Large-scale production of selected type A trichothecenes: the use of HT-2 toxin and T-2 triol as precursors for the synthesis of d 3-T-2 and d 3-HT-2 toxin.
The type A trichothecenes T-2 and HT-2 toxins are toxic secondary metabolites produced by fungi of the Fusarium genus. Their occurrence in cereals, especially in oats, implies health risks for the consumer. Therefore, it is an important task to develop selective and sensitive methods for the analysis of T-2 and HT-2 toxins, and to undertake further studies on their stability and toxicity. Although most toxins are commercially available, their high prices are the limiting factor on the realization of these experiments. Thus, we developed a method for large-scale production of T-2 and HT-2 toxin as well as T-2 triol and T-2 tetraol. T-2 toxin was obtained in gram quantities by biosynthetic production with cultures of F. sporotrichioides. As HT-2 toxin was only formed as a by-product, and T-2 triol and T-2 tetraol were not generated, these compounds were produced by alkaline hydrolysis of T-2 toxin. Separation and isolation of crude toxins was achieved by fast centrifugal partition chromatography (FCPC), which is an efficient tool for the large-scale purification of natural products. Using this fast and yield effective technique, several hundred milligrams of HT-2 toxin, T-2 triol, and T-2 tetraol were obtained. Subsequent, HT-2 toxin and T-2 triol were used for the large-scale synthesis of isotope-labeled T-2 and HT-2 toxin, respectively. Using these standards, an isotope dilution-(ID)-HPLC-MS/MS method for the quantification of T-2 and HT-2 toxin in different matrices was developed.


Irreversibly inhibitory kinetics of 3,5-dihydroxyphenyl decanoate on mushroom (Agaricus bisporus) tyrosinase.
3,5-Dihydroxyphenyl decanoate (DPD) is found to inhibit the diphenolase activity of tyrosinase from mushroom (Agaricus bisporus). The effects of DPD on the diphenolase activity of mushroom tyrosinase have been studied. The results show that the enzyme activity decreases very slowly with an increase in DPD concentrations at lower concentrations of DPD (between 5 and 60 microM). But at higher concentrations of DPD, DPD can strongly inhibit the diphenolase activity of the enzyme and the inhibition is irreversible. The IC50 value was estimated to be 96.5 microM. The inhibition mechanism of DPD has been investigated and the results show that DPD can bind to the free enzyme molecule and enzyme-substrate complex and lose the enzyme activity completely. The inhibition kinetics has been studied in detail by using the kinetic method of the substrate reaction described by Tsou. The microscopic rate constants of the enzyme inhibited by DPD at higher concentrations have been determined.


Topography and extent of pulmonary vagus nerve supply with respect to transthoracic oesophagectomy.
Pulmonary complications are frequently observed after transthoracic oesophagectomy. These complications may be reduced by sparing the vagus nerve branches to the lung. However, current descriptions of the regional anatomy are insufficient. Therefore, we aimed to provide a highly detailed description of the course of the pulmonary vagus nerve branches. In six fixed adult human cadavers, bilateral microscopic dissection of the vagus nerve branches to the lungs was performed. The level of branching and the number, calibre and distribution of nerve branches were described. Nerve fibres were identified using neurofilament immunohistochemistry, and the nerve calibre was measured using computerized image analysis. Both lungs were supplied by a predominant posterior and a smaller anterior nerve plexus. The right lung was supplied by 13 (10-18) posterior and 3 (2-3) anterior branches containing 77% (62-100%) and 23% (0-38%) of the lung nerve supply, respectively. The left lung was supplied by a median of 12 (8-13) posterior and 3 (2-4) anterior branches containing 74% (60-84%) and 26% (16-40%) of the left lung nerve supply, respectively. During transthoracic oesophagectomy with en bloc lymphadenectomy and transection of the vagus nerves at the level of the azygos vein, 68-100% of the right lung nerve supply and 86-100% of the inferior left lung lobe nerve supply were severed. When vagotomy was performed distally to the last large pulmonary branch, 0-8% and 0-13% of the nerve branches to the right middle/inferior lobes and left inferior lobe, respectively, were lost. In conclusion, this study provides a detailed description of the extensive pulmonary nerve supply provided by the vagus nerves. During oesophagectomy, extensive mediastinal lymphadenectomy denervates the lung to a great extent; however, this can be prevented by performing the vagotomy distal to the caudalmost large pulmonary branch. Further research is required to determine the feasibility of sparing the pulmonary vagus nerve branches without compromising the completeness of lymphadenectomy.


Stage-specific roles of Ezh2 and Retinoic acid signaling ensure calvarial bone lineage commitment.
Development of the skull bones requires the coordination of two stem progenitor populations, the cranial neural crest cells (CNCC) and head paraxial mesoderm (PM), to ensure cell fate selection and morphogenesis. The epigenetic methyltransferase, Ezh2, plays a role in skull bone formation, but the spatiotemporal function of Ezh2 between the CNCC- and PM-derived bone formation in vivo remains undefined. Here, using a temporally-inducible conditional deletion of Ezh2 in both the CNCC- and PM- derived cranial mesenchyme between E8.5 and E9.5, we find a reduction of the CNCC-derived calvarial bones and a near complete loss of the PM-derived calvarial bones due to an arrest in calvarial bone fate commitment. In contrast, deletion of Ezh2 after E9.5 permits PM-derived skull bone development, suggesting that Ezh2 is required early to guide calvarial bone progenitor commitment. Furthermore, exposure to all-trans Retinoic acid at E10.0 can mimic the Ezh2 mutant calvarial phenotype, and administration of the pan retinoic acid receptor (RAR) antagonist, BMS-453, to Ezh2 mutants partially restores the commitment to the calvarial bone lineage and PM-derived bone development in vivo. Exogenous RA signaling activation in the Ezh2 mutants leads to synergistic activation of the anti-osteogenic factors in the cranial mesenchyme in vivo. Thus, RA signaling and EZH2 can function in parallel to guide calvarial bone progenitor commitment by balancing the suppression of anti-osteogenic factors.


Water depth determines spatial and temporal phosphorus retention by controlling ecosystem transition and P-binding metal elements.
Shallow lakes are more susceptible to eutrophication than deep lakes. The geochemical and biogeochemical mechanisms controlling the vulnerability to eutrophication for deep lakes and shallow lakes remain unknown. Therefore, we investigated the combined Phosphorus (P) retention mechanism with P fractions, water depth, distribution of P-binding metal elements, and macrophytes coverage in a degrading ecosystem of Erhai Lake. We concluded that different mechanisms control the P retention in deep-water areas and shallow-water areas. In shallow areas covered by macrophytes, the biogeochemical process manipulates the P retention by changing the total organic carbon (TOC), calcium (Ca) distributions and turbulence. In deep areas without macrophyte coverage, the aluminum (Al) and iron (Fe) distributions control the P retention by a physicochemical process. Manganese (Mn) was found to be a potential proxy in tracking the kinetic release and readsorb of redox-sensitive P (BD-P) in deep areas. The historical record and core sample indicate that the hydrological engineering induced water depth variation is a vital factor changing the ecosystem of Erhai Lake by forming a large area of intermediate area where macrophytes could only survive at low water level. The uplift of water level in the 1990s gradually changed the ecosystem of Erhai Lake from macrophyte-dominated to algal-macrophyte concomitant that reduced the accumulation of stable P fractions and their binding metals. Macrophytes were capable to preserve P in biomass in the macrophyte-dominated ecosystem, which released 150% and 72% of more labile organic P (NaOH25-nrP) and BD-P in the sediment after the deterioration than before, respectively. Therefore, water depth is a prerequisite to restoring the P preservation capacity of sediment and the macrophyte ecosystem. Further hydraulic engineering projects should consider the effect of water-level-variation-induced ecosystem transition.


Comparing the in vivo sonodynamic effects of dual- and single-frequency ultrasound in breast adenocarcinoma.
Dual-frequency ultrasound has recently been shown to extensively enhance the acoustic cavitation yield in water. In this study, the in vivo antitumor effect of simultaneous, dual-frequency ultrasound at low-level intensity (I SATA = 2 W/cm(2) for 1 MHz and I SATA = 0.2 W/cm(2) for 150 kHz) in combination with an intravenous injection of 5 mg/kg hematoporphyrin (Hp) was investigated in a model of breast adenocarcinoma in Balb/c mice. Seventy-one tumor-bearing mice were divided into nine treatment groups: control, sham, Hp injection, and single- and dual-frequency sonication in the presence and absence of Hp. The tumor growth delay was then calculated based on the tumor volume at various times after treatment using the following parameters: relative volume percent, [Formula: see text] and [Formula: see text] times (to reach five and two times the initial volume), percent of tumor growth inhibition ratio, and survival period. Our results showed no significant difference between the 150 kHz and 1 MHz single-frequency groups when compared with the sham group after 9 days of treatment (p > 0.05). However, treatment with dual-frequency ultrasound significantly delayed tumor growth when compared with the sham group (p < 0.05) after 9 days of treatment. Furthermore, in vivo experiments showed that combined dual-frequency sonication controlled tumor growth more effectively than single-frequency sonication. Evaluation of the therapeutic effects of single- and dual-frequency ultrasound in sonodynamic therapy revealed that treatment with the combination of dual-frequency ultrasound and Hp resulted in a significant reduction in the relative volume percent of tumors after 3 days of treatment (p < 0.05) compared with the controls. Additionally, the T 5 time and the survival period in the group treated with the combination therapy was significantly longer than those in all the other groups (p < 0.05). These findings were further verified histopathologically. In conclusion, our results reveal that sonodynamic therapy using dual-frequency ultrasound is able to extend the survival time of animals compared with single-frequency sonication.


Chlamydia pneumoniae infection in IL-10 knock out mice: accelerated clearance but severe pulmonary inflammatory response.
In interleukin-10 knock out (IL-10 KO) mice, accelerated clearance of pulmonary Chlamydia pneumoniae infection was observed. On the other hand, the histopathological changes in lung tissue were more pronounced in IL-10 KO mice at all time points after infection and repeated infection than in the wild type mice. Both ex vivo induced antigen-specific proliferation as well as production of proinflammatory cytokines by splenocytes were higher in IL-10 KO mice than in WT mice. Also, intrapulmonary proinflammatory cytokine levels were higher in IL-10 KO mice than in the WT mice. The lack of anti-inflammatory action of IL-10 is likely to contribute to the enhanced clearance but severe inflammation in this experimental model.


