A histological assessment on the distribution of the osteocytic lacunar canalicular system using silver staining.
Giving the complexity that characterizes the mechanisms of bone remodeling and the number of events that have to be in absolute harmony for it to occur flawlessly, the postulation that temporospatial distribution of osteocytes and their lacunar canalicular system might influence and be influenced by bone remodeling can be regarded, at least, as feasible. In this study, using Schoen's silver staining, we have examined the distribution of the osteocytic lacunar canalicular system (OLCS) in bones of developing mice. Trabecular bones of 3-day-old, 2-week-old, and 3-week-old mice displayed osteocytic cytoplasmic processes without any perceptible alignment. Also, many plump osteocytes were embedded in the mineralized bone matrix in a disorderly manner. At 4 weeks of age, however, mice bones showed some osteocytic processes that reached the bone surface on a right angle, while other osteocytes displayed the same features seen on 3-week specimens. Samples at 8 weeks of age featured osteocytes with their usual spindle shape, organized so as to parallel the longitudinal axis of trabecular bone. They also extended their cytoplasmic processes perpendicularly to the bone surface. However, several osteocytes immersed in older bone, i.e., a residual mix of cartilage and bone matrices, still showed a random pattern of distribution of their cytoplasmic processes. Up to 12 weeks of age, the majority of the osteocytes became flattened and were shown to be aligned with their long axis paralleling the bone surface. This tendency for such a gradual arrangement was also observed in cortical bones. We have further demonstrated that 8-week-old osteoprotegerin-deficient mice, which demonstrated histological evidence of higher than average bone turnover, revealed a disorganized OLCS. Given the data gathered in this work, the OLCS appears to assume an organized, probably function-related spatial distribution as normal bone remodeling goes on.


Brachialis syndrome: a rare consequence of patient positioning causing postoperative median neuropathy.
Poor positioning of patients can result in devastating permanent neurologic deficits. We describe a previously unreported cause of median nerve compression that we have termed the brachialis syndrome, associated with patient positioning that results in permanent median nerve damage. We identified this condition affecting 6 median nerves. All patients underwent surgical decompression of the proximal median nerve at the level of the antecubital fossa. Five patients presented with symptoms of median nerve compression; 6 affected median nerves manifested brachialis syndrome after a lengthy index surgery. Every patient had a similar presentation characterized by a mixed sensory and motor deficit. Average time to symptom presentation postoperatively was 1 hour. Two patients had delayed time to decompression, one of 25 days and one of 92 days. In the additional patients, the average time to decompression was 19.7 hours. At median nerve decompression, the brachialis was found to have varying degrees of muscle necrosis. In the patients whose decompression was delayed, there was only partial neurologic recovery at follow-up to 1 year. In the patients expeditiously decompressed, full neurologic recovery occurred in 1 to 14 days. This is the first description of the brachialis syndrome. During surgery, arms were placed into full extension, compressing the brachialis against the trochlea. The brachialis reliably developed necrosis, resulting in swelling, compressing the median nerve against the lacertus fibrosus. Two patients with delayed decompression had poor neurologic outcomes. This supports modification of patient positioning, postoperative vigilance, and timely surgical management of brachialis syndrome.


Characterization of aerosol sources in León (Spain) using Positive Matrix Factorization and weather types.
A one-year aerosol sampling campaign, between 2016 and 2017, was conducted in a suburban area of León city, Spain. An association between the Positive Matrix Factorization (PMF) results and air masses through circulation weather types was carried out, through the construction of linear models from the PM10 concentrations and its chemical composition. The aerosol sources, identified by PMF six-factor solution, were: traffic (29%), aged sea salt (26%), secondary aerosols (16%), dust (13%), marine aerosol (7%) and biomass burning (3%). Traffic and secondary factors showed the highest PM10 contribution in the hybrid cyclonic types with wind component from the first and second quadrant. Anticyclonic types with wind component from the first quadrant exhibited high values of secondary, aged sea salt and dust factors. The highest contributions of the dust factor were also associated with northerly types. The linear models built for estimating the source apportionment of PM10, from aerosol chemical composition and geostrophic flow, showed positive coefficients for: westerly flows (WF) in marine factor, southerly flows (SF) in secondary and traffic factors, and shear southerly vorticities (ZS) in dust factor. Negative dependences were observed for ZS in aged sea salt factor and for SF in dust factor. The PM10 mass concentration calculated by the linear models and by the PMF model were strongly correlated. This can be very useful to determine the contribution of a specific source to PM10 in León, only by knowing some meteorological and chemical variables.


Chromosome aberrations, spontaneous SCE, and growth kinetics in PHA-stimulated lymphocytes of five cases with Sézary syndrome.
Cytogenetic studies of five patients with Sézary syndrome (SS) revealed clonal chromosome aberrations in all cases. In one patient, a del(8)(p21) was the sole abnormality, whereas the remaining cases had karyotypes with multiple chromosome changes. In three SS cases with hypodiploid chromosome numbers, structural rearrangements affecting regions 10q22-24 and 12p11-13, and aberrations leading to loss of material from 17p were found concurrently. Bands 14q11 and 14q32 were involved in structural rearrangements in one case each. Our results and review of 51 published previously SS cases that were analyzed with banding techniques indicate that the chromosomes most frequently involved in structural changes were chromosomes 1 and 2 (in 43% of cases), 6 (in 38%), 17 (in 34%), 14 (in 27%), 11 (in 25%), 13 (in 21%), and 9 (in 20%). In particular, the breakpoints tended to aggregate at 1p11, 1p36, 2p11-24, 6q, 9q, 11q, 13q11-14, 14q11, 14q32, and in the pericentric region of chromosome 17. The most common numerical change was loss of chromosome 10, detected in 32% of SS cases. In our studies of three SS cases, sister chromatid exchange frequencies were significantly higher in comparison to the normal control. Cell cycle kinetics analysis revealed that the cell cycle time in the malignant cells was significantly longer than in lymphocytes of normal individuals.


Estrous cycle-dependent changes in basal and ethanol-induced activity of cortical dopaminergic neurons in the rat.
The influence of the estrous cycle on dopamine levels in the rat medial prefrontal cortex under basal and ethanol-stimulated conditions was evaluated by microdialysis. The basal dopamine concentration in the dialysate varied markedly during the estrous cycle, being highest in estrus and lowest in proestrus. Furthermore, a challenge intraperitoneal administration of ethanol (0.5 g/kg) induced a significant increase in dopaminergic output (+50%) during estrus but had no effect in diestrus or proestrus. Ovariectomy or pretreatment with either finasteride (a 5alpha-reductase inhibitor) or clomiphene (an estrogen receptor antagonist) prevented this ethanol-induced increase in dopamine concentration. The effect of ethanol was restored in ovariectomized rats by pretreatment with estrogen but not by that with progesterone. Our results thus show that the basal levels of dopamine in the prefrontal cortex are dependent on the phase of the estrous cycle. Furthermore, this dependence appears to be attributable to the effects of ovarian steroid hormones and results in a differential sensitivity of the dopaminergic neurons to ethanol. The hormone-induced changes in the activity of these neurons might contribute to the differences in drug sensitivity and mood state apparent among phases of the estrous cycle and between the sexes.


Impact of baseline calibration on semiquantitative assessment of myocardial perfusion reserve by adenosine stress MRI.
In this study, we sought to investigate the impact of baseline calibration, which is used in quantitative cardiac MRI perfusion analysis to correct for surface coil inhomogeneity and noise, on myocardial perfusion reserve index (MPRI) and its contribution to previously reported paradoxical low MPRI < 1.0 in patients with unobstructed coronary arteries. Semiquantitative perfusion analysis was performed in 20 patients with unobstructed coronary arteries undergoing stress/rest perfusion CMR and in ten patients undergoing paired rest perfusion CMR. The following baseline calibration settings were compared: (1) baseline division, (2) baseline subtraction and (3) no baseline calibration. In uncalibrated analysis, we observed ~ 20% segmental dispersion of signal intensity (SI)-over-time curves. Both baseline subtraction and baseline division reduced relative dispersion of t0-SI (p < 0.001), but only baseline division corrected for dispersion of peak-SI and maximum upslope also (p < 0.001). In the assessment of perfusion indices, however, baseline division resulted in paradoxical low MPRI (1.01 ± 0.23 vs. 1.63 ± 0.38, p < 0.001) and rest perfusion index (RPI 0.54 ± 0.07 vs. 0.94 ± 0.12, p < 0.001), respectively. This was due to a reversed ratio of blood-pool and myocardial baseline-SI before the second perfusion study caused by circulating contrast agent from the first injection. In conclusion, baseline division reliably corrects for inhomogeneity of the surface coil sensitivity profile facilitating comparisons of regional myocardial perfusion during hyperemia or at rest. However, in the assessment of MPRI, baseline division can lead to paradoxical low results (even MPRI < 1.0 in patients with unobstructed coronary arteries) potentially mimicking severely impaired perfusion reserve. Thus, in the assessment of MPRI we propose to waive baseline calibration.


Biocatalysis of Steroids by Mycobacterium sp. in Aqueous and Organic Media.
Mycobacterium sp. can convert steroids such as β-sitosterol, campesterol, and cholesterol, by selective side-chain cleavage and oxidation of the C3 hydroxyl group to a ketone, into key intermediates that can be easily functionalized to yield commercially interesting pharmaceutical products. In aqueous systems, the biocatalysis is limited by the low solubility of the steroids in water. Several strategies have been introduced to tackle this limitation, e.g., formation of cyclodextrin-steroid complexes and generation of aqueous microdispersions with steroid particle size in the range of hundreds of nanometers. Still, the introduction of an organic phase acting as a substrate and/or product reservoir is a well-established and relatively easy to implement strategy to overcome the sparing water solubility of steroid molecules. However, the organic phase has to be carefully chosen to prevent tampering with the activity/viability of microbial cells.In this chapter, we describe the methodology for the biocatalysis of β-sitosterol to 4-androstene-3,17-dione (AD) and 1,4-androstadiene-3,17-dione (ADD), both in aqueous and organic:aqueous systems. In the latter case, both traditional organic solvents and green solvents are proposed.


