Co-existince of sickle cell disease and hemidiaphragm paralysis.
Sickle cell anemia is a disease caused by production of abnormal hemoglobin. Infection, acute splenic sequestration crisis, aplastic crises, acute chest syndrome, stroke, cholelithiasis, renal disease and pain are the major complications. Unilateral or bilateral diaphragm paralysis maybe seen following phrenic nerve injury and with a variety of motor-neuron diseases, myelopathies, neuropathies, and myopathies. Prominent right hemi-diaphragma elevation was observed on chest radiograph of a 14 years' old female patient with sickle cell disease. Her medical history yielded neither trauma nor intra-thoracic surgery. She didn't have either motor deficit or sensation disorder on any region of her body. Thorax CT yielded no lesion except the significantly elevated right diaphragm. Her cranial CT showed no lesion, too. Diagnosis of right hemidiaphragm paralysis was confirmed by positive Hitzenberg Sniff test on fluoroscopy. Although several pathophysiologic mechanisms are known to be involved and lead to central neurologic complications in sickle cell disease, involvement of peripheric nerves have not been reported. Here we present a 14 years' old female patient with sickle cell anemia and unilateral diaphragm paralysis, co-existence of which have not been reported so far.


Relationship between first-year blood donation, return rate for subsequent donation and demographic characteristics.
The aim of this study was to determine any relationships between return rates of first-time donors, number of donations in the first year and the donors' demographic characteristics. Data from 1,500 volunteer, first-time donors who donated blood at Shiraz Blood Transfusion Centre (Iran) were obtained from the donor database. The donors' demographic characteristics (sex, age, educational status) and donation histories were obtained for a 3-year period and the number of donations and interval between the first donation and the following donation within a 1-year period were recorded. We searched for correlations between return rate and demographic characteristics and the number of donations in the first, second and third years. The data were analysed with the chi-squared test, multiple logistic regression and Spearman's correlation tests. P values less than 0.05 were considered statistically significant. More than half of the first-time donors returned to donate again (n=776, 51.7%) during the 3 years after the first donation. The mean number of donations in the whole 3-year period was 0.69±0.49, while it was 1.33±0.62 (median 1) in the first year, 0.4±0.7 (median 0) in the second, and 0.3±0.65 (median 0) in the third year. The return rate was directly correlated with the number of donations in the first year (r=0.74, P<0.001). The return rate increased as the interval between the first and second donations decreased (P<0.05, OR=1.03). The return rate was higher for male donors and single donors (P<0.05), but was not related to age or educational level (P>0.05). In this study 51.7% of first-time donors returned to donate again, a high figure indicating the success of our recruitment strategy. Understanding the importance of the number of donations in the first year for donors' return rate and planning recruitment programs and more effective measures to encourage donors in the first year may help to increase return rates.


Do larvae from deep-sea hydrothermal vents disperse in surface waters?
Larval dispersal significantly contributes to the geographic distribution, population dynamics, and evolutionary processes of animals endemic to deep-sea hydrothermal vents. Little is known as to the extent that their larvae migrate vertically to shallower waters and experience stronger currents and richer food supplies. Here, we first provide evidence from early life-history traits and population genetics for the surface dispersal of a vent species. Planktotrophic larvae of a red blood limpet, Shinkailepas myojinensis (Gastropoda: Neritimorpha: Phenacolepadidae), were cultured to observe their swimming behavior and to evaluate the effects of temperature on survival and growth. In addition, the population structure was analyzed based on 1.2-kbp mitochondrial DNA sequences from 77 specimens that cover the geographic and bathymetric distributions of the species (northwest Pacific, 442-1,227 m in depth). Hatched larvae constantly swam upward at 16.6-44.2 mm/min depending on temperature. Vertical migration from hydrothermal vents to the surface, calculated to take ~4-43 d, is attainable given their lengthy survival time without feeding. Fed larvae best survived and grew at 25°C (followed by 20°C), which approximates the sea surface temperature in the geographic range of the species. Little or no growth was observed at the temperature of the vent habitat where adult limpets occur (≤15°C). Population genetic analyses showed no differentiation among localities that are <1,350 km apart. The larvae of S. myojinensis most likely migrate to the surface water, where high phytoplankton biomass and strong currents enable their growth and long distance dispersal over many months. Sea surface temperature may represent a critical factor in determining the geographic distribution of many vent endemic species with a planktotrophic early development, and in turn the faunal composition of individual vent sites and regions.