An immunohistochemical study of abnormal keratinocyte proliferation in molluscum contagiosum.
Molluscum contagiosum is a common cutaneous tumour that is characterized by usually spontaneous involution and self-limited spreading in immunocompetent individuals. We aimed to investigate the apoptosis and the expression of cell-cycle proteins in molluscum contagiosum lesions. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling (TUNEL)-based apoptotic index and the expression of the cell-cycle proteins Ki-67, p53, p21WAF and Bcl-2 were investigated in molluscum contagiosum lesions obtained from the trunk of 20 immunocompetent patients and in normal skin samples from the trunk of six healthy volunteers. Whereas molluscum contagiosum lesions displayed a TUNEL-based apoptotic index similar to that of normal skin, they exhibited an increased Ki-67 index, which was confined to the basal and first suprabasal layers (P < 0.001). Compared with normal non-sun-exposed skin, molluscum contagiosum lesions also exhibited increased p53 staining in basal cells (P < 0.01), increased p21WAF in suprabasal cells (P < 0.001) and loss of Bcl-2 expression. These results indicate that molluscum contagiosum lesions exhibit an increased proliferation rate of keratinocytes, which is likely to be partially counteracted by accumulation of p53.


Acute effect of electrical stimulation of the dorsal genital nerve on rectal capacity in patients with spinal cord injury.
Constipation and fecal incontinence are considerable problems for most individuals with spinal cord injury (SCI). Neurogenic bowel symptoms are caused by several factors including abnormal rectal wall properties. Stimulation of the dorsal genital nerve (DGN) can inhibit bladder contractions and because of common innervation inhibitory effects are anticipated in the rectum too. Therefore, DNG could have a future role in the treatment of neurogenic fecal incontinence. To study the effect of acute DGN stimulation on the rectal cross sectional area (CSA) in SCI patients. Seven patients with complete supraconal SCI (median age 50 years) were included. Stimulation was applied via plaster-electrodes using an amplitude of twice the genito-anal reflex threshold (pulse width: 200 μs; pulse rate: 20 Hz). A pressure controlled phasic (10, 20 and 30 cmH(2)O) rectal distension protocol was repeated four times with subjects randomized to stimulation during 1st and 3rd distension series or 2nd and 4th distension series. The rectal CSA and pressure were measured using impedance planimetry and manometry. All patients completed the investigation. Median stimulation amplitude was 51 mA (range 30-64). CSA was smaller during stimulation and differences reached statistical significance at distension pressures of 20 cmH(2)O (average decrease 9%; P = 0.02) and 30 cmH(2)O (average decrease 4%; P = 0.03) above resting rectal pressure. Accordingly, rectal pressure-CSA relation was significantly reduced during stimulation at 20 (P=0.03) and 30 cmH(2)O distension (P=0.02). DGN Stimulation in patients with supraconal SCI results in an acute decrease of rectal CSA and the rectal pressure-CSA relation.


Synthesis, characterization and luminescence property of N,N'-di(pyridine N-oxide-2-yl)pyridine-2,6-dicarboxamide and corresponding lanthanide (III) complexes.
A new ligand, N,N'-di(pyridine N-oxide-2-yl)pyridine-2,6-dicarboxamide (LH2) and its several lanthanide (III) complexes (La, Eu, Gd, Tb, Y) were synthesized and characterized in detail based on elemental analysis, conductivity measurements, IR, 1H NMR, MS (FAB) and UV spectra and TG-DTA studies. The results indicated that the composition of these binary complexes is [Ln(LH2)(NO3)2.H2O]NO3.nH2O (n=0-1); while the ligand has a good planar structure with strong hydrogen bonds. The fluorescence spectra exhibits that the Tb (III) complex and the Eu (III) complex display characteristic metal-centered fluorescence in solid state while ligand fluorescence is completely quenched. However, the Tb (III) complex displays more effective luminescence than the Eu (III) complex, which is attributed to especial effectivity in transferring energy from the lowest triplet energy level of the ligands (T) onto the excited state (5D4) of Tb (III) than that (5D1) of Eu (III).


Administration of BDNF in the ventral tegmental area produces a switch from a nicotine-non-dependent D1R-mediated motivational state to a nicotine-dependent-like D2R-mediated motivational state.
Brain-derived neurotrophic factor (BDNF) has been implicated in the transition from a non-dependent motivational state to a drug-dependent and drug-withdrawn motivational state. Chronic nicotine can increase BDNF in the rodent brain and is associated with smoking severity in humans; however, it is unknown whether this increased BDNF is linked functionally to the switch from a nicotine-non-dependent to a nicotine-dependent state. We used a place conditioning paradigm to measure the conditioned responses to nicotine, showing that a dose of acute nicotine that non-dependent male mice find aversive is found rewarding in chronic nicotine-treated mice experiencing withdrawal. A single BDNF injection in the ventral tegmental area (in the absence of chronic nicotine treatment) caused mice to behave as if they were nicotine dependent and in withdrawal, switching the neurobiological substrate mediating the conditioned motivational effects from dopamine D1 receptors to D2 receptors. Quantification of gene expression of BDNF and its receptor, tropomyosin-receptor-kinase B (TrkB), revealed an increase in TrkB mRNA but not BDNF mRNA in the VTA in nicotine-dependent and nicotine-withdrawn mice. These results suggest that BDNF signalling in the VTA is a critical neurobiological substrate for the transition to nicotine dependence. The modulation of BDNF signalling may be a promising new pharmacological avenue for the treatment of addictive behaviour.


PCBs stimulate laccase production and activity in Pleurotus ostreatus thus promoting their removal.
Pleurotus ostreatus degrades polychlorinated biphenyls (PCBs) with an increase of laccase activity. Laccases are well known for their detoxifying activity. We show, using reverse transcription polymerase chain reaction and a biochemical assay, that reduction in PCBs (di, tri, tetra, and penta) levels are correlated with an increase in laccase activity. P. ostreatus cultures were obtained from 0 to 30 days in the presence or absence of 7,100 mg/L PCBs (from transformer oil) and a surfactant. After each selected time cultures were withdrawn and remaining PCBs were determined, a maximal removal percentage of PCBs was obtained at 20 (63.5 ± 2.0) and 30 days (63.8 ± 4.6) post-induction. Also, the activity of the enzyme was analyzed and it was found to increase at 10 (6.9-fold) and 20 (6.77-fold) days post-induction in the presence of PCBs, as determined by its activity. Taken together, these data suggest that PCBs induce laccase expression and that laccase catalyzes PCBs removal.


Imperforate anus with long but apparent low fistula in females.
Despite the low entry of the rectum into the vagina in some females with imperforate anus, the fistula may be deceivingly long. This variation should alert the surgeon to measure the fistula prior to anoplasty. During surgery, biopsies of the mobilized segment should also be done to assure that cloacal-transitional lined structures have been removed and rectal mucosa anastamosed to the perineum.


Correlations between the CagA Antigen and Serum Levels of Anti-Helicobacter pylori IgG and IgA in Children.
We tested correlations between anti-Helicobacter pylori IgG and IgA levels and the urease test, anti-CagA protein antibody, degree of gastritis, and age. In total, 509 children (0-15 years) were enrolled. Subjects were stratified as 0-4 years (n = 132), 5-9 years (n = 274), and 10-15 years (n = 103) and subjected to the urease test, histopathology, ELISA, and western blot using whole-cell lysates of H. pylori strain 51. The positivity rate in the urease test (P = 0.003), the degree of chronic gastritis (P = 0.021), and H. pylori infiltration (P < 0.001) increased with age. The median titer for anti-H. pylori IgG was 732.5 IU/mL at 0-4 years, 689.0 IU/mL at 5-9 years, and 966.0 IU/mL at 10-15 years (P < 0.001); the median titer for anti-H. pylori IgA was 61.0 IU/mL at 0-4 years, 63.5 IU/mL at 5-9 years, and 75.0 IU/mL at 10-15 years (P < 0.001). The CagA-positivity rate was 26.5% at 0-4 years, 36.5% at 5-9 years, and 46.6% at 10-15 years for IgG (P = 0.036), and 11.3% at 0-4 years, 18.6% at 5-9 years, and 23.3% at 10-15 years for IgA (P < 0.001). Anti-H. pylori IgG and IgA titers increased with the urease test grade, chronic gastritis degree, active gastritis, and H. pylori infiltration. Presence of CagA-positivity is well correlated with a high urease test grade and high anti-H. pylori IgG/IgA levels.


Improved myocardial oxygen utilization following propranolol infusion in adolescents with postburn hypermetabolism.
The purpose of this study was to determine if propranolol (0.5 mg/kg and 1 mg/kg), administered intravenously (IV) at the height of the postburn hypermetabolic response, would decrease myocardial oxygen requirements, without adversely affecting overall oxygen delivery or total body oxygen consumption. To test this hypothesis, six nonseptic patients age 17 +/- 3 years with burns over 82% +/- 11% total body surface area were given propranolol with continuous hemodynamic monitoring. Propranolol was administered to these patients 20 +/- 15 days postburn. Two clinically derived indices of myocardial oxygen consumption, pressure-work index (PWI) and rate-pressure product (RPP), were used to estimate the energy expenditure of the working heart. Both PWI and RPP were significantly decreased from baseline after 0.5 mg/kg propranolol, 31% for PWI (P less than .001) and 30% for RPP (P less than .01). Similarly, a decrease from baseline was seen after 1.0 mg/kg propranolol, 32% for PWI (P less than .001) and 35% for RPP (P less than .01). Cardiac index (L/min/m2) demonstrated no significant change [7.4 +/- 1.1 (prepropranolol), 6.5 +/- 1.3 (after 0.5 mg/kg propranolol), and 6.8 +/- 1.0 (after 1.0 mg/kg propranolol)] and exceeded the upper limits of normal (hyperdynamic state) throughout the study. Oxygen delivery index (962 +/- 209 mL/min/m2) and oxygen consumption indices [(254 +/- 78 mL/min/m2 by Fick method and 236 +/- 78 mL/min/m2 by inspired and expired gases)] were elevated at baseline and unaffected by propranolol. The decrease in PWI and RPP was achieved mainly by propranolol's effect to lower both heart rate and BP.(ABSTRACT TRUNCATED AT 250 WORDS)


Exposure to and experiences of violence among adolescents in lower socio-economic groups in Johannesburg, South Africa.
We explored exposure to and experiences of violence and their risk factors amongst ethnically diverse adolescents from lower socio economic groups in Johannesburg. This cross-sectional study recruited a stratified sample of 16-18 year old adolescents from four low socio-economic suburbs in Johannesburg to reflect ethnic group clustering. We collected socio-demographic, sexual behaviour, alcohol and drug use and trauma events data. Proportions and risk factors were assessed by chi-square and logistic regression. Of 822 adolescents, 57% (n = 469) were female. Approximately 62% (n = 506) were Black, 13% (n = 107) Coloured, 13% (n = 106) Indian and 13% (n = 103) White. Approximately 67% (n = 552) witnessed violence to a non-family member, 28% (n = 228) experienced violence by a non-family member, and 10% (n = 83) reported sexual abuse. Multivariate analysis determined that witnessing violence in the community was associated with being Black (OR: 4.6, 95%CI: 2.7-7.9), Coloured (OR: 3.9, 95%CI: 2.0-7.4) or White (OR: 8.0, 95%CI:4.0-16.2), repeating a grade (OR: 1.5, 95%CI: 1.01-2.1), having more than one sexual partner (OR: 1.7, 95%CI: 1.1-2.5) and ever taking alcohol (OR: 2.1, 95%CI: 1.5-2.9). Witnessing violence in the family was associated with being female (OR: 1.8, 95%CI: 1.3-2.6), being Black (OR: 2.2, 95%CI: 1.1-4.1), or White (OR: 3.0, 95%CI: 1.4-6.4), repeating a grade (OR: 1.6, 95%CI: 1.1-2.2) and ever taking alcohol (OR: 2.9, 95%CI: 2.0-4.3). In low socio-economic areas in Johannesburg, Black, White and Coloured adolescents experience a high burden of violence. Interventions to mitigate the effects of violence are urgently required.


