 3(1/2)-year-old children. The study was a prospective cohort study of 1345 mothers enrolled during early pregnancy. Parents answered a questionnaire and children's behavioural difficulties were categorized into the following groups; "hyperactive-distractible", "hostile-aggressive" and "anxious-fearful" following a modified Behar scale. The male gender was found to be related to a description of all kinds of deviant behaviour. Having a family member experiencing employment difficulties was the factor most closely related to "hyperactive-distractible" behaviour. Familial stress due to lack of time, experience of divorce and mothers in excess of 35 years at childbirth were found to be related to "hostile-aggressive" behaviour, but male gender was the most closely related. Social disadvantage was not found to be related to "anxious-fearful" behaviour.


ASCL1 regulates neurodevelopmental transcription factors and cell cycle genes in brain tumors of glioma mouse models.
Glioblastomas (GBMs) are incurable brain tumors with a high degree of cellular heterogeneity and genetic mutations. Transcription factors that normally regulate neural progenitors and glial development are aberrantly coexpressed in GBM, conferring cancer stem-like properties to drive tumor progression and therapeutic resistance. However, the functional role of individual transcription factors in GBMs in vivo remains elusive. Here, we demonstrate that the basic-helix-loop-helix transcription factor ASCL1 regulates transcriptional targets that are central to GBM development, including neural stem cell and glial transcription factors, oncogenic signaling molecules, chromatin modifying genes, and cell cycle and mitotic genes. We also show that the loss of ASCL1 significantly reduces the proliferation of GBMs induced in the brain of a genetically relevant glioma mouse model, resulting in extended survival times. RNA-seq analysis of mouse GBM tumors reveal that the loss of ASCL1 is associated with downregulation of cell cycle genes, illustrating an important role for ASCL1 in controlling the proliferation of GBM.


Neobladder Obstruction: A Non-ischemic Cause for Hepatic Portal Venous Gas: Case Report.
Hepatic portal venous gas (HPVG) is a rare ominous radiological sign usually indicative of mesenteric ischemia. Increased detection of HPVG has been associated with a growing number of non-ischemic causes. A 64-year-old gentleman following radical cystectomy and neobladder formation developed clinical signs suggestive of bowel obstruction. HPVG was demonstrated on abdominal imaging. Urgent laparotomy revealed no evidence of ischemia. We hypothesize an obstructed neobladder permitted gas to enter the mesenteric circulation. The patient made a complete recovery with supportive management.


Two-component signal transduction in Corynebacterium glutamicum and other corynebacteria: on the way towards stimuli and targets.
In bacteria, adaptation to changing environmental conditions is often mediated by two-component signal transduction systems. In the prototypical case, a specific stimulus is sensed by a membrane-bound histidine kinase and triggers autophosphorylation of a histidine residue. Subsequently, the phosphoryl group is transferred to an aspartate residue of the cognate response regulator, which then becomes active and mediates a specific response, usually by activating and/or repressing a set of target genes. In this review, we summarize the current knowledge on two-component signal transduction in Corynebacterium glutamicum. This Gram-positive soil bacterium is used for the large-scale biotechnological production of amino acids and can also be applied for the synthesis of a wide variety of other products, such as organic acids, biofuels, or proteins. Therefore, C. glutamicum has become an important model organism in industrial biotechnology and in systems biology. The type strain ATCC 13032 possesses 13 two-component systems and the role of five has been elucidated in recent years. They are involved in citrate utilization (CitAB), osmoregulation and cell wall homeostasis (MtrAB), adaptation to phosphate starvation (PhoSR), adaptation to copper stress (CopSR), and heme homeostasis (HrrSA). As C. glutamicum does not only face changing conditions in its natural environment, but also during cultivation in industrial bioreactors of up to 500 m(3) volume, adaptability can also be crucial for good performance in biotechnological production processes. Detailed knowledge on two-component signal transduction and regulatory networks therefore will contribute to both the application and the systemic understanding of C. glutamicum and related species.


