Valpha14 NK T cell-triggered IFN-gamma production by Gr-1+CD11b+ cells mediates early graft loss of syngeneic transplanted islets.
Pancreatic islet transplantation is a highly promising approach for the treatment of insulin-dependent diabetes mellitus. However, the procedure remains experimental for several reasons, including its low efficiency caused by the early graft loss of transplanted islets. We demonstrate that Gr-1+CD11b+ cells generated by transplantation and their IFN-gamma production triggered by Valpha14 NKT cells are an essential component and a major cause of early graft loss of pancreatic islet transplants. Gr-1+CD11b+ cells from Valpha14 NKT cell-deficient (Jalpha281-/-) mice failed to produce IFN-gamma, resulting in efficient islet graft acceptance. Early graft loss was successfully prevented through the repeated administration of alpha-galactosylceramide, a specific ligand for Valpha14 NKT cells, resulting in dramatically reduced IFN-gamma production by Gr-1+CD11b+ cells, as well as Valpha14 NKT cells. Our study elucidates, for the first time, the crucial role of Gr-1+CD11b+ cells and the IFN-gamma they produce in islet graft rejection and suggests a novel approach to improving transplantation efficiency through the modulation of Valpha14 NKT cell function.


[B symptoms in unexplained mediastinal lymphadenopathy : Case report of a 72-year-old male patient with VEXAS syndrome].
The case of a 72-year-old male patient who presented to our centre for rare diseases with recurrent fever, night sweats and weight loss with initially confirmed mediastinal lymphadenopathy is reported. Investigation of lymph node material was unrevealing. As an additional finding, the patient had a myelodysplastic syndrome. VEXAS syndrome (vacuoles, E1 enzyme, X‑linked, autoinflammatory, somatic) could be confirmed on the basis of a bone marrow biopsy and genetic testing.


[Properties of alpha-chymotrypsin enclosed into polycarbonate microcapsules. Estimation of the diffusion effect].
The conditions for the microencapsulation of alpha-chymotrypsin into semi-permeable polycarbonate membranes are selected. The diameter of the microcapsules is 100-150 microns. The microencapsulation can be carried out at any value of pH. The effect of the diffusion in the hydrolysis of some esters by microcapsulated alpha-chymotrypsin was estimated. The thermostability of the microencapsulated enzyme was studied.


The nauticAttiva project: A mobile phone-based tool for the citizen science plastic monitoring in the marine and coastal environment.
Plastic pollution is involving large coastal areas of the Mediterranean Sea. Innovative methods of plastic monitoring can be addressed through the citizen science approaches integrated with mobile phones. On the other hand, the availability of mobile phones is increasing among several users. Mobile phones can be integrated with a web mobile app, which allows to collect a lot of data for extended areas and in a short temporal range. In this study, the web service of iNaturalist was applied to implement a mobile phone-based tool to collect pictures of plastic items. At present, the web mobile app has been used to collect pictures of plastic debris in the Mediterranean Sea. Results were compared with the Mediterranean hydrodynamic regime, to highlight the pathways and densities of the plastic items. The proposed mobile phone-based tool represented a citizen science approach useful for the acquisition of plastic observations in the marine and coastal environment.


Polyphenols in Liubao Tea Can Prevent CCl₄-Induced Hepatic Damage in Mice through Its Antioxidant Capacities.
The present study investigated the preventive effect of polyphenols in Liubao tea (PLT) on carbon tetrachloride (CCl₄)-induced liver injury in mice. The mice were initially treated with PLT, followed by induction of liver injury using 10 mL/kg CCl₄. Then liver and serum indices, as well as the expression levels of related messenger RNAs (mRNAs) and proteins in liver tissues were measured. The results showed that PLT reduces the liver quality and indices of mice with liver injury. PLT also downregulates aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglycerides (TGs), and malondialdehyde (MDA), and upregulates superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the sera of mice with liver injury. PLT also reduces serum levels of interleukin-6 (IL-6), interleukin-12 (IL-12), tumor necrosis factor- α (TNF- α ), and interferon- γ (IFN- γ ) cytokines in mice with liver injury. Pathological morphological observation also shows that PLT reduces CCl₄-induced central venous differentiation of liver tissues and liver cell damage. Furthermore, qPCR and Western blot also confirm that PLT upregulates the mRNA and protein expressions of Gu/Zn-SOD, Mn-SOD, catalase (CAT), GSH-Px, and nuclear factor of κ -light polypeptide gene enhancer in B-cells inhibitor- α (I κ B- α ) in liver tissues, and downregulates the expression of cyclooxygenase 2 (COX-2) and nuclear factor κ -light-chain-enhancer of activated B cells (NF- κ B). Meanwhile, PLT also raised the phosphorylated (p)-NF- κ B p65 and cytochrome P450 reductase protein expression in liver injury mice. The components of PLT include gallic acid, catechin, caffeine, epicatechin (EC), epigallocatechin gallate (EGCG), gallocatechin gallate (GCG), and epicatechin gallate (ECG), which possibly have a wide range of biological activities. Thus, PLT imparts preventive effects against CCl₄-induced liver injury, which is similar to silymarin.


Risk factors for febrile convulsions.
Little is known about the relative importance of genes and early environment in the etiology of febrile convulsions. We performed a follow-up study using data from two nationwide registers in Denmark, 1980-1998. The study population comprised 10,224 younger siblings of children who had had febrile convulsions, and 21,218 younger siblings of children who had never been hospitalized with febrile convulsions. The study provides three main findings. First, if a previous child had had a febrile convulsion, the risk was lower for the next child if either parent changed partners. Compared with full-siblings, the hazard ratio (HR) of febrile convulsions was 0.6 for paternal half-siblings and 0.7 for maternal half-siblings. In contrast, if there was no history of febrile convulsions in the previous child, a change in partner was associated with a slight increase in risk (1.2 among paternal half-siblings and 1.3 among maternal half-siblings). Secondly, the risk of febrile convulsion was strongly associated with the number of hospitalizations for febrile convulsions experienced by the older siblings, with a doubling of risk among those whose older sibling had had three or more hospitalizations. Thirdly, the risk of febrile convulsions increased with decreasing gestational age, birth weight, and birth weight ratio regardless of family history. Our data suggest that the etiology of febrile convulsions depends on a genetic susceptibility that can be transmitted through both parents, and corroborates the hypothesis that multiple febrile convulsions may constitute a separate etiological entity.


Temporal variations of soil NO and NO2 fluxes in two typical subtropical forests receiving contrasting rates of N deposition.
Soils have been widely acknowledged as important natural sources of nitric oxide (NO) and meanwhile sinks of nitric dioxide (NO2). High nitrogen deposition across South China could potentially result in large NO emissions from subtropical forests soils there. In this study, the dynamic chamber method was applied to monitor NO and NO2 fluxes at two subtropical forest sites in South China, namely "Qianyanzhou" (QYZ) and "Tieshanping" (TSP). Chronically higher N deposition occurred at TSP than that at QYZ. Besides soil water filled pore spaces (WFPS) and temperature, ambient NO concentration could also possibly be important in regulating temporal NO emissions, especially in the winter. For both sites, the optimum soil temperature was above 25 °C, while the optimum WFPS for NO release at QYZ was higher (65-70%) than that at TSP (<23%). Moreover, heavy rainfall could trigger NO emission pulses from moist soils at QYZ, while rainfall-induced NO pulses were only observed after a long drying period at TSP. Distinctly different contents of mineral nitrogen and soil moisture conditions between the two sites might induce the divergent preference of WFPS and responses to rainfall. The cumulative soil emission of NO reached 0.41 ± 0.01 and 0.76 ± 0.01 kg N ha-1 yr-1 at QYZ and TSP, contributing to 2.5% and 1.4% of the annual throughfall N input, respectively. At both sites, NO2 were mainly deposited to soils, accounting for 2% and 21% of soil-emitted NO at QYZ and TSP, respectively. The observed annual NO emissions at these two sites were larger than the median values observed for tropical and temperate forests and unfertilized croplands. Higher N deposition could induce larger NO emission potential, while soil temperature and pH might also be important in regulating regional soil NO emissions as N-loss from subtropical forests.


Should we consider the sex when estimating bone age from hand bone biometrics?
Recently, it was proposed to estimate age from the biometric information of hand bones. We observed that these estimations became less precise as children get older, especially from the age of 13-15 years. This study aimed to evaluate the influence of considering sex for age estimation based on hand bones biometrics. The study sample consisted of metacarpals and proximal phalanges measurements collected on 1003 medical images performed at Nancy and Marseille Hospitals of individuals aged under 21 years. This sample was divided into two subgroups delineated by the age of 13, as it is a relevant legal threshold for most European countries. First, the influence of sex on the hand bones biometrics and on the estimated age was evaluated. Then, based on these results, new sex-specific age estimation formulas were constructed from linear models and their precision was assessed using residual analysis, in comparison with previous global formulas. An influence of sex was only highlighted from the age of 13 and for the total study sample. Thus, new sex-specific age estimation formulas were built for the [1-21] global sample and the [13-21] subsample. Even though the differences with the previous formulas were minor, age was more accurately estimated when sex was considered. Considering sex in age estimation is relevant when relying on hand bone biometrics. A new tool was proposed to select the most appropriate age estimation formula, based on the discriminant analysis result and the a priori knowledge of the sex.


Effect of food restriction on energy budget in warm-acclimated striped hamsters.
The capacity of small mammals to sustain periods of food shortage largely depends on the adaptive regulation of energy budget in response to the decrease in food supply. In addition to food availability, ambient temperature (Ta) is an important factor affecting the rates of both energy intake and expenditure. To examine the effect of Ta on energy strategy and the capacity to sustain food shortage, striped hamsters were exposed to a warm condition (30°C) and were then restricted to 70% of ad libitum food intake. Body mass, energy intake and expenditure and physiological markers indicative of thermogenesis were measured. Warm exposure had no effect on body mass and digestibility, but decreased energy intake, basal metabolic rate and maximum nonshivering thermogenesis. The mitochondria protein content, cytochrome c oxidase activity and uncoupling protein 1 level of brown adipose tissue were significantly lower in hamsters at 30°C than at 21°C. Food restriction induced a significant decrease in body mass, but the decreased body mass was attenuated at 30°C relative to 21°C. This suggests that striped hamsters could not compensate for the limited food supply by decreasing daily energy expenditure at 21°C, whereas they could at 30°C. The significant reductions in the rates of metabolism and thermogenesis in warm-acclimated hamsters increase the capacity to cope with food shortage. Although, it remains uncertain whether this response represents some generalized evolutionary adaptation, the Ta-dependent adjustment in the capacity to survive food restriction may reflect that warm acclimation plays an important role in adaptive regulation of both physiology and behavior in response to the variations of food availability.


