A Novel Mechanism for Protein Delivery by the Type 3 Secretion System for Extracellularly Secreted Proteins.
The type 3 secretion system (T3SS) is essential for bacterial virulence through delivering effector proteins directly into the host cytosol. Here, we identified an alternative delivery mechanism of virulence factors mediated by the T3SS, which consists of the association of extracellularly secreted proteins from bacteria with the T3SS to gain access to the host cytosol. Both EspC, a protein secreted as an enteropathogenic Escherichia coli (EPEC) autotransporter, and YopH, a protein detected on the surface of Yersinia, require a functional T3SS for host cell internalization; here we provide biophysical and molecular evidence to support the concept of the EspC translocation mechanism, which requires (i) an interaction between EspA and an EspC middle segment, (ii) an EspC translocation motif (21 residues that are shared with the YopH translocation motif), (iii) increases in the association and dissociation rates of EspC mediated by EspA interacting with EspD, and (iv) an interaction of EspC with the EspD/EspB translocon pore. Interestingly, this novel mechanism does not exclude the injection model (i.e., EspF) operating through the T3SS conduit; therefore, T3SS can be functioning as an internal conduit or as an external railway, which can be used to reach the translocator pore, and this mechanism appears to be conserved among different T3SS-dependent pathogens.IMPORTANCE The type 3 secretion system is essential for injection of virulence factors, which are delivered directly into the cytosol of the host cells for usurping and subverting host processes. Recent studies have shown that these effectors proteins indeed travel inside an "injectisome" conduit through a single step of translocation by connecting the bacterium and host cell cytoplasms. However, all findings are not compatible with this model. For example, both YopH, a protein detected on the surface of Yersinia, and EspC, an autotransporter protein secreted by enteropathogenic E. coli, require a functional T3SS for host cell translocation. Both proteins have an intermediate extracellular step before their T3SS-dependent translocation. Here, we show an alternative delivery mechanism for these extracellularly secreted virulence factors that are then incorporated into the T3SS to enter the cells; this novel mechanism coexists with but diverges from the canonical injection model that involves the passage of the protein inside the injectisome.


Effects of voluntary attendance patterns on first-year medical students' wellness and academic performance during COVID-19.
There is increasing evidence that attendance is not a reliable predictor of academic performance, which invites the question of whether attendance may have alternative positive effects such as increased student wellness. While conducting an observational cohort study on the relationship of attendance to performance and wellness, the COVID-19 pandemic lockdown changed the ability of students to attend class. Matriculating medical students were recruited to participate at the beginning of their first year in August 2019. In-person attendance was collected prospectively until the lockdown began in March 2020. This was then correlated with performance on course final examinations and responses to two separate incidences of a survey on stress, burnout, social isolation, and loneliness. The first survey was deployed January through February 2020 (pre-COVID) and the second June 2020 (during COVID lockdown). Attendance declined across the year (76.0%-25.0%). There was a small positive correlation of attendance to performance in module 1 (r = 0.235, P = 0.035) with none in the remaining two modules (P = 0.870, P = 0.410). The high attenders at the time of the lockdown had no decline in their performance when attendance was no longer possible. Attendance negatively correlated with stress and loneliness but not with the remainder of the wellness metrics. There was no significant change in any of the wellness metrics between the pre-COVID and during COVID time points. While the data support the conclusion that attendance is not required to maintain performance in this population, these data suggest a small protective effect against stress and loneliness.


Quantitative determination of steroids in the fruiting bodies and submerged-cultured mycelia of Inonotus obliquus.
This study describes the method of quantitative determination of betulin, ergosterol, cholesterol, lanosterol, stigmasterol and sitosterol in the fruiting bodies and submerged-cultured mycelia of Inonotus obliquus. A high performance liquid chromatographic (HPLC) method was applied to separate these steroids. The procedure was carried out on a reversed-phase C, column, using a stepwise gradient of water-methanol as mobile phase with the following profile: 0-10 min, 10% water, 90% methanol; 10-40 min, 3% water, 97% methanol. The flow rate was 1.4 mL/min and the detection wavelength was 202 nm. The analysis was completed within 40 min. The results showed that this method has good reproducibility and satisfactory recoveries for the determination of steroids. The relative standard deviations of the peak areas were less than 2.94% (n = 5) for intraday assays. A good linear correlation was obtained in a range of 0.4-4.8 microg. The recoveries of betulin, ergosterol, cholesterol, lanosterol, stigmasterol, and sitosterol were 100.05%-100.72%, 99.31%-101.04%, 97.52%-101.63%, 96.61%-100.08%, 96.21%-100.76% and 100.04%-100.51%, respectively. This method can be applied to evaluate real samples, and it is rapid, accurate and suitable for the quantitative determination of steroids in the fruiting bodies and submerged-cultured mycelia of Inonotus obliquus.


Disrupted development and integrity of frontal white matter in patients treated for pediatric medulloblastoma.
Treatment of pediatric medulloblastoma is associated with known neurocognitive deficits that we hypothesize are caused by microstructural damage to frontal white matter (WM). Longitudinal MRI examinations were collected from baseline (after surgery but before therapy) to 36 months in 146 patients and at 3 time points in 72 controls. Regional analyses of frontal WM volume and diffusion tensor imaging metrics were performed and verified with tract-based spatial statistics. Age-adjusted, linear mixed-effects models were used to compare patient and control images and to associate imaging changes with Woodcock-Johnson Tests of Cognitive Abilities. At baseline, WM volumes in patients were similar to those in controls; fractional anisotropy (FA) was lower bilaterally (P < 0.001) and was associated with decreased Processing Speed (P = 0.014) and Broad Attention (P = 0.025) performance at 36 months. During follow-up, WM volumes increased in controls but decreased in patients (P < 0.001) bilaterally. Smaller WM volumes in patients at 36 months were associated with concurrent decreased Working Memory (P = 0.026) performance. Lower FA in patients with pediatric medulloblastoma compared with age-similar controls indicated that patients suffer substantial acute microstructural damage to supratentorial frontal WM following surgery but before radiation therapy or chemotherapy. Additionally, this damage to the frontal WM was associated with decreased cognitive performance in executive function 36 months later. This early damage also likely contributed to posttherapeutic failure of age-appropriate WM development and to the known association between decreased WM volumes and decreased cognitive performance.


[Use of an electronic barostat for measurement of motor response of the proximal stomach to feeding and different nervous stimuli in man].
Electronic barostat allows to measure volume variations of an intragastric air-filled bag maintained at a low constant pressure level, reflecting gastric tone variations. The aims of the present study were to record physiologic variations of gastric tone in humans a) in the fasting state, with and without different stimuli, and b) in the fed state. The bag was placed into the proximal stomach of healthy subjects and connected to the barostat. Volume variations were recorded in the basal state and in response to a) a balanced liquid meal (200 mL, 200 kcal, n = 8), b) sham feeding and sham feeding after cholinergic blockade (atropine 10 micrograms.kg-1 IV, n = 8), and c) graded antroduodenal distensions performed with a tandem balloon (n = 6). During fasting, the proximal stomach presented both a tonic and an intermittent phasic motor pattern (volume waves of 1.2 min maximal period). Meal ingestion was rapidly followed by a dramatic fall in proximal gastric tone (> 200% of preprandial volume within 11 +/- 5 min). After a meal, the intragastric bag returned to preprandial volume after 92 +/- 12 min (ranges: 75-110). Proximal gastric tone was not significantly modified by atropine or by sham feeding alone or after atropine. Antroduodenal distensions reproducibly induced volume dependent relaxations (r = 0.98, P < 0.001) with inhibition of the phasic activity. These results confirm the value of electronic barostat for the measurement of proximal gastric tone, and show that postprandial relaxation is not masked by a low volume liquid meal. Sham feeding does not appear to be an adequate stimulus to induce gastric relaxation. This suggests that vagal fibers involved in gastric relaxation are not the same (or have a higher stimulation threshold) than fibers involved in cephalic phase of gastric secretion.


Pharmacokinetics of ciprofloxacin in patients with liver cirrhosis.
Bacterial infections are common in patients with cirrhosis of liver and are frequently treated with ciprofloxacin. Literature on pharmacokinetics of ciprofloxacin in patients with cirrhosis of the liver is scanty. The present study compared the pharmacokinetics of ciprofloxacin in cirrhotic patients with that in healthy volunteers. In 20 patients with cirrhosis of liver (all Child-Pugh class B) and 10 healthy volunteers, plasma levels of ciprofloxacin were measured using high-performance liquid chromatography at several time points after a 500-mg oral dose. Various pharmacokinetic parameters were calculated. No significant differences were observed in maximum plasma levels reached (mean [SD] 2.6 [0.6] vs 2.6 [1.3] microg/ml), time taken for maximum plasma levels to be reached (1.3 [0.6] vs 1.5 [0.9] h), t1/2a (0.7 [0.3] vs 0.4 [0.9] h), elimination half-life (3.6 [1.2] vs 3.2 [1.8] h), and area under the curve (19.3 [3.8] vs 21.9 [4.5] microg/mL x h) in healthy volunteers and cirrhotic patients, respectively. Pharmacokinetics of ciprofloxacin is unaltered in patients with liver cirrhosis. Ciprofloxacin can be safely administered in the usual doses in such patients.


Differences between urban and rural suicides.
As part of a nation-wide psychological autopsy we examined the differences in DSM-III-R mental disorders, recent life events and other characteristics between urban (n = 143) and rural (n = 85) completed suicides in a random sample of 229 cases from the National Suicide Prevention Project in Finland for the period 1987-1988. Psychoactive substance use disorders (48% vs. 34%), cluster B personality disorders (24% vs. 9%) and psychiatric comorbidity (66% vs. 42%) were found more commonly among urban than rural suicides. Urban suicides were also more often reported to be preceded by a recent separation (25% vs. 8%), whereas rural suicide victims tended to have lacked a close companion of the opposite sex (36% vs. 18%) and to have had physical disorders (56% vs. 40%). Overall, urban and rural suicides may vary with regard to the prevalence of some mental disorders, their comorbidity, and physical disorders, as well as the preceding life situation. This variation may also imply the need for differences in strategies for suicide prevention in each setting.