The distribution of nerves supplying the testis, epididymis and accessory sex glands of Suncus murinus.
Chronic testicular pain remains an important challenge for urologists. Investigation of the innervation of male gonads thus becomes essential for deepening our understanding of their regulatory roles in male reproductive physiology and pathophysiology. Studies of testicular innervation are mainly limited to the intratesticular peptidergic nerves of the testis by immunohistochemical and acetylcholinesterase histochemical investigations in some animals. Little is known about the detailed, overall distribution in general experimental animal testis. In this study, the distribution of nerves supplying the testis, epididymis and accessory sex glands of Suncus murinus was investigated by whole mount immunohistochemistry staining using a neurofilament protein antibody. Testicular nerves arose through three routes: nerves deriving from the mesenteric and renal plexuses accompanied the testicular artery, entering into the testicular hilum through the superior ligament of the testis. The nerves originating from the hypogastric plexus then ran along the internal iliac artery, deferential artery, and passed through the mesoductus deferens or mesoepididymis, innervating the cauda and corpus of the epididymis, the vas deferens and the inferior pole of the testis. The third route arose from the pelvic plexus, distributed in the seminal vesicle and the prostate. The density of nerve fibers was higher in the cauda epididymidis than in the testis, and more abundant in the vas deferens. The different origins and distribution densities of testicular nerves in S. murinus may serve different neuronal regulatory functions, and, therefore, S. murinus may be an important model animal for understanding the different characteristics of testicular pain.


Limited benefit to genetically dystrophic chickens from a synthetic proteinase inhibitor: Ep475.
Chickens with inherited muscular dystrophy (Line 413) were treated in two separate trials with daily intraperitoneal injections of 10% DMSO-water solutions containing the proteinase inhibitors, Ep475 and E64. Drug therapy in each case significantly prolonged the functional ability of the treated chickens. Diluent control chickens around day 35 ex ovo characteristically reached a maximum ability to right from the supine position in a standardized functional test for muscle weakness. Subsequently, the control chickens were found to decline progressively in their ability to right. Treatment with the proteinase inhibitors had no effect on the typically elevated levels of plasma creatine kinase activity. In a histological analysis of the affected pectoralis major muscle, drug treatment had no effect on the relative distribution of degenerating, and vacuolated fibers, inflammatory cells, and abnormal fiber diameters. An exception was seen in decreased necrotic fibers of chickens treated with high doses of Ep475. Moreover, both inhibitors had positive effects on two biochemical abnormalities common to the dystrophic pectoralis muscle: increase in noncollagen protein, and reduction in total calcium.


Dried blood spots for the diagnosis and quantitation of HIV-1: stability studies and evaluation of sensitivity and specificity for the diagnosis of infant HIV-1 infection in Thailand.
Molecular methods for HIV-1 infection using dried blood-spot (DBS) for HIV-1 CRF01_AE subtypes have not been fully optimized. In this study assays for HIV-1 diagnosis or quantitation were evaluated using infant DBS from Thailand. Paired DBS and whole blood samples from 56 HIV-1 CRF01_AE or B'-infected infants were tested for infant diagnosis using modified Amplicor DNA PCR and NucliSens RNA NASBA and an in-house real-time PCR assay. The Amplicor Monitor viral load (VL) assay, with modifications for DBS, was also evaluated. DBS VL were hematocrit corrected. Stability studies were done on DBS stored at -70 degrees C to 37 degrees C for up to 1 year. The DBS diagnostic assays were 96-100% sensitive and 100% specific for HIV-1 diagnosis. DBS HIV-1 VL were highly correlated with plasma VL when corrected using the actual or an assumed hematocrit factor (r(c)=0.88 or 0.93, respectively). HIV-1 DNA in DBS appeared to be more stable than RNA and could be detected after up to 9 months at most temperatures. DBS VL could be consistently determined when stored frozen. These results show that DBS can be used accurately instead of whole blood for the diagnosis of HIV-1 infection and VL quantitation, particularly if samples are appropriately stored.