Expression of Inflammatory and Regenerative Genes in a Model of Liver Ischemia/Reperfusion and Partial Hepatectomy.
Hepatic ischemia/reperfusion (I/R) describes the paradox of additional tissue injury caused by reperfusion. The aim of this survey was to investigate the mRNA expression of genes exerting their inflammatory and regenerative reaction in a porcine model of I/R and extended hepatectomy. Twelve pigs were used, weighing 30-35 kg in average, which were allocated in two groups: the I/R group with eight pigs and the sham-operated (control) one with four pigs. The I/R group underwent portacaval anastomosis and Pringle maneuver followed by extended hepatectomy. The hepatoduodenal ligament was occluded for 150 min and the liver remnant was reperfused for 24 hours. Blood samples were steadily received throughout the surgical procedure, where hepatic biopsies were taken for pathological evaluation. Animals were sacrificed in 24 hours after the onset of reperfusion. Between the two groups, statistically significant differences were noticed in serum values of AST, ALT, ALP, and total bilirubin in the early and late phase of reperfusion. The mRNA expression of iNOS, IL-1b, and TGF-a did not increase significantly in the I/R group. Conversely, the mRNA modification of IL-6, STAT-3, and E-selectin demonstrated significantly increased expression in I/R animals. In the present survey, a new I/R swine model was proposed and specific parameters were analyzed, revealing differences between the study groups.


Discovery, biosynthesis, and rational engineering of novel enterocin and wailupemycin polyketide analogues.
The marine actinomycete Streptomyces maritimus produces a structurally diverse set of unusual polyketide natural products including the major metabolite enterocin. Investigations of enterocin biosynthesis revealed that the unique carbon skeleton is derived from an aromatic polyketide pathway which is genetically coded by the 21.3 kb enc gene cluster in S. maritimus. Characterization of the enc biosynthesis gene cluster and subsequent manipulation of it via heterologous expression and/or mutagenesis enabled the discovery of other enc-based metabolites that were produced in only very minor amounts in the wild type. Also described are techniques used to harness the enterocin biosynthetic machinery in order to generate unnatural enc-derived polyketide analogues. This review focuses upon the molecular methods used in combination with classical natural products detection and isolation techniques to access minor metabolites of the S. maritimus secondary metabolome.


Comparison of wound complications between one-stage and two-stage brachiobasilic arteriovenous fistula: A meta-analysis.
Research assessing the outcome of brachiobasilic arteriovenous fistulas (BB-AVF) after a one- and two-stage technique was conducted. A strict review of the comprehensive literature up to May 2023 was carried out using four databases-PubMed,Embase, Cochrane Library and web of science. Inclusion and exclusion criteria developed for the study were then applied to assess the quality of the literature, it was decided to review 12 interrelated studies.95% confidence intervals (Cl) and odds ratios (OR) were calculated using fixed effects models. The data were meta-analysed using RevMan 5.3 software. The results showed that two-stage technique with BB-AVF reduced surgical site haematoma (SSH) formation, (OR, 2.28; 95% CI, 1.24-4.17, p = 0.008), and also reduced surgical site wound infection (SSWI) (OR, 1.86; 95% CI, 1.17-2.94, p = 0.008). There are, however, several small sample sizes in the selected studies for this meta-analysis, so caution should be used when processing their values. There are more high-quality studies with large sample sizes that should be considered for future meta-analyses.


H-2-determined kinetic differences for the induction of nucleoside-specific suppression.
The kinetic and the H-2 requirements for the induction of nucleoside-specific suppression were examined in several strains of mice; specifically, whether adenosine (A)-coupled spleen cells given intravenously suppress the primary response to adenosine-KLH. The adenosine system was chosen because C57Bl/6 mice were originally found to be resistant to immune suppression when challenged 5 days after treatment with adenosine-coupled spleen cells. (Raps et al. J. Immunol. 126, 1542, 1981.) It was determined (i) whether A-specific nonresponsiveness is inducible in strains other than C57Bl/6; (ii) whether changes in hapten density on the A-conjugated spleen cells could alter C57Bl/6s ability to become nonresponsive, and (iii) whether there are interstrain differences in the time required to induce A-specific suppressor T cells (Ts). The results show that there are H-2-associated differences in the time required to induce A-specific immune suppression. While A-spleen cells failed to suppress the A-specific response in C67Bl/10 (H-2b), they did induce unresponsiveness in B10.D2 (H-2d on C57Bl/10 background). A 2.5-fold increase in epitope density of adenosine on cells did not influence the kinetics of suppression. C67Bl/6 were resistant to suppression on Day 5, but like the CB6F1, susceptible to unresponsiveness 10 days after treatment. Nonresponsiveness was T-cell-mediated and transferable across IgH-V barriers. Suppression induced by Balb/c donor mice is transferable to Igh-incompatible CAL-20 mice. These results are discussed in the context of genetic restrictions which regulate suppressor T-cell interactions.


Probing cellular protein targets of H2O2 with fluorescein-conjugated iodoacetamide and antibodies to fluorescein.
Recent studies suggest that H2O2, at subtoxic concentrations generated in response to the activation of a variety of cell surface receptors, functions as an intracellular messenger. However, the intracellular targets of H2O2 action have not been identified. A procedure to detect proteins with reactive cysteine residues susceptible to oxidation by intracellularly generated H2O2 is now described. This approach is based on the labeling of proteinaceous cysteine with 5-iodoacetamidofluorescein at pH 5.5 and immunoblot analysis of the labeled proteins with antibodies specific to fluorescein. With this procedure, many proteins in human A431 cells were shown to contain reactive cysteines and to be readily oxidized by H2O2 generated in response to cellular stimulation with epidermal growth factor. One of these H2O2-sensitive proteins was identified as protein tyrosine phosphatase 1B.


A stability-indicating reversed-phase high performance liquid chromatography method for simultaneous assay of two corticosteroids and estimation of their related compounds in a pharmaceutical injectable formulation.
Betamethasone Sodium Phosphate and Betamethasone Acetate are the two corticosteroids active pharmaceutical ingredients (APIs) that are present in the injectable formulation, Celestone Chronodose(®) Injection. It is extremely challenging to develop a Quality Control friendly HPLC method to separate all the potential impurities and degradation products of the two APIs from each other using a single HPLC method. A novel stability-indicating reversed-phase HPLC (RP-HPLC) method using two oxo-cyclic organic modifiers in the mobile phase was developed and validated. This method can separate a total of 32 potential impurities and degradation products from the two APIs and also from each other. Peak symmetry and separation efficiency were enhanced by using two chaotropic agents (trifluoroacetic acid and potassium hexafluorophosphate) in the mobile phases of this method. The stability-indicating capability of this method has been demonstrated by analyzing aged and stressed degraded stability samples of the drug product. This method uses an ACE 3 C18 (15 cm × 4.6 mm) HPLC column. The method was validated per ICH guidelines and proved to be suitable for routine QC use.


Preparing Anesthetists to Manage Cannot Intubate/Cannot Ventilate Situations.
Cannot intubate/cannot ventilate (CICV) situations during anesthesia are rare, potentially catastrophic to the patient, and difficult to predict. Widely adopted practice guidelines advocate an algorithmic approach to CICV situations in which the anesthetist: (a) recognizes the CICV situation, (b) calls for help, (c) steadily progresses through a variety of methods to ventilate the patient and secure the airway, (d) restores ventilation via an infraglottic airway if the patient cannot be safely awakened prior to becoming moribund. Despite widespread consensus that rapid progression to placement of an infraglottic airway is critical to the survival of the patient in a CICV situation, the rarity of CICV is a substantial barrier for anesthetists attempting to gain and maintain skill at placing infraglottic airways. Peer-reviewed literature reveals a number of themes relevant to training anesthetists in infraglottic airway placement. Specific training in infraglottic airway access consistently decreased the time required for anesthetists to decide to place an infraglottic airway. No one approach or method for placing an infraglottic airway was consistently faster or more successful. Model fidelity (i.e., the use of low-fidelity task trainers vs. high-fidelity simulated patients) during teaching did not affect the performance of anesthetists at placing infraglottic airways, and performance rapidly plateaued after five repetitions during teaching. Finally, skill at placing an infraglottic airway was sustained for 6 to 12 months after training. These findings have implications for how nurse anesthetists should learn and sustain the skill of infraglottic airway placement. Nurse anesthetists should undergo initial training that requires the repetitive placement of infraglottic airways, and receive sustainment training every 6 to 12 months thereafter. Although rarely used, the skill of an anesthetist in infraglottic airway placement can mean the difference between life and death for the patient in a CICV situation.


Lupus-like syndrome in a mother and newborn following administration of hydralazine; a case report.
A very-low-birth-weight infant died from pericardial effusion and cardiac tamponade confirmed by the post-mortem findings. The mother suffered from lupus-like syndrome consequent to hydralazine treatment for pregnancy-induced hypertension. The possible relationship between mother-infant pathology and hydralazine administration is discussed.


Expression and localization of apolipoprotein M in human colorectal tissues.
It has been well documented that apolipoprotein M (apoM) is principally expressed in the liver and kidney. However we found that there was weak apoM expression in other tissues or organs too, which could not be ignored. In the present study, we therefore examined apoM expression in human colorectal tissues including cancer tissues, cancer adjacent normal tissues, polyp tissues and normal mucosa as well as inflammatory mucosa. Tissue samples were collected from patients who underwent surgical resection or endoscopic examination. ApoM mRNA levels were determined by the real-time RT-PCR and apoM protein mass were examined by the immunohistochemistry. ApoM protein can be detected in all colorectal tissues. However, apoM protein mass were significantly lower in the cancer tissues than its matched adjacent normal tissues, polyp tissues, normal mucosa and inflammatory mucosa. In parallel, apoM mRNA levels in the colorectal cancer tissues (0.0536 ± 0.0131) were also significantly lower than those in their adjacent normal tissues (0.1907 ± 0.0563) (P = 0.033). Interestingly, apoM mRNA levels in colorectal cancer tissues were statistic significant higher in the patients with lymph node metastasis than the patients without lymph node metastasis (P = 0.008). Patients under Dukes' C and D stages had much higher apoM mRNA levels than patients under Dukes' A and B stages (P = 0.034). It is concluded that apoM could also be expressed in human colorectal tissues besides liver and kidney. ApoM mRNA levels in the colorectal cancer tissues were significantly increased in the patients with lymph node metastasis. Whether increased apoM expression in the patients with lymph node metastasis being related to patients' prognosis and the physiopathological importance of apoM expression in colorectal tissues need further investigation.


Three new cases with a supernumerary ring chromosome 1.
We report on three cases with a cytogenetically identical ring chromosome containing euchromatin from the long arm of chromosome 1 (r[1][::p11.1-->q21.1::]). Two cases were newborn males (Cases 1 and 2) and the third one was prenatally identified as female (Case 3). Mosaicism was present in all three cases in different degrees, i.e. 48%, 25% and 14% of the cells, respectively. Clinical signs and symptoms vary between the three cases. The results of our three cases are compared with those from the literature.