Trimodality of isoniazid elimination: phenotype and genotype in patients with tuberculosis.
The study was undertaken to show that polymorphic isoniazid elimination in humans is trimodal; that the acetylator genotype and eliminator phenotype of the individual patient are concordant; and that the differences in the pharmacokinetic parameters of fast, intermediate, and slow eliminator subgroups are statistically significant. Sixty adult patients of both sexes and of mixed race with tuberculosis participated in the trial. The apparent elimination rate constant (k, h(-1)) and the area under the isoniazid concentration-time curve (AUC, mg/L/h), over the interval 2 to 6 h after oral isoniazid were determined in all patients; NAT2 allele composition was determined in 47 patients. Serum INH concentrations were determined by HPLC and genotypes by PCR/restriction enzyme analysis. Three eliminator phenotypes could be distinguished, and concordance between the phenotype and the genotype of the individual could be demonstrated. The isoniazid concentration-time profiles of the three eliminator subgroups were significantly different (p < 0.05). The NAT2*12A allele, which codes for fast acetylation, has a high frequency in the population studied, the intermediate acetylator genotype is constituted of codominant fast and slow alleles, and the distribution of phenotypes/genotypes in the population is consistent with Hardy-Weinberg predictions. The therapeutic implications of polymorphic isoniazid metabolism are discussed.


Executive deficits in major depression.
Clinically depressed patients and control subjects were examined by means of different tests to assess executive functions (the Wisconsin Card Sorting Test, the Stroop Color Word Test and a test of verbal fluency). The results indicate that the depressed patients were generally slower than the controls, as reflected by longer retrieval times for both words and colours. Furthermore, the depressed patients showed impairment with regard to altering behaviour appropriately in response to feedback. However, there were no depression-related increases in perseverative responses, and the ability to inhibit irrelevant information was unaffected by depression. No relationship was found between specific depressive symptoms or the severity of depression and cognitive performance. The overall pattern of results suggests that depression may affect various executive functions in a differentiated manner.


Associations of leptin gene polymorphisms with production traits in pigs.
The associations of leptin (LEP) gene polymorphisms C798T, T2411C, T3266G and T3469C with production traits were investigated in a F2 pig population produced by divergent crosses. The statistical model included genotype, sex, batch and genotype by sex interaction as fixed effects and sire as random effect. Polymorphism C798T was associated with variation in total teat number (p < 0.02) and left teat number (p < 0.03), and polymorphism T3469C was associated with weight at 21 days (p < 0.03), 42 days (p < 0.05), 63 days (p < 0.02) and 77 days of age (p < 0.04) as well as feed intake (p < 0.01), average daily gain (p < 0.01), feed conversion (p < 0.01), bacon depth (p < 0.03) and slaughter weight (p < 0.03). Phenotypic associations were also performed by combining T3469C and C798T genotypes. Interaction between C798T genotypes and sex was observed for some traits. LEP genotypes had significant influence on performance traits, and can be considered as potential genetic markers for selection. However, these results have to be validated in commercial herds.


Improvement of a dynamic scanning force microscope for highest resolution imaging in ultrahigh vacuum.
We report on a modification of a commercial scanning force microscope (Omicron UHV AFM/STM) operated in noncontact mode (NC-AFM) at room temperature in ultrahigh vacuum yielding a decrease in the spectral noise density from 2757 to 272 fm/Hz. The major part of the noise reduction is achieved by an exchange of the originally installed light emitting diode by a laser diode placed outside the vacuum, where the light is coupled into the ultrahigh vacuum chamber via an optical fiber. The setup is further improved by the use of preamplifiers having a bandpass characteristics tailored to the cantilever resonance frequency. The enhanced signal to noise ratio is demonstrated by a comparison of atomic resolution images on CeO(2)(111) obtained before and after the modification.


Regional citrate anticoagulation in critically ill patients treated with plasma filtration and adsorption.
In high-risk bleeding conditions conventional systemic anticoagulation with heparin is a contraindication to renal replacement therapy. We evaluate the feasibility and safety of regional citrate anticoagulation in high-risk bleeding conditions during coupled plasma filtration adsorption (CPFA). Thirteen critically ill patients (9 severely burned, 4 polytraumas) with septic shock and acute renal failure treated with CPFA-CVVHD by using bicarbonate-based solutions (heparin-CPFA group, 58 sessions) or with CPFA-CVVHF using citrate (citrate-CPFA group, 36 sessions). Plasma flow and used cartridges showed no differences between the citrate-CPFA and heparin-CPFA groups, while lost clotted cartridges were significantly lower in the citrate-CPFA group. Blood ionized calcium (iCa2+), Ca2+ infusion, pH and bicarbonates remained constant during citrate-CPFA, with no difference between pre- and post-cartridge plasma citrate. A significant positive correlation between iCa2+ in blood and ultrafiltrate was present. These suits demonstrate the feasibility and safety of regional citrate anticoagulation in severely burned and polytrauma septic patients treated by CPFA.


Tunable 1D and 2D Polyacrylonitrile Nanosheet Superstructures.
Carbon superstructures are widely applied in energy and environment-related areas. Among them, the flower-like polyacrylonitrile (PAN)-derived carbon materials have shown great promise due to their high surface area, large pore volume, and improved mass transport. In this work, we report a versatile and straightforward method for synthesizing one-dimensional (1D) nanostructured fibers and two-dimensional (2D) nanostructured thin films based on flower-like PAN chemistry by taking advantage of the nucleation and growth behavior of PAN. The resulting nanofibers and thin films exhibited distinct morphologies with intersecting PAN nanosheets, which formed through rapid nucleation on existing PAN. We further constructed a variety of hierarchical PAN superstructures based on different templates, solvents, and concentrations. These PAN nanosheet superstructures can be readily converted to carbon superstructures. As a demonstration, the nanostructured thin film exhibited a contact angle of ∼180° after surface modification with fluoroalkyl monolayers, which is attributed to high surface roughness enabled by the nanosheet assemblies. This study offers a strategy for the synthesis of nanostructured carbon materials for various applications.


Cutaneous lesions associated with dual infection caused by canine distemper virus and orthopoxvirus in a domestic cat.
Within the context of an increased epidemiological pressure caused by canine distemper virus (CDV) in Switzerland together with a potential re-emergence of endemic pathogens such as orthopoxviruses (OPXV), dual infections are possible among susceptible species. To describe a case of concurrent CDV and OPXV infection in a cat. A 5-year-old, neutered male cat was presented with erythema, crusts and ulcerations around the left eye. High-grade pruritus and a severe conjunctivitis were also present. Formalin-fixed skin biopsy samples were obtained from lesional skin. Histopathology, CDV immunohistochemistry and CDV and OPXV RT-PCR were performed. Histopathological examination showed severe epidermal necrosis extending to the follicular walls and a dermal infiltration, predominantly eosinophilic. Intranuclear and intracytoplasmic eosinophilic inclusion bodies were visible in the wall of affected hair follicles, with occasional formation of syncytia. The RT-PCR revealed the contextual presence of both CDV and OPXV. Scattered cells stained positive for CDV by immunohistochemistry. Dual infections with CDV and OPXV, although rare, may occur and represent additional differential diagnoses for ulcerative skin lesions in cats.


Reductively Stable Hydrogen-Bonding Ligands Featuring Appended CF2-H Units.
We present the development of ligands featuring the unconventional hydrogen bond donor, -CF2H, within a metal's secondary coordination sphere. When metalated with palladium, o-CF2H-functionalized 1,10-phenanthroline provides highly directed H-bonding interactions with Pd-coordinated substrates. Spectroscopic and computational analyses with a series of X-type ligand acceptors (-F, -Cl, -Br, -OR) establish the H-bonding interaction strength for the -CF2H group (∼3 kcal/mol). The synthesis of Pd0/Ni0 complexes and subsequent coupling (Ni) highlight the unique reductive and base compatibility of the -CF2H hydrogen bond donor group.


Infectious diseases in systemic lupus erythematosus: risk factors, management and prophylaxis.
Infections are one of the leading causes of morbidity and mortality in patients with systemic lupus erythematosus (SLE). Therapeutic, disease-related and genetic factors all contribute to a lupus patient's increased susceptibility to infections. Although bacterial pathogens are the most common cause of infections, a wide variety of pathogens have been reported. In high-risk populations, identification and treatment of chronic infections such as tuberculosis, hepatitis B or human immunodeficiency virus, are important prior to the institution of immunosuppression to prevent reactivation or exacerbation of the infection. Judicious use of corticosteroids and cytotoxic drugs is critical in limiting infectious complications. Vaccines against influenza and pneumococcus appear to be safe and immunogenic in SLE patients and their routine administration should be encouraged.


Spinal antinociception of synthetic omega-conotoxin SO-3, a selective N-type neuronal voltage-sensitive calcium channel blocker, and its effects on morphine analgesia in chemical stimulus tests in rodent.
SO-3, a novel Omega-superfamily conotoxin derived from Conus striatus, selectively inhibits N-type neuronal voltage-sensitive calcium channels. In current study, antinociception of SO-3 compared with MVIIA or morphine and its effects on morphine analgesia were investigated in rodent chemical stimulus tests after acute or repeated intrathecal administration. In mice acetic acid writhing test, similar to MVIIA, SO-3 caused dose- and time-dependent spinal antinociception with ED(50) of 0.25 microg/kg and t(1/2) of 4h, which was more potent and longer-acting than morphine. In rat formalin test after intrathecal bolus injection, SO-3 produced dose- and time-dependent antinociception by suppressing acute (ED(50), 1.79 microg/kg) and tonic phases (ED(50), 0.41 microg/kg), which was similar to MVIIA and approximately 10-fold potency and twice longer-acting of morphine in blocking tonic phase responses. After repeated intrathecal injections twice daily for 5 consecutive days, SO-3 produced analgesia without loss of potency whereas morphine produced analgesia tolerance in rat formalin test; further, SO-3 still produced potent analgesia in morphine-tolerant rats. SO-3 co-administered with morphine left-shift the dose-response curve of morphine in mice acetic acid writhing test and significantly potentiated morphine analgesia in rat formalin test. No changes in motor function were seen in mice or rats receiving antinociceptive doses of SO-3 whereas MVIIA caused motor dysfunction at doses of 1.0-2.0 microg/kg in rats. This study showed that (1) novel SO-3 produced potent and long-acting spinal antinociception without observable motor dysfunction, (2) SO-3 significantly potentiated morphine analgesia, (3) After repeated intrathecal administration, SO-3 produced neither tolerance nor cross-tolerance to morphine analgesia.