Prognostic value of morphometry in papillary thyroid carcinoma.
The value of morphometric analysis in addition to standard prognostic indicators was studied in 28 cases of papillary thyroid carcinoma. Standard features included age, sex, lymph node status, tumor size, and encapsulation. The mean follow-up was 47 months (maximum, 140 months). Recurrences were documented in six patients at a mean time of 34 months; five patients recurred with distal metastases and one patient recurred with local disease. Univariate analysis most closely associated tumor recurrence with nuclear anisotropism (the standard deviation of the estimated nuclear area [ENASD]) and tumor size. With forward stepwise incremental analysis, the value of tumor size was lost and only the ENASD and the cellularity mean index (CMI), defined as the percentage of tumor volume composed of tumor cells, significantly correlated with recurrence. Fifty-five percent of patients with an ENASD greater than 17 microns2 and a CMI greater than 40% developed recurrence as compared with 5% of patients with lesser values (P = .0001). Morphometric analysis may significantly contribute to the role of histopathology in the evaluation of papillary thyroid carcinoma and may also provide information regarding prognosis not obtained by standard methods.


Power spectrum analysis in the study of ocular mechanisms.
The use of power-spectrum analysis in the study of ocular dynamics is becoming increasingly widespread. This paper summarizes the key points of signal acquisition and power-spectrum analysis, emphasizing aspects which we have found to be of relevance when the technique is applied to experiments in binocular accommodation. The considerations and conclusions are relevant to other areas of investigation into ocular dynamics and similar biological systems.


Evolution and pathogenicity of H6 avian influenza viruses isolated from Southern China during 2011 to 2017 in mice and chickens.
H6 subtype avian influenza viruses spread widely in birds and pose potential threats to poultry and mammals, even to human beings. In this study, the evolution and pathogenicity of H6 AIVs isolated in live poultry markets from 2011 to 2017 were investigated. These H6 isolates were reassortant with other subtypes of influenza virus with increasing genomic diversity. However, no predominant genotype was found during this period. All of the H6N2 and most of the H6N6 isolates replicated efficiently in lungs of inoculated mice without prior adaptation. All of the H6N2 and two H6N6 isolates replicated efficiently in nasal turbinates of inoculated mice, which suggested the H6N2 viruses were more adaptive to the upper respiratory tract of mice than the H6N6 viruses. One of H6N2 virus caused systemic infection in one out of three inoculated mice, which indicated that H6 avian influenza virus, especially the H6N2 viruses posed a potential threat to mammals. Five H6 strains selected from different genotypes caused no clinical signs to inoculated chickens, and their replication were limited in chickens since the viruses have been detected only from a few tissues or swabs at low titers. Our study strongly suggests that the H6 avian influenza virus isolated from live poultry markets pose potential threat to mammals.


Modeling facilitative sugar transporters: transitions between single and double ligand occupancy of multiconformational channel models explain anomalous kinetics.
The four-state simple carrier model (SCM) is employed to describe ligand translocation by diverse passive membrane transporters. However, its application to systems like facilitative sugar transporters (GLUTs) is controversial: unidirectional fluxes under zero-trans and equilibrium-exchange experimental conditions fit a SCM, but flux data from infinite-cis and infinite-trans experiments appear not to fit the same SCM. More complex kinetic models have been proposed to explain this "anomalous" behavior of GLUTs, but none of them accounts for all the experimental findings. We propose an alternative model in which GLUTs are channels subject to conformational transitions, and further assume that the results from zero-trans and equilibrium-exchange experiments as well as trans-effects corresponds to a single-occupancy channel regime, whereas the results from the infinite-cis and infinite-trans experiments correspond to a regime including higher channel occupancies. We test the plausibility of this hypothesis by studying a kinetic model of a two-site channel with two conformational states. In each state, the channel can bind the ligand from only one of the compartments. Under single-occupancy, for conditions corresponding to zero-trans and equilibrium-exchange experiments, the model behaves as a SCM capable of exhibiting trans-stimulations. For a regime including higher degrees of occupancy and infinite-cis and infinite-trans conditions, the same channel model can exhibit a behavior qualitatively similar to a SCM, albeit with kinetic parameters different from those for the single-occupancy regime. Numerical results obtained with our model are consistent with available experimental data on facilitative glucose transport across erythrocyte membranes. Hence, if GLUTs are multiconformational channels, their particular kinetic properties can result from transitions between single and double channel occupancies.