A magnetic hydrophilic molecularly imprinted material with multiple stimuli-response properties for efficient recognition of bisphenol A in beverages.
A magnetic hydrophilic molecularly imprinted material (MIM) with multiple stimuli-responses was prepared for efficient recognition of bisphenol A (BPA). MIM consisted of a magnetic core, an inner silica dioxide shell and an outer hydrophilic polymer shell, responsive for rapid magnetic separation, protection of the magnetic core and pH-/thermo-sensitivity to BPA, respectively. Controlled adsorption and release of BPA by pH/temperature regulation were realized through the reversible components of poly[N-isopropylacrylamide-co-2-(dimethylamino)ethyl methacrylate (NIPAM-DMAEMA)] (PND). The resultant MIM possessed superior affinity, selectivity and kinetics to BPA in aqueous solution. MIM was successfully applied to detect BPA in beverages via dispersive solid-phase extraction (d-SPE) coupled with high performance liquid chromatography (HPLC), exhibiting satisfactory recoveries of 80.70-108.18% with relative standard deviations (RSD) below 6.08%. The limit of detection of BPA was obtained as low as 3.75 nmol/L. The versatile MIM could be a promising alternative for extraction/removal of BPA in complicated samples by multiple-stimuli regulations.


The study aimed to validate a whole-genome sequencing-based NIPT laboratory method and our recently developed NIPTmer aneuploidy detection software with the potential to integrate the pipeline into prenatal clinical care in Estonia. In total, 424 maternal blood samples were included. Analysis pipeline involved cell-free DNA extraction, library preparation and massively parallel sequencing on Illumina platform. Aneuploidies were determined with NIPTmer software, which is based on counting pre-defined per-chromosome sets of unique k-mers from sequencing raw data. SeqFF was implemented to estimate cell-free fetal DNA (cffDNA) fraction. NIPTmer identified correctly all samples of non-mosaic trisomy 21 (T21, 15/15), T18 (9/9), T13 (4/4) and monosomy X (4/4) cases, with the 100% sensitivity. However, one mosaic T18 remained undetected. Six false-positive (FP) results were observed (FP rate of 1.5%, 6/398), including three for T18 (specificity 99.3%) and three for T13 (specificity 99.3%). The level of cffDNA of <4% was estimated in eight samples, including one sample with T13 and T18. Despite low cffDNA level, these two samples were determined as aneuploid. We believe that the developed NIPT method can successfully be used as a universal primary screening test in combination with ultrasound scan for the first trimester fetal examination.


Influence of surface coverage with poly(ethylene oxide) on attachment of sterically stabilized microspheres to rat Kupffer cells in vitro.
The attachment to rat Kupffer cells of polymeric microspheres, sterically stabilized with different amounts of pendant poly(ethylene oxide) (PEO), was assessed in vitro. Four types of copolymer polystyrene (PS) microspheres were synthesized by variation of four possible monomer ratios that included styrene, methoxy-PEO-methacrylate (750 and 2000 mol. wt PEO) and allylurea. This produced poly(styrene-(methoxy-PEO)methacrylate) microspheres with hydrophilic side-groups of either urea (PS-U-PEO) and/or mixed molecular weight (750/2000 mol. wt) PEO (PS-U-M-PEO, PS-M-PEO), or single molecular weight (2000) PEO (PS-PEO) at their surfaces. The hypothesis was tested that increasing the total content of PEO comprising the steric barrier reduces attachment to cell surfaces. Attachment of PEO microspheres bearing the urea spacer and/or mixed molecular weight PEO was found to be intermediate between charge stabilized control PS and PEO (2000 mol. wt) bearing particles. Post-adsorption of different Poloxamer (PEO-poly(propylene oxide)-PEO) surfactants to the microspheres further decreased attachment. Significant negative linear correlations between surface PEO content, measured by electron spectroscopy for chemical analysis (ESCA), and attachment to Kupffer cells were demonstrated. Decreases in attachment also resulted with all graft PEO particles bearing adsorbed sodium dodecyl sulphate (SDS), whilst the attachment of SDS-treated PS control particles increased. It is proposed that trains of adsorbed graft PEO are displaced by the SDS to increase the effective fraction of graft PEO within the steric layer. Overall, increasing the amount of hydrophilic PEO in the steric layer, from graft and adsorbed sources, reduces the attachment of these particles to Kupffer cells in vitro.


Effects of Multi-metal (Cu, Zn, Cd, Cr, and Mn) Mixtures on the Reproduction of Freshwater Rotifer Brachionus calyciflorus.
In the field, organisms are usually exposed to mixtures of various metals. However, the effects of multi-metal mixtures on growth and reproduction of rotifers remain unknown. In the present study, effects of multi-metal mixtures (Cu, Zn, Cd, Cr, and Mn) on reproduction of the freshwater rotifer Brachionus calyciflorus were assessed by determining various endpoints, including the ratio of ovigerous females to nonovigerous females, the ratio of mictic to amictic females, the mictic rate, the fertilization rate, the population growth rate, and the resting eggs production. The results demonstrated that reproduction of rotifers was significantly affected by all multi-metal mixtures assessed. Moreover, the ratio of mictic to amictic females was the most sensitive endpoint and might be suitable to evaluate effects of multi-metal mixtures to rotifers.


Characteristics of sorbitol uptake in rat glial primary cultures.
Uptake of [U-14C]sorbitol was studied in astrogliarich rat primary cultures. Initial rate of sorbitol uptake is proportional to sorbitol concentration between 20 microM and 400 mM. Sorbitol transport is not inhibited by glucose, fructose, and a variety of structurally related polyols, or by cytochalasin B, an inhibitor of glucose transport. Phloretin, phlorizin, filipin, and n-hexanol, all compounds that alter the properties of biological membranes, and the sulfhydryl reagent p-chloromercuribenzoate inhibit sorbitol uptake to various degrees. Variation in the concentrations of extracellular Na+ and K+ does not affect transfer of sorbitol across the cell membrane. It is concluded that sorbitol is taken up into glial cells by a diffusion process, not involving a carrier and probably not through the lipid bilayer, but through a proteinaceous channel-like structure.


Effectiveness of a telehealth patient education intervention for breast cancer awareness and screening uptake among Afghan refugee women: a cross-sectional survey and feasibility study.
Refugee populations have low levels of cancer awareness, low cancer screening rates, and a high rate of advanced or metastatic cancer at diagnosis. Educational interventions to improve cancer awareness and screening have been successful in other nationality refugee populations but have never been implemented in Afghan refugee populations. We aimed to estimate the level of breast cancer awareness among Afghan refugee women and test the feasibility of a telehealth breast cancer educational intervention to increase breast cancer awareness in this population. A cross-sectional survey of Afghan refugee women residing in Istanbul, Türkiye who had no personal history of breast cancer and who presented to outpatient primary clinics for care between August 1, 2022, and February 10, 2023 was performed. Participant awareness of breast cancer (BC) was assessed using the validated BC awareness tool, Breast Cancer Awareness Measure (BCAM), during a telehealth encounter. After this baseline assessment, a BC educational intervention was administered to each participant during the telehealth encounter. Six months after the initial assessment and education, a follow-up BCAM was administered via telehealth in order to determine the effectiveness of the education intervention. One hundred participants were accrued to the study. Median age was 49 years (range: 40-64). All participants had no formal education, were married, and were not employed. Prior to the educational intervention, BC awareness was low; none of the participants were able to identify some common signs/symptoms and risk factors for BC. Prior to the educational intervention, zero participants had ever had a mammogram or seen a physician for a breast-related concern. Six months after the educational intervention, up to 99 percent of participants (99 of 100 participants) were able to correctly identify common signs or symptoms and risk factors for BC. Six months after the educational intervention, all one hundred participants had accepted the offer of a screening mammogram. A telehealth BC education intervention meaningfully increased BC awareness in Afghan refugee women. This increase in BC awareness was associated with a strong increase in completion of BC screening. Further implementation of educational interventions is warranted in order to increase participant awareness and improve screening rates. Dana Farber Cancer Institute Jay Harris Junior Faculty Research Grant.


Mutational analysis of disulfide bridges in the type C atrial natriuretic peptide receptor.
The type C receptor (ANP-C or NPR-C) for the natriuretic peptides was demonstrated, by site-directed mutagenesis, to have an immunoglobulin-like disulfide bonding pattern that is very similar to that of the cytokine receptor superfamily. The mature form of ANP-C has a disulfide-linked homodimeric structure and contains 5 conserved cysteine residues per subunit, all in the extracellular domain. To identify the cysteine residue involved in the dimerization and further to determine the intramolecular disulfide bridges and their functional roles, cysteine to serine mutations of the 5 cysteine residues were constructed. An analysis of the mutant receptors expressed in COS-1 cells by 125I-ANP binding assay and by measuring difference in their electrophoretic mobilities on sodium dodecyl sulfate-polyacrylamide gels indicated that 1) the first 4 cysteine residues are joined sequentially, forming the Cys104-Cys132 and Cys209-Cys257 loops of 29 and 49 residues, respectively; 2) the two disulfide-linked loops are essential for the ligand binding activity; 3) the 5th cysteine residue Cys469 is used in the formation of covalently linked dimers; and 4) the covalent association of the subunit through the disulfide bond involving Cys469 has no apparent influence on ligand-receptor interactions. The intramolecular disulfide bond Cys104-Cys132 was also confirmed by direct protein sequencing of tryptic fragments of purified ANP-C receptor. The secondary structural features revealed here will be useful in understanding the structure and function relationships of not only the dimeric ANP-C receptor, which has only a short cytoplasmic tail, but also the ANP-A (GC-A) and ANP-B (GC-B) receptor subtypes, which have a guanylate cyclase domain in their long cytoplasmic tail and have recently been shown to possess an oligomeric structure, since they have similarly spaced cysteine residues in their extracellular domains.