Against the Odds: Massive Lumbar Intradural Disk Herniation in the Elderly.
Study Design Case report. Objective Presentation of an unusual case of an elderly patient with massive intradural disk herniation at the L2-L3 level. Methods Clinical and imaging data are presented after obtaining informed consent from the patient. Results A 90-year-old man suffering from sudden-onset neurogenic bladder dysfunction and lower back pain but no further neurologic deficits initially presented with magnetic resonance imaging and laboratory values suggestive of an intraspinal infection. However, intraoperative inspection proved the unexpected finding of a large intradural lumbar disk herniation at the L2-L3 level. Conclusions Lumbar soft disk herniation to the intradural space is a rare event and has never been described in a patient over the age of 75. This case of a 90-year-old man with acute-onset bladder dysfunction underlines the necessity to consider this as a differential diagnosis in the case of a newly diagnosed intradural mass.


Treatment of rupture of suspensory ligament and superficial flexor tendon in a bull.
Rupture of the suspensory ligament at the insertions on the proximal sesamoid bones, and of the superficial flexor tendon of the left fore limb, occurred in an adult Angus bull as a result of fighting. There was severe hyperextension of the metacarpophalangeal (MCP) joint with the dewclaws almost touching the ground. Radiographs revealed severe hyperextension of the MCP joint with the sesamoid bones aligned directly distal to the metacarpus. Initially, a full length fiberglass cast was applied with the limb partially flexed within the cast and the heels elevated. The cast was replaced twice. The cast was removed after 136 days and the bull was bearing full weight on the limb. Prolonged immobilisation of the limb produced new bone in the area (a normal response in cattle) to cause ankylosis of the traumatized MCP joint and partial ankylosis of the carpus. The bull was being used for pasture breeding one year after the injury.


Does PET SUV Harmonization Affect PERCIST Response Classification?
Pre- and posttreatment PET comparative scans should ideally be obtained with identical acquisition and processing, but this is often impractical. The degree to which differing protocols affect PERCIST classification is unclear. This study evaluates the consistency of PERCIST classification across different reconstruction algorithms and whether a proprietary software tool can harmonize SUV estimation sufficiently to provide consistent response classification. Eighty-six patients with non-small cell lung cancer, colorectal liver metastases, or metastatic melanoma who were scanned for therapy monitoring purposes were prospectively recruited in this multicenter trial. Pre- and posttreatment PET scans were acquired in protocols compliant with the Society of Nuclear Medicine and Molecular Imaging and the European Association of Nuclear Medicine (EANM) acquisition guidelines and were reconstructed with a point spread function (PSF) or PSF + time-of-flight (TOF) for optimal tumor detection and also with standardized ordered-subset expectation maximization (OSEM) known to fulfill EANM harmonizing standards. After reconstruction, a proprietary software solution was applied to the PSF ± TOF data (PSF ± TOF.EQ) to harmonize SUVs with the OSEM values. The impact of differing reconstructions on PERCIST classification was evaluated. For the OSEMPET1/OSEMPET2 (OSEM reconstruction for pre- and posttherapeutic PET, respectively) scenario, which was taken as the reference standard, the change in SUL was -41% ± 25 and +56% ± 62 in the groups of tumors showing a decrease and an increase in 18F-FDG uptake, respectively. The use of PSF reconstruction affected classification of tumor response. For example, taking the PSF ± TOFPET1/OSEMPET2 scenario increased the apparent reduction in SUL in responding tumors (-48% ± 22) but reduced the apparent increase in SUL in progressing tumors (+37% ± 43), as compared with the OSEMPET1/OSEMPET2 scenario. As a result, variation in reconstruction methodology (PSF ± TOFPET1/OSEMPET2 or OSEM PET1/PSF ± TOFPET2) led to 13 of 86 (15%) and 17 of 86 (20%) PERCIST classification discordances, respectively. Agreement was better for these scenarios with application of the propriety filter, with κ values of 1 and 0.95 compared with 0.79 and 0.72, respectively. Reconstruction algorithm-dependent variability in PERCIST classification is a significant issue but can be overcome by harmonizing SULs using a proprietary software tool.


Water slug formation and motion in gas flow channels: the effects of geometry, surface wettability, and gravity.
Water emerging from ∼100 μm pores into millimeter-size gas flow channels forms drops that grow and become slugs which span the flow channel. Flowing gas causes the slugs to detach and move down the channel. The effect of channel geometry, surface wettability, and gravity on the formation and motion of water slugs has been analyzed using high-speed video images of the drops and differential pressure-time traces. Drops grow and appear, assuming a sequence of shapes that minimize the total interfacial energy of the gas-liquid and liquid-solid interfaces. The drops are initially spherical caps centered on the pore (the liquid contacts one wall). Above a certain size, the drops move to the corner, forming "corner drops" (the liquid contacts two walls). Corner drops grow across the channel, evolving into partial liquid bridges (drops confined by three walls), and finally the drops span the channel cross-section forming slugs (contacting all four walls). Smaller slugs are formed in channels with hydrophobic walls than in channels with hydrophilic walls. Smaller slugs are formed in channels with curved walls than in square or rectangular channels. Slugs move when the differential gas pressure overcomes the force to move the advancing and receding gas-liquid-solid contact lines of the slugs. Residual water left behind in corners by moving slugs reduces the barriers for drops to form slugs, causing the steady-state slug volumes to be smaller than those seen at start-up in dry channels.


Prevalence of molecular markers of resistance to sulfadoxine-pyrimethamine before and after community delivery of intermittent preventive treatment of malaria in pregnancy in sub-Saharan Africa: a multi-country evaluation.
The effectiveness of community delivery of intermittent preventive treatment (C-IPT) of malaria in pregnancy (IPTp) with sulfadoxine-pyrimethamine has been evaluated in selected areas of the Democratic Republic of the Congo, Madagascar, Mozambique, and Nigeria. We aimed to assess the effect of C-IPTp on the potential development of Plasmodium falciparum resistance to sulfadoxine-pyrimethamine, since it could threaten the effectiveness of this strategy. Health facility-based cross-sectional surveys were conducted at baseline and 3 years after C-IPTp implementation in two neighbouring areas per country, one with C-IPTp intervention, and one without, in the four project countries. Dried blood spots from children under five years of age with clinical malaria were collected. Sulfadoxine-pyrimethamine resistance-associated mutations of the P falciparum dhfr (Asn51Ile/Cys59Arg/Ser108Asn/Ile164Leu) and dhps (Ile431Val/Ser436Ala/Ala437Gly/Lys540Glu/Ala581Gly/Ala613Ser) genes were analysed. 2536 children were recruited between June 19 and Oct 10, 2018, during baseline surveys. Endline surveys were conducted among 2447 children between July 26 and Nov 30, 2021. In the Democratic Republic of the Congo, the dhfr/dhps IRNI/ISGEAA inferred haplotype remained lower than 10%, from 2% (5 of 296) at baseline to 8% (24 of 292) at endline, and from 3% (9 of 300) at baseline to 6% (18 of 309) at endline surveys in intervention and non-intervention areas respectively with no significant difference in the change between the areas. In Mozambique, the prevalence of this haplotype remained stable at over 60% (194 [64%] of 302 at baseline to 194 [64%] of 303 at endline, and 187 [61%] of 306 at baseline to 183 [61%] of 301 in endline surveys, in non-intervention and intervention areas respectively). No isolates harbouring the dhps ISGEAA genotype were found in Nigeria. In Madagascar, only five isolates with this haplotype were found in the non-intervention area (2 [>1%] of 300 at baseline and 3 [1%] of 300 at endline surveys). No isolates were found carrying the dhps ISGEGA genotype. C-IPTp did not increase the prevalence of molecular markers associated with sulfadoxine-pyrimethamine resistance after three years of programme implementation. These findings reinforce C-IPTp as a strategy to optimise the control of malaria during pregnancy, and support the WHO guidelines for prevention of malaria in pregnancy. UNITAID [2017-13-TIPTOP].


Arctic charr brain transcriptome strongly affected by summer seasonal growth but only subtly by feed deprivation.
The Arctic charr (Salvelinus alpinus) has a highly seasonal feeding cycle that comprises long periods of voluntary fasting and a short but intense feeding period during summer. Therefore, the charr represents an interesting species for studying appetite-regulating mechanisms in fish. In this study, we compared the brain transcriptomes of fed and feed deprived charr over a 4 weeks trial during their summer feeding season. Despite prominent differences in body condition between fed and feed deprived charr at the end of the trial, feed deprivation affected the brain transcriptome only slightly. In contrast, the transcriptome differed markedly over time in both fed and feed deprived charr, indicating strong shifts in basic cell metabolic processes possibly due to season, growth, temperature, or combinations thereof. The GO enrichment analysis revealed that many biological processes appeared to change in the same direction in both fed and feed deprived fish. In the feed deprived charr processes linked to oxygen transport and apoptosis were down- and up-regulated, respectively. Known genes encoding for appetite regulators did not respond to feed deprivation. Gene expression of Deiodinase 2 (DIO2), an enzyme implicated in the regulation of seasonal processes in mammals, was lower in response to season and feed deprivation. We further found a higher expression of VGF (non-acronymic) in the feed deprived than in the fed fish. This gene encodes for a neuropeptide associated with the control of energy metabolism in mammals, and has not been studied in relation to regulation of appetite and energy homeostasis in fish. In the Arctic charr, external and endogenous seasonal factors for example the increase in temperature and their circannual growth cycle, respectively, evoke much stronger responses in the brain than 4 weeks feed deprivation. The absence of a central hunger response in feed deprived charr give support for a strong resilience to the lack of food in this high Arctic species. DIO2 and VGF may play a role in the regulation of energy homeostasis and need to be further studied in seasonal fish.


Calcific uremic arteriolopathy in peritoneal dialysis populations.
Calciphylaxis or calcific uremic arteriolopathy is an infrequent complication of end stage kidney disease. It is characterized by arteriolar medial calcification, thrombotic cutaneous ischemia, tissue necrosis often leading to ulceration, secondary infection and increased mortality rates. Current, multimodality treatment involves local wound care, well-controlled calcium, phosphate and parathyroid hormone levels and combination therapy with sodium thiosulfate and hyperbaric oxygen therapy. This combination therapy may be changing the historically poor prognosis of calcific uremic arteriolopathy reported in the literature. Peritoneal dialysis is considered a risk factor based on limited publications, however this remains to be proven. Clinical presentation, diagnosis, pathogenesis and treatment of calcific uremic arteriolopathy in these patients are no different from other patients manifesting with this condition.