Clinical neurological examination of infants and children.
A thorough but focused history and neurological examination remain the most important initial elements of neurological diagnosis at all ages. Advances over the past two decades in clinical neurophysiology, neuroimaging, genetics, and neuropathological examination of tissue have at times appeared to predominate over traditional history and physical exam, but no laboratory studies can provide the focus and clues to diagnosis that clinical findings offer. History taking and the techniques of neurological examination are skills to be learned by the student, refined by the resident, and practiced and perfected throughout the career of a pediatric neurologist. Examination must be specifically modified to correspond to age and with the expectation of developmental skills achieved at various ages, in addition to the localizing value of particular signs that may apply at all ages. Hypotonia, extensor plantar responses, and lack of visual fixation may be normal in a preterm infant but abnormal at several months of age. "Primitive" reflexes disappear at a certain age, but really are only suppressed or inhibited and may become re-expressed with disinhibition many decades later. Finally, the pediatric neurologist needs to have a firm foundation in normal development, neuroembryology, and changes in the expression of diseases at various stages of maturation of the nervous system.


A giant cystic pheochromocytoma of the adrenal gland.
Adrenal pheochromocytomas are rare catecholamine-secreting tumors that originate from chromaffin cells in the adrenal medulla, and giant pheochromocytomas with cystic changes are particularly rare. We report a case of a 46-year-old man who presented with episodic hypertension and headache. Radiographic studies showed an 18-cm cystic mass in the left upper quadrant of the abdomen; further workups, which included light microscopy, immunohistochemical, and electron microscopic analysis, revealed a pheochromocytoma of the left adrenal gland. Cytogenetic analysis and genetic mutation analyses for von-Hippel-Lindau (VHL), rearranged during transfection (RET), and succinate dehydrogenase complex subunit B (SDHB) genes were also performed but failed to reveal any abnormalities within the tumor cells.


Chemotherapy combined with antiangiogenic drugs as salvage therapy in advanced melanoma patients progressing on PD-1 immunotherapy.
This study aimed to evaluate the effect of salvage therapy with nab-paclitaxel (nab-p) or temozolomide (TMZ) combined with antiangiogenic drugs in programmed death 1 (PD-1) inhibitor-resistant patients with unresectable metastatic melanoma. We conducted a retrospective review of 69 metastatic melanoma patients who received nab-p or TMZ combined with antiangiogenic drugs after developing PD-1 inhibitor resistance and were treated at the Beijing Cancer Hospital between 2016 and 2019. The disease control rate (c-DCR) and progression-free survival (c-PFS) of salvage CA (chemotherapy combined with antiangiogenic drugs) regimens were investigated. Univariate and multivariate analyses were performed to evaluate the clinical pathological factors affecting the outcomes. Then, a nomogram was formulated to predict the probability of 3-month and 6-month c-PFS based on the multivariate analysis results. The c-DCR was 63.8%, and the median c-PFS was 3.0 months. In the univariate analysis, factors associated with the c-DCR were included the melanoma subtype, baseline platelet-to-lymphocyte ratio (PLR) and best response status to PD-1 inhibitors. Factors influencing c-PFS included age, baseline lactic dehydrogenase, PLR, neutrophil-to-lymphocyte ratio (NLR), PFS duration of anti-PD-1 therapy (p-PFS), and the best response and progression pattern of PD-1 inhibitors. In the multivariate analysis, age <65 years, heterogeneous progression pattern and baseline PLR<200 were significantly associated with improved c-PFS. The concordance index (C-index) of the nomogram was equal to 0.65 (95% CI 0.566-0.734). CA regimens demonstrated promising effects in PD-1 inhibitor-resistant patients. The nomogram could be a valuable predictive module for salvage therapy choice in PD-1 inhibitor-resistant patients.


Modelling the Mediterranean Sea ecosystem at high spatial resolution to inform the ecosystem-based management in the region.
Cumulative pressures are rapidly expanding in the Mediterranean Sea with consequences for marine biodiversity and marine resources, and the services they provide. Policy makers urge for a marine ecosystem assessment of the region in space and time. This study evaluates how the whole Mediterranean food web may have responded to historical changes in the climate, environment and fisheries, through the use of an ecosystem modelling over a long time span (decades) at high spatial resolution (8 × 8 km), to inform regional and sub-regional management. Results indicate coastal and shelf areas to be the sites with highest marine biodiversity and marine resources biomass, which decrease towards the south-eastern regions. High levels of total catches and discards are predicted to be concentrated in the Western sub-basin and the Adriatic Sea. Mean spatial-temporal changes of total and commercial biomass show increases in offshore waters of the region, while biodiversity indicators show marginal changes. Total catches and discards increase greatly in offshore waters of the Western and Eastern sub-basins. Spatial patterns and temporal mean changes of marine biodiversity, community biomasses and trophic indices, assessed in this study, aim at identifying areas and food web components that show signs of deterioration with the overall goal of assisting policy makers in designing and implementing spatial management actions for the region.


Relational surrogacies excluded from the French bioethics model: a euro-american perspective in the light of Marcel Mauss and Louis Dumont.
In the French context of prohibition of surrogacy by a legislative framework established in 1994, couples are using surrogacy abroad to create their family. Why does surrogacy not find room in the landscape of donor-conceived families in France? Based on a survey among French intended parents using surrogacy in the USA and Belgium, and a 2-year ethnography on medical practice in a fertility centre in Belgium, this study shows that surrogacy is, in fact, a particular type of gift: the gift of gestational capacity. The preconceptional journey in Belgium or in the USA is a relational process that allows complementary places and statuses to be acquired. This process will transform applicants into intended parents (recipients), and candidates into surrogates (donors). The relationships created by the gift have the particularity of being woven around responsibility towards the fetus. It is the hierarchy of encompassing and encompassed responsibilities in relation to the fetus that organizes the relationships and actions of each protagonist: parents, grandparents, surrogate, surrogate's partner and children, etc. The article thus shows that surrogacy, because it is a gift of a particular type, has no place in the French bioethics model, which is, in fact, built entirely on the notion of 'donation without a donor' in a therapeutic and medicalized view of reproductive donations.


Unconventional Hall response in the quantum limit of HfTe5.
Interacting electrons confined to their lowest Landau level in a high magnetic field can form a variety of correlated states, some of which manifest themselves in a Hall effect. Although such states have been predicted to occur in three-dimensional semimetals, a corresponding Hall response has not yet been experimentally observed. Here, we report the observation of an unconventional Hall response in the quantum limit of the bulk semimetal HfTe5, adjacent to the three-dimensional quantum Hall effect of a single electron band at low magnetic fields. The additional plateau-like feature in the Hall conductivity of the lowest Landau level is accompanied by a Shubnikov-de Haas minimum in the longitudinal electrical resistivity and its magnitude relates as 3/5 to the height of the last plateau of the three-dimensional quantum Hall effect. Our findings are consistent with strong electron-electron interactions, stabilizing an unconventional variant of the Hall effect in a three-dimensional material in the quantum limit.


On the selection of an opioid for local skin analgesia: Structure-skin permeability relationships.
Recent studies demonstrated that post-herpetical and inflammatory pain can be locally managed by morphine gels, empirically chosen. Aiming to rationalize the selection of the most suitable opioid for the cutaneous delivery, we studied the in vitro penetration through human epidermis of eight opioids, evidencing the critical modifications of the morphinan core. Log P, log D, solid-state features and solubility were determined. Docking simulations were performed using supramolecular assembly made of ceramide VI. The modifications on position 3 of the morphinan core resulted the most relevant in determining both physicochemical characteristics and diffusion pattern. The 3-methoxy group weakened the cohesiveness of the crystal lattice structure and increased the permeation flux (J). Computational studies emphasized that, while permeation is essentially controlled by molecule apolarity, skin retention depends on a fine balance of polar and apolar molecular features. Moreover, ChemPLP scoring the interactions between the opioids and ceramide, correlated with both the amount retained into the epidermis (Qret) and J. The balance of the skin penetration properties and the affinity potency for μ-receptors evidenced hydromorphone as the most suitable compound for the induction of local analgesia.


Cardiac toxicity of 5-ring polycyclic aromatic hydrocarbons is differentially dependent on the aryl hydrocarbon receptor 2 isoform during zebrafish development.
Petroleum-derived compounds, including polycyclic aromatic hydrocarbons (PAHs), commonly occur as complex mixtures in the environment. Recent studies using the zebrafish experimental model have shown that PAHs are toxic to the embryonic cardiovascular system, and that the severity and nature of this developmental cardiotoxicity varies by individual PAH. In the present study we characterize the toxicity of the relatively higher molecular weight 5-ring PAHs benzo[a]pyrene (BaP), benzo[e]pyrene (BeP), and benzo[k]fluoranthene (BkF). While all three compounds target the cardiovascular system, the underlying role of the ligand-activated aryl hydrocarbon receptor (AHR2) and the tissue-specific induction of the cytochrome p450 metabolic pathway (CYP1A) were distinct for each. BaP exposure (40μM) produced AHR2-dependent bradycardia, pericardial edema, and myocardial CYP1A immunofluorescence. By contrast, BkF exposure (4-40μM) caused more severe pericardial edema, looping defects, and erythrocyte regurgitation through the atrioventricular valve that were AHR2-independent (i.e., absent myocardial or endocardial CYP1A induction). Lastly, exposure to BeP (40μM) yielded a low level of CYP1A+ signal in the vascular endothelium of the head and trunk, without evident toxic effects on cardiac function or morphogenesis. Combined with earlier work on 3- and 4-ring PAHs, our findings provide a more complete picture of how individual PAHs may drive the cardiotoxicity of mixtures in which they predominate. This will improve toxic injury assessments and risk assessments for wild fish populations that spawn in habitats altered by overlapping petroleum-related human impacts such as oil spills, urban stormwater runoff, or sediments contaminated by legacy industrial activities.