Measurements of serum-free thyroid hormone concentrations by ultrafiltration--a comparison with equilibrium dialysis and mathematical calculation.
An ultrafiltration method (UF) for measuring free thyroxine (FT4) and free triiodothyronine (FT3) using the Diaflow YM membrane (Centricon-10) is described. The results are compared with those by equilibrium dialysis (ED) and also by mathematical calculations derived from T4, T3, and binding protein concentrations. The precision with the UF method was excellent. The normal ranges of FT4 and FT3 by the three methods are all comparable. There was a high degree of correlation of FT4 or FT3 results by UF with those by ED and by calculation (r = 0.940-0.974, n = 161, P less than 0.001). FT4 and FT3 by all methods agreed well for hyperthyroidism, hypothyroidism, and for patients with low T4-binding globulin. The mean FT3 in pregnancy was lower than the normal value for all methods, and FT4 concentrations by UF and calculation also decreased in late pregnancy. The mean FT4 by UF and ED in low T3 syndrome were significantly higher than in the normal controls, while the calculated FT4 was lower. The FT3 in low T3 syndrome distributed normal to subnormal in all methods. These results indicate that a) the UF method is a reliable reference method for measuring FT4 and FT3 concentrations; b) the UF results agree well with those by ED and also with theoretically derived values in subjects with thyroid diseases and TBG abnormalities; c) for patients with low T3 syndrome, the FT4 results obtained by UF and ED are similarly discrepant from the calculated results, implying the existence of binding inhibitor(s) which affect both UF and ED measurements.


Suspension Trapping-Based Sample Preparation Workflow for In-Depth Plant Phosphoproteomics.
Plant phosphoproteomics provides a global view of phosphorylation-mediated signaling in plants; however, it demands high-throughput methods with sensitive detection and accurate quantification. Despite the widespread use of protein precipitation for removing contaminants and improving sample purity, it limits the sensitivity and throughput of plant phosphoproteomic analysis. The multiple handling steps involved in protein precipitation lead to sample loss and process variability. Herein, we developed an approach based on suspension trapping (S-Trap), termed tandem S-Trap-IMAC (immobilized metal ion affinity chromatography), by integrating an S-Trap micro-column with a Fe-IMAC tip. Compared with a precipitation-based workflow, the tandem S-Trap-IMAC method deepened the coverage of the Arabidopsis (Arabidopsis thaliana) phosphoproteome by more than 30%, with improved number of multiply phosphorylated peptides, quantification accuracy, and short sample processing time. We applied the tandem S-Trap-IMAC method for studying abscisic acid (ABA) signaling in Arabidopsis seedlings. We thus discovered that a significant proportion of the phosphopeptides induced by ABA are multiply phosphorylated peptides, indicating their importance in early ABA signaling and quantified several key phosphorylation sites on core ABA signaling components across four time points. Our results show that the optimized workflow aids high-throughput phosphoproteome profiling of low-input plant samples.


'Man up': Medical students' perceptions of gender and learning in clinical practice: A qualitative study.
Gender-related inequality and disparity hinders efforts to develop a medical workforce that facilitates universal access to safe, just and equitable health care. Little is known about how medical students perceive the impact of their gender on their learning in clinical practice. Our aim in this study was to address this gap, establishing students' perceptions of the impact of their gender on learning in the clinical context as part of the wider medical education community of practice. We undertook a qualitative study that simultaneously gathered data through narrative individual interviews and online case reports from male and female students (n = 31) from different academic cohorts with prior experience of clinical practice in a Russell Group University medical school in the UK. Interviews were transcribed and analysed thematically alongside case report data. The participants revealed that there was a culture in clinical practice where their gender influenced how they were taught and supported by senior medical and surgical colleagues. Gender was also said to determine the clinical learning opportunities afforded to students, especially with regards to the care of patients of a different gender. The mentorship and support for learning provided to students in clinical practice was also said to be influenced by the medical student's gender. Our findings suggest that students undergo a gendered clinical apprenticeship within what are in effect gendered communities of practice with some distinct features. These findings underscore the imperative for further work to establish how medical students of all genders can be supported to fulfil their potential in clinical practice.


Ion-swimming speed variation of Vibrio cholerae cells.
In the present work we report the variation in swimming speed of Vibrio cholerae with respect to the change in concentration of sodium ions in the medium. We have also studied the variation in swimming speed with respect to temperature. We find that the swimming speed initially shows a linear increase with the increase of the sodium ions in the medium and then plateaus. The range within which the swimming speed attains saturation is approximately the same at different temperatures.


A role for ATP-sensitive potassium channels in male sexual behavior.
ATP-sensitive potassium (K(+)(ATP)) channels regulate cell excitability and are expressed in steroid-responsive brain regions involved in sexual behavior, such as the preoptic area (POA) and medial basal hypothalamus (MBH). We hypothesized that K(+)(ATP) channels serve as a mechanism by which testosterone can control the electrical activity of neurons and consequently elicit male sexual responsiveness. RT-PCR analysis indicated that castration induces, while testosterone inhibits, mRNA expression of the K(+)(ATP) channel subunit Kir6.2 in both the POA and MBH of adult male rats. Intracerebral infusion of the pharmacological K(+)(ATP) channel inhibitor tolbutamide increased the proportion of long-term castrates displaying sexual behavior and restored mount frequency, intromission frequency, and copulatory efficacy to values observed in testes-intact animals. Infusions of tolbutamide, but not vehicle, also decreased latencies to mount and intromit in castrated males. Unilateral tolbutamide infusion directly into the POA significantly reduced mount latency of castrates; however, it did not affect other copulatory measures, suggesting that blockade of K(+)(ATP) channels in additional brain regions may be necessary to recover the full range of sexual behavior. These data indicate that blockade of K(+)(ATP) channels is sufficient to elicit the male sexual response in the absence of testosterone. Our observations are consistent with the hypothesis that testosterone modulates male sexual behavior by regulating K(+)(ATP) channels in the brain. Decreased channel expression or channel blockade may increase the excitability of androgen-target neurons, rendering them more sensitive to the hormonal, chemical, and somatosensory inputs they receive, and potentially increase secretion of neurotransmitters that facilitate sexual behavior.


Cardiological disease in the Armed Forces: a clear and present danger.
Despite a younger cohort of individuals than typically found in other large organisations the Armed Forces are not free from cardiac disease. Indeed some conditions are more frequently found and the high risk occupations of individuals within the Armed Forces makes it imperative that all cardiac conditions are identified rapidly and correctly. By illustrating actual cases, we highlight the need for service personnel to have rapid access to expert cardiological expertise, from individuals with an in depth occupational knowledge of the Armed Forces, 365 days per year.


Total syntheses of tambjamines C, E, F, G, H, I and J, BE-18591, and a related alkaloid from the marine bacterium Pseudoalteromonas tunicata.
The acetate salts of tambjamines C, E, and F (2-4, respectively), as well as those of the related alkaloids BE-18591 (5) and 6, have been prepared by treatment of bipyrrole aldehyde 16 with the relevant amine in the presence of acetic acid. The 5'-bromo-analogue, 30, of compound 16 has also been prepared and used to obtain the acetate salts of tambjamines G, H, I, and J (8-11 respectively).


Circulating tumour DNA profiling reveals heterogeneity of EGFR inhibitor resistance mechanisms in lung cancer patients.
Circulating tumour DNA (ctDNA) analysis facilitates studies of tumour heterogeneity. Here we employ CAPP-Seq ctDNA analysis to study resistance mechanisms in 43 non-small cell lung cancer (NSCLC) patients treated with the third-generation epidermal growth factor receptor (EGFR) inhibitor rociletinib. We observe multiple resistance mechanisms in 46% of patients after treatment with first-line inhibitors, indicating frequent intra-patient heterogeneity. Rociletinib resistance recurrently involves MET, EGFR, PIK3CA, ERRB2, KRAS and RB1. We describe a novel EGFR L798I mutation and find that EGFR C797S, which arises in ∼33% of patients after osimertinib treatment, occurs in <3% after rociletinib. Increased MET copy number is the most frequent rociletinib resistance mechanism in this cohort and patients with multiple pre-existing mechanisms (T790M and MET) experience inferior responses. Similarly, rociletinib-resistant xenografts develop MET amplification that can be overcome with the MET inhibitor crizotinib. These results underscore the importance of tumour heterogeneity in NSCLC and the utility of ctDNA-based resistance mechanism assessment.


Alpha 2-adrenoceptor density in forebrain areas of Zucker rats: no effect of genetic obesity or antiglucocorticoid treatment.
The density of alpha 2-adrenoceptors was determined in different forebrain regions of lean and obese Zucker rats by para-[125I]iodoclonidine ([125I]PIC) receptor autoradiography. No significant differences were observed between the Zucker phenotypes in the density of alpha 2-adrenoceptors in any brain areas examined. Four days treatment with the antiglucocorticoid mifepristone (RU 38486) did not affect [125I]PIC-binding. It is concluded that the hyperphagia and abnormal corticosterone secretion of genetically obese Zucker rats do not appear to be associated with changes in alpha 2-adrenoceptor density in brain.


Role of isozyme group-specific sequence 4 in the isozyme-specific properties of human aldolase C.
To assess which regions of the aldolase C molecule are required for exhibiting isozyme-specific kinetic properties, we have constructed nine chimeric enzymes of human aldolases A and C. Kinetic studies of these chimeric enzymes revealed that aldolase C absolutely required its own isozyme group-specific sequences (IGS), particularly IGS-4, for exhibiting the characteristics of aldolase C which differ significantly from those of isozymes A and B (Kusakabe T, Motoki K, Hori K. Human aldolase C: characterization of the recombinant enzyme expressed in Escherichia coli. J Biochem (Tokyo) 1994;115:1172-7). Whereas human aldolases A and B required their own isozyme group-specific sequences-1 and -4 (IGS-1 and -4) as the main determinants of isozyme-specific kinetic properties (Motoki K, Kitajima Y, Hori K. Isozyme-specific modules on human aldolase A molecule. J Biol Chem 1993;268:1677-83; Kusakabe T, Motoki K, Sugimoto Y, Takasaki Y, Hori K. Human aldolase B: liver-specific properties of the isoenzyme depend on type B isozyme group-specific sequence. Prot. Eng. 1994;7:1387-93), the present studies indicate that the IGS-1 is principally substitutable between aldolases A and C. The kinetic data also suggests that the connector-2 (amino acid residues 243-306) may modulate the interaction of IGS units with the alpha/beta barrel of the aldolase molecule.


[Analysis of indications of the acupoints in the same nervous segment on back and abdomen].
To explore the indication law of fourteen channels acupoint that is located in the same nervous segment of T1-T12 on the back and abdomen. By retrieving indications of fourteen channels acupoint that is located in the dominating areas of T1 - T12 in LIN Zhao-geng 's New Collection of Acupuncture-moxibustion and SHEN Xue-yong's Science of meridian-collateral and acupoint, indications of fourteen channels acupoint in the same nervous segment of skin and muscle were statistically managed, respectively. There was an obvious nervous segmental law of acupoint in the skin and muscle. The acupoint indication of neighboring nervous segment was similar, which was closely related to corresponding internal organs. The acupoint indication is decided by the space of nervous segment which dominats their related organ. From aspect of nervous structure, this article confirms that acupoint indications focus on nervous segment to carry out the regulation effect of acupoint on internal organs function, which means it has superior regulation effect on internal organs disease that is located in the same or neighboring nervous segment of acupoint.