Abundance of badgers (Meles meles) in England and Wales.
The European badger (Meles meles) is of considerable interest in the UK as it is both a protected species and the main wildlife reservoir for bovine tuberculosis infection in cattle. While there have been three national badger surveys in the 1980s, 1990s and 2011-13, using the number of badger main setts as a proxy for the abundance of badger social groups, none has combined contemporary data on social group size at landscape and national scales. We estimated social group size by genotyping hair samples collected at 120 main setts across England and Wales and employing a capture-mark-recapture method based on genotypes. The estimated mean social group size in England and Wales was 6.74 (±0.63) badgers. There was considerable variation in badger social group size among Land Class Groups (LCGs), with a low of 2.67 in LCG3 and a high of 7.92 in LCG4. Combining these results with the recent Badger Sett Survey of England and Wales, we estimate there are approximately 485,000 badgers (95% confidence intervals 391,000-581,000) in England and Wales. Although direct comparison with previous estimates is not ideal owing to methodological differences, our results are consistent with a marked increase in the badger population of England and Wales since the 1980s.


Natural cotton fiber-supported liquid extraction for convenient protein-rich aqueous sample preparation: Determination of glucocorticoids in milk and plasma as a proof-of-concept study.
Protein-rich aqueous samples such as milk and plasma usually require complex sample preparation steps prior to instrumental analysis. This study proposed a novel cotton fiber-supported liquid extraction (CF-SLE) method for convenient sample preparation. Natural cotton fiber was directly loaded into a syringe tube to conveniently construct the extraction device. No filter frits were required due to the fibrous feature of the cotton fibers. The cost of the extraction device was less than 0.5 CNY, and the costly syringe tube could be easily reused to decrease the cost further. Extraction used a simple two-step protocol: protein-rich aqueous sample loading and elution. Emulsification and centrifugation steps involved in the classic liquid-liquid extraction were avoided. As a proof-of-concept study, the glucocorticoids in milk and plasma were extracted with satisfactory extraction recoveries. Coupled with liquid chromatography-tandem mass spectrometry, a sensitive quantification method was established with excellent linearity (R2 > 0.991) as well as good accuracy (85.7-117.3%) and precision (<14.3%). This system is simple, low-cost, reproducible, and easy to automate. Thus, the proposed CF-SLE method is promising for the routine sample preparation of protein-rich aqueous samples prior to instrumental analysis.


A need for careful evaluation of endotoxin contents in acellular pertussis-based combination vaccines.
Two batches each of diphtheria-tetanus-acellular pertussis vaccine (DTaP) and that combined with inactivated polio vaccine purchased from foreign markets were tested by mouse body weight decreasing (BWD) toxicity test and Limulus amaebocyte lysate (LAL) test. Three out of the four imported vaccine batches showed the levels of BWD toxicity even comparable to that of DT-whole cell pertussis vaccine. BWD toxicity test is based on endotoxin dose-dependent weight loss of mice and has been used for controlling endotoxin in DTaP. Although of the strong BWD toxicity of the imported vaccines, there was no marked difference in LAL test results between the imported vaccines and Japanese DTaP. However, one imported DTaP batch showed very strong interference with LAL activity of spiked lipopolysaccharide (LPS). The batch interfered not only with LAL activity but also with pyrogenicity and prostaglandin E(2) induction activity. However, the pyrogenicity of the spiked LPS could be recovered from the precipitated fraction of the batch by treating with phosphate buffer to suggest the possibility of recovering in vivo toxicity. As an adequate in vitro test method could not be identified for controlling the safety of the interfering batch, an appropriate in vivo test would be required for testing such vaccines.


Single balloon valvotomy for rheumatic mitral restenosis.
Four male patients aged 22 to 35 years (mean 29 +/- 5.6) with mitral restenosis following closed mitral valvotomy 6 to 12 years (mean 8.5 +/- 2.5) ago were subjected to percutaneous single balloon valvotomy. There was significant increase in the area of the mitral valve accompanied by a substantial reduction in the transmitral gradient (P less than 0.01). Mitral regurgitation of grade +1 developed in one patient. Balloon valvotomy may be safely and effectively performed in selected patients with mitral restenosis after surgical commissurotomy.


Linezolid and the risk of QT interval prolongation: A pharmacovigilance study of the Food and Drug Administration Adverse Event Reporting System.
Few studies have investigated linezolid (LZD)-associated cardiotoxicity. This study explored the potential association between LZD and QT interval prolongation. Adverse event reports of QT interval prolongation associated with LZD from the Food and Drug Administration Adverse Event Reporting System from January 2013 to December 2021 were analysed and the reporting odds ratio (ROR) with 95% confidence intervals were calculated. A total of 6738 adverse event reports of LZD as the primary and secondary suspected drug were obtained from the database, including 192 reports with electrocardiogram QT prolonged (QTp), and the ROR value was 26.1 (95% CI = 22.6-30.2). There were 8 reports of long QT syndrome, ROR 14.2 (95% CI = 7.1-28.5); 5 reports of torsade de pointes, ROR 3.2 (95% CI = 1.3-7.6); and 5 reports of ventricular tachycardia, ROR 1.9 (95% CI = 0.8-4.5). Subgroup analysis revealed that patients with tuberculosis treated with LZD had a higher reporting rate among all QTp reports, exhibiting an odds ratio of 330.0 (95% CI = 223.1-488.1). The odds ratios of QTp associated with LZD treatments in patients with and without tuberculosis were 4.2 (95% CI = 3.4-5.3) and 1.2 (95% CI = 0.8-1.6), respectively. The study showed an association between LZD and QT interval prolongation. In the report on patients with tuberculosis, the incidence of QTp was higher when treated with LZD.


1,3-Diethylurea-enhanced Mg-ATPase activity of skeletal muscle myosin with a converse effect on the sliding motility.
We investigate the effects of urea and its derivatives on the ATPase activity and on the in vitro motility of chicken skeletal muscle actomyosin. Mg-ATPase rate of myosin subfragment-1 (S1) is increased by 4-fold by 0.3M 1,3-diethylurea (DEU), but it is unaffected by urea, thiourea, and 1,3-dimethylurea at ≤1M concentration. Thus, we further examine the effects of DEU in comparison to those of urea as reference. In in vitro motility assay, we find that in the presence of 0.3M DEU, the sliding speeds of actin filaments driven by myosin and heavy meromyosin (HMM) are significantly decreased to 1/16 and 1/6.6, respectively, compared with the controls. However, the measurement of the actin-activated ATPase activity of HMM shows that the maximal rate, Vmax, is almost unchanged with DEU. Thus, the myosin-driven sliding motility of actin filaments is significantly impeded in the presence of 0.3M DEU, whereas the cyclic interaction of myosin with F-actin occurs during the ATP turnover, the rate of which is close to that without DEU. In contrast to DEU, 0.3M urea exhibits only modest effects on both actin-activated ATPase and sliding motility of actomyosin. Thus, DEU has the effect of uncoupling the sliding motility of actomyosin from its ATP turnover.


[The Fuenlabrada study: smoking habits in children and adolescents. Effect on various cardiovascular risk factors].
We studied 1,274 healthy children of both sexes from 10 until 18 years of age, in Fuenlabrada, Madrid. We knew the tobacco consumption by means of direct asking to the children, without the presence of their parents. We studied the smoking habit of their parents by means of inquiries. The percentage of smoker children was 30% (24% of them had smoker parents, and 6% did not). We studied the tobacco influence in several parameters of cardiovascular risk; it was found that C-HDL levels in the smoker children were 5 mg/dl lower than the non-smokers, and the ratio C-LDL/C-HDL in the smokers was 1,2 times greater than in the non-smokers.


Long-term ailments due to anal sphincter rupture caused by delivery--a hidden problem.
Questionnaires concerning ailments were sent postpartum (mean two years) to 62 women with anal sphincter ruptures (ASR), who were compared with a matched control population. The frequency of anal sphincter rupture at the hospital during delivery in the period, 1978-82, was 0.7% (n = 63). Primiparity, instrumental deliveries, abnormal presentation, large babies and oxytocin stimulation were all risk factors. Of 59 women answering the questionnaire 37 (63%) stated that they had had ailments three months postpartum, mainly with pain and involuntary passage of flatus but also with dyspareunia and occasional incontinence of faeces. Long-term symptoms were noted by 28 (48%) of the women, mainly with involuntary passage of flatus but also perineal pain, dyspareunia and occasional incontinence of faeces. Long-term symptoms occurred in 7 (88%) of women with ASR also involving the anal mucosa, but only in 21 (39%) of those with ASR only. Three of the patients subsequently underwent reconstructive surgery, and three complained of psychological problems.


Responding to a crying infant - you do not learn it overnight: a phenomenological study.
to examine the views and practices of first-time and experienced mothers in response to infant crying during the first 12 weeks post birth. longitudinal, qualitative study using an interpretive, phenomenological approach. postnatal hospital and home settings in Switzerland. maximum variation sampling of 15 new mothers of diverse parity and educational background who had given birth to a full-term healthy neonate. participant observations in the postnatal ward and two narrative interviews at participants' homes at 6-8 and 12-14 weeks post partum. Data analysis used interpretive approaches of case analysis, thematic analysis and exemplars. first-time mothers showed some soothing skills from the beginning, but fine-tuned their practices of handling the crying infant and managing their own reactions. With growing experience mothers acquired a differentiated understanding of the crying's reason and urgency and used more successful soothing techniques. At the same time they learned to assess and mitigate their own stress reactions by self-soothing and adopting realistic expectations of normal infant behaviour. Experienced mothers knew the infant's frequent crying would diminish after a while whereas first-time mothers coped without this positive expectation. with increasing child-care experience mothers' skills and attitudes towards crying changed, leading to a calmer and less escalating response to their crying infant. inexperienced mothers need information on neonatal crying behaviour and on parents' stress response. They should be taught how to recognise and respond to the new-born's signals, and how to cope with their own stress. Postnatal care should provide novice mothers to learn from experienced role models.