[Comparative analysis of semiotic shifts, established by LCS of blood plasma from random samples of studied subjects from the zone of the Chernobyl accident, "Ural Radiation Trace", and collaborators from St. Petersburg Institute of Nuclear Physics of the Russian Academy of Sciences].
Using the developed "semiotic" classifier of laser correlation spectra of blood plasma the authors have carried out the verification of organism states of patients from the zone of Chernobyl accident, "Ural radiation trace" and collaborators from Sanct-Petersbourg Institute of Nuclear Physics. An analysis of results obtained using accidental selections which differed as to the character of radiation injury evidences for high informativeness of "semiotic" classifier of laser correlation spectra of blood plasma.


Behavioral hyperreactivity in the spontaneously hypertensive rat compared to its normotensive progenitor.
The spontaneously hypertensive rat (SHR) is an excellent model of essential hypertensive disease. Hyperreactivity has been postulated as a contributing factor in the development of high blood pressure in the SHR and in man. In the maturing organism recurring hypertensive stesss responses may promote permanent vascular changes and result in a fixed hypertension. Simple behavioral activity and emotionality rating scales were used to compare a large number of SHR with equally large groups of closely-related and distantly-related normotensive rats. As predicted, the SHR were clearly more active and emotional than their ancestral Wistar Kyoto (WKY) strain. However, the distantly-related normotensive Wistars did not differ from the SHR in either activity or emotionality. These results indicate that behavior and hypertension are not necessarily related in the rat. Nonetheless, the behavioral differences between the SHR and their closest genetic match, the WKY, suggest that arousal and blood pressure levels may be causally linked in this case of naturally occurring hypertension.


Comparative transcriptomic profiling in the pulp and peel of pitaya fruit uncovers the gene networks regulating pulp color formation.
Pitaya (genus Hylocereus) is a popular fruit. To develop pitaya fruit with greater marketability and high nutritional value, it is important to elucidate the roles of candidate genes and key metabolites that contribute to the coloration of the pitaya pulp and peel. By combining transcriptome and biochemical analyses, we compared and analyzed the dynamic changes in the peel and pulp of H. undatus (white pulp) and H. polyrhizus (red pulp) fruits at four key time points during ripening. Differential expression analysis and temporal analysis revealed the difference regulation in pathways of plant hormone signal transduction, phenylpropanoid biosynthesis, and betalain biosynthesis. Our results suggest that color formation of purple-red peel and pulp of pitaya is influenced by betalains. Increased tyrosine content and fluctuation in acylated betalain content may be responsible for pulp color formation, while some of the key genes in this network showed differential expression patterns during ripening between white pulp and red pulp fruits. The data and analysis results of this study provide theoretical basis for the red color formation mechanism of pitaya, which will facilitate future work to improve pitaya fruit physical appearance and marketability.


Interaction Between Innate Lymphoid Cells and the Nervous System.
The interaction between the immune system and the nervous system remains an intriguing enigma. Recent studies indicate that innate lymphoid cells (ILCs), a unique family of innate effector cells, participate in intense cross talk with the nervous system. In the mucosal barrier sites, ILCs have been found to co-localize with neurons, nerves, glial cell projectors, and neuroendocrine cells. The cross talk between ILCs and peripheral nervous system orchestrates mucosal homeostasis and immunity. In addition, the barrier tissues of the central nervous system (CNS) also provide conductive microenvironment for ILC development and maintenance. Activities of CNS-associated ILCs impact the outcome of various CNS disorders. In this chapter, we review and discuss the intricate and bidirectional interaction between ILCs and nervous system.


An individual double-blind crossover trial for assessing methylphenidate response in children with attention deficit disorder.
To obtain more objective data on response to therapy and to overcome parents', teachers', psychologists', and physicians' concerns about potential overuse, I instituted individual double-blind crossover trials of methylphenidate for children with attention deficit disorder (ADD). Each child is given 0.3 mg/kg/dose twice daily for 2 weeks and placebo for 2 weeks in random order and in double-blind fashion. Parents and teachers record observations of behavior and academic performance. Analysis of 70 trials and follow-up clinical data demonstrated that (1) 51 children showed improvement in one of the 2-week periods and that period corresponded with methylphenidate therapy in 48 (69%); (2) 6 of the 22 who did not respond to methylphenidate experienced worsening of function while taking the drug; (3) history and physical findings were not helpful in predicting methylphenidate response; (4) there were no serious side effects during the trial; and (5) all but three of the responders took methylphenidate for at least 1 year with sustained improvement in behavioral function, academic function, or both. Individual double-blind crossover trials can be used in an office setting to identify objectively which children with ADD respond to treatment with methylphenidate. Because the trial demonstrates to parents, teachers, psychologists, and physicians that methylphenidate is or is not beneficial to a particular child, this clinical tool is associated with a high level of confidence that the drug is being appropriately prescribed.


Fetal growth in Peru: comparisons with international fetal size charts and implications for fetal growth assessment.
To compare fetal biometry measurements obtained in a Peruvian population with reference fetal size charts obtained in Peruvian and non-Peruvian populations. Fetal biometry measurements collected prospectively in 195 uncomplicated pregnancies were included in the presented analysis. At 20, 24, 28, 32, 36 and 38 weeks' gestation, fetal head circumference, abdominal circumference and femur diaphysis length were measured. Fetal biometry measurements were compared with fetal size charts obtained from another Peruvian and two non-Peruvian populations from North America and Europe. When compared with ultrasound-based reference fetal size charts obtained from North American and European populations, fetuses from the studied population appeared to grow more slowly with advancing gestational age. This trend was not observed when a Peruvian population, similar to the one studied here, was used as a reference. The results suggest that fetal growth in this Peruvian population may not be adequately assessed by using reference charts obtained from other populations and have implications for the use of growth standards in antenatal management.


Reaction Kinetics of CaOH with H and O2 and O2CaOH with O: Implications for the Atmospheric Chemistry of Meteoric Calcium.
The ablation of cosmic dust particles entering the Earth's upper atmosphere produces a layer of Ca atoms around 90 km. Here, we present a set of kinetic experiments designed to understand the nature of the Ca molecular reservoirs on the underside of the layer. CaOH was produced by laser ablation of a Ca target in the fast flow tube and detected by non-resonant laser-induced fluorescence, probing the D(2Σ+) ← X(2Σ1) transition at 346.9 nm. The following rate constants were measured (at 298 K): k(CaOH + H → Ca + H2O) = (1.04 ± 0.24) × 10-10 cm3 molecule-1 s-1, k(CaOH + O → CaO + OH) < 1 × 10-11 cm3 molecule-1 s-1, and k(CaOH + O2 → O2CaOH, 1 Torr) = (5.9 ± 1.8) × 10-11 cm3 molecule-1 s-1 (uncertainty at the 2σ level of confidence). The recycling of CaOH from reaction between O2CaOH and O proceeds with an effective rate constant of keff(O2CaOH + O → CaOH + products, 298 K) = 2.8-1.2+2.0 × 10-10 cm3 molecule-1 s-1. Master equation modeling of the CaOH + O2 kinetics is used to extrapolate to mesospheric temperatures and pressures. The results suggest that the formation of O2CaOH slows the conversion of CaOH to atomic Ca via reaction with atomic H, and O2CaOH is likely to be a long-lived reservoir species on the underside of the Ca layer and a building block of meteoric smoke particles.


Prognostic Analysis of Primary Breast Signet Ring Cell Carcinoma and Mucinous Breast Adenocarcinoma: A SEER Population-Based Study.
Primary breast signet ring cell carcinoma (SRCC) is a rare type of breast cancer with typical morphological characteristics, high aggressiveness, and poor prognosis. SRCC is different from mucinous breast adenocarcinoma (MBC). However, only a few studies have explored the clinicopathological features and prognosis of SRCC and MBC. Data retrieved from the Surveillance, Epidemiology, and End-Results (SEER) database (2004-2015) were used to explore the prognostic effect of clinicopathological features and treatment modalities on survival outcomes of SRCC and MBC patients. Kaplan-Meier plot analysis, multivariate Cox proportional risk model, propensity score matching (PSM), and subgroup analysis were performed. A total of 167 patients with SRCC and 11,648 patients with MBC were included in the study. SRCC patients exhibited higher histological grade (p < 0.001), larger tumor volume (p < 0.001), higher rate of lymph node metastasis (p < 0.001), and higher frequency of distal metastasis (p < 0.001) compared with MBC patients. Cox proportional hazards regression analysis showed that SRCC patients had lower overall survival (OS) and breast cancer-specific survival (BCSS) compared with MBC patients. Subgroup survival analysis showed that the SRCC patients had lower OS and BCSS in subgroups including younger than 60 years old, white race, married, without chemotherapy, and received radiotherapy compared with the MBC patients in these subgroups. In addition, the SRCC patients had lower BCSS in subgroups including other races (including Asian or Pacific Islander and American Indian/Alaska Native), without surgery, and lymph node metastasis. The findings showed that primary breast SRCC patients have unique clinical characteristics and worse prognosis compared with MBC patients. Notably, different treatment methods resulted in different prognosis for SRCC and MBC types; therefore, SRCC patients should be distinguished from MBC patients to improve efficacy of treatment.


Sphingosine 1-phosphate antagonizes the effect of all-trans retinoic acid (ATRA) in a human colon cancer cell line by modulation of RARβ expression.
All-trans retinoic acid (ATRA) is a promising therapeutic agent, but exhibits low efficacy against human cancers. We investigated the effect of sphingosine-1-phosphate (S1P) on ATRA activity in human colon cancer HT-29 cells. S1P antagonized ATRA activity on HT-29 cell proliferation and retinoic acid receptor beta (RARβ) expression. S1P treatment or transient co-transfection with SphK2 expression vector antagonized ATRA-induced RARβ promoter activity. Proteasome inhibition prevented S1P-induced modulation of ATRA activity. Overall, S1P antagonized ATRA's inhibitory effects by down-regulating RARβ expression, likely via the proteasome-dependent pathway. Decreasing S1P production or inhibiting SphK2 activity could enhance the efficacy of retinoids in cancer treatments.


Nutrient loading of a forest soil : A manipulative approach using rooted experimental chambers.
During July 1988 rooted and non-rooted experimental chambers were established in a Norway spruce (Picea abies. Karst) stand in south Devon U.K. Replicates were supplemented with ammonium and nitrate. The leachates were analysed to monitor the release of mineral-N species and cations over the 17-week experimental period. Ammonium treatments leached 300% more calcium and magnesium than controls. The onset of nitrification resulted in a decrease in sodium losses from ammonium treatments reflecting a decrease in the exchanging capacity of the soil solution. These results are discussed in relation to mineral ion leaching in soils subjected to increesed N-loading, and the ability of soils to buffer these perturbations.