Multiparametric identification of subclinical atrial fibrillation after an embolic stroke of undetermined source.
Subclinical atrial fibrillation (SCAF) may represent a cause of embolic stroke of undetermined source (ESUS) and its detection has important implications for secondary prevention with anticoagulation. Indications to implantable cardiac monitors (ICM) include SCAF detection. The aims of this study were to (1) evaluate the frequency of ICM-detected SCAF; (2) determine predictors of SCAF; and (3) identify patients who would benefit most from ICM implantation. Between February 2017 and November 2020, all consecutive patients referred for ICM implantation after a diagnosis of ESUS and without previous history of atrial fibrillation or atrial flutter were included in this study. SCAF was diagnosed if the ICM electrogram demonstrated an episode of irregularly irregular rhythm without distinct P waves lasting > 2 min. We enrolled 109 patients (age 66, SD = 13 years; 36% females). During a median follow-up of 19.2 (IQR 11.0-27.5) months, SCAF episodes were detected in 36 (33%) patients. Only abnormal P wave terminal force in lead V1, left atrial end-systolic indexed volume > 34 ml/m2, and BMI > 25 kg/m2 were independently associated with an increased risk of SCAF (HR 2.44, 95% CI 1.14-5.21, p = 0.021; HR 2.39, 95% CI 1.11-5.13, p = 0.026; and HR 2.64, 95% CI 1.06-6.49, p = 0.036 respectively). The ROC curve showed that the presence of all three parameters had the best accuracy (74%) to predict SCAF detection (sensitivity 39%, specificity 91%). A multiparametric evaluation has the best accuracy to predict SCAF in ESUS patients and may help identifying those who would benefit most from ICM.


Room temperature phosphorescence from natural wood activated by external chloride anion treatment.
Producing afterglow room temperature phosphorescence (RTP) from natural sources is an attractive approach to sustainable RTP materials. However, converting natural resources to RTP materials often requires toxic reagents or complex processing. Here we report that natural wood may be converted into a viable RTP material by treating with magnesium chloride. Specifically, immersing natural wood into an aqueous MgCl2 solution at room temperature produces so-called C-wood containing chloride anions that act to promote spin orbit coupling (SOC) and increase the RTP lifetime. Produced in this manner, C-wood exhibits an intense RTP emission with a lifetime of ~ 297 ms (vs. the ca. 17.5 ms seen for natural wood). As a demonstration of potential utility, an afterglow wood sculpture is prepared in situ by simply spraying the original sculpture with a MgCl2 solution. C-wood was also mixed with polypropylene (PP) to generate printable afterglow fibers suitable for the fabrication of luminescent plastics via 3D printing. We anticipate that the present study will facilitate the development of sustainable RTP materials.


Cumulative effect of T-2 toxin and vitamin C on chromosomal abnormalities in the bone marrow cells of mice (Mus musculus).
A regular dose of 10 mg/kg body wt/day of vitamin C as a part of the daily diet markedly reduced T-2 toxin-induced abnormalities in the chromosomal cells in mice (Mus musculus). The preventive effect on individual types of change in chromosomes such as breakages, chromatid gaps, ring formations and widespread fragmentation, was increased.


Surgical treatment of aneurysms of the thoracic aorta with the aid of partial cardiopulmonary bypass: an analysis of 95 patients.
We retrospectively evaluated the surgical results in 95 patients with aneurysm of the thoracic aorta who were surgically treated using partial cardiopulmonary bypass (CPB) as an adjunctive method during the past 10 years. The cause of the aneurysm was atherosclerosis in 52% and dissection in 41%. Fifty-eight percent of the patients had an aneurysm of the entire descending thoracic aorta and 14 of these patients had a thoracoabdominal aneurysm. Emergency operation was performed in 16 patients (17%). There were 14 early deaths (14.7%) within one month after operation. Postoperative complications included renal dysfunction, partial paraplegia, and hemorrhage. Renal dysfunction occurred in 7 (7.8%) of the operative survivors; 2 of the 7 required hemodialysis. Partial paraplegia was observed in 2 patients undergoing total replacement of the thoracoabdominal aorta. Neither renal dysfunction nor paraplegia was related to the duration of aortic cross-clamping. Postoperative hemorrhage necessitating reopening of the chest occurred in 8 (8.9%) of the operative survivors. Partial CPB is useful in reducing the incidence of postoperative complications among patients undergoing aortic cross-clamping for a long period.


Production of auto-anti-idiotype antibody during the normal immune response. X. Response to TNP-Ficoll in the chicken.
The spontaneous production of auto-anti-idiotype (Id) was demonstrated after injection of chickens with trinitrophenylated Ficoll (TNP-F) by: (a) the presence of hapten-augmentable plaque-forming cells (PFC), (b) the ability of serum and of hapten eluates from immune spleen cells to cause hapten-reversible inhibition of anti-TNP plaque formation, and (c) an enzyme-linked immunosorbent assay (ELISA). Tests for anti-Id using the ELISA and hapten-reversible inhibition of PFC correlated very well. As in the mouse, the incidence of hapten-augmentable PFC was reduced by thymectomy and increased by the transfer of TNP-F-immune spleen cells. Hapten-augmentable PFC were also observed during the immune response of chickens to p-azobenzene arsonate-conjugated Brucella abortus.


Effective treatment at a Danish trauma centre.
The trauma centre at Aarhus University Hospital, Denmark was established in 1999 and has continuously tried to improve its efficiency through regular training of personnel and auditing of selected trauma cases. The purpose of the present study was to assess the efficiency of the trauma teams to perform the initial evaluation using the time spent in the emergency room after arrival and the time to the first chest X-ray as measures for effectiveness. This was a prospective cohort study conducted from January 2000 to December 2008 and which included all trauma patients admitted to the hospital. The results are based on 4,493 admissions, of which 1,102 patients (24%) had an injury severity score > 15. The median time spent in the trauma room was 50 minutes in 2000, which was steadily reduced throughout the period reaching a median time of 27 minutes in 2008 (p < 0.0001, Kruskal-Wallis test). The median time to the first chest X-ray was reduced from seven minutes in 2001 to five minutes in 2008 (p < 0.024, Kruskal-Wallis test). Utilisation of a standardised protocol for initial evaluation and treatment of trauma patients and continuous training of trauma teams may considered some of the main factors responsible for these findings. not relevant. not relevant.


Usability of Nasal Glucagon Device: Partially Randomized Caregiver and Third-Party User Experience Trial with Simulated Administration at a Japanese Site.
Glucagon is the only approved medicine for severe hypoglycemia available for caregivers of people with diabetes. Nasal glucagon (NG) was recently approved in the USA as a needle-free, ready-to-use alternative to injectable glucagon. This simulated user experience study in Japan compared NG and intramuscular glucagon (IMG) administration by caregivers, and NG administration by untrained third parties. This was an open-label, single-center, partially randomized crossover, simulated user experience trial conducted in Japan (October 2018 to December 2018). Caregivers who live with and care for a relative with diabetes were randomized (1:1, stratified by patient diabetes type 1 or 2) to one of two simulated administration sequences (group 1: NG then IMG; group 2: IMG then NG). Caregivers received training on each device 2 weeks before simulated administration of the device. Third parties received no training and only conducted simulated NG administration. Outcome measures included the percentage of successful administrations (based on critical step completion and dose; primary outcome), time to complete administration, and user satisfaction/preferences. In caregivers (N = 19), the percentage of successful administrations was greater (89.5% vs 26.3%, P < 0.001) and mean time to complete administration was shorter (23.9 vs 207.3 s, P < 0.001) with NG than with IMG. In third parties (N = 20), 95% of NG administration attempts were successful (mean time to complete administration, 55.5 s). All caregivers and 80% of third parties reported that the NG device was easy to use. All caregivers and 70% of third parties were confident and willing to use the device in a real emergency, and more than 80% of caregivers preferred the NG device to IMG. This simulated user experience study confirmed that glucagon administration using a nasal delivery device was quicker, easier, and had a higher success rate than intramuscular administration in Japan, where the glucagon injection kit is not available. Eli Lilly. Plain language summary available for this article.


The orbital pulley system: a revolution in concepts of orbital anatomy.
Magnetic resonance imaging (MRI) now enables precise visualization of the mechanical state of the living human orbit. Resulting insights have motivated histological re-examination of human and simian orbits, providing abundant consistent evidence for the active pulley hypothesis, a re-formulation of ocular motor physiology. Each extraocular muscle (EOM) consists of a global layer (GL) contiguous with the tendon and inserting on the eyeball, and a similar-sized orbital layer (OL) inserting on a connective tissue ring forming the EOM pulley. The pulley controls the EOM path and serves as the EOM's functional origin. Activity of the OL positions the pulley along each rectus EOM to assure that its pulling direction shifts by half the change in ocular orientation, the half-angle behavior characteristic of a linear ocular motor plant. Half-angle behavior is equivalent to Listing's law of ocular torsion, and makes 3-D ocular rotations effectively commutative. Pulleys are configured to maintain oblique EOM paths orthogonal to half-angle behavior, and violate Listing's law during the vestibulo-ocular reflex. Rectus pulley positions shift during convergence, facilitating stereopsis. Innervations, fiber types, and metabolism of the OL and GL differ, consistent with the elastic loading of the former, and viscous loading of the latter. Disorders of the location and stability of rectus pulleys are associated with predictable patterns of incomitant strabismus that may mimic cranial nerve palsies. Surgical interventions improve defective pulley function. Understanding of ocular motor control requires characterization of the behavior of the EOM pulleys as well as knowledge of angular eye orientation.