Proteome analysis and identification of symbiosis-related proteins from Medicago truncatula Gaertn. by two-dimensional electrophoresis and mass spectrometry.
Time-course analysis of root protein profiles was studied by two-dimensional gel electrophoresis and silver staining in the model plant Medicago truncatula, inoculated either with the arbuscular mycorrhizal fungus Glomus mosseae or with the nitrogen fixing bacterium Sinorhizobium meliloti. Protein modifications in relation to the development of both symbioses included down- and upregulations, as well as newly induced polypeptides. Matrix assisted laser desorption/ionization-time of flight-mass spectrometry after trypsin digestion clearly identified one polypeptide induced in nodulated roots as a M. truncatula leghemoglobin. Internal sequencing with a quadrupole time-of-flight mass spectrometer and database searches confirmed the induction of proteins previously described in root symbioses, and revealed the implication of other proteins. In nodulated roots, one polypeptide was identified as an elongation factor Tu from S. meliloti, while another one could not be assigned a function. In mycorrhizal roots, analyzed proteins also included a protein of unknown function, as well as a glutathione-S-transferase, a fucosidase, a myosin-like protein, a serine hydroxymethyltransferase and a cytochrome-c-oxidase. These results emphasize the usefulness of proteome analysis in identifying molecular events occurring in plant root symbioses.


Colon capsule endoscopy leading to gastrointestinal stromal tumor (GIST) diagnosis after colonoscopy failure.
Gastrointestinal stromal tumors are a subtype of mesenchymal tumors. In recent years a significant progress was made in their diagnosis and treatment which led to significant improvement of their prognosis. Endoscopy remains one of the main diagnostic methods. In the rare instance of colonoscopy failure, different approaches are available: different endoscope, computed tomography colonography, capsule endoscopy, use of an enteroscope. We present the case of a 75-year-old Caucasian man admitted for abdominal pain, diarrhea and weight loss. Two colonoscopy attempts failed in a different center and a decision to use colon capsule endoscopy was made. This exam revealed a submucosal mass located in the sigmoid colon. Surgery was performed and a local invading gastrointestinal stromal tumor was removed. This is the first image of a colonic gastrointestinal stromal tumor seen on capsule endoscopy. Colon capsule is a useful diagnostic tool in selected patients after colonoscopy failure or contraindication.


Upregulation of interferon-induced genes in infants with virus-associated acute bronchiolitis.
To determine whether there is an airway IFN response in infants with acute bronchiolitis and to establish whether the rate of such a response is related to the severity of illness, the expression of some IFN-induced genes was measured in nasopharyngeal washes from 39 infants with acute bronchiolitis. The results indicate that in infants with a virus-associated acute bronchiolitis there is a strong activation of IFN system and that the severity of illness is inversely related to the level of expression of IFN-induced genes. This suggests that the IFN response plays an important role in determining virus-associated respiratory disease in early life.


Differences between the in vitro combinations of secretory component (SC) and immunoglobulin polymers (Ig) by enumeration of SC epitopes.
The enumeration of the SC epitopes has been established on 125I-labelled free and combined SC, by binding to anti-SC coated beads, then by addition of 3H-labelled anti-SC Fab' fragments of various specificities. The number of moles of Fab' fragments found on the beads increases in relation to the introduced amount. The extrapolation to an infinite concentration of added Fab' fragments gives the maximal theoretical accessible number of SC epitopes. The number of hidden epitopes (cryptotopes) is established by subtracting the total number found on sIgA, IgA-SC and IgM-SC from those found for free SC. These values are confirmed with Fab' fragments specific for the inaccessible determinant of SC. There are 4 cryptotopes in the case of sIgA, 3 for IgA1-SC, 2 for dimer IgA-SC and only 1 for IgM-SC (polyclonal or monoclonal). Thus the in vitro combinations of SC with polyclonal IgA dimers are different from the in vitro combinations with polyclonal or monoclonal IgM. The structural implications of these differences are discussed.


Repulsive double many-body expansion potential energy surface for the reactions N(4S)+H2<-->NH(X3Sigma-)+H from accurate ab initio calculations.
A single-sheeted DMBE potential energy surface is reported for the reactions N(4S)+H2<-->NH(X3Sigma-)+H based on a fit to accurate multireference configuration interaction energies. These have been calculated using the aug-cc-pVQZ basis set of Dunning and the full valence complete active space wave function as reference, being semi-empirically corrected by scaling the two-body and three-body dynamical correlation energies. The topographical features of the novel global potential energy surface are examined in detail, including a conical intersection involving the two first 4A'' potential energy surfaces which has been transformed into an avoided crossing in the present single-sheeted representation.


Invasion fitness, inclusive fitness, and reproductive numbers in heterogeneous populations.
How should fitness be measured to determine which phenotype or "strategy" is uninvadable when evolution occurs in a group-structured population subject to local demographic and environmental heterogeneity? Several fitness measures, such as basic reproductive number, lifetime dispersal success of a local lineage, or inclusive fitness have been proposed to address this question, but the relationships between them and their generality remains unclear. Here, we ascertain uninvadability (all mutant strategies always go extinct) in terms of the asymptotic per capita number of mutant copies produced by a mutant lineage arising as a single copy in a resident population ("invasion fitness"). We show that from invasion fitness uninvadability is equivalently characterized by at least three conceptually distinct fitness measures: (i) lineage fitness, giving the average individual fitness of a randomly sampled mutant lineage member; (ii) inclusive fitness, giving a reproductive value weighted average of the direct fitness costs and relatedness weighted indirect fitness benefits accruing to a randomly sampled mutant lineage member; and (iii) basic reproductive number (and variations thereof) giving lifetime success of a lineage in a single group, and which is an invasion fitness proxy. Our analysis connects approaches that have been deemed different, generalizes the exact version of inclusive fitness to class-structured populations, and provides a biological interpretation of natural selection on a mutant allele under arbitrary strength of selection.


Stress development during emergency remote teaching in higher education.
Higher education includes e-learning in addition to on-site learning. Still, the shift to Emergency Remote Teaching (ERT) as reaction to the Covid-19 pandemic in the summer semester 2020, presented a challenging situation for students. Cross-sectional studies pointed towards higher stress levels of students. However, only a few studies addressed the development of students' stress across several dimensions (joy, worry, tension, demands) within one semester. The current study analyzed trajectories of stress in ERT in relation to age, gender, digital readiness, and experience of loneliness, based on a sample of N = 2795 German students. Latent Growth Curve Models (LGCM) revealed a significant increase in demands, tension and worries and a decrease in joy during the summer term 2020. The development of tension and demands was influenced by age, gender, digital readiness, and loneliness. The decrease in joy and increase in worries could be primarily attributed to digital readiness and loneliness.


Age-associated reduction of the count and functional activity of stromal precursor cells can be caused by both true reduction (exhaustion) of cell pool and regulatory effects of the organism.
The study was carried out on CBA mice using the method of heterotopic transplantation. A fragment of the femoral bone marrow (1/2) or spleen (1/5 of the organ) was transplanted under the renal capsule of a recipient. The following donor-recipient cross-transplantation variants were studied: young-young (Y-Y), young-old (Y-O), old-old (O-O), and old-young (O-Y). Cell suspensions were prepared from 2-month transplants inoculated in monolayer cultures and the cloning efficiency (ECF-F) of stromal precursor cells (CFC-F) was evaluated. The bone marrow transplant ECF-F and the count of CFC-F in the O-O group were 8-fold lower than in the Y-Y group. In the O-Y group, ECF-F was 3-fold higher than in the O-O group, but by 2.5 times lower than in the Y-Y group. ECF-F in Y-O group was 2-fold lower than in Y-Y group. The ECF-F and CFC-F count in spleen transplants in the O-O group were 4- and 6-fold lower, respectively, than in Y-Y group. However, in O-Y group ECF-F was 7-fold higher than in O-O group and higher than even in Y-Y group. The weight of induced ectopic bone tissue after transplantation of the osteoinductor (fragments of the allogenic urinary bladder mucosa) was 2-fold lower in the O-O vs. Y-Y group. However, comparison of the ectopic bone tissue weights in different experimental groups showed that osteoinductor activity of the bladder epithelium did not decrease, but increased 3-fold with age (O-Y:Y-Y). A 5-fold reduction of this proportion in groups where the osteoinductor was transplanted from old donors to old and young recipients (O-Y:O-O) could be attributed to age-specific reduction of the count of inducible osteogenic precursor cells (IOPC). The data in general suggest that age-specific reduction of the stromal precursor count and functional activity could be caused by the true reduction (exhaustion) of cell pool (bone marrow CFC-F; presumably, IOPC) and by the regulatory effects of the organism (bone marrow and splenic CFC-F, IOPC). These data seem to be significant for understanding of the role of osteogenic stromal precursor cells in the development of age-associated bone tissue defects, for example, senile osteoporosis.


The correlation between specific protein synthesis and tumoricidal function in murine peritoneal macrophages.
Macrophages from the lipopolysaccharide (LPS)-responsive C3H/HeN mouse strain and the closely related LPS-nonresponsive C3H/HeJ strain were compared for tumoricidal activation and protein synthetic changes following in vivo and in vitro stimulation, utilizing two-dimensional polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins. Peritoneal macrophages elicited from C3H/HeN mice with heat-killed Propionibacterium acnes exhibited tumoricidal activity in a 16-hr cytolytic assay and expressed cytoplasmic levels of a 23.5-kDa protein during 48 hr of culture. The inability to detect persistent expression of p23.5 in P. acnes-stimulated C3H/HeJ macrophages correlated with the cytolytic impotence of those cells in the 16-hr chromium release assay. C3H/HeN macrophage populations lacking tumoricidal capacity could be rendered lytic, as could P. acnes-elicited C3H/HeJ macrophages, following in vitro stimulation with bacterial lipopolysaccharide. Concomitant with the LPS-induced expression of new functional activity was the appearance of augmented levels of several macrophage-specific proteins, including p23.5. This effect was dependent upon the lipid A moiety of LPS as the effects of LPS could be blocked by inclusion of polymyxin B sulfate in the culture medium. However, neither tumoricidal function nor protein modulation could be readily induced in C3H/HeJ proteose peptone-elicited or resident macrophages. These results identify biochemical responses to stimuli which may be requisite to acquisition or execution of cytolytic activity.


Differences between individuals in high-molecular weight glycoproteins from mammary epithelia of several species.
Milk fat globules are secreted by envelopment in plasma membrane of the lactating cell. SDS-gel electrophoresis of proteins from this membrane has revealed differences between milk donors in two mucin-like glycoproteins. One of these glycoproteins resolves in 3% acrylamide stacking gel and the other in 4% running gel. The proteins vary in number of bands (one or two) and band mobilities. This polymorphism arises, at least in part, from expression of hypervariable genes. In this study, gel electrophoretic evidence of similar polymorphism in glycoproteins from cow, chimpanzee, horse and human milks is presented. In distinction to the other species, the cow expressed only one of these proteins which was detected in the running gel at Mr 180,000 to 200,000. The electrophoresis pattern for this protein from six cows was highly varied with respect to number (one or two) and position of bands. Peanut agglutinin, wheat germ agglutinin and concanavalin A all were bound specifically by bands of the bovine glycoprotein. Binding of concanavalin A distinguishes the bovine protein from the two human glycoproteins. Further studies of species differences should help shed light on the evolution of these unique glycoproteins and their possible functions in mother and young.