Endogenous inhibitors of cyclic adenosine 3',5'-monophosphate-phosphodiesterase in rat epididymis.
Stabilization of cyclic adenosine 3',5'-monophosphate (cAMP)-phosphodiesterase (PDE) in 50% glycerol made possible the removal of endogenous inhibitors from tissue extracts by dialysis and the storage of the extracts at -20 degrees C without loss of PDE activity. Dialysates of heat-inactivated epididymal extracts were fractionated by liquid chromatography, and 4 fractions-F2, F5, F7, and F12-were found to contain endogenous inhibitors of PDE. The masses of the fractions required to inhibit low-Km PDE activity by ca. 50% in 430-microliter incubation mixtures were F2, 89 micrograms; F5, 23 micrograms; F7, 275 micrograms; and F12, 1.2 mg. The mechanisms of inhibition of low-Km PDE by the endogenous inhibitors were investigated by kinetic analysis of enzyme-inhibitor interaction. F2 and F12 inhibited PDE competitively; F5 and F7 decreased both apparent Km and Vmax, suggesting an uncompetitive mechanism of inhibition. The high potency of F5 in low concentration suggests that it may be a physiological modulator of low-Km cAMP-PDE activity.


[Calcium involved in the signaling pathway of jasmonic acid induced stomatal closure of Vicia faba L].
Ca2+, an ubiquitous second messenger in the signal transudation pathway, is required for various physiological and developmental processes in plant. Jasmonic acid (JA) has been known to induce the stomatal closure. By monitoring the changes of [Ca2+]cyt with fluorescent probe Fluo-3 AM under the confocal microscopy, we observed that exogenous JA increased [Ca2+]cyt in guard cells of Vicia faba L. while the control and linolenic acid (LA), which is a precursor of JA, could hardly affect the change of [Ca2+]cyt. EGTA, a chelator of Ca2+ completely blocked JA-induced stomatal closure. After epidermis pretreated with EGTA, JA failed to result in [Ca2+]cyt increasing. Ruthenium red that blocked Ca2+ released from intracellular Ca2+ store could not significantly change JA-induced stomatal closure, while JA still increased [Ca2+]cyt. Furthermore, Ca2+ channel inhibitor of nifedipine (NIF) reduced the effectiveness of JA-induced stomatal closure and JA-induced increasing fluorescent intensity in guard cells. The results demonstrated that Ca2+ is involved in the signal transduction of JA induced stomatal closure, and the source of [Ca2+]cyt increasing in guard cells induced by JA might derive mainly from the external stores.


A rationale for identification of cases of drug abuse.
To overcome the blanket of secrecy developed by many drug-abusing populations, special strategies have been used to generate more reliable and valid data on the extent and nature of the problem. The epidemiological rationale for eight strategies to identify cases of drug abuse are reviewed along with their limitations and examples of their application. The choice of strategies in any given setting will depend upon the drug-use patterns and legal framework of the particular society, the form of social organization developed by drug abusers and the types of contact they have with formal institutions such as law enforcement and treatment agencies.


Laparoscopic uterine artery occlusion before cervical curettage in cervical ectopic pregnancy: Safe and effective for preventing massive bleeding.
Cervical ectopic pregnancy is associated with high risk for massive bleeding conditions. Cervical ectopic pregnancy can usually be treated by methotrexate injection or surgery. We present 4 cases of cervical ectopic pregnancy that were treated successfully with different uterine-conserving methods. By comparing our experience of 4 cases managed in different ways, we found that laparoscopic uterine artery occlusion before cervical curettage is more effective method for preventing massive bleeding.


Selection bias in occupational sperm studies.
Participation rates in sperm studies are typically 25-50%, and therefore it is a matter of concern whether the men who provide semen samples truly reflect the parent population. The authors analyzed data from three Danish occupational sperm studies and evaluated the relation between age and occurrence of subfertility to participation and provision of semen samples. Age and subfertility were found to be rather strong determinants of participation. Willingness to provide semen samples was greater among men aged <40 years (odds ratio (OR) = 1.8, 95% confidence interval (CI) 1.3-2.6) and among men who had experienced an infertile period (OR = 1.7, 95% CI 1.3-2.1). Furthermore, the effect of infertility was modified by occupational exposure status, thus resulting in a tendency to differential selection and possibly biased risk estimates. In a cross-sectional study, the authors recommend that priority be given to a high participation rate and that data on the basic variables be collected from the entire study population, so that it is possible to make a nonresponder analysis and evaluate bias. Such an analysis can be carried out by means of questions enquiring about infertility, genital disorders, and earlier seminal examinations. Although the longitudinal study design is without many of the limitations of the cross-sectional study, a longitudinal study is usually not feasible.


Antimicrobial and Physicochemical Characterization of Biodegradable, Nitric Oxide-Releasing Nanocellulose-Chitosan Packaging Membranes.
Biodegradable composite membranes with antimicrobial properties consisting of nanocellulose fibrils (CNFs), chitosan, and S-nitroso-N-acetyl-d-penicillamine (SNAP) were developed and tested for food packaging applications. As a nitric oxide donor, SNAP was encapsulated into completely dispersed chitosan in 100 mL of 0.1 N acetic acid and was thoroughly mixed with CNFs to produce a composite membrane. The fabricated membranes had a uniform dispersion of chitosan and SNAP within the CNFs, which was confirmed through scanning electron microscopy (SEM) micrographs and a chemiluminescence nitric oxide analyzer. The membranes prepared without SNAP showed lower water vapor permeability than that of the membranes with SNAP. The addition of SNAP resulted in a decrease in Young's modulus for both two- and three-layer membrane configurations. Antimicrobial property evaluation of SNAP-incorporated membranes showed an effective zone of inhibition against bacterial strains of Enterococcus faecalis, Staphylococcus aureus, and Listeria monocytogenes and demonstrated its potential applications for food packaging.


Systemic virus infection results in CD8 T cell recruitment to the retina in the absence of local virus infection.
During recent years, evidence has emerged that immune privileged sites such as the CNS and the retina may be more integrated in the systemic response to infection than was previously believed. In line with this, it was recently shown that a systemic acute virus infection leads to infiltration of CD8 T cells in the brains of immunocompetent mice. In this study, we extend these findings to the neurological tissue of the eye, namely the retina. We show that an acute systemic virus infection in mice leads to a transient CD8 T cell infiltration in the retina that is not directed by virus infection inside the retina. CD8 T cells were found throughout the retinal tissue, and had a high expression of CXCR6 and CXCR3, as also reported for tissue residing CD8 T cells in the lung and liver. We also show that the pigment epithelium lining the retina expresses CXCL16 (the ligand for CXCR6) similar to epithelial cells of the lung. Thus, our results suggest that the retina undergoes immune surveillance during a systemic infection, and that this surveillance appears to be directed by mechanisms similar to those described for non-privileged tissues.


Mean nuclear volume of bladder cancer: stereological estimation and its clinical value.
Mean nuclear volumes (MNVs) of bladder cancer cells from the specimens of 100 patients were estimated by stereological procedure. The value of MNVs of bladder cancer were ranged widely, from 114.6 to 1,041.9 microns3. However, strong correlation was observed between MNV and histopathological grade and stage. Furthermore, the MNV predicts the biological behavior including the malignant potential and prognosis of the patient. Patients with MNV below 315 microns3 had significantly better prognosis than those with MNV above this value (p < 0.01). The 5- and 10-year survival rates for patients with MNV below 315 micron3 were both 97%, while those for patients with MNV above 315 microns3 were 65 and 58%, respectively. MNVs of the recurrent tumors were significantly increased, compared with those of the initial tumors (p < 0.01). MNV of extirpated tumor specimens following chemotherapy consisting of cisplatin and Adriamycin through balloon-occluded arterial infusion (BOAI) was significantly decreased, compared with that before BOAI (p < 0.01). We conclude that the MNV is an excellent predictor for the prognosis of the patient and the tumor recurrence. Furthermore, the stereological estimation of nuclear volume is a simple, quick, inexpensive, and reliable method to assess quantitative analysis of the histological and biological character of bladder cancer.


Idiopathic ventricular tachycardia. A review.
Idiopathic or unexplained VT occurs in a small but important subset of patients without clinically evident heart disease. The majority of these patients appear to have a structurally normal heart. The cause of the arrhythmias in these individuals is unclear and may never be recognized. Other patients with this condition may have minor abnormalities not sufficient to impair overall cardiac function. The significance of these abnormalities to the genesis of the arrhythmia is uncertain. Whether patients with minor abnormalities are more likely to harbor covert heart disease such as myocarditis or a focal defect is not known, nor is it resolved whether such patients warrant a more aggressive search for a structural cause. The question that remains in any patient not subjected to surgical or pathological exploration is whether undetermined heart disease is responsible for the arrhythmia. Continued correlation between functional (electrophysiological) and structural (pathological) data will provide meaningful information concerning the pathophysiology (substrate) of these arrhythmias. Because of the preservation of normal cardiac function, these arrhythmias are generally well-tolerated. Although the condition is usually associated with a favorable prognosis, the occasional deaths reported in patients with apparently idiopathic ventricular arrhythmias may not permit calling this condition benign. It would be important to know the extent to which unrecognized abnormalities play a role in the genesis of these tachycardias, and whether such patients are more predisposed to fatal arrhythmias or have a different natural history. If cases involving undetermined or covert heart disease were excluded from consideration, then a relatively homogeneous disease-free group may be identified with a truly benign condition and a uniformly favorable prognosis. In these cases, a primary electrical abnormality may prove to be the basis for the arrhythmia. These issues remain to be elucidated in future studies.


Liver Fibrosis Scores and Coronary Artery Disease: Novel Findings in Patients with Metabolic Dysfunction-Associated Fatty Liver Disease.
Metabolic dysfunction-associated fatty liver disease (MAFLD) is a recently proposed term as a more appropriate definition for nonalcoholic fatty liver disease (NAFLD). Previous studies have shown an association between liver fibrosis scores and cardiovascular disease (CVD) in patients with NAFLD. In this study, we aimed to investigate the relationship between liver fibrosis scores and coronary artery disease (CAD) severity in patients with MAFLD. This study was conducted on 1346 patients with MAFLD at the Second Hospital of Dalian Medical University between January 2018 and December 2021. We calculated the liver fibrosis scores, including the fibrosis 4 (FIB-4) score, nonalcoholic fatty liver disease fibrosis score (NFS), and aspartate aminotransferase-to-platelet ratio index (APRI). We divided the participants into three groups based on the degree of coronary artery stenosis assessed using coronary computed tomography angiography (CCTA): CAD (≥50%), non-obstructive (1-49%), and normal (no stenosis). An increased FIB-4 score and NFS were significantly associated with CAD severity in patients with MAFLD. The percentage of patients with a high FIB-4 score was higher in the CAD group than in the other two groups (5.80%, 4.31%, and 2.24%, respectively; p<0.001), as was the percentage of patients with NFS (11.12%, 5.19%, and 0.93%, respectively; p<0.001). Carotid atherosclerosis, creatinine levels, and CAC scores were significant predictors of CAD. The FIB-4 score and NFS were independently associated with CAD even after adjusting for sex and well-known cardiovascular risk factors. The APRI was not a significant factor for CAD in any model. In the bivariate correlation analysis, the FIB-4 score and NFS were directly correlated with CAC scores. Non-invasive liver fibrosis scores (FIB-4 and NFS) were significantly associated with the CAD severity and CAC scores in patients with MAFLD. Screening for CAD may be beneficial for subjects with high liver fibrosis risk MAFLD.