Long-term prognosis of childhood absence epilepsy.
Childhood absence epilepsy (CAE) is considered easily manageable with medication provided that a strict patient classification system is employed. It accounts for 10% of all childhood epilepsy cases starting before the age of 15 and it is most frequent in school-aged girls. The aim of this study is to analyse long-term outcomes of patients diagnosed with CAE according to the Loiseau and Panayiotopoulos criteria and treated during childhood. We conducted a retrospective study including 69 patients with CAE who are currently older than 11; data were gathered from medical histories, EEG records, and telephone questionnaires. 52 patients met the Loiseau and Panayiotopoulos criteria. Mean age is now 17.16 years. Female-to-male ratio was 1.65:1; mean age at onset was 6 years and 2 months; mean duration of treatment was 3 years and 9 months. A family history of epilepsy was present in 30.8% of the patients and 7.7% had a personal history of febrile convulsions. Absence seizures were simple in 73.5% of the patients and complex in 26.5%. Response rates to first-line treatment were as follows: valproic acid, 46.3%; and valproic acid plus ethosuximide, 90.9%. The rate of response to second-line therapy (ethosuximide or lamotrigine) was 84.2%; 4% of the patients experienced further seizures after treatment discontinuation, 78.8% achieved seizure remission, and 25% needed psychological and academic support. Our data show that epileptic patients should be classified according to strict diagnostic criteria since patients with true CAE have an excellent prognosis. The relapse rate was very low in our sample. Despite the favourable prognosis, psychological and academic support is usually necessary.


Prevention of iron deficiency by milk fortification. II. A field trial with a full-fat acidified milk.
In a longitudinal study from age 3 to 15 mo, 276 term, healthy, spontaneously weaned infants received a full-fat acidified milk fortified with 15 mg of elemental Fe as ferrous sulfate and 100 mg of ascorbic acid/100 g of powder and 278 control infants received milk without additives. At ages 9 and 15 mo significant differences were encountered in all measures of Fe nutriture in favor of the fortified group (p less than 0.001). Anemia (Hg less than 110 g/L) was present in 25.7% of unfortified infants compared with only 2.5% in those fortified at age 15 mo. Saturation of transferrin less than 9% was present in 33.8% and serum ferritin less than 10 micrograms/L in 39.1% of the nonfortified infants. The figures for the fortified group were 7 and 8.5% respectively. The efficiency of the fortified acidified milk in eradicating Fe deficiency in the infants while discouraging use by other family members make this milk a useful targeted product in programs of supplementary food distribution in the underdeveloped world.


[The brachial plexus lesion. Management, consequences of palsy and reconstructive operations].
The treatment of traumatic brachial plexus lesions follows sequential steps. After acute therapy (phase I), neurological diagnostics (phase II), neurosurgery and postoperative treatment (phase III/IV), reconstructive operations (phase V) can be indicated and performed. In most cases an insufficient grade of muscle power remains. Within 6 months after injury, neurosurgery must be performed in patients with brachial plexus palsy. After malfunction of the muscles, taking into account the individual neuromuscular defects, passive joint function and bony deformities, different procedures such as muscle transposition, arthrodesis and corrective osteotomy can be performed to improve the function of the upper extremity. The treatment of patients suffering from brachial plexus lesion requires interdisciplinary teamwork.


Mitigating Financial Toxicity in Breast Cancer From Diagnosis to Treatment and Reconstruction.
Breast cancer is the most common cancer affecting women in United States with a significant patient and familial financial burden. Limited resources offer financial support to patients in the form of cost support for bills, health insurance payments, childcare, and post-treatment recovery. Financial resources are often limited to patients of certain geographic regions or low socioeconomic status as determined by the federal poverty level, which may systematically exclude patients in need.


Association between the clastogenic effect in peripheral lymphocytes and human exposure to arsenic through drinking water.
We describe the association between structural chromosome aberrations (CAs) and parameters of exposure to arsenic among 42 individuals exposed to arsenic through well waters in Finland. The median concentration of arsenic in the wells was 410 microg/l, the total arsenic concentrations in urine (As-tot) was 180 microg/l, and in hair 1.3 microg/g, for current users (n = 32) of contaminated wells. Urinary arsenic species and CAs were also analyzed in eight control individuals from the same village who consumed water which contained arsenic <1.0 microg/l (detection limit). Increased arsenic exposure, indicated best by increased concentrations of arsenic species (inorganic arsenic, methylarsonic acid (MMA), dimethylarsinic acid (DMA)) in urine, was associated with increased frequency of CAs. The increased urinary ratio of MMA/As-tot and the decreased ratio of DMA/As-tot were associated with increased CAs when all aberration types, including gaps, were considered. Associations between CAs and arsenic exposure indicators were stronger among current users than among persons who had stopped using the contaminated well water for 2-4 months before sampling (ex-users, n = 10). Furthermore, there was a positive but not statistically significant association between CAs and arsenic in hair among the current users, but not among the ex-users, who still had relatively high arsenic concentrations in hair. The results suggest that the effect observed in the present study reflects relatively recent arsenic exposure.


Biosurfactants of Rhodococcus erythropolis IMV Ас-5017: synthesis intensification and practical application.
Intensification of the surfactant synthesis by Rhodococcus erythropolis IMV Ac-5017 on different substrates, including industrial waste, as well as the use of surfactant preparations for oil degradation were studied. It was established that the addition of fumarate (0.2 %) and citrate (0.1 %) into the medium with ethanol, n-hexadecane, or glycerol (1-2 %) was accompanied by an increase of conditional surfactant concentration by 1.5-1.7 times compared to the indexes in the medium without organic acids. The intensification of surfactant synthesis in the presence of fumarate and citrate is caused by the increased activity of isocitrate lyase (by 1.2-15-fold) and enzymes of the surfactant biosynthesis (by 2-4.8-fold) compared to their activity in the medium without precursors. The possibility of surfactant synthesis intensification (by 3-4-fold) while cultivating of R. erythropolis IMV Ac-5017 in the medium with oil containing substrates (2 %) and glucose (0.1 %) was shown. The introduction of 0.01 mM Cu(2+) in the exponential growth phase of strain IMV Ac-5017 in the medium with ethanol accompanied by the increasing conditional surfactant concentration by 1.9 times. The highly efficient remediation (92-95 %) of oil (2-2.6 g/L) and Cu(2+) polluted water after treatment with surfactant preparations (native cultural liquid) at low concentrations (5 %) was determined.


Exploiting gene-environment independence in haplotype-based inferences for population-based case-control studies with complex sampling.
The use of complex sampling in population-based case-control studies is becoming more common. Although most single nucleotide polymorphism-based association studies with complex sampling account for the design complications, many of haplotype-based genetic association studies with complex sampling tend to ignore them when estimating haplotype frequencies, regression coefficients, or both. In this article, we develop innovative one-step and two-step statistical methods that account for the design complications in haplotype-based association studies when cases and/or controls are sampled with complex sampling. Attracted by the efficiency advantage of the retrospective method, we explore the assumptions of Hardy-Weinberg equilibrium and gene-environment independence in the underlying population. Results of our simulation studies demonstrate superior performance of the proposed methods over selected existing methods under various complex sampling designs. An application of the proposed methods is illustrated using a population-based case-control study of kidney cancer.


[Expression of mitogen-activated protein kinase and its upstream regulated signal in human hepatocellular carcinoma].
To detect protein expression of ERK(1), ERK(2), JNK(1), p38 and MEK(1), MEK(2) in human hepatocellular carcinoma and adjacent non-neoplastic liver. In 16 surgically resected hepatocellular carcinoma and para-carcinoma tissues, Western blotting was used to detect expression of ERK(1), ERK(2), JNK(1), p38 and MEK(1), MEK(2). In all cases, ERK(1), ERK(2), p38 expression in hepatocellular carcinoma was significantly higher than that in para-carcinoma: integral optic density (IOD) of ERK(1) was 300 +/- 98 in carcinoma and 98 +/- 48 in para-carcinoma tissues (t = 2.519, P < 0.01); IOD of ERK(2) was 587 +/- 83 in carcinoma and 232 +/- 96 in para-carcinoma tissues (t = 2.745, P < 0.01); IOD of p38 was 270 +/- 85 in carcinoma and 107 +/- 88 in para-carcinoma tissues (t = 2.491, P < 0.01). JNK(1) expression in hepatocellular carcinoma was significantly lower than that in para-carcinoma; IOD of JNK(1) was 111 +/- 93 in carcinoma and 292 +/- 109 in para-carcinoma tissues (t = 2.473, P < 0.01). Protein levels of MEK(1) and MEK(2) in carcinoma were significantly higher than in para-carcinoma. IOD of MEK(1) was 1 418 +/- 244 in carcinoma and 806 +/- 90 in para-carcinoma tissues (t = 2.546, P < 0.01). IOD of MEK(2) was 1 041 +/- 122 in carcinoma and 468 +/- 40 in para-carcinoma tissues (t = 2.861, P < 0.01). ERK(1), ERK(2), MEK(1) and MEK(2) in the signal transduction pathway for cell proliferation are significantly overexpressed and the expression of JNK(1) is lower in hepatocellular carcinoma. Their unbalance is one of the important reasons for the over growth and infinite proliferation of the hepatocellular carcinoma cell. The p38 and JNK(1) may be activated by different pathway.


Are there promising biologic therapies for osteoarthritis?
A large unmet need exists for desirable pharmacologic therapeutic interventions for osteoarthritis (OA). Developers of these interventions should be mindful of the current status of therapeutic interventions and focus on the mechanics of the joint and tissues of symptom origin. Much of the pathophysiology of the joint is still poorly understood, particularly as it relates to tissues other than hyaline articular cartilage. Improved knowledge of the role of these tissues (especially bone, fat, and synovium) in OA pathophysiology will likely yield treatment breakthroughs. This review discusses promising biologic therapies for the treatment of OA. Investigators are developing promising therapies, but we need to pay heed to the lessons learned thus far if new therapies are to be effective.


Acute appendicitis in childhood: a feasibility study of computer-assisted diagnosis.
The case records of 169 children admitted to hospital with suspected appendicitis were subjected to Bayesian analysis in order to determine whether computer-assisted diagnosis would be likely to reduce the negative laparotomy rate. The results suggest that the negative laparotomy rate could be reduced from 27% to 18% if computer-assisted diagnosis was used.


Metal-Free Addition of Alkyl Bromides to Access 3,3-Disubstituted Quinoxalinones Enabled by Visible-Light Photoredox Catalysis.
A metal-free addition of unactivated alkyl bromides to quinoxalin-2(1H)-ones is described. This method enables the construction of valuable 3,3-disubstituted dihydroquinoxalin-2(1H)-ones bearing quaternary carbon centers under mild, visible-light photoredox catalysis. High functional group tolerance is observed in both the quinoxalinone and alkyl bromide partners. The ability to scale up this method was demonstrated under photo-flow conditions to enable gram-scale synthesis.