Effects of vitamin C on characteristics retaining of in vitro-cultured mouse adipose-derived stem cells.
Adipose-derived stem cells (ADSCs), a subset of mesenchymal stem cells, have promising potential for regenerative medicine applications. However, the efficient culture of mouse adipose-derived stem cells (mADSCs) is complicated or impracticable and many properties of mADSCs are still unknown. Here, we report that vitamin C (Vc) is available for the long-term culture of mADSCs in vitro. Compared with that cultured without Vc, mADSCs growing in Vc-added media proliferate faster. The occurrence of replicative senescence and transformation of Vc-treated mADSCs is also postponed. Vc also enhanced the secretory activity of collagen and adipogenic differentiation ability of mADSCs. Moreover, the expression of CD44, Sca-1, and CD105 is higher in Vc-treated mADSCs than nontreated ones. We also found that genes related to proliferation, secretion, and pluripotency are all promoted in Vc-treated mADSCs. However, the adipogenesis ability and expression of CD44, Sca-1, and CD105 decreased when passage increased from very low passages, in parallel to the downregulation of closed-related gene expression. Our results indicate that Vc is essential for the maintenance of original properties of mADSCs in vitro; additional insights into the function of Vc on mADSCs are provided. Furthermore, as the passage increased in six passages, the characteristics of mADSCs with Vc addition were also revealed.


Evaluation of automated glucose oxidase methods for serum glucose: comparison to hexokinase of a colorimetric and an electrometric method.
Two automated glucose oxidase methods have been evaluated with respect to accuracy, precision, recovery, linearity and various potential interferences. Trinder's method on an AutoAnalyzer II had a between-day coefficient of variation (C.V.) of 2.6% (mean 228 mg/dl), was linear to 500 mg/dl, and produced a mean recovery of 99.7%. Comparison of Trinder's method with a manual, blanked hexokinase method yielded the regression equation: TR=--1.95 + HEX (1.04); Spearman's rho correlation coefficient was: 0.974. The Beckman System I glucose method had a between-day C.V. of 1.6% (mean 198 mg/dl), was linear to 500 mg/dl, and recovered an average of 98.0% of added glucose. Comparison with the same hexokinase method yielded: SYI = 1.27 + HEX (1.02: Spearman's rho = 0.991. None of the possible interfering compounds tested caused significant deviation of results by either method within the range of concentrations encountered physiologically. Trinder's method on the AutoAnalyzer II and the System I method are accurate, precise methods and are highly recommended for routine use in the clinical laboratory.


The Geriatric Syndrome and Oral Health: Navigating Oral Disease Treatment Strategies in the Elderly.
Poor oral health in the geriatric population is being framed as a potentially new geriatric syndrome; an oral and maxillofacial geriatric syndrome. As such, the treatment of oral diseases will require a comprehensive approach that considers the multimorbidity of disease, and polypharmacy that is precipitated/exacerbated by oral and maxillofacial dysfunction. This is challenging because oral diseases are not 1 discrete systemic illness but a combination of many with common causes. This article presents a roadmap approach to evaluate symptoms and apply therapeutic strategies for 5 common oral and maxillofacial dysfunctions seen in the elderly.


Detection of Voltage-Gated Sodium Channel (VGSC) Mutation in Deltamethrin-Resistant Blattella germanica L. from Indonesia.
Background and Objective: German cockroach (Blattella germanica L.) is one of the most common residential pests in Indonesia. Controlling the population face obstacles due to insecticide resistance, especially to deltamethrin. This research investigated the resistance status and the possibility of a Voltage-Gated Sodium Channel (VGSC) knockdown resistance mutation (L1014F) in two field strains of German cockroaches collected from two cities in Indonesia (Bukittinggi, named RMKN-BKT and Bandung, named KRSA-BDG) with VCRU-WHO as the standard strain. Materials and Methods: This study started with a bioassay test to determine the lethal dose of 50% (LD50) for each strain and followed by a molecular test for mutation detection. Results: The results showed that the RMKN-BKT and KRSA-BDG strains were highly resistant to deltamethrin with RR50 values of 80,090 times and 73,272 times, respectively. Only the RMKN-BKT strain was shown to carry L1014F kdr mutation which lead to an amino acid replacement from leucine (TTG) to phenylalanine (TTC). Two silent mutations were also found in both field strains at codons 983 (TGC/cysteine→TGT/cysteine) and 984 (GGG/ glycine→GGA/glycine) which were suggested as polymorphism phenomena. The absence of the L1014F mutation in the Bandung strain does not exclude the possibility of the presence of the VGSC mutation at other points. Conclusion: It requires subsequent investigation in mutation detection at other points and the possible presence of other resistance mechanisms to get a precise solution in the population control. Bioinsecticides may stand as a breakthrough so that the strategy will no longer focus on insecticides.


Anti-alpha-fetoprotein imaging is useful for staging hepatoblastoma.
Liver transplantation (Tx) has become an alternative treatment of malignant childhood liver tumors, and the importance of careful pretransplantation evaluation has been emphasized. Anti-alpha-fetoprotein (AFP) imaging has been suggested for evaluation of adult patients with AFP-positive tumors. Antibody imaging utilizing Tc-99 m-labeled monoclonal anti-AFP Fab' fragments was used to demonstrate pathologic uptake in hepatoblastoma (HB). Radical operation or liver Tx was not possible after four cycles of chemotherapy in a child with HB because of a single extrahepatic metastasis. Chemotherapy was continued, and reevaluation with anti-AFP imaging demonstrated a pathologic uptake only in the liver. Subsequently, a right liver lobe resection was performed. Along with a new rise in serum AFP, repeated anti-AFP imaging revealed active liver tumor but no metastases. A liver Tx was performed, and the child is well with a normal serum AFP level 18 months after the operation. This is the first case of pediatric HB in which anti-AFP imaging has been successfully used for patient management.


The effects of a specific growth hormone antagonist on overnight insulin requirements and insulin sensitivity in young adults with Type 1 diabetes mellitus.
Growth hormone hypersecretion in Type 1 diabetes could exacerbate insulin resistance and contribute to declining glycaemic control. Our aim was to determine the effects of specific growth hormone blockade on insulin sensitivity and lipolysis in young adults with Type 1 diabetes. We studied the effects of two doses of a specific growth hormone antagonist (B2036-PEG; Somavert, Pharmacia Corporation, Milton Keynes, UK) on insulin sensitivity in seven young adults (17-22 yrs, 3M) with Type 1 diabetes. Subjects received 5 and 10 mg B2036-PEG, in random order for 3 weeks, with a 3-week washout. At baseline and following each treatment block, an overnight (03:00 to 08:00 h) insulin infusion for euglycaemia (5 mmol/l), followed by two-step hyperinsulinaemic euglycaemic clamp, using [6,6 2H2] glucose and 2H5 glycerol to measure glucose and glycerol turnover was performed. Compared to baseline, overnight insulin requirements decreased with both doses: (means+/-SEM) 0.34+/-0.02 mU/Kg/min vs 0.25+/-0.01 (5 mg) (p=0.04), and 0.24+/-0.01 (10 mg) (p=0.004). IGF-I (ng/ml) decreased following 10 mg [223.5+/-23.9 vs 154.6+/-28.1 (p=0.005], but not 5 mg. Mean overnight non esterified fatty acid concentrations (mmol/l) decreased with 10 mg [0.51+/-0.04 vs 0.38+/-0.04 (p=0.03)], as did beta-hydroxybutyrate (mmol/l); [0.31+/-0.04 vs 0.15+/-0.02 (p=0.004)]. Glycerol production rate, an index of lipolysis, was lower following 10 mg (p=0.04), but insulin sensitivity during the clamp did not change with either dose. Treatment with both doses of B2036-PEG reduced overnight insulin requirements. The 10 mg dose suppressed lipolysis and reduced IGF-I. Failure to show enhanced insulin sensitivity during the clamp with the 10 mg dose could reflect opposing actions of growth hormone and IGF-I.


Mechanistic Insights into Regulation of the ALC1 Remodeler by the Nucleosome Acidic Patch.
Upon DNA damage, the ALC1/CHD1L nucleosome remodeling enzyme (remodeler) is activated by binding to poly(ADP-ribose). How activated ALC1 recognizes the nucleosome, as well as how this recognition is coupled to remodeling, is unknown. Here, we show that remodeling by ALC1 requires a wild-type acidic patch on the entry side of the nucleosome. The cryo-electron microscopy structure of a nucleosome-ALC1 linker complex reveals a regulatory linker segment that binds to the acidic patch. Mutations within this interface alter the dynamics of ALC1 recruitment to DNA damage and impede the ATPase and remodeling activities of ALC1. Full activation requires acidic patch-linker segment interactions that tether the remodeler to the nucleosome and couple ATP hydrolysis to nucleosome mobilization. Upon DNA damage, such a requirement may be used to modulate ALC1 activity via changes in the nucleosome acidic patches.


Psychological impact of stroke: a recovering nurse's story.
The author, both a nurse and patient, suffered a massive stroke. Included are the internal and external struggles and how to cope. Derived are lessons learned for nurses, to more fully understand a person with a stroke.


Surfactant abnormalities and adult respiratory failure.
Surfactant abnormalities have been implicated in the development of the acute respiratory distress syndrome in adults. Experimental studies show that surfactant inhibition by protein-leak into the alveolar space is of major importance under these circumstances. Fibrin(ogen)-surfactant-interaction appears to contribute to disturbances of surfactant function with subsequent alveolar instability and ventilation-perfusion-mismatch. In a prospective study in severely injured patients, the surfactant in serially obtained bronchoalveolar lavage fluids was investigated. An early leakage of plasma proteins into the alveolar space was noted in those patients, who developed severe ARDS. Moreover, deterioration of surfactant function was markedly more pronounced in those patients than in trauma victims who developed only mild pulmonary dysfunction. In addition to the protein-leakage, a progressive decrease of the surfactant-specific dipalmitoyl-phosphatidylcholine was noted, significantly correlated with the deterioration of surfactant function and the severity of respiratory failure. In conclusion, experimental and clinical studies show surfactant abnormalities in the adult respiratory distress syndrome. Plasma protein-leakage and progressive alteration of alveolar type II surfactant secretion appear to be of major importance.