Serum bactericidal activity of ceftriaxone plus metronidazole against common intra-abdominal pathogens.
The activity of ceftriaxone plus metronidazole against pathogens usually involved in intra-abdominal infections was studied. Metronidazole 1 g and ceftriaxone 1 g (as the sodium salt) were simultaneously administered i.v. over 30 minutes every 24 hours to 12 healthy volunteers for three doses. Serum samples were collected at baseline, just before the last dose, and 12, 16, 20, 22, and 24 hours after the start of infusion of the last dose. Serum bactericidal titers (SBTs) were performed in duplicate for each serum sample from 12 hours on. Serum ceftriaxone and metronidazole concentrations were determined by high-performance liquid chromatography, and the elimination rate and half-life were calculated for each antimicrobial in each volunteer. The minimum inhibitory concentrations (MICs) of each antibiotic for two strains each of Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, and Bacteroides fragilis were determined by microdilution. Eleven volunteers completed the study. Ceftriaxone and metronidazole maintained SBTs of at least 1:4. Serum ceftriaxone concentrations remained above the MICs for E. coli, P. mirabilis, and K. pneumoniae, and serum metronidazole concentrations remained above the MIC for B. fragilis throughout the study. Ceftriaxone combined with metronidazole resulted in intense and prolonged activity against E. coli, P. mirabilis, K. pneumoniae, and B. fragilis.


Nobiletin suppresses cholangiocarcinoma proliferation via inhibiting GSK3β.
Background: Cholangiocarcinoma (CCA) is a type of hepatobiliary cancer characterized by uncontrolled cell proliferation, with a poor prognosis and high mortality. Nobiletin (NBT) is a promising anti-tumor compound derived from the peels of oranges and other citrus plants, citrus plant. But the effect of NBT on CCA remains unknown. Results: Our data showed that NBT suppressed CCA cell proliferation in vitro and in vivo. Colony formation and Edu assay indicated that NBT inhibited cell proliferation. Cell cycle analysis showed that NBT arrested the cell cycle in G0/G1 phase. Target prediction showed that GSK3β was a direct target. Western blot and immunofluorescence confirmed that NBT reduced the phosphorylation of GSK3β. The antiproliferative effect of NBT was intercepted in GSK3β knockdown CCA cells. The cellular thermal shift assay (CETSA) showed NBT directly bound to GSK3β. Finally, NBT showed an anti-proliferative effect in tumor-bearing mice with no hepatotoxicity. Conclusion: NBT could inhibit CCA proliferation, and the pharmacological activity of NBT in CCA was attributed to its direct binding to GSK3β. We suggested that NBT might be a potential natural medicine in CCA treatment.


Medical ethics in its American context. An historical survey.
Until the 1950's, moral aspects of clinical practice were handled in the USA within the medical profession. Over the last 30 years, these issues have become subjects for public debate, and have changed the public perception of medicine, in four steps. In the 1950's, moral theologians questioned the implications of medical technology at the edges of life. In the late '60s and '70s, these theologians were joined by political activists, whose zeal provoked a counter-reaction from physicians. In the late '70s and early '80s, the debate became largely theoretical; but in the late '80s it is once again "clinical", though respecting the rights of patients, their families, and other nonphysicians to participate in the relevant moral decisions. In part, these four steps reflect the special feature of American social history in the last 30 years; but in part they also had counterparts in Britain and elsewhere. Either way, the monopoly control over the ethics of medical practice exercised by doctors before the 1950s is unlikely to return.


A systematic forest survey showing an association of Saccharomyces paradoxus with oak leaf litter.
Although we understand the genetics of the laboratory model yeast Saccharomyces cerevisiae very well, we know little about the natural ecology and environment that shaped its genome. Most isolates of Saccharomyces paradoxus, the wild relative of S. cerevisiae, come from oak trees, but it is not known whether this is because oak is their primary habitat. We surveyed leaf litter in a forest in Northern Germany and found a strong correlation between isolation success of wild Saccharomyces and the proximity of the nearest oak. We compared the four most common tree genera and found Saccharomyces most frequently in oak litter. Interestingly, we show that Saccharomyces is much more abundant in oak leaf litter than on oak bark, suggesting that it grows in litter or soil rather than on the surfaces of oaks themselves. The distribution and abundance of Saccharomyces over the course of a year shows that oak leaf litter provides a stable habitat for the yeast, although there was significant tree-to-tree variation. Taken together, our results suggest that leaf litter rather than tree surfaces provide the better habitat for wild Saccharomyces, with oak being the preferred tree genus. 99.5% of all strains (633/636) isolated were S. paradoxus.


Design and initial operation of multichord soft x-ray detection arrays on the STOR-M tokamak.
Two miniature pinhole camera arrays for spatially and temporally resolved measurements of soft x-ray emission have been designed and installed on the STOR-M tokamak. Each array consists of a photodiode array, with one array viewing vertically and one viewing horizontally through a plasma cross section. Preamplifiers with fixed gains of 10(5) VA and custom built amplifiers with variable gains are used for signal amplification. Digitizers with 14 bit resolution and 3 MSs sampling rate are used for data acquisition. In the initial operation, an Al foil with a thickness of 1.8 microm installed for one array and Be filter of 7.6 microm installed for the other array are used to test signal strength. Initial tests have identified sawtooth oscillations and 20 kHz fluctuations, which are also detected by Mirnov coils, superimposed on the sawtooth oscillations.


Genetic diversity of the dengue vector Aedes aegypti in Australia and implications for future surveillance and mainland incursion monitoring.
In February 2004, the discovery of an incursion of the dengue vector Aedes aegypti into the town of Tennant Creek in the Northern Territory caused concern for the Northern Territory health authorities who proceeded to implement a Commonwealth-funded eradication program. To determine the origin of the incursion, we performed a genetic analysis on Ae. aegypti from several Queensland and overseas localities. A comparison of DNA sequences from the mitochondrial cytochrome oxidase 1 gene indicated that the incursion was probably from Cairns or Camooweal. This genetic marker was also useful in identifying a separate Townsville haplotype population and another population on Thursday Island in the Torres Strait that was genetically divergent to the mainland populations. The possible use of this marker as a surveillance tool for identifying the origins of local and overseas incursions is discussed.


Can ischemia-modified albumin be used to differentiate between generalized seizures and pseudoseizures?
In recent years ischemia-modified albumin (IMA) has been suggested as a marker that can be used in differentiating nonconvulsive conditions from epilepsy. The purpose of this study was to investigate changes in IMA levels caused by generalized clonic tonic (GTC) seizures. A total of 114 children presenting to the Karadeniz Technical Pediatric Emergency Polyclinic with GTC seizures were included in the study. Sixteen cases meeting the inclusion criteria were included in the study and sixteen healthy children were enrolled as the control group. The patients' IMA, albumin, and IMA/albumin values at hours 0 and 1 following the episode were compared with control group values. IMA levels in the patient group were significantly higher at hour 1 compared to hour 0, and were also significantly higher than those of the control group levels at hour 1. In addition, the patient group IMA/albumin index value at hour 1 was significantly higher than the baseline value. IMA levels increased significantly with length of seizure. Although there were no markers of hypoxia in patients undergoing GTC seizures in this study, hypoxia was observed to develop, and this caused serum IMA levels to rise in line with seizure duration.


Switchavidin: reversible biotin-avidin-biotin bridges with high affinity and specificity.
Switchavidin is a chicken avidin mutant displaying reversible binding to biotin, an improved binding affinity toward conjugated biotin, and low nonspecific binding due to reduced surface charge. These properties make switchavidin an optimal tool in biosensor applications for the reversible immobilization of biotinylated proteins on biotinylated sensor surfaces. Furthermore, switchavidin opens novel possibilities for patterning, purification, and labeling.


Studies of the reductive biotransformation of selected carbonyl compounds by whole cells and extracts of baker's yeast, Saccharomyces carevisiae.
The progress of reductive biotransformations of a variety of earbonyl compounds by whole cells of baker's yeast was monitored with time. Biotransformations rates ranged from 0.11 to 112.12 mg product formed per g dry yeast per h. While rapid biotransformations of citronellal and ethyl benzoylformate were observed, complete conversion of substrate to product did not occur. Reductive conversions of ethyl- and methyl-acetoacetate went to completion in 6 and 12 h respectively. Ethyl mandelate was produced stereoselectively, favoring the (R)- stereoisomer and ethyl and methyl-3-hydroxybutyrate were produced with (S)-enantiospecificity. Yeast crude extract and resuspended presence of NAD(P)H. Ethyl benzoylformate and methyl-and ethyl-acetoacetate were preferentially reduced by yeast crude extract as compared to resuspended pellet and, in the case of the former two substrates, the reaction manifested a preference for NADPH over NADH.


Syndesmosis procedure: a non-osteotomy approach to metatarsus primus varus correction.
Hallux valgus and metatarsus primus varus deformities usually are the result of failure of the supporting soft tissues rather than bone deformities. Since soft-tissue procedures have been shown to only be suitable for mild deformities, first metatarsal osteotomy has become an integral part of correcting moderate to severe deformities. A soft-tissue technique referred to as the "syndesmosis procedure" by the author was evaluated for its effectiveness in correcting metatarsus primus varus in feet with hallux valgus. This is a retrospective clinical and radiographic study of six patients (11 feet) an average of 6 years and 10 months after the "syndesmosis procedure." A cerclage technique using absorbable suture (PDS) was used for the first metatarsal realignment, and its long-term maintenance depended on a syndesmosis (fibrous) bonding between the first and second metatarsal bases. The average preoperative hallux valgus angle of 29.5 (21 to 43) degrees improved to 13.5 (-2 to 24) degrees and the average preoperative metatarsus primus varus angle of 13.6 (12 to 16) degrees improved to 5.2 (2 to 8) degrees at an average of 85.7 (33 to 128) months. All patients were satisfied with their results and were able to return to sports and wearing high-heeled shoes as desired. Complications were few and mild. Followup American Orthopaedic Foot and Ankle Society Hallux scores averaged 93 points. This small but long-term retrospective study showed encouraging results for the correction of metatarsus primus varus deformity and a high patient satisfaction.