Internal heavy atom effect of Au(III) and Pt(IV) on hypocrellin A for enhanced in vitro photodynamic therapy of cancer.
Hypocrellin A (HA), an a natural perylene quinine photosensitizers (PSs), can chelate with heavy metal ions, including Au(III) and Pt(IV), to form a 1:2 complex, which exhibits enhanced (1)O2 generation quantum yield through the increased intersystem crossing efficiency mediated by internal heavy atom effect. Besides, the chelate process greatly improved the water solubility of HA. Comparative studies with HA and complexes have demonstrated that the heavy-atom effect on HA molecules enhances the efficiency of in vitro photodynamic (PDT) efficacy.


Health care reform 2010: a fresh view on tort reform.
We reviewed the state of medical malpractice tort reform in the context of a new political climate and the current debate over comprehensive health care reform. Specifically we asked whether medical malpractice tort reform is necessary, and evaluated the strengths and weaknesses of contemporary reform proposals. The medical, legal and public policy literature related to medical malpractice tort reform was reviewed and synthesized. We include a primer for understanding the current structure of medical malpractice law, identify the goals of the current system and analyze whether these goals are presently being met. Finally, we describe and evaluate the strengths and weaknesses of the current reform proposals including caps on damages, safe harbors and health care courts. Medical malpractice tort law is designed to improve health care quality and appropriately compensate patients for medical malpractice injuries, but is failing on both fronts. Of the 3 proposed remedies, caps on damages do little to advance the quality and compensatory goals, while safe harbors and health care courts represent important advancements in tort reform. Tort reform should be included in the current health policy debate because the current medical malpractice system is not adequately achieving the basic goals of tort law. While safe harbors and health care courts both represent reasonable remedies, health care courts may be preferred because they do not rely on jury determination in the absence of strong medical evidence.


Yellow Nail Syndrome With Dramatic Improvement of Nail Manifestations After Endoscopic Sinus Surgery.
Yellow nail syndrome (YNS) is a rare disease of unknown cause characterized by the triad of yellow nails, respiratory manifestations, and lymphedema. Although several therapies for YNS have been reported, there is no common consensus in the treatment. In this case report, we present a case of 56-year-old woman with YNS, whose nail manifestation was dramatically improved after endoscopic sinus surgery for the treatment of chronic rhinosinusitis. Endoscopic sinus surgery involving middle meatal antrostomy was performed for the case of YNS with chronic rhinosinusitis and bronchiectasis resistant to antibacterial drugs. A month after the surgery, the patient's nails eventually showed dramatic improvement. Otorhinolaryngologists should recognize that chronic rhinosinusitis can be a symptom of YNS, and that the aggressive treatment including surgical approach for chronic rhinosinusitis may be a useful in the control of nail manifestation in YNS.


Determination of an immunosuppressive substance in serum by latex particle electrophoresis.
The level of immunosuppressive substance (IS), which increases in the serum of patients with cancer, was determined by an assay based on particle electrophoresis. Polystyrene latex particles (PLP) were coated with IS, which was extracted from the ascitic fluid of patients with cancer. The IS was a glycoprotein with a molecular weight of about 52,000, and an isoelectric point in the range pH 2.7-3.3. When the IS on the surface of the PLP reacted with the anti-IS antibody, the mean electrophoretic mobility of the PLP changed from -3.16 to +0.21 micron.s-1.V-1.cm in the medium of pH 7.2 and ionic strength I = 0.0154. After preincubation of anti-IS antiserum and tested serum, the PLP coated with IS were added to this solution. It was incubated again and then the surface charge of the PLP was measured by an automatic cell-electrophoretic instrument. This method was used to determine the IS concentration in the serum of cancer patients and pregnant women. When compared to healthy controls, the serum IS level was significantly higher in patients with cancer, and lower in pregnant women. The assay based on latex-particle electrophoresis proved to be a sensitive and rapid method for determining the IS level in serum.


Bronchial adenoma--benign or malignant?
Bronchial adenomas grow slowly and are potentially malignant. Of the 45 cases reviewed, 36 were carcinoids, six were adenocystic carcinomas, two were mixed tumors, and one was mucoepidermoid carcinoma. The carcinoid group is subdivided into histologically typical 27(75%), atypical 9 (25%), and metastasizing 9 (25%) adenomas. There were 32 lobectomies, seven pneumonectomies, one sleeve resection, and two tracheal resections for adenocystic carcinoma. Ten-year survival rate was 88% for typical carcinoids, 66% for atypical carcinoids, and 44% for metastasizing carcinoids. In the cylindroma group, one patient was lost to follow-up, one died postoperatively, and one developed recurrence of the tumor eight years later. The only patient with mucoepidermoid carcinoma is alive without evidence of recurrence nine years after operation.


Electron cyclotron resonance plasma production by using pulse mode microwaves and dependences of ion beam current and plasma parameters on the pulse condition.
We measure the ion beam current and the plasma parameters by using the pulse mode microwave operation in the first stage of a tandem type ECRIS. The time averaged extracted ion beam current in the pulse mode operation is larger than that of the cw mode operation with the same averaged microwave power. The electron density n(e) in the pulse mode is higher and the electron temperature T(e) is lower than those of the cw mode operation. These plasma parameters are considered to cause in the increase of the ion beam current and are suitable to produce molecular or cluster ions.


Comprehensive genome based analysis of Vibrio parahaemolyticus for identifying novel drug and vaccine molecules: Subtractive proteomics and vaccinomics approach.
Multidrug-resistant Vibrio parahaemolyticus has become a significant public health concern. The development of effective drugs and vaccines against Vibrio parahaemolyticus is the current research priority. Thus, we aimed to find out effective drug and vaccine targets using a comprehensive genome-based analysis. A total of 4822 proteins were screened from V. parahaemolyticus proteome. Among 16 novel cytoplasmic proteins, 'VIBPA Type II secretion system protein L' and 'VIBPA Putative fimbrial protein Z' were subjected to molecular docking with 350 human metabolites, which revealed that Eliglustat, Simvastatin and Hydroxocobalamin were the top drug molecules considering free binding energy. On the contrary, 'Sensor histidine protein kinase UhpB' and 'Flagellar hook-associated protein of 25 novel membrane proteins were subjected to T-cell and B-cell epitope prediction, antigenicity testing, transmembrane topology screening, allergenicity and toxicity assessment, population coverage analysis and molecular docking analysis to generate the most immunogenic epitopes. Three subunit vaccines were constructed by the combination of highly antigenic epitopes along with suitable adjuvant, PADRE sequence and linkers. The designed vaccine constructs (V1, V2, V3) were analyzed by their physiochemical properties and molecular docking with MHC molecules- results suggested that the V1 is superior. Besides, the binding affinity of human TLR-1/2 heterodimer and construct V1 could be biologically significant in the development of the vaccine repertoire. The vaccine-receptor complex exhibited deformability at a minimum level that also strengthened our prediction. The optimized codons of the designed construct was cloned into pET28a(+) vector of E. coli strain K12. However, the predicted drug molecules and vaccine constructs could be further studied using model animals to combat V. parahaemolyticus associated infections.


Further electrophoretic studies of erythrocyte glutathione peroxidase.
A new, inexpensive technique has been devised for the detection of glutathione peroxidase after electrophoresis. Surveys of several racial groups have revealed new variant phenotypes at polymorphic frequencies in African and Lebanese populations.


Risk of Human Papillomavirus (HPV) Infection and Cervical Neoplasia after Pregnancy.
Parity is well established as a risk factor for cervical cancer. It is not clear, however, how pregnancy influences the natural history of HPV infection and cervical neoplasia. Our objective was to study the risk of HPV infection and cervical squamous intraepithelial lesions (SIL) after pregnancy. We used the Ludwig-McGill cohort study which includes 2462 women recruited in Sao Paulo, Brazil in 1993-97 and followed for up to 10 years. Cellular specimens were collected every 4-6 months for Pap cytology and HPV detection and genotyping by a polymerase chain reaction protocol. Study nurses recorded pregnancy occurrence during follow-up. HPV and Pap results from pregnant women were available before and after, but not during pregnancy. The associations between pregnancy and post-partum HPV infection/SIL were studied using generalized estimating equation models with logistic link. Adjusted odds ratios (OR) were estimated with empirical adjustment for confounding. We recorded 122 women with a history of pregnancy during follow-up. Of these, 29 reintegrated the cohort study after delivery. No association between HPV and pregnancy was found. A single SIL case (high grade SIL) occurred post-partum. Likewise, there was no association between pregnancy and risk of low grade SIL or any-grade SIL at the next visit (adjusted OR = 0.84, 95 % CI: 0.46-15.33) after controlling for confounders. No associations were found between pregnancy and HPV or LSIL. The single observed case of HSIL post-partum was more than would be expected based on the rate of these abnormalities among non-pregnant women. As this association was found with only one case, caution is required in the interpretation of these results.


Non-compliance to social distancing during COVID-19 pandemic: A comparative cross-sectional study between the developed and developing countries.
Despite mass vaccination campaigns, the world has seen a steady rise in the number of SARS-CoV-2 cases, with 178,765,626 cases and 3,869,994 COVID-19 related deaths by June 19th, 2021. Therefore, it is important to enforce social distancing to control its spread. With the variation observed in the severity of the pandemic in different countries, it is also imperative to study the social distancing behaviors amongst the population in developed and developing countries. In this cross-sectional study, a total of 384 participants from 14 different countries were surveyed via an online REDCap form. In this study, it was highlighted that despite adequate knowledge, the overall compliance to COVID-19 related preventive measures remains poor, the lowest being in the senior age group (≥ 65 years), and the highest being in adults aged between 25-64 years (p=0.003). Population from the developing countries were more compliant to all preventative measures against COVID-19 spread, except for handwashing, where the difference between the two populations remained insignificant (p=0.038, p<0.001, p=0.016) Socioeconomic status, prior history of COVID-19 infection or presence of comorbidities did not significantly affect compliance rates, however, participants with no prior history of this infection were found to be more compliant to donning a mask in public as compared to those with a positive history (p=0.044). Since compliance remains subpar in both the developing and the developed countries, mass campaigns about COVID-19 related preventive measures remain essential in controlling the disease spread.


Prolonged disease-free survival in bronchogenic carcinoma associated with HLA-Aw19 and HLA-B5. A two-year prospective study.
Seventy patients with squamous carcinoma or adenocarcinoma of the lung were HLA typed at the time of their diagnosis and initial therapy. No abnormal HLA antigen frequencies were found. However, the possession of HLA-Aw19 or HLA-B5 was significantly correlated with two year disease-free survival. Twelve of 21 patients (57%) with either Aw 19 or B5 were disease-free at two years. In contrast, only six of 48 patients (13%) not having either antigen were disease-free (p less than .001). One patient was lost to follow-up. It is possible that Aw 19 and/or B5 confer resistance to progression of bronchogenic carcinoma on their possessors.