A clinical study of Yigu capsule in treating postmenopausal osteoporosis.
To observe the efficacy and safety of Yigu capsule (YGC), a Chinese herbal compound preparation, in treating postmenopausal osteoporosis (PMO) and to explore its possible mechanism. The clinical study was conducted in a prospective, randomized, double blinded method lasting for 6 months with placebo and positive control. Two hundred and ten PMO patients with confirmed diagnosis were assigned into the YGC group, the calciferol group and the placebo group. Besides being administered element calcium, they were treated with YGC, calciferol capsule and placebo capsule respectively. And such symptoms as newly found fracture and ostealgia, bone mineral density (BMD) of the 2nd-4th lumbar vertebrae (L(2-4)) and upper femur, blood and urinary indexes for bone metabolism, sex hormone level and adverse reaction that occurred in patients were observed. In the YGC group, the total effective rate was 95.50%, with no new occurrence of fractures, which was significantly better than that in the other two groups (P < 0.05). Moreover, in the YGC group, the increase rate of BMD was 9.83% in L(2-4), 4.09% in femoral neck, 4.60% in Wards triangle, 3.00% in greater trochanter, which was also better than that in the placebo group (P < 0.05, P < 0.01). As compared with the placebo group, levels in the YGC group of urinary oxyproline hydroxyproline/creatinine, urinary calcium/creatinine were significantly lower, serum and bone alkaline phosphatase, osteocalcin, estradiol and estradiol/testosterone were significantly higher, but no difference was shown in the comparison of testosterone level. In the observation period, no abnormality in blood or urine routine, liver or renal function was found. Only mild, transient gastro-intestinal response occurred in individual patients, but it did not affect the treatment. YGC could treat PMO effectively, as it could obviously increase the BMD of lumbar vertebrae and coxafemoral bone, elevate the alleviating rate of ostealgia and incessant motion time, yet causing no newly found compressive fracture of vertebrae, or and any related adverse reaction. YGC could not only promote the formation, but also inhibit the absorption of bone as well as increase the sex hormone level. Therefore, it is a pure Chinese herbal compound preparation worthy of further research and development.


Identification Accuracy of Safety-Relevant Environmental Sounds in Adult Cochlear Implant Users.
Examine cochlear implant (CI) users' ability to identify safety-relevant environmental sounds, imperative for safety, independence, and personal well-being. Twenty-one experienced adult CI users completed an Environmental Sound Identification (ESI) test consisting of 42 common environmental sounds, 28 of which were relevant to personal safety, along with 14 control sounds. Prior to sound identification, participants were shown sound names and asked to rate the familiarity and, separately, relevance to safety of each corresponding sound on a 1-5 scale. Overall ESI accuracy was 57% correct for the safety-relevant sounds and 55% correct for control sounds. Participants rated safety-relevant sounds as more important to safety and more familiar than the non-safety sounds. ESI accuracy significantly correlated with familiarity ratings. The present findings suggest mediocre ESI accuracy in postlingual adult CI users for safety-relevant and other environmental sounds. Deficits in the identification of these sounds may put CI listeners at increased risk of accidents or injuries and may require a specific rehabilitation program to improve CI outcomes. 4 Laryngoscope, 133:2388-2393, 2023.


Interleukin-12 inhibits the survival of human colon cancer stem cells in vitro and their tumor initiating capacity in mice.
Interleukin-12 (IL-12) is a potent immunomodulatory cytokine with unknown direct effect on the property of cancer stem cells (CSCs). In this study, we investigated the capacity of IL-12 to regulate the self-renewal and differentiation of human colon CSCs in vitro, as well as the effect of IL-12 on the growth of tumors initiated by CSCs in mice. After over-expression of IL-12 with lentiviral transfection, CSCs exhibited reduced invasiveness and tumorsphere formation in association with increased apoptosis in vitro. After injection into NOD/SCID mice, tumors initiated by CSCs transfected with IL-12 showed markedly reduced rate of growth. Mechanistic studies revealed that over-expression of IL-12 reduced the expression of IL-4 and STAT6 in CSCs. Thus, our study demonstrates a potentially beneficial role of IL-12 in directly limiting the malignant phenotype of CSCs.


Testing a mobile app for child abuse treatment: A mixed methods study.
This study provides a preliminary evaluation of the usability and acceptability of a mobile application (sexual assault care algorithm, SACA). An explanatory sequential mixed methods research was used. A quantitative survey was followed up by a qualitative study. A convenience sample of participants (n = 4) was recruited. The research was conducted on a one-on-one basis. In the quantitative phase, a random assignment technique was used to divide four participants into two groups of two participants each. Post-Study System Usability Questionnaire(PSSUQ) and Acceptability e-Scale were used to collect quantitative data. In the qualitative phase, interview, observation, and documentation were used to collect qualitative data. Data were analyzed both quantitative and qualitatively. The qualitative data were linked with the initial quantitative data to determine how the follow-up qualitative data helped explain the initial quantitative results. The quantitative results suggested that SACA has high usability (5.05 ± 1.83) and acceptability (3.81 ± 1.22). The qualitative results further indicate that the participants thought SACA was easy to use and useful, and most of them would recommend it to others. Areas of improvement include adding features that would calculate and validate the elapsed time since the sexual assault, adding explanations to some buttons, and providing training. Our findings highlight the value of using a mixed methods research design to conduct a usability and acceptability test. Nurses are more likely to adopt a new technology for their evidence-based practice when the technology is easy to use and useful and requires less time to find the right piece of guideline evidence. Individualized training needs to be designed based on users' characteristics.


Can supportive parenting protect against school delay amongst violence-exposed adolescents in South Africa?
Exposure to multiple forms of violence is common amongst adolescents from socioeconomically disadvantaged communities in South Africa. Adolescents' exposure to violence at home, in school and in their communities can lead to detrimental outcomes in education. In particular, adolescents who are more frequently exposed to multiple forms of violence are at risk of school delay. This paper investigates the potential for supportive parenting to protect against adolescents' school delay in this context. With this aim, this paper applies structural equation modelling to a sample of 503 adolescents exposed to multiple forms of violence from 40 socioeconomically disadvantaged communities. Adolescents' self-report data on child abuse in the family, school and community, and adolescents' perceptions of positive parenting, consistent discipline, good monitoring, parental involvement and social support were analyzed. Results showed that perceptions of more positive parenting and consistent discipline moderated the relationship between more frequent exposure to multiple forms of violence and school delay. Our findings suggest that supportive parenting has the potential to protect against school delay for poly-victimized adolescents in South Africa.


Ntau- and Npi-histidinoalanine: naturally occurring cross-linking amino acids in calcium-binding phosphoproteins.
Two isomeric amino acid cross-links, Ntau-histidinoalanine and Npi-histidinoalanine have been isolated from calcium-binding phosphoprotein particles derived from the extrapallial fluid of the estuarine clam Rangia cuneata. The cross-links were identified and compared by 13C and 1H NMR spectroscopy and mass spectroscopy. In the phosphoprotein particles, 6% of the amino acid residues are involved in cross-linkages. This is the first demonstration of the occurrence of the isomer Npi-histidinoalanine.


Exploring structure-selectivity relationships of biogenic amine GPCR antagonists using similarity searching and dynamic compound mapping.
We design and analyze compound selectivity sets of antagonists with differential selectivity against seven biogenic amine G-protein coupled receptors. The selectivity sets consist of a total of 267 antagonists and contain a spectrum of in part closely related molecular scaffolds. Each set represents a different selectivity profile. Using these com- pound sets, a systematic computational analysis of structure-selectivity relationships is carried out with different 2D similarity methods including fingerprints, recursive partitioning, clustering, and dynamic compound mapping. Screening calculations are performed in a background database containing nearly four million molecules. Fingerprint searching and compound mapping are found to enrich target-selective antagonists over family-selective ones. Dynamic compound mapping effectively discriminates database compounds from GPCR antagonists and consistently retains target-selective antagonists during the final dimension extension levels. Furthermore, the widely used MACCS key fingerprint displays a strong tendency to distinguish between target- and family-selective GPCR antagonists. Taken together, the results indicate that different types of 2D similarity methods are capable of distinguishing closely related molecules having different selectivity. The reported compound benchmark system is made freely available in order to enable selectivity-oriented analyses using other computational approaches.


Activation of brain tryptophan hydroxylase by a phosphorylating system.
A significant activation of tryptophan hydroxylase (TPH) was achieved by the addition of 1 mM ATP and 10 mM MgCl2 to 100,000g supernatant prepared from mouse midbrain. The activation produced an increase of enzyme activity by 50-70% above control. Cyclic AMP was not a necessary component in the activation as initially suspected; neither was the involvement of a cyclic AMP-dependent protein kinase detected. The enzyme activation by Mg+2-ATP was totally retained after dialysis, thus excluding the possibility of an allosteric effect. In contrast to the activation, TPH was found to be sensitive to inactivation by acid phosphatase, a dephosphorylating enzyme. These findings are consistent with the proposal that the enzyme activation may involve a phosphorylation process. Furthermore, the activation by Mg+2-ATP in the brain preparations was only minimally observed from neonatal mice and could be abolished from adult mice by administration of hydrocortisone. It appears that the phosphorylation process, as postulated here, may be under developmental and hormonal influences.


Automatic Detection and Association Analysis of Multiple Surface Defects on Shield Subway Tunnels.
The surface defects on a shield subway tunnel can significantly affect the serviceability of the tunnel structure and may compromise operation safety. To effectively detect multiple surface defects, this study uses a tunnel inspection trolley (TIT) based on the mobile laser scanning technique. By conducting an inspection of the shield tunnel on a metro line section, various surface defects are identified with the TIT, including water leakage defects, dislocation, spalling, cross-section deformation, etc. To explore the root causes of the surface defects, association rules between different defects are calculated using an improved Apriori algorithm. The results show that: (i) there are significant differences in different association rules for various surface defects on the shield tunnel; (ii) the average confidence of the association rule "dislocation & spalling → water leakage" is as high as 57.78%, indicating that most of the water leakage defects are caused by dislocation and spalling of the shield tunnel in the sections being inspected; (iii) the weakest rule appears at "water leakage → spalling", with an average confidence of 13%. The association analysis can be used for predicting the critical defects influencing structural reliability and operation safety, such as water leakage, and optimizing the construction and maintenance work for a shield subway tunnel.