Optical microbubble resonator.
We develop a method for fabricating very small silica microbubbles having a micrometer-order wall thickness and demonstrate the first optical microbubble resonator. Our method is based on blowing a microbubble using stable radiative CO(2) laser heating rather than unstable convective heating in a flame or furnace. Microbubbles are created along a microcapillary and are naturally opened to the input and output microfluidic or gas channels. The demonstrated microbubble resonator has 370 microm diameter, 2 microm wall thickness, and a Q factor exceeding 10(6).


A tailored Internet-delivered intervention for smoking cessation designed to encourage social support and treatment seeking: usability testing and user tracing.
While Internet technologies show promise for changing behavior, new methods for engaging individuals are needed to maximize effectiveness. The aim of this study is to design and evaluate an Internet-delivered intervention for smoking cessation that encouraged seeking support from family and treatment from doctors. To evaluate different introductions to the Internet site. We conducted usability testing and analyzed server logs to trace user participation in the website. Two groups of users (current smokers) were recruited using Google advertisements. In Phase 1, 58% (75/126) of users accessed the self-management strategies, but few users accessed the social support (28%) and treatment-seeking modules (33%). Then, a brief motivational introduction was added, stating the proven effectiveness of content in the unused modules, low use of these modules, and recommendations by two doctors to use all modules. Compared with Phase 1, in Phase 2 the mean time spent on the website per session increased (8 to 18 min, p = 0.01) and use of the social support (50%) and treatment seeking modules (56%) increased (both p < 0.01). At 1-month follow-up, reports of talking to family about smoking cessation also increased from 84% to 100% (p = 0.038). Changing the rationale and context of Web-based health information using a motivational introduction can change user behavior.


Transportation cost as a barrier to contraceptive use among women initiating treatment for HIV in Tanzania.
Transportation cost is a barrier to HIV treatment, yet no studies have examined its association with contraceptive use among women living with HIV. We analyzed cross-sectional data from women attending three public healthcare facilities in Shinyanga, Tanzania where they initiated antiretroviral therapy for HIV infection in the previous 90 days; all facilities offered free contraception. Women self-reported current contraceptive use and the round-trip cost of transportation to the facility. Among 421 women aged 18-49, 86 (20.4%) were using any modern contraceptive method, of which half were using modern methods other than condoms. Women who paid more than 2,000 Tanzanian shillings for transportation had a significantly lower prevalence of any modern method use than women who paid nothing (9.1% vs. 21.3%; adjusted difference: -12.9; 95% confidence interval: -21.3, -4.4). A similar difference was observed for non-condom modern method use. We conclude that high transportation cost may impede contraceptive use even among women accessing HIV treatment.


Protective effect of N-acetyl-L-cysteine (L-NAC) against styrene-induced cochlear injuries.
Styrene exposure causes hair cell death through both apoptotic and necrotic pathways and treatment with N-acetyl-L-cysteine (L-NAC) reduces styrene ototoxicity. Exposure to styrene causes hearing loss and hair cell death in the middle frequency region in the cochlea. The current study was designed to examine the cell death pathways and the protective effect of L-NAC against styrene-induced cochlear injuries. Seventeen rats were exposed to styrene by gavage at 400 mg/kg 5 days per week for 3 weeks. Nine of the styrene-treated rats received L-NAC by intraperitoneal injection (325 mg/kg), and the remaining eight rats received saline injections as controls. The styrene-induced hearing loss was assessed by auditory brainstem responses (ABRs). Apoptotic, necrotic, and missing hair cells were quantified using combined methods, including nuclear staining with propidium iodide, F-actin staining with FITC-phalloidin, and the TUNEL assay. The styrene exposure caused a threshold shift of 15±4.3 dB. Both apoptosis and necrosis were involved in the pathogenesis of the cochlear lesion, but apoptosis appeared to be the major cell death pathway leading to the styrene ototoxicity. Treatment with L-NAC reduced the number of missing and dying outer hair cells (OHCs) and reduced the styrene-induced hearing loss.


Increasing maize seed methionine by mRNA stability.
The amino acid methionine is a common protein building block that is also important in other cellular processes. Plants, unlike animals, synthesize methionine de novo and are thus a dietary source of this nutrient. A new approach for using maize as a source of nutrient methionine is described. Maize seeds, a major component of animal feeds, have variable levels of protein-bound methionine. This variability is a result of post-transcriptional regulation of the Dzs10 gene, which encodes a seed-specific high-methionine storage protein. Here we eliminate methionine variability by identifying and replacing the cis-acting site for Dzs10 regulation using transgenic seeds. Interestingly, two different mechanisms affect mRNA accumulation, one dependent on and the other independent of the untranslated regions (UTRs) of Dzs10 RNA. Accumulation of chimeric Dzs10 mRNA was not reduced in hybrid crosses and was uncoupled from genomic imprinting by Dzr1, a regulator of Dzs10. Uniform high levels of Dzs10 protein were maintained over five backcross generations of the transgene. The increased level of methionine in these transgenic seeds allowed the formulation of a useful animal feed ration without the addition of synthetic methionine.


Impact of health conditions on food intakes among older adults.
Even well older adults may experience a range of age-related physiological changes and chronic health conditions and may become increasingly sedentary--key factors that could affect appetite and hunger and lead to changes in diet composition. The present article reviews recent literature on the impact of prevalent health conditions on dietary choice. Research shows some evidence that older adults make positive dietary changes following the onset of certain chronic health conditions. However, most chronic diseases appear to lead to dietary restrictions that compromise nutritional status. The nature of the health condition and its effect on physiological function will drive the dietary change process. The extent to which health care providers counsel their patients to incorporate changes into their diet and the mindset and belief system of each individual may also have a strong impact on food choices. A recurring theme in the research examined in this review was older adults' desire to improve their diet, coupled with their frustration at not being offered sufficient counseling by their health care providers. The role of health care professionals, particularly nutritionists, in guiding older adults toward an improved diet cannot be overstated.


Severe COVID-19 patients display hyper-activated NK cells and NK cell-platelet aggregates.
COVID-19 is characterised by a broad spectrum of clinical and pathological features. Natural killer (NK) cells play an important role in innate immune responses to viral infections. Here, we analysed the phenotype and activity of NK cells in the blood of COVID-19 patients using flow cytometry, single-cell RNA-sequencing (scRNA-seq), and a cytotoxic killing assay. In the plasma of patients, we quantified the main cytokines and chemokines. Our cohort comprises COVID-19 patients hospitalised in a low-care ward unit (WARD), patients with severe COVID-19 disease symptoms hospitalised in intensive care units (ICU), and post-COVID-19 patients, who were discharged from hospital six weeks earlier. NK cells from hospitalised COVID-19 patients displayed an activated phenotype with substantial differences between WARD and ICU patients and the timing when samples were taken post-onset of symptoms. While NK cells from COVID-19 patients at an early stage of infection showed increased expression of the cytotoxic molecules perforin and granzyme A and B, NK cells from patients at later stages of COVID-19 presented enhanced levels of IFN-γ and TNF-α which were measured ex vivo in the absence of usual in vitro stimulation. These activated NK cells were phenotyped as CD49a+CD69a+CD107a+ cells, and their emergence in patients correlated to the number of neutrophils, and plasma IL-15, a key cytokine in NK cell activation. Despite lower amounts of cytotoxic molecules in NK cells of patients with severe symptoms, majority of COVID-19 patients displayed a normal cytotoxic killing of Raji tumour target cells. In vitro stimulation of patients blood cells by IL-12+IL-18 revealed a defective IFN-γ production in NK cells of ICU patients only, indicative of an exhausted phenotype. ScRNA-seq revealed, predominantly in patients with severe COVID-19 disease symptoms, the emergence of an NK cell subset with a platelet gene signature that we identified by flow and imaging cytometry as aggregates of NK cells with CD42a+CD62P+ activated platelets. Post-COVID-19 patients show slow recovery of NK cell frequencies and phenotype. Our study points to substantial changes in NK cell phenotype during COVID-19 disease and forms a basis to explore the contribution of platelet-NK cell aggregates to antiviral immunity against SARS-CoV-2 and disease pathology.


Control of the gyrotron filament power supplies for the experimental advanced superconducting tokamak.
The electron cyclotron heating system including four gyrotrons is being developed in the Institute of Plasma Physics, Chinese Academy of Sciences. The filament is an important part of the gyrotron, which is used to heat the cathode. The gyrotron output power can be controlled by adjusting the filament power. We have developed the filament power control and measurement system using Labview. The filament characteristics were tested using this power control system. The test results show that the filament current can be fitted with the e-exponential function of the filament voltage. It can be seen that as the voltage increases, the filament resistance gradually increases. We have developed a new "burst" function to prevent the beam current from dropping too fast, which is conducive to long pulse stable operation of the gyrotrons.


Single-wire pressure and flow velocity measurement for quantifying microvascular dysfunction in patients with coronary vasospastic angina.
Endothelial and vascular smooth muscle dysfunction of epicardial coronary arteries play a pivotal role in the pathogenesis of vasospastic angina (VSA). However, coronary microvascular (MV) function in patients with VSA is not fully understood. In the present study, subjects without coronary obstruction were divided into two groups according to the acetylcholine provocation test: VSA group (n = 29) and non-VSA group (n = 21). Hyperemic MV resistance (hMR) was measured using a dual-sensor (Doppler velocity and pressure)-equipped guidewire, and guidewire-derived hemodynamic parameters were compared. There were no between-group differences in clinical demographics, including potential factors affecting MV function (e.g., diabetes). Although coronary flow velocity reserve was similar between the two groups [2.4 ± 1.0 (VSA group) vs. 2.4 ± 0.9 (non-VSA group); P = 0.8], coronary vessel resistance and hMR were significantly elevated in the VSA group compared with the non-VSA group (2.6 ± 3.1 vs. 1.2 ± 0.8, P = 0.04; 1.9 ± 0.6 vs. 1.6 ± 0.5, P = 0.03, respectively). Coronary vasospasm, older age, E/e', and estimated glomerular filtration rate were significantly associated with MV dysfunction [defined as ≥ median value of hMR (1.6)] in univariate analysis. Coronary vasospasm most strongly predicted higher hMR in multivariate logistic regression analysis (odds ratio, 4.61; 95% confidence interval, 0.98-21.60; P = 0.053). In conclusion, coronary MV resistance is impaired in patients with VSA compared with non-VSA patients, whereas coronary flow velocity reserve is maintained at normal levels in both groups. In vivo assessment of hMR might be a promising index of coronary MV dysfunction in patients with VSA.