Design and validation of a phantom for transcranial ultrasonography.
Commercial medical ultrasound phantoms are highly specific as they simulate particular clinical scenarios. This makes them expensive to use in multi-target research and training. General approaches to human tissue and organ modeling are described in the manufacturing methodology, access to which is restricted by the manufacturer's trade secret. Our aim is to propose a reproducible methodology to design a head phantom for transcranial ultrasound training and research from widely available materials and to validate its applicability. To create an anthropomorphic phantom, we used data from real patients obtained by CT and MRI scans. We combined FDM and LCD 3D printing to achieve the desired acoustic performance and ergonomics of the phantom. We fabricated the phantom using polyvinyl chloride plastisol, photopolymer, and PLA to simulate brain tissue, temporal acoustic windows, and acoustically opaque parts of the skull, respectively. Notably, the phantom fabrication method uses only readily available materials and is easy to reproduce. We developed a basic one and anatomical one versions of the head phantom. The basic version contains a simplified brain: tissue-mimicking material is poured into the skull with needles inserted, which specific pattern is easy to recognize in B-mode images. The anatomical version has an anatomically correct brain dummy extracted from MRI data and contains multiple randomly distributed small metal, plastic, and bony objects ranging in size from 1 to 3 mm each. The proposed methodology allows producing head phantoms for transcranial ultrasound training and research. The anatomical accuracy of the model is proved by ultrasonography and CT studies. Both versions of the phantom comprise the skull and the brain and are intended for ultrasound imaging through the temporal bone acoustic window. Needles and small objects serve as navigation targets during the training procedure. The basic version helps learning basic navigation skills, while the anatomical one provides a realistic setting to perform the diagnostic procedure.


Too packed to change: side-chain packing and site-specific substitution rates in protein evolution.
In protein evolution, due to functional and biophysical constraints, the rates of amino acid substitution differ from site to site. Among the best predictors of site-specific rates are solvent accessibility and packing density. The packing density measure that best correlates with rates is the weighted contact number (WCN), the sum of inverse square distances between a site's C α and the C α of the other sites. According to a mechanistic stress model proposed recently, rates are determined by packing because mutating packed sites stresses and destabilizes the protein's active conformation. While WCN is a measure of C α packing, mutations replace side chains. Here, we consider whether a site's evolutionary divergence is constrained by main-chain packing or side-chain packing. To address this issue, we extended the stress theory to model side chains explicitly. The theory predicts that rates should depend solely on side-chain contact density. We tested this prediction on a data set of structurally and functionally diverse monomeric enzymes. We compared side-chain contact density with main-chain contact density measures and with relative solvent accessibility (RSA). We found that side-chain contact density is the best predictor of rate variation among sites (it explains 39.2% of the variation). Moreover, the independent contribution of main-chain contact density measures and RSA are negligible. Thus, as predicted by the stress theory, site-specific evolutionary rates are determined by side-chain packing.


Chronic toxicity of diclofenac, carbamazepine and their mixture to Daphnia magna: a comparative two-generational study.
The chronic toxicity of diclofenac (DCF) and carbamazepine (CBZ) as separate substances and in conjunction with their mixture on Daphnia magna was assessed in the parental (F0) and first filial (F1) generations. The second (F1-B2) and fifth (F1-B5) broods of F1 offspring were investigated and compared. Both drugs and their mixture were exposed to each generation of Daphnia magna for 21 days with life history, behavioural and gene expressions as measured endpoints. After the parental exposure, offspring from these two broods were transferred to a clean medium for a 21-day recovery. Exposure to diclofenac, carbamazepine and their mixture significantly inhibited growth, reproduction, swimming activities, heart rate, thoracic limb activities, reproductive and antioxidant-related genes in the parental as well as the first filial generations. These effects were relatively greater in the F1 generation. This indicates that Daphnia magna's sensitivity improved while its fitness declined over the two generations, which is an indicator of greater energy requirements for maintenance. Besides, the significant inhibition in the antioxidant-related genes implies that oxidative stress occurred in Daphnia magna under the exposure to these drugs. The significant reduction in the reproductive output, moulting frequency and cyp314 gene expression as a result of exposure to CBZ simultaneously obtained herein may indicate that this drug could act as an endocrine disruptor. Most of these significant effects were not recoverable after the 21-day recovery period. The findings reported herein highlight the necessity to include maternal effects in environmental risk assessment processes, considering that pollutant effects are underestimated during single-generational exposure.


Network modelling reveals the mechanism underlying colitis-associated colon cancer and identifies novel combinatorial anti-cancer targets.
The connection between inflammation and tumourigenesis has been well established. However, the detailed molecular mechanism underlying inflammation-associated tumourigenesis remains unknown because this process involves a complex interplay between immune microenvironments and epithelial cells. To obtain a more systematic understanding of inflammation-associated tumourigenesis as well as to identify novel therapeutic approaches, we constructed a knowledge-based network describing the development of colitis-associated colon cancer (CAC) by integrating the extracellular microenvironment and intracellular signalling pathways. Dynamic simulations of the CAC network revealed a core network module, including P53, MDM2, and AKT, that may govern the malignant transformation of colon epithelial cells in a pro-tumor inflammatory microenvironment. Furthermore, in silico mutation studies and experimental validations led to a novel finding that concurrently targeting ceramide and PI3K/AKT pathway by chemical probes or marketed drugs achieves synergistic anti-cancer effects. Overall, our network model can guide further mechanistic studies on CAC and provide new insights into the design of combinatorial cancer therapies in a rational manner.


Structural insights at acidic pH of dye-decolorizing peroxidase from Bacillus subtilis.
Dye-decolorizing peroxidases (DyPs), a type of heme-containing oxidoreductase enzymes, catalyze the peroxide-dependent oxidation of various industrial dyes as well as lignin and lignin model compounds. In our previous work, we have recently reported the crystal structures of class A-type DyP from Bacillus subtilis at pH 7.0 (BsDyP7), exposing the location of three binding sites for small substrates and high redox-potential substrates. The biochemical studies revealed the optimum acidic pH for enzyme activity. In the present study, the crystal structure of BsDyP at acidic pH (BsDyP4) reveals two-monomer units stabilized by intermolecular salt bridges and a hydrogen bond network in a homo-dimeric unit. Based on the monomeric structural comparison of BsDyP4 and BsDyP7, minor differences were observed in the loop regions, that is, LI (Ala64-Gln71), LII (Glu96-Lys108), LIII (Pro117-Leu124), and LIV (Leu295-Asp303). Despite these differences, BsDyP4 adopts similar heme architecture as well as three substrate-binding sites to BsDyP7. In BsDyP4, a shift in Asp187, heme pocket residue discloses the plausible reason for optimal acidic pH for BsDyP activity. This study provides insight into the structural changes in BsDyP at acidic pH, where BsDyP is biologically active.


Multiple electrodes for detecting spikes in partial complex seizures.
The contribution of various electroencephalographic electrodes in detecting spikes from patients with seizures of suspected anterior temporal origin was prospectively studied with a standard protocol. The following electrodes were studied: International Standard 10-20 positions F7-8 and A1-2, sphenoidal (SP), nasopharyngeal (NP), anterior temporal (T1-2), mandibular notch surface (MNS), and mandibular notch subdermal (MNSD). Twenty patients were recorded of whom 16 demonstrated anterior temporal spikes. There was no difference in the number of spikes detected by SP, MNS, MNSD, or T1-2 electrodes (p less than 0.05); however these electrodes detected significantly more spikes than NP, F7-8, or A1-2. The SP electrode recorded spikes of highest amplitude (p less than 0.05). We conclude that for patients suspected of having seizures of anterior temporal origin, (1) a substantial number of spikes will be missed if only the International Standard electrode system is employed; (2) in comparison to SP electrodes the non-invasive and easily applied MNS or T1-2 electrodes will detect almost all spikes and should be used in outpatient EEG recordings; (3) NP electrodes provide no information that cannot be obtained by more reliable and better tolerated electrodes.


Quantification of collagen organization in histopathology samples using liquid crystal based polarization microscopy.
A number of histopathology studies have utilized the label free microscopy method of Second Harmonic Generation (SHG) to investigate collagen organization in disease onset and progression. Here we explored an alternative label free imaging approach, LC-PolScope that is based on liquid crystal based polarized light imaging. We demonstrated that this more accessible technology has the ability to visualize all fibers of interest and has a good to excellent correlation between SHG and LC-PolScope measurements in fibrillar collagen orientation and alignment. This study supports that LC-PolScope is a viable alternative to SHG for label free collagen organization measurements in thin histology sections.


CdGAP/ARHGAP31, a Cdc42/Rac1 GTPase regulator, is critical for vascular development and VEGF-mediated angiogenesis.
Mutations in the CdGAP/ARHGAP31 gene, which encodes a GTPase-activating protein for Rac1 and Cdc42, have been reported causative in the Adams-Oliver developmental syndrome often associated with vascular defects. However, despite its abundant expression in endothelial cells, CdGAP function in the vasculature remains unknown. Here, we show that vascular development is impaired in CdGAP-deficient mouse embryos at E15.5. This is associated with superficial vessel defects and subcutaneous edema, resulting in 44% embryonic/perinatal lethality. VEGF-driven angiogenesis is defective in CdGAP(-/-) mice, showing reduced capillary sprouting from aortic ring explants. Similarly, VEGF-dependent endothelial cell migration and capillary formation are inhibited upon CdGAP knockdown. Mechanistically, CdGAP associates with VEGF receptor-2 and controls VEGF-dependent signaling. Consequently, CdGAP depletion results in impaired VEGF-mediated Rac1 activation and reduced phosphorylation of critical intracellular mediators including Gab1, Akt, PLCγ and SHP2. These findings are the first to demonstrate the importance of CdGAP in embryonic vascular development and VEGF-induced signaling, and highlight CdGAP as a potential therapeutic target to treat pathological angiogenesis and vascular dysfunction.


Syntheses and pharmacokinetic studies of prodrug esters for the development of oral carbapenem, L-084.
We discovered an orally active carbapenem, L-084, through pharmacokinetic studies on various prodrug esters of (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-l-methyl-2-[1-(1,3-thiazolin-2-yl)azetidin-3-yl]thio-1-carbapen-2-em-3-carboxylic acid (LJC11,036). L-084 showed a strong antimicrobial activity against Gram-positive and Gram-negative bacteria and exhibited the highest intestinal absorption among synthesized prodrugs of LJC11,036.