Preclinical evaluation of adult stem cell engraftment and toxicity in the CNS of rhesus macaques.
Congenital neurodegenerative diseases exhibit progressive postnatal neurologic impairment leading to premature death and are intractable to systemic therapies such as bone marrow transplantation. We injected bone marrow-derived mesenchymal stem cells (MSCs) into the CNS of young adult rhesus macaques to evaluate their safety and feasibility as vectors for direct intervention of neurologic disorders. Levels of engrafted male, donor MSCs were quantified in the CNS of female transplant recipients by real-time PCR using an SRY gene-specific probe. Analysis of coronal brain slices encompassing one-third of the total brain volume revealed engraftment levels ranging from 0.026 x 10(-3) to 0.163 x 10(-3)% of the total DNA content of brain tissue. Fine-mapping revealed male DNA distributed within specific anatomic structures along the neuraxis where label-retaining MSCs were visualized in histological sections by immunohistochemistry. Double labeling of sections confirmed that engrafted donor cells lacked expression of the macrophage marker CD68, the astrocytes marker GFAP, and neuronal markers NeuN and MAP2. MSC engraftment had no adverse effects on animal health, behavior, postural and locomotor patterns, or upper limb motor performance evaluated over a 6-month period posttransplantation. Therefore, MSC-based therapies represent a safe alternative for clinical intervention of CNS disorders.


Relationship between pulmonary embolism, airway obstruction and hypersensitivity of the airways in dogs: influence of partial vagus nerve blockade.
Experiments in dogs showed an increase in deltaPoes/100 ml TV after embolization which returns to normal after blocking the vagus nerves, indicating an increase in airway resistance. A marked sensitization to ACh inhalation by embolism with both blood clots and glass beads was demonstrated. This hypersensitivity is vagus-dependent.


Temperature-dependent biosynthesis of glucose monomycolate and its recognition by CD1-restricted T cells.
Mycolic acids are long chain fatty acids that constitute the lipid-rich cell wall framework of mycobacteria. Upon infection, mycobacteria begin to synthesize glucose monomycolate (GMM), a glucosylated species of mycolic acids, by utilizing host-derived glucose as sugar source. Accordingly, GMM production serves as a good indicator for local invasion of mycobacteria, and its detection by the host immune system would favor efficient monitoring of mycobacterial infection. Here, we found that GMM was produced abundantly at 30 degrees C rather than at 37 degrees C and recognized by a GMM-specific, CD1-restricted T cell line that was isolated from mycobacteria-infected human skin. Since the common portal sites for mycobacterial infection include ventilating alveoli of the lung and the externally exposed skin that often render invading microbes survive at reduced temperature, sampling GMM by CD1 lipid antigen-presenting molecules may allow the host to detect mycobacterial infection at its early phases.


A variant of the Klippel-Trenaunay-Weber syndrome with temporal lobe astrocytoma.
An 8-year-old boy with a variant of the Klippel-Trenaunay-Weber syndrome (KTW syndrome) is described. The hemangiomatous tissue located to the right half of his trunk and extremities was hypotrophic. On the same side, on his face and gingivae the tissue appeared hypertrophic and dental abnormalities were present. Moreover, the patient suffered from psychomotor epilepsy caused by a right-sided temporal astrocytoma. The connection between the KTW syndrome and the neurocutaneous syndromes is discussed.


Characteristics of defective phage particles of Pectobacterium carotovorum ZM1.
It is shown for the first time that the expression products of defective prophages are typical of defective lysogenic systems of phytopathogenic Pectobacterium carotovorum. It is established that virus-like particles (LP) such as phage capsids are packing bacterial DNA which size is determined by pulse field gel electrophoresis separation. Based on data about capsid structures which are formed by the virulent mutant ZF40/421, there is made a suggestion about the forming mechanism of defective virions of P carotovorum.


Laser-induced etching of few-layer graphene synthesized by Rapid-Chemical Vapour Deposition on Cu thin films.
The outstanding electrical and mechanical properties of graphene make it very attractive for several applications, Nanoelectronics above all. However a reproducible and non destructive way to produce high quality, large-scale area, single layer graphene sheets is still lacking. Chemical Vapour Deposition of graphene on Cu catalytic thin films represents a promising method to reach this goal, because of the low temperatures (T < 950°C-1000°C) involved during the process and of the theoretically expected monolayer self-limiting growth. On the contrary such self-limiting growth is not commonly observed in experiments, thus making the development of techniques allowing for a better control of graphene growth highly desirable. Here we report about the local ablation effect, arising in Raman analysis, due to the heat transfer induced by the laser incident beam onto the graphene sample.


Predictors of Maintained Transitions Between Robustness and Prefrailty in Community-Dwelling Older Spaniards.
We aimed to explore predictors of sustained transitions (those that are maintained for an extra follow-up) between robustness and prefrailty in both directions. Longitudinal population-based cohort. Community-dwelling Spaniards 65 years or older from the Toledo Study of Healthy Ageing. The Fried's frailty phenotype was measured over 3 waves (2006-2009, 2011-2013, and 2014-2017). Multiple logistic regressions compared individuals following the pattern robust-prefrail-prefrail with those who remained robust across waves, and those following the pattern prefrail-robust-robust with those who remained prefrail, for sociodemographic, clinical, life-habits, dependency for activities of daily living, upper and lower extremities' strength variables. The Fried's items of those who remained prefrail and those who became robust were compared. Mean age was 72.3 years (95% CI: 71.8-72.8) and 57.9% (52.7%-63.0%) were women. After multivariate adjustment, predictors (apart from age) of the sustained transition robustness-prefrailty were as follows: number of drugs taken (odds ratio: 1.37; 95% CI: 1.14-1.65), not declaring the amount of alcohol consumed (8.32; 1.78-38.88), and grip strength (0.92 per kg; 0.86-0.99). Predictors of the sustained transition prefrailty-robustness were as follows: drinking alcohol (0.2; 0.05-0.83), uricemia (0.67; 0.49-0.93), number of chair stands in 30 seconds (1.14; 1.01-1.28), and grip strength (1.12; 1.05-1.2). Low grip strength was associated with a lower probability of regaining robustness. Prediction of sustained transitions between the first stages of frailty development can be achieved with a reduced number of variables and noting whether the Fried's item leading to a diagnosis of prefrailty is low grip strength. Our results suggest the need to intensify interventions on deprescription, quitting alcohol, and strengthening of upper and lower limbs.


Clinical developments in the treatment of relapsed or relapsed and refractory multiple myeloma: impact of panobinostat, the first-in-class histone deacetylase inhibitor.
Panobinostat is a new agent for the treatment of relapsed and refractory multiple myeloma (rrMM) as part of a combination regimen. This article presents an overview of the mechanism of action, pharmacokinetics, safety, efficacy, patient care strategies, and role of the agent in treating rrMM patients. Panobinostat belongs to the class of drugs known as histone deacetylase inhibitors, and has high activity against Class I, II, and IV nonhistone deacetylases and histone deacetylases. It represents the first of its class to receive approval for use in MM, and received priority review and orphan drug status in both US and Europe, when used in combination with bortezomib and dexamethasone in the treatment of rrMM. Approval of panobinostat was based on subgroup analysis of Phase III data obtained in the PANORAMA trial program for evaluation of the combination of panobinostat, bortezomib, and dexamethasone. Additional clinical trials have continued to explore optimal dosing regimens and novel combination regimens to further clarify the optimal role of panobinostat in the arsenal of drugs for rrMM. Panobinostat has shown a manageable safety profile characterized primarily by hematologic toxicities (thrombocytopenia, neutropenia, lymphopenia, and anemia), gastrointestinal toxicities, notably diarrhea and nausea, as well as fatigue/asthenia, electrolyte abnormalities, and less commonly cardiac toxicities. Panobinostat represents an important addition to the treatment armamentarium for patients with rrMM, and studies are underway evaluating its optimal dosing strategy and role in combination with other drugs used to treat this patient population.


The role of microRNAs in the development and progression of chemical-associated cancers.
Human exposure to certain natural and man-made chemical carcinogens is one of the major risk factors for cancer development. The effect of chemical carcinogens on genetic and epigenetic alterations and their significance in the development of cancer has been well-established. In contrast, the role of microRNAs (miRNAs) in the etiology of chemical-associated cancers remains relatively unexplored despite extensive reports on changes in miRNA expression upon carcinogen exposure. This review summarizes the current knowledge for the role of miRNAs as drivers of chemical-induced carcinogenesis by bridging the gap between carcinogen exposure and cancer development through functional studies. It also emphasizes the potential for miRNA changes as early indicators of the carcinogenic process, markers for carcinogen exposure, and identification of chemical carcinogenic hazards.


Phase-Transformation-Induced Extra Thermal Expansion Behavior of (SrxBa1-x)TiO3/Cu Composite.
The properties of metal matrix composites (MMCs) can be optimized effectively through adjusting the type or the volume fraction of reinforcement. Generally, the coefficient of thermal expansion (CTE) of MMCs can be reduced by increasing the volume fraction of the reinforcement with lower CTE than metal matrix. However, it is great challenge to fabricate low CTE MMCs with low reinforcement volume fraction because of the limitation of reinforcement CTEs. SrxBa1-xTiO3 (SBT) powder presents negative thermal expansion behavior during the phase transformation from tetragonal to cubic phase. Here, we demonstrate that the phase transformation of SBT can be utilized to reduce and design the thermal expansion properties of SBT particle-reinforced Cu (SBT/Cu) composite, and ultralow CTE can be obtained in SBT/Cu composite. The X-ray diffraction analysis on heating indicates that the temperature range of phase transformation is extended greatly, therefore, the low CTE can be achieved within wide temperature range. Landau-Devonshire theory study on the phase transformation behaviors of SBT particles in the composite indicates that thermal mismatch stress significantly affects the Curie temperature of SBT particles and the CTE of the composite. The results given in the present study provide a new approach to design the MMCs with low CTE.


Indirect evolutionary rescue: prey adapts, predator avoids extinction.
Recent studies have increasingly recognized evolutionary rescue (adaptive evolution that prevents extinction following environmental change) as an important process in evolutionary biology and conservation science. Researchers have concentrated on single species living in isolation, but populations in nature exist within communities of interacting species, so evolutionary rescue should also be investigated in a multispecies context. We argue that the persistence or extinction of a focal species can be determined solely by evolutionary change in an interacting species. We demonstrate that prey adaptive evolution can prevent predator extinction in two-species predator-prey models, and we derive the conditions under which this indirect evolutionary interaction is essential to prevent extinction following environmental change. A nonevolving predator can be rescued from extinction by adaptive evolution of its prey due to a trade-off for the prey between defense against predation and population growth rate. As prey typically have larger populations and shorter generations than their predators, prey evolution can be rapid and have profound effects on predator population dynamics. We suggest that this process, which we term 'indirect evolutionary rescue', has the potential to be critically important to the ecological and evolutionary responses of populations and communities to dramatic environmental change.