Biomechanical differences between left- and right-handed baseball pitchers.
Left-handed baseball pitchers are thought to have a number of theoretical advantages compared to right-handed pitchers; however, there is limited scientific research detailing differences in the pitching mechanics of right- and left-handed pitchers. Therefore, this study sought to understand whether any kinematic and kinetic differences existed between right- and left-handed baseball pitchers. A total of 52 collegiate pitchers were included in this study; 26 left-handed pitchers were compared to 26 age-, height-, weight- and ball velocity-matched right-handed pitchers. Demographic information, passive shoulder range of motion and kinematic and kinetic data were obtained for each pitcher participating in the study. Results indicated that left-handed pitchers did not have a glenohumeral internal rotation deficit as compared to right-handed pitchers. Kinematic analysis indicated that elbow flexion, horizontal glenohumeral abduction and wrist coronal plane motion were significantly different between the two study cohorts. It was also noted that left-handed pitchers had increased elbow varus moments. The findings of this study suggest that pitching coaches should be aware that there are biomechanical differences between left- and right-handed pitchers.


[Titration and dosage of gabapentin].
Gabapentin (GBP) is a new anti epileptic drug which is indicated in partial epilepsy as either monotherapy or additional therapy. Unlike most anti epileptic drugs, GBP can be rapidly titrated. A recent double blind trial has shown that treatment with GBP may start with 900mg from the first day (300 mg every 8 hours). This dose is sufficient to be effective as monotherapy in patients with partial epilepsy of recent onset. It is also known that in patients with drug resistant epilepsy high doses of anti epileptic drugs are often necessary. In the clinical trials carried out before gabapentin was put on the market, the doses used were generally low (almost always below 1,800 mg/day). However, recent experience has shown that doses of up to 4,800 mg/day markedly increase the efficacy without significant increase in the rate of adverse effects. Recent data suggest that GBP may be rapidly titrated and high doses of GBP show greater efficacy with no increase in adverse effects, so they are advisable in drug resistant patients.


The perils, pitfalls and opportunities of using high sensitivity cardiac troponin.
Cardiac troponins (cTn) are considered to be the 'gold standard' biomarker for the diagnosis of acute coronary syndrome (ACS); a pathological spectrum which includes cardiac ischemia, angina, myocardial infarction and ultimately cardiac failure. The growing evidence base for the diagnostic and prognostic use of cTn in ACS has resulted in a universal redefinition of acute myocardial infarction (AMI). Recently a number of immunoassays with claims of superior sensitivity have been produced. The analytical and clinical performance of these assays require appropriate evaluation. Sensitive assays can be used for diagnosis in the first few hours after an ischemic episode. Early elevations in cTn are prognostic. A single time point for cTn testing may be useful for rule out, however such a strategy does not detect the rising and falling pattern required for diagnosis as suggested in the universal definition of AMI. The newer assays demonstrate low level cTn positivity in apparently healthy people. In addition, the sensitive assays detect more cTn positive patients who do not have a final diagnosis of ACS. It is unknown if such mild elevations in cTn detected by sensitive assays are of clinical concern. What is certain is that AMI remains a clinical not a biochemical diagnosis and interpretation of cTn concentrations should be made according to the clinical context. This review highlights the development of the sensitive assays, documents their analytical and clinical performance and reviews the usefulness of cTn elevation in non-ACS conditions.


Initiation by RNA polymerase II and formation of runoff transcripts containing unblocked and unmethylated 5' termini.
Transcription of cloned adenovirus, beta-globin, and retrovirus long terminal repeat DNAs in HeLa whole-cell lysate was inhibited by S-adenosylhomocysteine. However, full-length 1.7-kilobase transcripts made on adenovirus 2 late promoter DNA contained 5'-terminal GpppA, consistent with specific initiation and runoff synthesis in the absence of product methylation. Formation of runoff transcripts including retrovirus RNAs that normally contain 5'-m7GpppGmpC was not decreased by replacing GTP with non-hydrolyzable analogs, and Rous-associated virus-2 runoff products made in the presence of GTP-gamma-S contained 5'-terminal gamma-S-pppGpC. The results indicate that capping and specific transcript synthesis by RNA polymerase II are not obligatorily linked in HeLa whole-cell lysate. Accurate initiation is dependent on ATP hydrolysis, and in contrast to GTP, replacement of ATP by 5'-adenylyl-imidodiphosphate blocked specific initiation of transcripts that start with either GTP (Rous-associated virus-2, Rous-associated virus-0) or ATP (beta-globin, adenovirus).


[A modified culture method for astrocytes from rat cortical tissue in vitro].
To evaluate the efficiency of a modified culture method for rat cerebral cortical astrocytes in vitro. The astrocytes derived from the cerebral cortex of 3-day-old Sprague-Dawley rats were first purified as described previously, then the cells were replanted at a low density. The culture flask was changed after 1 hour and substratum was replaced after 24 hours. Cells were syncretized to a monolayer, followed by cell passage. After three passages the cells were cultured in DMEM medium containing 10% fetal serum for a long period. The derivation of the cells was identified by immunofluorescent staining with anti-GFAP polyclonal antibodies. A variety of morphologically distinct astrocytes with many long processes and small cell bodies were obtained. Finally an astrocytic network occurred through cellular process connections. The immunofluorescent staining demonstrated the percentage of GFAP-positive cells was above 98%. The modified culture method for astrocytes from rat cerebral tissue is reliable, with a high purity. The cultured astrocytes have a similar morphological development to those in vivo.


Tumor-suppressive miR-34a induces senescence-like growth arrest through modulation of the E2F pathway in human colon cancer cells.
Accumulating evidence suggests a role for microRNAs in human carcinogenesis as novel types of tumor suppressors or oncogenes. However, their precise biological role remains largely elusive. In the present study, we aimed to identify microRNA species involved in the regulation of cell proliferation. Using quantitative RT-PCR analysis, we demonstrated that miR-34a was highly up-regulated in a human colon cancer cell line, HCT 116, treated with a DNA-damaging agent, adriamycin. Transient introduction of miR-34a into two human colon cancer cell lines, HCT 116 and RKO, caused complete suppression of cell proliferation and induced senescence-like phenotypes. Moreover, miR-34a also suppressed in vivo growth of HCT 116 and RKO cells in tumors in mice when complexed and administered with atelocollagen for drug delivery. Gene-expression microarray and immunoblot analyses revealed down-regulation of the E2F pathway by miR-34a introduction. Up-regulation of the p53 pathway was also observed. Furthermore, 9 of 25 human colon cancers (36%) showed decreased expression of miR-34a compared with counterpart normal tissues. Our results provide evidence that miR-34a functions as a potent suppressor of cell proliferation through modulation of the E2F signaling pathway. Abrogation of miR-34a function could contribute to aberrant cell proliferation, leading to colon cancer development.


Health problems of college students.
College health professionals deal with a range of medical problems and risky behaviors. Some medical conditions occur more frequently in the college-age population, but college health is not unique because of the types of medical problems seen. Community providers welcome the opportunity to deliver primary care to this relatively healthy population, with less emphasis on screening, intervention, mental health, social well-being, and altering unhealthy behaviors. Young people have been recognized as experiencing higher rates of morbidity, disability, and mortality from various developmental, environmental, and behavioral risk factors than the general population. These risk factors are so interrelated that successful efforts to change them require a more comprehensive approach that extends beyond the health of individuals to the wellness of an entire campus community. On the continuum of health and well-being, college health must move away from focusing on disease and move toward community wellness.


Changes to pubic hair removal practices during COVID-19 restrictions and impact on sexual intimacy.
Pubic hair grooming (hair removal) has been linked to body image and sexual behaviour and, given the impacts of COVID-19 restrictions on access to sexual partners and beauty salons, we explored these associations. Data derive from a cross-sectional online survey. Participants were aged 18 years or over and in Australia. All genders were included. We explored factors associated with changes to hair removal using logistic regression and analysed free-text responses using content analysis. Of 256 participants, 89 (34.8%) had changed their grooming practices. Multivariable analysis found that female participants had 4.5 times increased odds of changed grooming (95%CI 1.8, 11.0; p = 0.001) compared with males. Individuals who reported 'more' or 'less' oral sex compared with the previous year were more likely to report increased odds of changed grooming (AOR 3.2; 95%CI: 1.4, 7.7; p = 0.009 and AOR: 3.3; 95%CI:1.6, 6.7; p = 0.001, respectively). Most who changed their grooming practices said it had reduced, attributing this to lack of partners, inability to access beauticians, and the mental health effects of the pandemic. For most, this change had no impact on sexual behaviour or body image; however, some spoke of reduced sexual desire and willingness to receive oral sex. Partner expectations influenced whether reduced grooming had negative consequences, and women were more affected than men, highlighting a gendered impact.


Social network of family caregivers of disabled and dependent patients.
Cross-sectional study that used the Social Network Index and the genogram to assess the social network of 110 family caregivers of dependent patients attended by a Home Care Service in São Paulo, Brazil. Data were analyzed using the test U of Mann-Whitney, Kruskal-Wallis and Spearman correlation. Results were considered statistically significant when p<0,05. Few caregivers participated in activities outside the home and the average number of people they had a bond was 4,4 relatives and 3,6 friends. Caregivers who reported pain and those who had a partner had higher average number of relatives who to trust. The average number of friends was higher in the group that reported use of medication for depression. Total and per capita incomes correlated with the social network. It was found that family members are the primary caregiver's social network.


Successful outcome of triangle tilt as revision surgery in a pediatric obstetric brachial plexus patient with multiple previous operations.
Introduction. Obstetric brachial plexus injury (OBPI) occurs during the process of labor and childbirth. OBPI has been reported to be associated with shoulder dystocia, macrosomia, and breech delivery. Its occurrence in uncomplicated delivery is possible as well. Case Presentation. The patient in the present report is a 6.5-year-old girl, who suffered a severe brachial plexus injury at birth and had many reconstructive surgical procedures at an outside brachial plexus center before presenting to us. Discussion. The traditional surgical treatments by other surgical groups were unsuccessful and therefore the patient came to our clinic for further treatment. She had triangle tilt surgery with us, as a salvage procedure. Conclusion. The OBPI patient in this study clearly showed noticeable clinical and functional improvements after triangle tilt surgical management. The posture of the arm at rest was greatly improved to a more normal position, and hand to mouth movement was improved as well. Triangle tilt surgery should be conducted as a first choice treatment for medial rotation contracture of the shoulder in OBPI patients.