Latent fingermark development using low-vacuum vaporization of ninhydrin.
The vacuum technique is a method of vaporizing a solid material to its gas phase, helping deposit reagents gently on target surfaces to develop latent fingermarks. However, this application is rarely reported in the literature. In this study, a homemade fume hood with a built-in vacuum control system and programmable heating system designed by the Taiwan Criminal Investigation Bureau is introduced. Factors that affect the instrument's performance in developing fingermarks are discussed, including the quantity of chemicals for vaporization, heating program arrangement, and paper of different materials. The results show that fingermarks are effectively developed by vaporizing solid ninhydrin. This would be an alternative application in selecting a solvent-free method for protecting the environment and reducing health hazards in the lab. In terms of the heating program, the result indicates that under a low-vacuum condition (50 mTorr), 80-90 °C is a suitable temperature range for ninhydrin vaporization, allowing ninhydrin to be vaporized without bumping and waste. In terms of the performance on different material papers, this instrument demonstrates its capacity by developing latent fingermarks on thermal paper without discoloration or damaging the original writing, and the same results are also observed on Taiwan and United States banknotes. However, a coherent result could be hardly obtained using the same vaporization setting because different banknotes have their own surface features and water absorption ability or other unique factors may influence the effect of ninhydrin deposition. This study provides a reliable application for developing latent fingermarks without using solvents, and it is also expected to contribute to environmental protection along with the trend of green chemistry technology.


[Blood requirements and transfusion practice evolution in surgery].
During the years 1990, the focus was on transfusion hazards and blood savings strategies. This resulted in a decrease in labile blood products consumption. For a few years, there is a reverse tendency, with a reincrease in blood products consumption. In the same time, the limit of saving strategies has been attained, by identifying the risk of lack of transfusion (or too low transfusion threshold). A so-called multimodal blood savings strategy must be set. Vigilance is mandatory concerning the blood savings strategies that are detailed by scientific societies consensus conferences. Those strategies include a good anticipation of blood requirement, improvement of surgical techniques, pre- and intra-operative autologous transfusion, blood clotting pharmacological improvements, and anesthesia techniques. Transfusion today is relatively safe. Blood requirements must be precisely assessed to avoid going from Charybdis to Scylla i.e. from over- to under-transfusion.


Direction of threat attention bias predicts treatment outcome in anxious children receiving cognitive-behavioural therapy.
A bias to selectively direct attention to threat stimuli is a cognitive characteristic of anxiety disorders. Recent studies indicate that individual differences in pre-treatment threat attention bias predict treatment outcomes from cognitive-behavioural therapy (CBT) in anxious individuals. However, there have been inconsistent findings regarding whether attention bias towards threat predicts better or poorer treatment outcome. This longitudinal study examined treatment outcomes in 35 clinically-anxious children following a 10-week, group-based CBT program, as a function of whether children showed a pre-treatment attention bias towards or away from threat stimuli. The effect of CBT on attention bias was also assessed. Both groups showed significant improvement after receiving CBT. However, anxious children with a pre-treatment attention bias towards threat showed greater reductions not only in anxiety symptom severity, but also in the likelihood of meeting diagnostic criteria for anxiety disorders at post-treatment assessment, in comparison with anxious children who showed a pre-treatment attention bias away from threat. Children who had a pre-treatment bias away from threat showed a reduction in this bias over the course of CBT. Findings suggest that pre-existing differences in the direction of attention towards versus away from threat could have important implications for the treatment of anxious children.


Immunological aspects of Werner's syndrome: an analysis of 17 patients.
Several immunological assessments of the Werner syndrome are described, including detection of antibodies frequently found in autoimmune diseases and analysis of age-related changes of cell subpopulations. By using sensitive techniques such as fluorescence-activated cell sorting and solid-phase radioimmunoassay, anti-lymphocyte antibodies and anti-DNA antibodies were frequently detected in the sera from patients with the Werner syndrome. Anti-nuclear antibodies were also detected in a conventional manner; however, the titers of these antibodies were very low. Examination of cell populations revealed marked decreases in the T cell subsets reactive to both antilymphocytic antibodies and anti-brain associated T cell antigen antibodies.


Ultrastructure and synaptic targets of tectothalamic terminals in the cat lateral posterior nucleus.
The recent appreciation of the fact that the pulvinar and lateral posterior (LP) nuclei receive two distinct types of cortical input has sparked renewed interest in this region of the thalamus. A key question is whether the primary or "driving" inputs to the pulvinar/LP complex originate in cortical or subcortical areas. To begin to address this issue, we examined the synaptic targets of tectothalamic terminals within the LP nucleus. Tectothalamic terminals were labeled using the anterograde transport of biotinylated dextran amine (BDA) or Phaselous leucoagglutinin placed in the superior colliculus or using immunocytochemical staining for substance P, a neurotransmitter found to be used by the tectothalamic pathway (Hutsler and Chalupa [ 1991] J. Comp. Neurol. 312:379-390). Our results suggest that most tectothalamic terminals are large and occupy a proximal position on the dendritic arbor of LP relay cells. In the medial LP, tectothalamic terminals labeled by the transport of neuronal tracers or substance P immunocytochemistry can form tubular clusters that surround the proximal dendrites of relay cells. In a rostral and lateral subdivision of the lateral LP nucleus (LPl-2), tectothalamic terminals form more typical glomerular arrangements. When compared with existing physiological data, these results suggest that a unique integration of tectal and cortical inputs may contribute to the response properties of LP neurons.


Progress of preoperative and postoperative radiotherapy in gastric cancer.
Gastric carcinoma, a highly common malignant tumor, is treated mainly by surgery. Meanwhile, radiotherapy is attracting increased attention as a crucial locoregional therapy. However, the application of radiotherapy in gastric carcinoma is still limited and radiation standards remain debatable. The use of preoperative radiotherapy for treating gastroesophageal junction cancer has advanced. However, additional phase III clinical trials are needed to further verify the therapeutic value of preoperative radiotherapy for gastric cancer. Patients with D1 or D1 plus lymphadenectomy can benefit from postoperative radiotherapy obviously, and postoperative radiotherapy may be effective for patients with D2 lymphadenectomy with a high N stage. The target volume delineation of preoperative and postoperative radiotherapy should be based on clinical experience and the characteristics of lymphatic drainage. With the advancement of radiotherapy technology, preoperative and postoperative radiotherapy are becoming increasingly accepted as important auxiliary treatments for gastric cancer.


Andropause.
Age-related decline in serum testosterone level may present as a constellation of signs and symptoms of hypogonadism known as andropause. However, low testosterone can also be indicative of an underlying pituitary disorder; therefore, careful workup should be performed to distinguish between benign and pathologic conditions in individual men. This article presents an overview of the etiology and pathophysiology of hypogonadism, and offers guidelines for the evaluation of men who should have further testing. The pros and cons of various testosterone formulations are discussed.


Simplified immunonephelometric quantitation of apolipoprotein B in hyperlipoproteinemic serum.
An immunonephelometric procedure for determination of apolipoprotein B in human serum [Clin. Chem, 25, 221-226 (1979)] was modified by using the detergent octadecenylamine-polyoxyethylene, which may bind selectively to the phospholipid moiety of lipoproteins. Preincubation of hyperlipemic serum samples with lipases could then be omitted.


Single-cell transcriptomics and cell-specific proteomics reveals molecular signatures of sleep.
Every day, we sleep for a third of the day. Sleep is important for cognition, brain waste clearance, metabolism, and immune responses. The molecular mechanisms governing sleep are largely unknown. Here, we used a combination of single-cell RNA sequencing and cell-type-specific proteomics to interrogate the molecular underpinnings of sleep. Different cell types in three important brain regions for sleep (brainstem, cortex, and hypothalamus) exhibited diverse transcriptional responses to sleep need. Sleep restriction modulates astrocyte-neuron crosstalk and sleep need enhances expression of specific sets of transcription factors in different brain regions. In cortex, we also interrogated the proteome of two major cell types: astrocytes and neurons. Sleep deprivation differentially alters the expression of proteins in astrocytes and neurons. Similarly, phosphoproteomics revealed large shifts in cell-type-specific protein phosphorylation. Our results indicate that sleep need regulates transcriptional, translational, and post-translational responses in a cell-specific manner.


Myogenic lineage differentiated mesenchymal stem cells enhance recovery from dextran sulfate sodium-induced colitis in the rat.
Although mounting evidence implicates mesenchymal stem cells (MSCs) in intestinal tissue repair, uncertainty remains concerning the distribution, function, and fate of repopulating MSCs in recipient colonic tissues. Therefore, we investigated the role of transplanted MSCs in the repair phase of DSS colitis. LacZ-labeled rat MSCs were transplanted into rats with colitis induced by 4% DSS on day 2. Regular water replaced the DSS solution on day 6. Therapeutic effect was evaluated on day 9 by clinicopathologic and growth factor/cytokine expression profiles. We analyzed the Notch signaling pathway by Western blotting and characterized immunofluorescence of lacZ-labeled MSCs with confocal laser microscopy. In vivo differentiation of MSC was confirmed by transmission electron microscopy (TEM). Recovery of colitis was modestly but significantly promoted by MSC transplantation due to proceeding cell cycle and inhibiting apoptosis in the epithelia. Tgfa mRNA expression increased significantly, while Notch signaling was inhibited in the colonic tissues with MSC transplantation. β-Galactosidase-positive cells, which expressed α-SMA, desmin, and vimentin, were infrequently detected in the lamina propria stroma. DSS exposure in vitro proved to be the most potent inducer for α-SMA in MSCs where TEM demonstrated myogenic lineage differentiation. We found that MSCs transplantation modestly promoted the repair of DSS colitis. The donor-derived MSCs were likely reprogrammed to differentiate to myogenic lineage cells by cues from the micro milieu. Further characterization of these cells is warranted as a basis for applying cell-based therapy for inflammatory bowel disease.


Isolation of insulin degradation products from endosomes derived from intact rat liver.
Rats were injected with [125I]iodoinsulin labeled at either the A14 or B26 tyrosine, and the animals were killed and livers subcellularly fractionated to yield light (early or neutral) endosomes and heavy (late or acidic) endosomes. 125I-Labeled material was extracted from endosomes and analyzed by Sephadex G-50 filtration and high performance liquid chromatography (HPLC). Radiolabeled material in both types of endosomes is comprised of high molecular weight, insulin-sized, and low molecular weight components, with B chain-labeled small molecular weight material in two peaks, one corresponding to iodotyrosine and one to small peptides (Mr less than 1500). As compared with A chain label, however, less of the B chain material appears in the degradation components (both high and low molecular weight fractions) suggesting that a fragment of B chain containing the B26 residue is lost from the endosomes. Analysis on HPLC shows that significant amounts of the insulin-sized and high molecular weight material have proteolytic cleavage(s) in the B chain with an intact A chain. The B chain-derived labeled peptides elute from HPLC identically with products generated by insulin protease. These results therefore show substantial insulin degradation occurring in light endosomes prior to endosomal acidification and to receptor dissociation, suggesting receptor-bound insulin is a substrate for insulin protease.