Evidence for stable transformation of Pseudomonas aeruginosa with a pUC-based plasmid.
Pseudomonas aeruginosa was transformed with pUC8:16, a pUC-based plasmid bearing the gene (vgb) encoding Vitreoscilla (bacterial) hemoglobin (VHb). Transformation was initially indicated by an increase in ampicillin resistance from 1500 to 2500 mg l(-1). Presence of the plasmid in P. aeruginosa was confirmed by amplification of a portion of vgb from and detection of VHb in the transformant but not the untransformed host. Southern blot analysis further indicated that pUC8:16 existed as an autonomous plasmid rather than integrated into the chromosome of the P. aeruginosa transformant.


Increased Hepatotoxicity Associated with Sequential Immune Checkpoint Inhibitor and Crizotinib Therapy in Patients with Non-Small Cell Lung Cancer.
Immune checkpoint inhibitors (ICIs) are standard therapies in advanced NSCLC. Although genotype-directed tyrosine kinase inhibitors represent the standard of care for subsets of oncogene-driven NSCLC, patients may receive ICIs during their disease course. The impact of sequential ICI and tyrosine kinase inhibitor therapy on the risk of hepatotoxicity has not been described. Patients with advanced ALK receptor tyrosine kinase (ALK)-driven, ROS1-driven, or MET proto-oncogene, receptor tyrosine kinase (MET)-driven NSCLC treated with crizotinib, with or without preceding ICI therapy, were identified. The cumulative incidences of crizotinib-associated grade 3 or higher increases in transaminase level (per the Common Terminology Criteria for Adverse Events, version 4.0) were compared. We identified 453 patients who had NSCLC with an oncogenic alteration in ALK receptor tyrosine kinase gene (ALK), ROS1, or MET proto-oncogene, receptor tyrosine kinase gene (MET) and were treated with crizotinib (11 with and 442 without prior ICI therapy). Among the 11 patients treated with an ICI followed by crizotinib, five (cumulative incidence 45.5% [95% confidence interval (CI): 14.9-72.2]) experienced development of a grade 3 or 4 increase in alanine transaminase level and four (cumulative incidence 36.4% [95% CI: 10.0-64.2]) experienced development of a grade 3 or 4 increase in aspartate transaminase level. In comparison, among the 442 patients who received crizotinib only, a grade 3 or 4 increase in alanine transaminase level occurred in 34 patients (cumulative incidence 8.1% [95% CI: 5.7-11.0, p < 0.0001]) and a grade 3 or 4 increase in aspartate transaminase level occurred in 14 (cumulative incidence 3.4% [95% CI: 1.9-5.5, p < 0.0001]). There were no grade 5 transaminitis events. All cases of hepatotoxicity after sequential ICI and crizotinib use were reversible and nonfatal, and no case met the Hy's law criteria. Sequential ICI and crizotinib treatment is associated with a significantly increased risk of hepatotoxicity. Careful consideration and monitoring for hepatotoxicity may be warranted in patients treated with crizotinib after ICI therapy.


Design and instrumentation of new devices for performing appendectomy at colonoscopy (with video).
There is increasing interest in natural orifice surgery (NOS). Because the lumen of the appendix is connected to the cecum, a minimally invasive method for removing the appendix by colonoscopy may be feasible. Our purpose was to design, develop, and test new devices for inverting and removing the appendix by colonoscopy. Prospective prototype development program. University-based study in 25 colons from adult human cadavers. Various prototypes were evaluated by inserting them into the appendiceal orifice to its luminal tip, with the intent to invert the appendix in a controlled fashion into the lumen of the cecum. The advantage of using a tubular structure as a counterforce to aid inversion of the appendix was evaluated. When inversion was incomplete, the growing tissue strain was relieved by endoluminal incision of the mesenteric side of the appendix. Closure methods with endoloops or ligating loops were studied. Appendiceal resection was completed by snare diathermy, leaving an inverted intraluminal stump. Ability to invert the appendix into the cecum. The mean appendix length and luminal diameter were 84 +/- 23 mm and 4.9 +/- 1.2 mm, respectively. It was possible to advance various types of inversion devices to the tip of the appendiceal lumen. Partial inversion of the appendix was successful in 22 of 25 tests. Mesenteric tissue tension, tissue volume, and device slippage were the main reasons for incomplete inversion. The complete inversion was achieved with a combination of vacuum, tip grip, counterforce at the appendix base, and eventually endoluminal incision. The inversion of the human appendix by colonoscopy seems feasible and may be an alternative approach to conventional appendectomy.


Invasive Candida kefyr infection presenting as pyelonephritis in an ICU hospitalized COVID-19 patient: Case report and review of the literature.
Candida kefyr (Kluyveromyces marxianus), an ascomycetous environmental yeast, occasionally isolated from dairy products, represents an uncommon but emerging pathogen in immunocompromised patients. Herein, we present a case of C. kefyr pyelonephritis in a 41-year-old, previously immunocompetent, patient who was hospitalized in an COVID-19 ICU. Pyelonephritis was associated with caliectasis and obstruction due to possible fungus ball formation. Predisposing factors included ICU stay, use of broad spectrum antibiotics and steroids, central venous catheterization, mechanical ventilation and urologic manipulation. Susceptibility testing revealed high MIC values to amphotericin B. Infection was effectively controlled by prolonged administration of fluconazole without further surgical intervention. COVID-19 complicated with invasive candidiasis is an increasingly observed clinical situation that warrants high suspicion index and careful evaluation of laboratory data.


Apoptotic death of human lympholeukemia HL-60 cells resultant from combined effect of cobalt octa-4,5-carboxyphthalocyanine propylenglycol ether and ascorbate.
Cobalt octa-4,5-carboxyphthalocyanine propylenglycol ether proposed for antitumor therapy potentiates the cytotoxic effect of ascorbate on HL-60 human leukemia cells. Combination of these substances caused the formation of H2O2 in the medium and initiated apoptotic death of cells. Catalase abolished the cytotoxic effect of this combination. The results indicate that binary catalytic system of this combination can be regarded as a potential antitumor agent.


Prevalence of Human Hydatidosis Based on Hospital Records in Hamadan West of Iran from 2006 to 2013.
The reservoir and intermediate hosts of Echinococcus granulsus and human hydatidosis are more prevalent in the Alborz and Zagros Mountains slop due to rich pastures for sheep raising. Many cases of hydatid cysts operate in local hospitals annually. The present study aimed to review the epidemiologic characteristics of the hydatidosis patients. In this descriptive study, the medical files of 182 hydatidosis patients from main public and private hospitals in Hamadan Province, western Iran from 2006 to 2013 were reviewed. The data collected from eight general hospitals including demographic data, clinical and diagnostic measures, surgical approaches, and outcome were entered into the prepared checklist and analyzed by descriptive statistics. All patients were diagnosed and operated giving an average of 26 cases per year, or 1.5 cases per 100000 inhabitants. The site of cysts was as follows: liver 70.9%, lung 24.7%, and in both these organs 2.2%. The ratio of male and female patients was approximately 1:1. Mean age of patients was 44.5±21.5 yr at range of 3 to 91 yr. The majority the patients were illiterate (32.2%) and resided in the rural areas (61.7%), and their occupations were housewives (36.8%). Almost 90% of patients diagnosed by imaging methods and 8% had history of surgery for cyst. Hydatidosis is a major health problem still in this region and more extensive epidemiological investigations of CE is necessary to better determine the prevalence, economic impact and risk factors for the disease control.


Association between Molecular Genetic Markers of DNA Repair and Cell Cycle Control Genes and Response to Platinum-Based Chemotherapy in Ovarian Cancer Patients.
We analyzed associations of polymorphic markers of DNA repair genes (XRCC1, ERCC2), cell cycle control genes (TP53, MDM2, and CDKN1A), methylation of promoter region, and mutation 5382insC of BRCA1 gene in ovarian cancer with effectiveness of platinumbased chemotherapy assessed by the median of progression-free survival time for markers of DNA repair genes and by relapse risk for all studied markers. An increase in the median of progression-free survival time for carriers of the Gln allele (р=0.025) and Gln/Gln genotype (р=0.022) of the Gln399Arg XRCC1 was observed during the 19-months period after chemotherapy. In carriers of C/C genotype of 5382insC mutation of BRCA1 gene (n=6), no relapses were observed (р=0.035), while 17 of 49 patients without this mutation developed relapses. Of 14 patients with BRCA1 gene function inactivation due to promoter methylation or the presence of the C/C genotype of 5382insC, one relapse was observed (p=0.033). Multivariate analysis revealed an association of markers of the XRCC1, TP53, MDM2 genes, BRCA1 gene inactivation, and type of surgery with the risk of relapse during the follow-up period up to 19 months after the end of chemotherapy (р≤0.0007).


Structural anisotropy of cyanido-bridged {CoII9WV6} single-molecule magnets induced by bidentate ligands: towards the rational enhancement of an energy barrier.
Pentadecanuclear {CII9[W(V)(CN)8]6} clusters were combined with bidentate 2,2'-bipyridine N,N'-dioxide (2,2'-bpdo) ligands resulting in two distinct molecules, {Co9W6(2,2'-bpdo)7} (cluster A) and {Co9W6(2,2'-bpdo)6} (cluster B), capped by seven and six 2,2'-bpdo ligands, respectively. They crystallize within a single {Co9W6(2,2'-bpdo)7}·{Co9W6(2,2'-bpdo)6}·solvent (1) supramolecular network, and reveal single-molecule magnet behaviour with an enhanced energy barrier, a ΔE/kB of 30.0(8) K, which was tentatively ascribed to seven-capped axially deformed cluster A.


Atom-efficient assembly of 1,5-oxygen-bridged medium-sized carbocycles by sequential combination of a Ru-catalyzed alkyne-alkene coupling and a Prins-type cyclization.
Ruthenium-catalyzed coupling of readily accessible 1-trimethylsilyl-1-alkyn-3-ols to allyl ethyl ether followed by in situ ketalization and Lewis-acid induced cyclization affords 1,5-oxygen-bridged eight- and nine-membered carbocycles, products which are difficult to construct with previously published strategies.