A taxometric analysis of cognitive and neuromotor variables in children at risk for schizophrenia.
A taxometric model was applied to detect a subgroup or taxon of children conjectured to be at highest risk for developing schizophrenia or related disorders in a sample of offspring of schizophrenic, depressed, and normal parents. Measures of cognitive and neuromotor performance in childhood were used as indicator tests in the analyses. A taxon consisting chiefly of children of schizophrenic parents was detected. Forty-seven percent of those children were assigned to the taxon, compared with 16% of the children of depressed parents and 4% of the children of normal parents. Assignment to the taxon is assessed in relation to the current functional status of the subjects in young adulthood.


[Atopic dermatitis. Psychosomatic and psychotherapeutic treatment].
Patients with chronic relapsing atopic dermatitis often suffer from stress-related exacerbations, exhaustion, depression, anxiety and helplessness. They then may feel they cannot influence the course of their disease, and often develop problems in social interaction. These case reports focus on the different possibilities in addition to classical dermatologic therapy--empathic psychosomatic-orientated conversations, strategies of behavioral medicine and the effect of long-term psychodynamic psychotherapy.


A collaborative study to establish the second national standard for hepatitis B immunoglobulin in Korea.
This study aimed to establish a second national standard for hepatitis B immunoglobulin (HBIG) that can be used for potency assays of hepatitis B and normal immunoglobulin. The candidate material was manufactured using a process approved as Good Manufacturing Practice. The freeze-dried candidate preparation was tested for physicochemical and biological properties, including pH, residual moisture, molecular size distribution, and potency. A collaborative study was performed involving four laboratories, including the National Institute of Food and Drug Safety Evaluation, as an official national control laboratory in Korea and manufacturers. The potency was calibrated against the second international standard for HBIG using two enzyme immunoassays: enzyme-linked immunosorbent assay and electrochemiluminescence immunoassay. Results from 240 assays were obtained from four laboratories, and combined potency estimates were obtained by calculating the geometric means. Intra- and inter-laboratory variability showed acceptable geometric coefficients of variation of 1.3-6.0 and 3.2-3.6%, respectively. The candidate preparation showed satisfactory stability in accelerated thermal degradation and real-time stability tests. Based on these results, the potency value of 105 IU/vial was assigned (95% confidence intervals: 100.0-109.2 IU/vial), and it was deemed suitable to serve as the Korean national standard for HBIG.


Paramagnetic viral nanoparticles as potential high-relaxivity magnetic resonance contrast agents.
In order to compensate for the inherent high threshold of detectability of MR contrast agents, there has been an active interest in the development of paramagnetic nanoparticles incorporating high payloads of Gd(3+) with high molecular relaxivities. Toward this end, the protein cage of Cowpea chlorotic mottle virus (CCMV), having 180 metal binding sites, is being explored. In vivo CCMV binds Ca(2+) at specific metal binding sites; however, Gd(3+) can also bind at these sites. Using fluorescence resonance energy transfer we have characterized the binding affinity of Gd(3+) to the metal binding sites by competition experiments with Tb(3+). The measured dissociation constant (K(d)) for Gd(3+) bound to the virus is 31 microM. The T(1) and T(2) relaxivities of solvent water protons in the presence of Gd(3+)-bound CCMV were 202 and 376 mM(-1) s(-1), respectively, at 61 MHz Larmor frequency. The unusually high relaxivity values of the Gd(3+)-CCMV are largely a result of the nanoparticle virus size and the large number of Gd(3+) ions bound to the virus. These preliminary results should encourage further investigations into the use of viral protein cages as a new platform for MR contrast agents.


Effect of complement depletion on O2 supply and consumption in ischemic dog myocardium.
The purpose of this study was to determine whether depletion of serum complement can decrease the severity of an ischemic episode by improving regional O2 supply and consumption parameters in the ischemic region of the heart. Fourteen anesthetized dogs with serum complement intact or depleted (100 U/kg cobra venom factor given 8 hrs before) were subjected to left anterior descending coronary artery (LAD) occlusion for 6 hrs. Myocardial blood flows were determined before and 6 hrs after LAD occlusion using radioactive microspheres. Regional arterial and venous O2 saturations were determined using microspectrophotometry. In control animals, flow decreased from 122 +/- 42 to 13 +/- 14 ml/min/100 g (mean +/- SD) in the occluded LAD region. With complement depletion, LAD occlusion resulted in a flow reduction in the ischemic region (38 +/- 29 ml/min/100 g), but to a lesser degree than seen in the same region in control animals, especially in the subendocardium. O2 consumption was decreased in the ischemic region of both treatment groups, though O2 consumption was higher in this region in complement depleted animals compared to the values in control animals. The O2 supply/consumption ratio was decreased similarly in the ischemic region of control and complement depleted groups. Thus, with complement depletion, flow to the ischemic zone was improved but this region was still flow restricted. The flow increase during complement depletion was sufficient to allow an increased O2 utilization in the ischemic region.