A survey of present levels of radiocesium in Swedish pulp mill liquors and the implications for wood radiocesium transfer factors.
Assessments of the impact from radioactive fallout on the pulp industry and the accumulation processes for radionuclides in such industries, using wood and water as primary raw materials, have been conducted in previous studies. Hitherto, there is a general lack of data on present levels of radioactivity in pulp mill liquors apart from the few mills studied by Nilsson (1992); Ravila and Holm (1992); Manjón et al. (1996); Krosshavn et al. (1997). A survey was therefore initiated to describe and compare the present activity levels in pulp mill liquors from various mill locations in Sweden. The 137Cs activity in pulp mill liquors were compared with the mill location and the deposition pattern of the Chernobyl fallout and nuclear weapons fallout. The large input of wood (approx. 2500 m3 day-1) to an average Nordic Kraft mill and the relatively long-term retention time for radiocesium in the Kraft mill recovery cycle enables representative sampling of substances directly related to the activity concentration in wood.


Computational image analysis of the baseplate-tail complex of O1 ElTor vibriophage M4.
We performed an in-depth computational image analysis of the baseplate-tail complex of the M4 vibriophage and identified seven major densities in its baseplate, which notably share structural similarities with baseplate modules of a number of other bacteriophages belonging to different species. Employing computational analysis, we explained the helical organization of the sheath protein, wrapping the tail tube. Based on the results obtained in this work along with the proteomics information published previously, we are able to decipher the plausible roles assigned to the different components of the M4 baseplate during infection of the host.


Transapical aortic valve implantation: experiences and survival analysis up to three years.
We determined our 30-day results after transapical aortic valve implantation (TA-AVI) according to Valve Academic Research Consortium criteria, analyzed midterm outcome, and summarize our institutional learning experience. From February 2008 to July 2011, 150 high-risk patients underwent TA-AVI. Endpoints of this retrospective analysis were safety as indicated by morbidity and 30-day mortality and midterm survival with a follow-up period up to 3.4 years (mean follow-up 14.1 months). In addition we analyzed our institutional learning curve by comparing the outcome of our first 50 patients (group 1) to the following 100 patients (group 2). Procedural success was 98% (147 patients). All-cause and cardiovascular cause 30-day mortality was 11.3% (n = 17) and 7.3% (n = 11), respectively. The cumulative survival rates were 78.7% at one year, 62.8% at two years, and 50.8% at three years. As compared to group 1, there was a significantly reduced incidence of relevant bleeding complications (0% vs. 14%[n = 7]; p < 0.001) and a reduced incidence of acute kidney injury (35%[n = 35] versus 56% (n = 28); p < 0.05) in group 2, resulting in a combined safety endpoint at 30 days of 22% in group 2 versus 40% in group 1 (p < 0.05). One-year mortality (group 2, n = 20 [20%] versus group 1, n = 10 [20%]; p = 1) and midterm survival (p = 0.998; Hazard ratio 1.001; 95% CI 0.5141 to 1.949) did not differ significantly. Although the incidence of technical complications decreased significantly over time, 30-day and midterm mortality were unaltered, most likely due to patients' comorbidities. The development of more accurate risk scores may improve future outcome.


Cerebellar activation during leg withdrawal reflex conditioning: an fMRI study.
The aim of the present study was to examine cerebellar areas related to conditioning of the nociceptive leg withdrawal reflex using event-related functional magnetic resonance imaging (fMRI). Because of the aversive nature of the unconditioned stimulus effects of accompanying fear conditioning were expected. In 20 healthy adult subjects leg withdrawal reflex conditioning was performed using a standard delay protocol during MR-scanning. Electromyographic recordings from the anterior tibial and biceps femoris muscles were used to quantify conditioned responses. Fear-related changes of heart rate were assessed. In the group of all subjects a significant increase of cerebellar activation was found in the anterior and posterior vermis. In the group of subjects (n=9) who showed conditioned leg withdrawal responses cerebellar activation was more pronounced in parts of the anterior vermis, which correspond to the known leg representation. In the group of subjects (n=11) who did not develop conditioned responses cerebellar activation was more pronounced in the posterolateral hemispheres. Changes of heart rate, however, did not significantly differ between groups. Results suggest that areas within the anterior vermis are involved in conditioning of the leg withdrawal response. The present results, however, do not allow to differentiate between motor performance, learning or timing-related processes. Areas in the posterior vermis and cerebellar hemispheres may be related to concomitant fear conditioning. Results of the present event-related fMRI study suggest involvement of the human cerebellum in conditioning of the nociceptive leg withdrawal response.


A useful endovascular technique for treating modular limb disconnection in a bifurcated stent-graft.
To report a technique for treating a type III endoleak owing to disconnection of an iliac graft limb from a modular aortic stent-graft. A hydrophilic guidewire is introduced via a homolateral femoral access and passed through the thrombosed iliac extension. Once the errant graft limb is recanalized, wire exchange for an extra stiff guidewire facilitates passage of an angioplasty balloon into the stent. Inflated under low pressure, the balloon is used to push the thrombosed limb retrograde into the aneurysm sac. With the aortoiliac axis restored, a new covered stent can be deployed. This approach is one of several that may be used to treat a type III endoleak arising from a disconnected and occluded stent-graft limb. It avoids the bleeding complications of thrombolysis and restores the aortoiliac axis so that another stent-graft may be implanted to re-exclude the aneurysm.


Endometrial cancer patients and compliance with genetic counseling: room for improvement.
Immunohistochemistry (IHC) for the presence or absence of DNA mismatch repair (MMR) proteins in tumor samples is a quick and relatively inexpensive method to screen for Lynch syndrome (LS). At our institution we have found that >20% of patients lack expression of MMR proteins and >10% would be expected to benefit from genetic counseling (GC). However, compliance with referral to GC is poor. Therefore, we set out to analyze the reasons for noncompliance, hypothesizing that it could be due to a perception of a low risk for developing other cancers. All patients with endometrial cancer between 2007 and 2009 were identified. Patients with absence of MMR protein expression who would be expected to benefit from GC were identified. These patients were sent a questionnaire assessing the utilization of GC services as well as their perception of LS and Lynch-associated cancers. Forty-seven patients out of 384 were identified who would have been expected to benefit from GC. Of the responders, 20 patients (77%) reported that they were referred by their physician for GC, of which 9 saw a genetic counselor, 8 had genetic testing, and 3 mutations were detected. The primary reason for not seeing a genetic counselor was no insurance coverage/cost of the visit followed by anxiety for the results. Nine patients stated that the patient/family did not want to know information regarding cancer risk. As compared to the general population 35% thought their risk of having LS was higher, 12% thought it was the same, 15% lower, and 38% was unsure. Forty-six percent of patients thought the risk of colon cancer was higher than background risk, and 26% thought it was the same. Most patients underestimate their risk of LS associated heritable cancer. Physicians should pay more attention to family history and IHC in order to refer patients appropriately. In addition, more verbal and written information may enable patients to accurately assess their cancer risk. This could further improve compliance with genetic counseling and detection of Lynch syndrome.


Effectiveness of a Bundled Payments for Care Improvement Program for Chronic Obstructive Pulmonary Disease.
The Medicare Bundled Payments for Care Improvement (BPCI) program reimburses 90-day care episodes post-hospitalization. COPD is a leading cause of early readmissions making it a target for value-based payment reform. Evaluate the financial impact of a COPD BPCI program. A single-site retrospective observational study evaluated the impact of an evidence-based transitions of care program on episode costs and readmission rates, comparing patients hospitalized for COPD exacerbations who received versus those who did not receive the intervention. Mean episode costs and readmissions. Between October 2015 and September 2018, 132 received and 161 did not receive the program, respectively. Mean episode costs were below target for six out of eleven quarters for the intervention group, as opposed to only one out of twelve quarters for the control group. Overall, there were non-significant mean savings of $2551 (95% CI: - $811 to $5795) in episode costs relative to target costs for the intervention group, though results varied by index admission diagnosis-related group (DRG); there were additional costs of $4184 per episode for the least-complicated cohort (DRG 192), but savings of $1897 and $1753 for the most complicated index admissions (DRGs 191 and 190, respectively). A significant mean decrease of 0.24 readmissions per episode was observed in 90-day readmission rates for intervention relative to control. Readmissions and hospital discharges to skilled nursing facilities were factors of higher costs (mean increases of $9098 and $17,095 per episode respectively). Our COPD BPCI program had a non-significant cost-saving effect, although sample size limited study power. The differential impact of the intervention by DRG suggests that targeting interventions to more clinically complex patients could increase the financial impact of the program. Further evaluations are needed to determine if our BPCI program decreased care variation and improved quality of care. This research was supported by NIH NIA grant #5T35AG029795-12.


Covalent organic framework membranes for efficient separation of monovalent cations.
Covalent organic frameworks (COF), with rigid, highly ordered and tunable structures, can actively manipulate the synergy of entropic selectivity and enthalpic selectivity, holding great potential as next-generation membrane materials for ion separations. Here, we demonstrated the efficient separation of monovalent cations by COF membrane. The channels of COF membrane are decorated with three different kinds of acid groups. A concept of confined cascade separation was proposed to elucidate the separation process. The channels of COF membrane comprised two kinds of domains, acid-domains and acid-free-domains. The acid-domains serve as confined stages, rendering high selectivity, while the acid-free-domains preserve the pristine channel size, rendering high permeation flux. A set of descriptors of stage properties were designed to elucidate their effect on selective ion transport behavior. The resulting COF membrane acquired high ion separation performances, with an actual selectivity of 4.2-4.7 for K+/Li+ binary mixtures and an ideal selectivity of ~13.7 for K+/Li+.


Experience in managing splenic hilar vascular injury during laparoscopic total gastrectomy.
Splenic hilar vascular injury may occur during laparoscopic radical total gastrectomy for splenic hilar lymph node dissection and often causes massive hemorrhage, requiring conversion to laparotomy and splenectomy. Surgeons treating splenic hilar vascular injuries need a way to stop bleeding promptly and accurately. Herein, we report a case of splenic hilar vascular injury during laparoscopic total gastrectomy in which we successfully managed to stop the bleeding and preserve the spleen.