Schizencephaly.
Schizencephaly means clefts in the brain and refers to that condition of dysmorphogenesis in which there are bilateral, usually symmetrical, clefts in the region of the central sulcus. This condition is readily demonstrated by computed tomography and is associated with severe mental retardation. We add three case studies to those previously recorded in the literature.


Differences in reading performance of patients with Drusen maculopathy and subretinal fibrosis after CNV.
To evaluate differences in reading performance and contrast sensitivity on patients with drusen maculopathy and subretinal fibrosis after CNV (choroidal neovascularisation). 136 patients (60 with drusen (D), 76 with fibrosis (F)) were studied. Patients were classified according to type of maculopathy and best-corrected visual acuity into groups D1 and F1 (LogMAR 0.2-0.4), groups D2 and F2 (LogMAR>0.4-0.7), and group F3 (LogMAR>0.7-1.3). Reading acuity (in LogRAD) and speed were examined with the Radner Reading charts and compared to the reading speed measured with the long paragraphs of the Zuercher Reading Test. Contrast sensitivity was measured with Pelli-Robson charts. The patients' distance visual acuity was comparable between the drusen and fibrotic CNV groups (D1 versus F1, D2 versus F2). The reading acuity of the corresponding groups D1 and F1 was also comparable, but F2 patients showed a statistically lower reading acuity than D2 patients (P=0.03). All reading speed measurements of the groups F1 and F2 were significantly worse than those of the corresponding groups D1 and D2 (P-values: 0.0005-0.02). The correlation of reading speed between the Radner and Zuercher texts was very high (r=0.73-0.94). The contrast sensitivity was significantly lower in all groups compared with group D1 (P<0.001), but comparable for groups F1, F2, and F3. Despite comparable results in distance visual acuity, patients with subretinal fibrosis after CNV had a greater reduction in reading ability than the patients with drusen. The distance visual acuity measurements alone, therefore, underestimate the real-life conditions and impact of AMD.


Phase behavior of the microemulsions and the stability of the chloramphenicol in the microemulsion-based ocular drug delivery system.
Microemulsion systems composed of Span20/80+Tween20/80+n-butanol+H2O+isopropyl palmitate (IPP)/isopropyl myristate (IPM) were investigated as model systems of drug carriers for eye drops. Effects of chloramphenicol, normal saline, sodium hyaluronate and various oils on the phase behavior were studied. The phase transition was investigated by the electrical conductivity measurements. The electrical conductivity of the microemulsion was affected by the encapsulation of the drug into the system, and the addition of normal saline and sodium hyaluronate. The chloramphenicol is used to treat the diseases such as trachoma and keratitis. However, this drug in the common eye drops hydrolyzes easily. The main product of the hydrolysis is glycols. Here, the chloramphenicol was trapped into the oil-in-water (o/w) microemulsions and its stability was investigated by the high performance liquid chromatography (HPLC) assays in the accelerated experiments of 3 months. Its location in the microemulsion formulations was determined by means of 1H NMR spectroscopy. The results of HPLC revealed that the contents of the glycols in the microemulsion formulations were much lower than that in the commercial eye drops at the end of the accelerated experiments. It implied that the stability of the chloramphenicol in the microemulsion formulations was increased remarkably. The NMR experiments confirmed that the chloramphenicol molecules should be trapped into the hydrophilic shells of the microemulsion drops, which was composed of many oxyethylene groups. The nitro-groups of the chloramphenicol molecules were near the alpha2-CH2 of the surfactant molecules and the benzene rings of the chloramphenicol molecules were near the oxyethylene groups of the surfactant molecules. It was this reason that enabled the chloramphenicol molecules in the microemulsions to be screened from the bulk water and its stability to be increased remarkably.


Detection of mutations in JAK2 exons 12-15 by Sanger sequencing.
The Janus kinase (JAK)2 p.V617F gain-of-function mutation is a hallmark of BCR-ABL1-negative myeloproliferative neoplasms (MPNs). This study analyzed JAK2 mutations in 1811 patients tested between 2010 and 2013. Exons 12-15 of JAK2 were sequenced in 1706 samples, and patients harboring mutations were clinically evaluated. Of 271 patients (16%) with JAK2 mutations, 148 (54.6%) were female and 123 (45.4%) were male; 103 (38%) were local and 168 (62%) were referred; and 13 (5%) had additional genetic abnormalities. The median patient age was 54 years, and there was only one pediatric patient. In agreement with previous reports, 262 patients (96.7%) were positive for the JAK2 p.V617F mutation. Non-p.V617F JAK2 mutations were detected in the remaining nine (3.3%) patients: five (1.8%) had a p.G571S mutation, and one (0.3%) each had p.E543_D544del, p.Y570Y silent, p.R541_E543delinsK, and p.I540_N542delinsM mutations. Diagnosis of 103 (38%) in-house cases revealed a predominance of MPN patients (87 cases, or 84.4%). JAK2 p.V617F was the most prevalent mutation detected among patients in this study. Non-p.V617F JAK2 mutations were identified in exons 12 and 13 corresponding to recently reported mutations, except for the novel p.I540_N542delinsM.


The relationship between immunization and circulating antigen-specific plaque-forming cells.
The relationship between the appearance of cells producing antibody to tetanus toxoid (TT) in the circulation and the serum titers of anti-TT IgG following booster immunization has been studied. It was found that cells producing anti-TT antibody can be detected in the circulation in a hemolytic plaque assay using sheep red blood cells (SRBC) coated with TT by the chromic chloride method. In symmetric inhibition studies using cells from TT or keyhole limpet hemocyanin (KLH)-immune donors, the homologous antigen inhibited 100% of the PFC with no cross-inhibition. Thus, the plaque-forming cells (PFC) detected in this assay are specific for the immunizing antigen. No evidence of polyclonal B-cell activation in response to TT was found, as shown by a failure to detect any PFC against unmodified or KLH or human serum albumin-treated SRBC. In addition, the increase in total Ig-secreting cells observed in a staphylococcal protein A reverse hemolytic plaque assay was always accounted for by the number of anti-TT antibody-producing cells observed. The peak number of anti-TT antibody-producing cells varied between donors, but the kinetics of their appearance was highly reproducible--none before Day 5, peak numbers between Days 6 and 8, and a sharp decline with only rare anti-TT Ig-secreting cells in the circulation by Day 15 postimmunization. Anti-TT antibody-producing cells appeared in the circulation prior to any detectable increase in serum anti-TT antibody titers, and following the disappearance of PFC from the circulation, there was no further increase in serum IgG anti-TT levels. These observations demonstrate a marked specificity of B-cell activation on boosting with a recall antigen, and a parallelism between the appearance of activated B cells in the circulation and of IgG anti-TT synthesis by the subject as a whole.


Hazy lesion in a ventricular tachycardia patient: stents or drugs?
Intravascular ultrasound (IVUS) is superior to coronary angiographyin guiding and optimizing complicated PCI. Here we report a successful treatment of a patient with ventricular tachycardia induced by treadmill test. Coronary angiography showed a hazy lesion in left anterior descending artery. IVUS revealed a 73% segment stenosis in the proximal segment of the LAD. Finally, a 3.5*21 mm drug eluting stent was implanted in the LAD and an excellent final angiographic result was achieved.


Macroscopic and molecular approaches of enrofloxacin retention in soils in presence of Cu(II).
The co-adsorption of copper and the fluoroquinolone antibiotic enrofloxacin (ENR) at the water-soil interface was studied by means of batch adsorption experiments, and extended X-ray absorption fine structure (EXAFS) spectroscopy. The system was investigated over a pH range between 6 and 10, at different contact times, ionic strengths, and ENR concentrations. Adsorption coefficient - Kd - was determined at relevant environmental concentrations and the value obtained in water at a ionic strength imposed by the soil and at soil natural pH was equal to 0.66Lg(-1). ENR adsorption onto the soil showed strong pH dependence illustrating the influence of the electrostatic interactions in the sorption processes. The simultaneous co-adsorption of ENR and Cu(II) on the soil was also investigated. The presence of Cu(II) strongly influenced the retention of the antibiotic, leading to an increase up to 35% of adsorbed ENR amount. The combined quantitative and spectroscopic results showed that Cu(II) and ENR directly interacted at the water-soil interface to form ternary surface complexes. Cu K-edge EXAFS data indicated a molecular structure where the carboxylate and carbonyl groups of ENR coordinate to Cu(II) to form a 6-membered chelate ring and where Cu(II) bridges between ENR and the soil surface sites. Cu(II) bonds bidentately to the surface in an inner-sphere mode. Thus, the spectroscopic data allowed us to propose the formation of ternary surface complexes with the molecular architecture soil-Cu(II)-ENR.


Probing cholesterol binding and translocation in P-glycoprotein.
P-glycoprotein (Pgp) is a biomedically important member of the ABC transporter superfamily that mediates multidrug resistance in various cancer types. Substrate binding and transport in Pgp are modulated by the presence of cholesterol in the membrane. Structural information on cholesterol binding sites and mechanistic details of its redistribution are, however, largely unknown. In this study, a set of 40 independent molecular dynamics (MD) simulations of Pgp embedded in cholesterol-rich lipid bilayers are reported, totaling 8 μs, enabling extensive sampling of cholesterol-protein interactions in Pgp. Clustering analyses of the ensemble of cholesterol molecules (∼5740) sampled around Pgp in these simulations reveal specific and asymmetric cholesterol-binding regions formed by the transmembrane (TM) helices TM1-6 and TM8. Notably, not all the putative cholesterol binding sites identified by MD can be predicted by the primary sequence based cholesterol-recognition amino acid consensus (CRAC) or inverted CRAC (CARC) motifs, an observation that we attribute to inadequacy of these motifs to account for binding sites formed by remote amino acids in the sequence that can still be spatially adjacent to each other. Binding of cholesterol to Pgp occurs more frequently through its rough β-face formed by the two protruding methyl groups, whereas the opposite smooth α-face prefers packing alongside the membrane lipids. One full and two partial cholesterol flipping events between the two leaflets of the bilayer mediated by the surface of Pgp are also captured in these simulations. All flipping events are observed in a region formed by helices TM1, TM2, and TM11, featuring two full and two partial CRAC/CARC motifs, with Tyr49 and Tyr126 identified as key residues interacting with cholesterol during this event. Our study is the first to report direct observation of unconventional cholesterol translocation on the surface of Pgp, providing a secondary transport model for the known flippase activity of ABC exporters of cholesterol. This article is part of a Special Issue entitled: Molecular biophysics of membranes and membrane proteins.