Poly (ADP-ribose) polymerase-1 haplotypes are associated with coeliac disease.
In coeliac disease (CD) there is an inflammatory status of the intestinal mucosa because of a high expression of proinflammatory mediators. The nuclear protein poly (ADP-ribose) polymerase-1 (PARP-1) has been implicated in the initial inflammatory response by modulating transcription of inflammation-related genes. The aim of this work was to investigate the role of PARP-1 gene promoter region haplotypes in relation to coeliac disease susceptibility. We analysed a coeliac population consisting of a case-control panel with 120 CD patients and 311 healthy blood donors. A CA microsatellite, as haplotype-defining variant of the whole PARP-1 promoter, was typed using a polymerase chain reaction (PCR) method combined with fluorescence technology. We considered two promoter haplotypes: A defined by short CA alleles (83-87 bp) and B defined by long CA alleles (89-101 bp). Haplotype A was significantly increased within the coeliac patients group (P = 0.007 OR 1.6 95%CI 1.12-2.35). Additionally, we observed a significant dose effect, showing homozygous individuals for haplotype A higher risk for CD susceptibility (P = 0.007, OR 1.79 95%CI 1.14-2.82). Our results provide the first evidence that PARP-1 haplotypes are related with coeliac disease susceptibility.


Myofibrils put the squeeze on nuclei.
During muscle development, nuclei travel from the centre of the myofibre to the periphery, a process defective in certain diseases. A new study reveals that this movement is due to centripetal forces imposed on nuclei by the crosslinking and contraction of myofibrils.


Effect of phosphonic analogues of glutamic acid on glutamate decarboxylase.
Among the phosphonic analogues of glutamic acid, only 4-amino-4-phosphono butyric acid, the compound which shows the highest affinity for pyridoxal phosphate, inhibits competitively both Escherichia coli and rat brain glutamate decarboxylases. Phosphinothricin, 2-amino-4-(methylphosphino)butyric acid, is a strong inhibitor of the mammalian enzyme.


Hyperinflation of the left anterior basal segment: a new sign of pectus excavatum severity on CT.
To describe a new finding in patients with pectus excavatum - left lower lobe anterior basal segment hyperinflation [LABSH]. A secondary objective is to determine the frequency of the new finding and association with pectus excavatum severity. The study population included 52 children who underwent preoperative computed tomography (CT) for the evaluation of pectus excavatum. A control group of 50 children was obtained after evaluating 137 CT examinations performed for other reasons. Patient age in both groups ranged from 12 to 20 years. The Haller index was calculated for all patients. LABSH was evaluated by visual inspection of lung windows. The difference in mean radiodensity measurements in regions of interest in the left and right anterior basal segments [ΔHU] was calculated. Spirometry was performed in 44 of the patients and the results were compared to Haller index severity and the presence of LABSH. Echocardiography reports were available for 50 children in the pectus excavatum group. LABSH was identified by visual inspection in 15 patients [29%] and was significantly associated with a Haller index >4.0 (p=0.001). ΔHU for the patients with LABSH was 90.2 HU (standard deviation [SD]=37.7) and for the non-hyperinflated group -5.51 (SD=44.63), which was significant (p<0.0001). There was a significant association of LABSH with the pectus excavatum group as compared to the control group. The difference in mean Haller index for children with normal spirometry (4.4, SD=2.7) was not significantly different (p=0.9899) than for children with obstructive disease (4.5, SD=1). There was mild cardiac compression on two echocardiograms. LABSH is a new sign associated with pectus excavatum. The sign suggests segmental bronchial compression caused by chest deformity results in segmental air trapping.