Preventing intellectual disability during pregnancy: what are the potentially high yield targets?
To identify risk factors among children with unknown cause intellectual disability (ID) and to estimate the population-attributable risk (PAR) associated with these factors. This was a retrospective cohort study of maternal and child pairs born between 2004 and 2010 in South Carolina, and information was obtained from Medicaid billing records, birth certificates, and other administrative data. The data included 123,922 children and logistic generalized estimating equations (GEE) regression models were used to estimate the association of maternal risk factors and ID. We estimated models with and without birth weight as a covariate, since low birth weight is known to be a mediator of the association between some risk factors and ID in children. The prevalence of ID in the children was 3.85% and the associations between risk factors and ID were similar for female and male children. We found that the odds of having ID were increased if a child's mother had a diagnosis of major depression; for male children, the odds ratio (OR) was 1.34 (95% confidence interval [CI] 1.14-1.59, PAR 2.17%); and for females, the OR was 1.59 (95% CI 1.30-1.95, PAR 4.70%). The odds of having ID were also increased for children of women with bipolar disorder (males - OR 1.95, 95% CI 1.53-2.48, PAR 2.85%; females - OR 1.63, 95% CI 1.20-2.22, PAR 2.05%). Major maternal depression and bipolar disorder were each significantly associated with the odds of ID in children, each accounting for approximately 2-5% of the PAR, when controlling for covariates.


Interindividual variability in P450-dependent generation of neoantigens in halothane hepatitis.
Halothane hepatitis occurs because susceptible patients mount immune responses to trifluoroacetylated protein antigens, formed following cytochrome P450-mediated bioactivation of halothane to trifluoroacetyl chloride. In the present study, an in vitro approach has been used to investigate the cytochrome P450 isozyme(s) which catalyze neoantigen formation and to explore the protective role of non-protein thiols (cysteine and reduced glutathione). Significant levels of trifluoroacetyl protein antigens were generated when human liver microsomes, and also microsomes from livers of rats pre-treated with isoniazid, phenobarbital or beta-naphtoflavone, were incubated with halothane plus a nicotinamide adenine dinucleotidephosphate (NADPH) generating system. Immunoblotting studies revealed that the major trifluoroacetyl antigens expressed in vitro exhibited molecular masses of 50-55 kDa and included 60 and 80 kDa neoantigens recognized by antibodies from patients with halothane hepatitis. Much lower concentrations of halothane were required to produce maximal antigen generation in isoniazid-induced rat microsomes, as compared with phenobarbital or isosafrole-induced microsomes (0.5 vs 12.5 microl/ml). In isoniazid-induced microsomes, antigen generation was inhibited > 90% by the nucleophiles cysteine and glutathione and by the CYP2E1-selective inhibitors diallylsulfide and p-nitrophenol, but was unaffected by inhibitors of other P450 isozymes (furafylline, sulfaphenazole or triacetyloleandomycin). Neoantigen formation in six human liver microsomal preparations was inhibited in the presence of diallylsulfide, but not by furafylline, sulfaphenazole or triacetyloleandomycin, and exhibited marked variability which correlated with CYP2E1 levels. These results suggest that the balance between metabolic bioactivation by CYP2E1 and detoxication of reactive metabolites by cellular nucleophiles could be an important metabolic risk factor in halothane hepatitis.


How to measure the treatment response in progressive multiple sclerosis: Current perspectives and limitations in clinical settings'.
New treatment options are available for active progressive multiple sclerosis (MS), including primary and secondary progressive forms. Several pieces of evidence have recently suggested a "window of beneficial treatment opportunities," principally in the early stages of progression. However, for progressive MS, which is characterised by an inevitable tendency to get worse, it is crucial to redefine the "response to treatment" beyond the concept of "no evidence of disease activity" (NEDA-3), which was initially conceived to evaluate disease outcomes in relapsing-remitting form, albeit it is currently applied to all MS cases in clinical practice. This review examines the current perspectives and limitations in assessing the effectiveness of DMTs and disease outcomes in progressive MS, the current criteria applied in defining the response to DMTs, and the strengths and limitations of clinical scales and tools for evaluating MS evolution and patient perception. Additionally, the impact of age and comorbidities on the assessment of MS outcomes was examined.


Enzymatic degradation of azo dyes methylene blue and congo red with peroxidase purified from cauliflower using affinity chromatography technique: Kinetic study, optimization and metal binding activity.
The effective results of the enzymatic decolorization of industrial azo dyes found in wastewater, which cause serious health and environmental problems, with peroxidases have recently increased the interest in these enzyme sources. Redox-mediated decolorization of Methylene Blue and Congo Red azo dyes with cauliflower (Brassica oleracea var. botrytis L.) peroxidase (CPOD) purified in one step using 4-amino 3-bromo 2-methyl benzohydrazide molecule was investigated for the first time. The inhibition effect of this molecule, which is used as a ligand in affinity chromatography, on the CPOD enzyme was investigated. The Ki and IC50 values for this enzyme were calculated as 0.113 ± 0.012 mM and 0.196 ± 0.011 mM, respectively. With the affinity gel obtained by binding to the Sepharose-4B-l-tyrosine matrix of this molecule, which shows a reversible inhibition effect, the purification values of CPOD enzyme were determined as 562-fold with a specific activity of 50,250 U mg-1. The purity of the enzyme was checked by the SDS-PAGE technique and its molecular weight was determined. A single band at 44 kDa was observed for the CPOD enzyme. In dye decolorization studies, the effects of dye, enzyme, and hydrogen peroxide concentrations as well as time, pH, and temperature were investigated. The profiles of the optimum conditions for both dyes were similar, and the percentages of decolorization of Methylene Blue and Congo Red under these conditions were 89% and 83%, respectively, at the end of the 40 min reaction time. Again, when examining the effect of metal ions on enzyme activity, it was found that there was no significant negative change in CPOD.


Pattern of SMARCB1 (INI1) and SMARCA4 (BRG1) in poorly differentiated endometrioid adenocarcinoma of the uterus: analysis of a series with emphasis on a novel SMARCA4-deficient dedifferentiated rhabdoid variant.
The role of the switch/sucrose nonfermenting chromatin remodeling complex in the initiation and progression of cancer is emerging. In the female genital tract, only ovarian small cell carcinoma, hypercalcemic type harbors recurrent inactivating SMARCA4 mutations. Otherwise, only rare case reports documented SMARCB1 involvement in endometrial cancer. We analyzed 24 grade 3 uterine endometrioid adenocarcinomas and 2 undifferentiated carcinomas for immunohistochemical expression of SMARCB1 and SMARCA4. All tumors showed high-grade nuclear features with a predominance of solid growth pattern. All cases showed intact nuclear SMARCB1 expression in all tumor cells. However, 1 case of a 78-year-old woman showed complete loss of SMARCA4 in 90% of the tumor with retained expression in 10% of the tumor. The SMARCA4-intact component was a moderate-to-poorly differentiated endometrioid adenocarcinoma. The SMARCA4-deficient dominating component showed solid growth of highly anaplastic undifferentiated large cells with prominent rhabdoid features. None of the 25 SMARCA4-intact cases showed rhabdoid cell morphology. To our knowledge, this is the first systematic study of SMARCB1 and SMARCA4 expression in endometrioid adenocarcinoma of uterus and the first description of a novel SMARCA4-deficient variant of dedifferentiated/undifferentiated endometrial carcinoma. The presence of a differentiated SMARCA4-intact endometrioid component points to a novel pathway of dedifferentiation in endometrioid adenocarcinoma as a consequence of a "second hit." This case further underlines the close link between the "rhabdoid phenotype" and the SWI/SNF pathway.


Vascular alterations impede fragile tolerance to pregnancy in type 1 diabetes.
To determine the impact of autoimmunity in the absence of glycemic alterations on pregnancy in type 1 diabetes (T1D). Because nonobese diabetic (NOD) mice experience autoimmunity before the onset of hyperglycemia, we studied pregnancy outcomes in prediabetic NOD mice using flow cytometry and enzyme-linked immunosorbent assays. Once we determined that adverse events in pregnancy occurred in euglycemic mice, we performed an exploratory study using electronic health records to better understand pregnancy complications in humans with T1D and normal hemoglobin A1c levels. University Medical Center. Nonobese diabetic mice and electronic health records from Vanderbilt University Medical Center. Nonobese diabetic mice were administered 200 μg of an anti-interleukin 6 (IL-6) antibody every other day starting on day 5 of gestation. Changes in the number of abnormal and reabsorbed pups in NOD mice and odds of vascular complications in pregnancy in T1D in relation to A1c. Prediabetic NOD mice had increased adverse pregnancy outcomes compared with nonautoimmune mice; blockade of IL-6, which was secreted by endothelial cells, decreased the number of reabsorbed and abnormal fetuses. Similarly, vascular complications were increased in pregnant patients with T1D across all A1c values. The vascular secretion of IL-6 drives adverse pregnancy outcomes in prediabetic NOD mice. Pregnant patients with T1D have increased vascular complications even with normal hemoglobin A1cs, indicating a potential effect of autoimmunity on the placental vasculature.


Active regulation of the epidermal calcium profile.
A distinct calcium profile is strongly implicated in regulating the multi-layered structure of the epidermis. However, the mechanisms that govern the regulation of this calcium profile are currently unclear. It clearly depends on the relatively impermeable barrier of the stratum corneum (passive regulation) but may also depend on calcium exchanges between keratinocytes and extracellular fluid (active regulation). Using a mathematical model that treats the viable sublayers of unwounded human and murine epidermis as porous media and assumes that their calcium profiles are passively regulated, we demonstrate that these profiles are also actively regulated. To obtain this result, we found that diffusion governs extracellular calcium motion in the viable epidermis and hence intracellular calcium is the main source of the epidermal calcium profile. Then, by comparison with experimental calcium profiles and combination with a hypothesised cell velocity distribution in the viable epidermis, we found that the net influx of calcium ions into keratinocytes from extracellular fluid may be constant and positive throughout the stratum basale and stratum spinosum, and that there is a net outflux of these ions in the stratum granulosum. Hence, the calcium exchange between keratinocytes and extracellular fluid differs distinctly between the stratum granulosum and the underlying sublayers, and these differences actively regulate the epidermal calcium profile. Our results also indicate that plasma membrane dysfunction may be an early event during keratinocyte disintegration in the stratum granulosum.


Basic principles of noninvasive blood pressure measurement in infants.
Correct measurement of blood pressure in infants is essential because blood pressure is an important indicator of cardiovascular status. This article provides a review of the fundamental principles of blood pressure measurement. Formulas to estimate mean arterial pressure are discussed along with an overview of pulse pressure. The advantages, challenges, and accuracy of the widely used oscillometric method of noninvasive blood pressure measurement are discussed. A step-by-step guide to noninvasive blood pressure measurement is outlined along with pragmatic strategies to ensure appropriate technique, thereby improving accuracy.