Correlation Between Presbylarynx and Laryngeal EMG.
The normal aging process affects many human functions profoundly. The voice is no exception, and some aging effects on the voice are obvious including vocal fold atrophy, bowing, stiffness, more prominent vocal fold processes, and glottic insufficiency. The study's primary aim was to determine the correlation between presbylarynx and laryngeal electromyography (LEMG) results and to compare these electromyography (EMG) results with young and old voice patients without evidence of presbylarynx changes on strobovideolaryngoscopy. Medical records for voice patients seen in the senior author's (RTS) practice between 2015 and 2019 were reviewed retrospectively. Patients with presbylarynx confirmed by strobovideolaryngoscopy were identified and compared to two control groups. The first control group included patients with ages and American Society of Anesthesiologists scores similar to the experimental group but no presbylarynx. The second control group included younger patients (ages ranging between 18 and 35 years with mean age of 24.57 years) with vocal fold paresis. All groups had undergone LEMG because of suspected paresis clinically. Social and medical factors reviewed and compared between groups included smoking history, alcohol consumption, occupation (voice demanding versus not voice demanding occupation), Voice Handicap Index score at the initial office visit, medical comorbidities, medications, and LEMG results. There was no significant difference in the recruitment results for three pairs of muscles (cricothyroid, thyroarytenoid and posterior cricoarytenoid) between the presbylarynx group and both control groups. Rate of asthma was higher in presbylarynx group compared with the first control group. Rates of corticosteroid inhaler use, thyroid medications, smoking, and Voice Handicap Index score were higher in presbylarynx group compared with the second control group. There was no significant difference in occupation type and alcohol consumption between groups. There was no significant difference found in the neuromuscular function based on laryngeal EMG between presbylarynx patients and both old and young patients with vocal fold paresis but without presbylarynx. Corticosteroid inhaler is associated with atrophic changes seen in presbylarynx. There still could be differences in the neuromuscular function which weren't detected. More research is needed to confirm or refute these findings.


The effect of ketoconazole on whole blood and skin ciclosporin concentrations in dogs.
Ciclosporin (CSA) is approved for the treatment of canine atopic dermatitis. Ciclosporin is metabolized by liver cytochrome P450 enzymes, a process inhibited by ketoconazole (KTZ). The aims of this study were to determine skin and blood CSA concentrations when CSA was administered alone at 5.0 (Treatment 1) or 2.5 mg/kg (Treatment 2) and when CSA was administered at 2.5 mg/kg concurrently with KTZ at 5 (Treatment 3) or 2.5 mg/kg (Treatment 4). We hypothesized that skin and blood CSA concentrations in Treatment 1 would not differ from those obtained with T3 or T4. In a randomized cross-over study, six healthy research dogs received each of the treatments (Treatment 1, 2, 3 and 4) once daily for 7 days. After the first, fourth and seventh dose for each treatment, a peak and trough skin punch biopsy sample and whole blood sample were collected and analysed with high-performance liquid chromatography-tandem mass spectrometry. Data were analysed using a repeated measures approach with PROC MIXED in SAS. Pairwise comparisons were performed with least squares means and Tukey-Kramer adjustment for multiple comparisons. Mean blood CSA concentrations in Treatment 1 were not different from those in Treatment 2 or 4, but were less than in Treatment 3. Mean skin CSA concentrations in Treatment 1 were greater than in Treatment 2, not different from those in Treatment 4, and less than those in Treatment 3. Administration of CSA and KTZ concurrently at 2.5 mg/kg each may be as effective as CSA alone at 5.0 mg/kg for treatment of canine atopic dermatitis.


Sp8 regulates inner ear development.
A forward genetic screen of N-ethyl-N-nitrosourea mutagenized Xenopus tropicalis has identified an inner ear mutant named eclipse (ecl). Mutants developed enlarged otic vesicles and various defects of otoconia development; they also showed abnormal circular and inverted swimming patterns. Positional cloning identified specificity protein 8 (sp8), which was previously found to regulate limb and brain development. Two different loss-of-function approaches using transcription activator-like effector nucleases and morpholino oligonucleotides confirmed that the ecl mutant phenotype is caused by down-regulation of sp8. Depletion of sp8 resulted in otic dysmorphogenesis, such as uncompartmentalized and enlarged otic vesicles, epithelial dilation with abnormal sensory end organs. When overexpressed, sp8 was sufficient to induce ectopic otic vesicles possessing sensory hair cells, neurofilament innervation in a thickened sensory epithelium, and otoconia, all of which are found in the endogenous otic vesicle. We propose that sp8 is an important factor for initiation and elaboration of inner ear development.


Socioeconomic factors and risk of hospitalization with infectious diseases in 0- to 2-year-old Danish children.
Although the association between low socioeconomic status and illness in childhood is well known, the impact of socioeconomic factors on risk and frequency of hospitalizations for infectious diseases, the most frequent disease category, during the first 2 years of life has scarcely been studied. Through linkage of records drawn from public administrative and health registries, we conducted a population-based cohort study of 5024 Danish children born in 1997 to examine the frequency of hospitalization for infectious diseases in very young children. The main exposure variables, adjusted for potential confounding factors, were mother's education level, household income, and cohabitation status. The outcome was number of hospital admissions (0, 1-2, or 3+) for infectious diseases. A total of 737 children (14.7%) were admitted to hospital 1-2 times, and 83 (1.6%) were admitted 3-10 times. The risk of hospitalization was increased in children of mothers with a low level of education compared with vocational education (1-2 admissions: adjusted odds ratio (OR) 1.3 (95% confidence interval [CI]: 1.1-1.6). Children from low-income families had an increased risk of 3 or more admissions (low cf middle income: adjusted OR 2.0 [95% CI: 0.6-6.0]). Children of single mothers had an increased risk of hospitalization (1-2 admissions: adjusted OR 1.7 [95% CI: 1.1-2.6]. We found the highest risk of hospitalization for infectious diseases in children of mothers with only basic schooling, and particularly among those coming from single parent homes with a low income.


Secretion of tumor necrosis factor-alpha from human placental tissues induced by hypoxia-reoxygenation causes endothelial cell activation in vitro: a potential mediator of the inflammatory response in preeclampsia.
Preeclampsia is a hypertensive complication of human pregnancy characterized by generalized maternal endothelial cell activation. Circulating pro-inflammatory cytokines derived from the placenta are thought to play a key role. We recently demonstrated that hypoxia-reoxygenation (H/R) of placental tissues in vitro causes equivalent oxidative stress to that seen in preeclampsia. Our aim was to determine whether H/R also increases production of tumor necrosis factor-alpha (TNF-alpha), and whether conditioned media from samples exposed to H/R causes activation of human umbilical vein endothelial cells (HUVECs). Concentrations of mRNA encoding TNF-alpha were significantly higher in placental tissues subjected to H/R compared to hypoxic or normoxic controls. Although there was no difference in the concentrations of TNF-alpha protein in tissue homogenates, levels of TNF-alpha protein in the medium were significantly higher after H/R compared to controls, indicating increased secretion. Furthermore, conditioned medium from samples subjected to H/R caused increased expression of E-selectin by HUVECs, and the addition of anti-TNF-alpha antibodies significantly reduced that activation. These results are consistent with our hypothesis that intermittent perfusion of the placenta, secondary to reduced trophoblast invasion, causes increased secretion of TNF-alpha, and that this contributes to the activation of maternal endothelial cells that characterizes preeclampsia.


Role for cyclic adenosine monophosphate in the synergistic interaction between oxytocin and corticotrophin-releasing factor in isolated human gestational myometrium.
We wished to investigate the role of cAMP in the synergistic effect of corticotrophin-releasing factor and oxytocin on human myometrial contractility. Isolated human gestational myometrium obtained from Caesarean sections at term was studied in vitro. Static incubation techniques as well as tension recordings were applied to the tissue obtained. The subjects were healthy pregnant women undergoing lower segment Caesarean section at term, prior to labour. Specimens obtained were immediately dissected into small strips and either incubated in multi-well trays (strip weight 2.75 mg) or superfused and used for tension recordings (strip weight 2.00 mg). cAMP accumulation was measured after incubation with oxytocin (0.1-10 nM), corticotrophin-releasing factor (1 nM) or a combination of both peptides. Tension generated by the muscle strips was recorded isometrically and response to oxytocin (0.01-10 nM), corticotrophin-releasing factor (1 nM) and forskolin (10 nM) expressed in force per gram wet tissue (N/g). Oxytocin (0.1 nM) causes a statistically significant dose-related decrease in cAMP when combined with 1 nM corticotrophin-releasing factor (P less than 0.001), as compared with cAMP stimulation by corticotrophin-releasing factor alone. Time course studies suggest a maximal effect at 1 minute. The hypothesis that an intracellular reduction of cAMP is a prerequisite for the synergistic response in contraction force was tested with tension recordings. Prevention of a decrease in cAMP in the tissue by addition of 10 nM forskolin to the superfusate abolished the potentiation between oxytocin and corticotrophin-releasing factor. These results indicate that a fall in cAMP concentration plays a vital mediating role in the synergistic interaction between oxytocin and corticotrophin-releasing factor.


[Incidence of endophthalmitis after intravitreal Avastin injection with and without postoperative topical antibiotic application].
Intravitreal injection of vascular endothelial growth factor inhibitors is safe as a standardized procedure. In a retrospective study, we examined the incidence of intraocular inflammation and endophthalmitis when intravitreal Avastin was given on both an outpatient and an inpatient basis. Two centers treated a total of 1844 patients with intravitreal Avastin injection (IVA), 1.25 mg/0.05 ml. The medication was given under aseptic conditions with a sterile drape and lid speculum and after an incubation of 5% povidone-iodine for 60 s in the conjunctival fornix. Center A performed all applications as an outpatient procedure with no postoperative topical antibiotics. Center B performed all IVAs prior to a 1-day hospital admission and gave topical antibiotics for 3 days. In both centers the patients were told to return to the hospital immediately if visual disturbance, pain, or redness of the eyes was noticed. Center A did 984 IVAs. Postoperatively no patients had endophthalmitis. On the 2nd postoperative day, two patients developed a mild intraocular uveitis, which was treated with local steroid ointments. Center B did 860 IVAs. Five days after discharge from the hospital, two patients presented with endophthalmitis and underwent vitrectomy when their vision dropped to counting fingers. There was no positive microbiological result of either vitreous probe. Although equal treatment guidelines were established, the perioperative hospital admission and postoperative application of topical antibiotic ointment could not reduce the occurrence of mild to severe intraocular endophthalmitis.


A randomized, controlled trial comparing compression bandaging and cold therapy in postoperative total knee replacement surgery.
To examine the difference between compression bandaging and cold therapy after total knee arthroplasty. Eighty-four postoperative, unilateral, total knee replacement, surgical clients. Clients were randomized into two groups: those receiving compression bandaging, and those receiving cryo-pad technology. Subjects were assessed for total length of stay, blood loss, blood transfusion, swelling, flexion, pain, and opiate use. Unlike other studies, the results of these data showed no significant differences between groups on the measured outcomes. A simple cost benefit analysis shows that the compression bandage is cheaper and more labor efficient than the cold therapy as delivered by cryo-pad technology.