Initial Symptoms of Langerhans Cell Histiocytosis: A Case Series.
Langerhans cell histiocytosis (LCH) is a rare childhood hematopoietic disease, and hence, there are few reports summarizing the course leading to the diagnosis. We described the initial symptoms and the clinical course of LCH. We carried out a retrospective review of charts from a single medical center, and 21 patients with the diagnosis of LCH were enrolled. The initial symptoms of 16 cases were caused by bone lesions; of these cases, there were 8 instances of soft tissue swelling as the initial symptom (38%) and 8 instances of bone pain without swelling (38%). Among the cases of bone lesion, 4 of 6 cases of skull lesion were painless while all vertebral body lesions and long bone lesions were accompanied by pain. LCH bone lesions caused various symptoms depending on the site of the lesion and this makes the diagnosis difficult. A detailed physical examination and imaging studies are recommended for early diagnosis.


[Somatomedin activity of the blood: hormonal metabolic and immunological shifts in cancer patients].
Blood somatomedin activity observed in patients with cancer of the corpus uteri, breast, colon and rectum, pancreas, ovary and lung proved to exceed those in control by 140-300%. The said parameter was found to depend upon endocrine-metabolic changes, relatively higher levels being registered in overweight patients and those with hypertriglyceridemia. Changes of varying degree in somatomedin activity and growth hormone levels produced by insulin and glucose in patients with breast and uterine cancer pointed to hormonal dependence of the parameter. Also, the effect is a circumstantial evidence for negative feedback between growth hormone and somatomedin production. In patients with cancer of the breast, uterus and lung, a significant correlation was established between somatomedin activity, on the one hand, and monocyte. B-lymphocyte and T-active lymphocyte levels, on the other, matched by an inverse one with T-lymphocyte level and blastogenic reaction of lymphocytes.


Pemphigus Vulgaris and Pemphigus Foliaceus: Differences in Epidemiology and Mortality.
Little is known about differences in epidemiological features and prognosis between pemphigus vulgaris (PV) and pemphigus foliaceus (PF). The objective of this study was to compare PV and PF patients regarding ethnic variations and mortality rates. Mortality of PV and PF patients was compared with age- and sex-matched control subjects in the general population. The study cohort comprised 207 patients with PV and 30 with PF diagnosed during the period 2000 to 2015. The incidence rate of PV among Jews was 3.6-fold higher than among Arabs (p<0.001), whereas no ethnic predisposition to PF was noted (p = 0.379). The risk of death for patients with PV was almost 3-fold higher than in the general population (standardized mortality ratio (SMR) 2.6). For patients with PF, the risk of mortality was not significantly increased relative to the general population (SMR 1.4). There is a racial predisposition to PV, whereas PF is sporadic. Mortality among patients with PV is higher compared with PF and the general population.


Reactive oxygen species via redox signaling to PI3K/AKT pathway contribute to the malignant growth of 4-hydroxy estradiol-transformed mammary epithelial cells.
The purpose of this study was to investigate the effects of 17-β-estradiol (E2)-induced reactive oxygen species (ROS) on the induction of mammary tumorigenesis. We found that ROS-induced by repeated exposures to 4-hydroxy-estradiol (4-OH-E2), a predominant catechol metabolite of E2, caused transformation of normal human mammary epithelial MCF-10A cells with malignant growth in nude mice. This was evident from inhibition of estrogen-induced breast tumor formation in the xenograft model by both overexpression of catalase as well as by co-treatment with Ebselen. To understand how 4-OH-E2 induces this malignant phenotype through ROS, we investigated the effects of 4-OH-E2 on redox-sensitive signal transduction pathways. During the malignant transformation process we observed that 4-OH-E2 treatment increased AKT phosphorylation through PI3K activation. The PI3K-mediated phosphorylation of AKT in 4-OH-E2-treated cells was inhibited by ROS modifiers as well as by silencing of AKT expression. RNA interference of AKT markedly inhibited 4-OH-E2-induced in vitro tumor formation. The expression of cell cycle genes, cdc2, PRC1 and PCNA and one of transcription factors that control the expression of these genes - nuclear respiratory factor-1 (NRF-1) was significantly up-regulated during the 4-OH-E2-mediated malignant transformation process. The increased expression of these genes was inhibited by ROS modifiers as well as by silencing of AKT expression. These results indicate that 4-OH-E2-induced cell transformation may be mediated, in part, through redox-sensitive AKT signal transduction pathways by up-regulating the expression of cell cycle genes cdc2, PRC1 and PCNA, and the transcription factor - NRF-1. In summary, our study has demonstrated that: (i) 4-OH-E2 is one of the main estrogen metabolites that induce mammary tumorigenesis and (ii) ROS-mediated signaling leading to the activation of PI3K/AKT pathway plays an important role in the generation of 4-OH-E2-induced malignant phenotype of breast epithelial cells. In conclusion, ROS are important signaling molecules in the development of estrogen-induced malignant breast lesions.


Socioeconomic status and waiting times for health services: An international literature review and evidence from the Italian National Health System.
In the absence of priority criteria, waiting times are an implicit rationing instrument where the absence or limited use of prices creates an excess of demand. Even in the presence of priority criteria, waiting times may be unfair because they reduce health care demand of patients in lower socio-economic conditions due to high opportunity costs of time or a decay in their health level. Significant evidence has shown a relationship between socioeconomic status and the length of waiting time. The first phase of the study involved an extensive review of the existent literature for the period of 2002-2016 in the main databases (Scopus, PubMed and Science Direct). Twenty-eight met the eligibility criteria. The 27 papers were described and classified. The e mpirical objective of this study was to determine whether socioeconomic characteristics affect waiting time for different health services in the Italian national health system. The services studied were specialist visits, diagnostics tests and elective surgeries. A classification tree and logistic regression models were implemented. Data from the 2013 Italian Health National Survey were used. The analysis found heterogeneous results for different types of service. Individuals with lower education and economic resources have a higher risk of experiencing excessive waiting times for diagnostic and specialist visits. For elective surgery, socioeconomic inequalities are present but appear to be lower.


Psychosocial study about the consequences of work in hospital nurses as human resource management.
To analyze the relation between psychosocial and sociodemographic variables in nursing professionals. To discover the levels of job satisfaction, psychosomatic symptoms and psychological well-being in nurses. The research was conducted in a sample of 476 nurses / as from nine hospitals in the region of Murcia (Spain). An occupational health protocol was distributed among the sample participants for completion, which contained tools for measuring and describing psychosocial and sociodemographic variables. Although there is no high risk for psychological problems among the sample participants, psychosomatic symptoms, job dissatisfaction, and low psychological wellbeing are present. The results contribute to research on the subject by identifying the presence of psychosocial disorders in nursing professionals, associated with job conditions and certain psychosocial and sociodemographic variables.


Effect of adipose-derived nucleated cell fractions on tendon repair in horses with collagenase-induced tendinitis.
To assess the potential of adipose-derived nucleated cell (ADNC) fractions to improve tendon repair in horses with collagenase-induced tendinitis. 8 horses. Collagenase was used to induce tendinitis in the superficial digital flexor tendon of 1 forelimb in each horse. Four horses were treated by injection of autogenous ADNC fractions, and 4 control horses were injected with PBS solution. Healing was compared by weekly ultrasonographic evaluation. Horses were euthanatized at 6 weeks. Gross and histologic evaluation of tendon structure, fiber alignment, and collagen typing were used to define tendon architecture. Biochemical and molecular analyses of collagen, DNA, and proteoglycan and gene expression of collagen type I and type III, decorin, cartilage oligomeric matrix protein (COMP), and insulin-like growth factor-I were performed. Ultrasonography revealed no difference in rate or quality of repair between groups. Histologic evaluation revealed a significant improvement in tendon fiber architecture; reductions in vascularity, inflammatory cell infiltrate, and collagen type III formation; and improvements in tendon fiber density and alignment in ADNC-treated tendons. Repair sites did not differ in DNA, proteoglycan, or total collagen content. Gene expression of collagen type I and type III in treated and control tendons were similar. Gene expression of COMP was significantly increased in ADNC-injected tendons. ADNC injection improved tendon organization in treated tendons. Although biochemical and molecular differences were less profound, tendons appeared architecturally improved after ADNC injection, which was corroborated by improved tendon COMP expression. Use of ADNC in horses with tendinitis appears warranted.


Mutational analysis of the HER2 gene in lung tumors from Caucasian patients: mutations are mainly present in adenocarcinomas with bronchioloalveolar features.
Activating mutations in the tyrosine kinase domain of the HER2 gene have recently been reported in lung adenocarcinomas, mainly in East Asian patients. Our study was devised to evaluate the prevalence and nature of HER2 mutations in lung adenocarcinomas from Caucasian patients. The mutational status of the HER2 gene was evaluated in 403 lung adenocarcinomas by PCR-single strand conformation polymorphism analysis and direct sequencing of Exons 19 and 20. We found HER2 mutations in 9 (2.2%) cases. Seven (78%) of the mutations were in frame duplications/insertions at codons 776-779 (YVMA), the other 2 were base substitutions resulting in aminoacid changes. The hotspot mutation at bases 776-779 was previously found to be the most frequent HER2 mutation in Asiatic patients. The distribution of mutations was significantly different between conventional lung adenocarcinomas (CLAs) and lung adenocarcinomas with bronchioloalveolar features (ABAFs). Seven (6.2%) of 113 ABAFs and 2 (0.7%) of 290 CLA were mutated (p = 0.0025). In addition, the frequency of HER2 mutations was slightly higher in females (4.1%) than in males (1.8%) and in never smokers (3.1%) than in smokers (1.9%), but differences were not statistically significant. This series of tumors was also investigated for EGFR and K-ras mutations. EGFR mutations were observed in 43 (10.7%) cases, and K-ras mutations in 110 (27.3%) cases. EGFR, HER2 and K-ras mutations were found to be mutually exclusive events. The presence of HER2 mutations in a subset of patients with lung adenocarcinoma raise hope to treat these patients with HER2 specific kinase inhibitors.


Different human tenascin-C variants in the extracellular matrix of cultured human fibroblasts.
Using an immunoadsorbent prepared with a monoclonal antibody specific for the high molecular mass isoform of human tenascin-C, we purified tenascin-C molecules containing at least one large subunit from the extracellular matrix of cultured normal human fibroblasts. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblotting analyses have shown that both high and low molecular mass subunits are present in these tenascin-C preparations. Because the monoclonal antibody used is able to bind only the high molecular mass isoform, the present data show that part of the tenascin-C present in the fibroblast extracellular matrix is made up of heterohexameric molecules.