Regulation of the expression of proinflammatory cytokines induced by SARS-CoV-2.
An outbreak of a novel coronavirus was reported in Wuhan, China, in late 2019. It has spread rapidly through China and many other countries, causing a global pandemic. Since February 2020, over 28 countries/regions have reported confirmed cases. Individuals with the infection known as coronavirus disease-19 (COVID-19) have similar clinical features as severe acute respiratory syndrome first encountered 17 years ago, with fever, cough, and upper airway congestion, along with high production of proinflammatory cytokines (PICs), which form a cytokine storm. PICs induced by COVID-19 include interleukin (IL)-6, IL-17, and monocyte chemoattractant protein-1. The production of cytokines is regulated by activated nuclear factor-kB and involves downstream pathways such as Janus kinase/signal transducers and activators transcription. Protein expression is also regulated by post-translational modification of chromosomal markers. Lysine residues in the peptide tails stretching out from the core of histones bind the sequence upstream of the coding portion of genomic DNA. Covalent modification, particularly methylation, activates or represses gene transcription. PICs have been reported to be induced by histone modification and stimulate exudation of hyaluronic acid, which is implicated in the occurrence of COVID-19. These findings indicate the impact of the expression of PICs on the pathogenesis and therapeutic targeting of COVID-19.


The comprehensiveness of computer-assisted searches of the medical literature.
Bibliographic searches using MEDLINE, the National Library of Medicine computerized database, can usually be done in less time and with greater specificity than searches using Index Medicus. To use a computer-assisted bibliographic retrieval system to full advantage, it is necessary to understand the indexing system. The study reported here compares both the number and the relevance of references retrieved using various search terms for two clinical questions. Based on the outcomes of these searches, recommendations are made for clinicians who plan to use MEDLINE services.


Association of the Placement of a Perirectal Hydrogel Spacer With the Clinical Outcomes of Men Receiving Radiotherapy for Prostate Cancer: A Systematic Review and Meta-analysis.
Perirectal spacers are intended to lower the risk of rectal toxic effects associated with prostate radiotherapy. A quantitative synthesis of typical clinical results with specific perirectal spacers is limited. To evaluate the association between perirectal hydrogel spacer placement and clinical outcomes of men receiving radiotherapy for prostate cancer. A systematic search was performed of the Cochrane Central Register of Controlled Trials, MEDLINE, and Embase for articles published through September 2019. Studies comparing men who received a hydrogel spacer vs men who did not receive a spacer (controls) prior to prostate radiotherapy. Via random-effects meta-analysis, group comparisons were reported using the weighted mean difference for continuous measures and the risk ratio for binary measures. Procedural results, the percentage volume of rectum receiving at least 70 Gy radiation (v70), early (≤3 months) and late (>3 months) rectal toxic effects, and early and late changes in bowel-related quality of life on the Expanded Prostate Cancer Index Composite (minimal clinically important difference, 4 points). The review included 7 studies (1 randomized clinical trial and 6 cohort studies) involving 1011 men (486 who received a hydrogel spacer and 525 controls), with a median duration of patient follow-up of 26 months (range, 3-63 months). The success rate of hydrogel spacer placement was 97.0% (95% CI, 94.4%-98.8% [5 studies]), and the weighted mean perirectal separation distance was 11.2 mm (95% CI, 10.1-12.3 mm [5 studies]). Procedural complications were mild and transient, occurring in 0% to 10% of patients within the studies. The hydrogel spacer group received 66% less v70 rectal irradiation compared with controls (3.5% vs 10.4%; mean difference, -6.5%; 95% CI, -10.5% to -2.5%; P = .001 [6 studies]). The risk of grade 2 or higher rectal toxic effects was comparable between groups in early follow-up (4.5% in hydrogel spacer group vs 4.1% in control group; risk ratio, 0.82; 95% CI, 0.52-1.28; P = .38 [6 studies]) but was 77% lower in the hydrogel spacer group in late follow-up (1.5% vs 5.7%; risk ratio, 0.23; 95% CI, 0.06-0.99; P = .05 [4 studies]). Changes in bowel-related quality of life were comparable between groups in early follow-up (mean difference, 0.2; 95% CI, -3.1 to 3.4; P = .92 [2 studies]) but were greater in the hydrogel spacer group in late follow-up (mean difference, 5.4; 95% CI, 2.8-8.0; P < .001 [2 studies]). For men receiving prostate radiotherapy, injection of a hydrogel spacer was safe, provided prostate-rectum separation sufficient to reduce v70 rectal irradiation, and was associated with fewer rectal toxic effects and higher bowel-related quality of life in late follow-up.


Neoadjuvant Chemotherapy: A New Standard for Muscle Invasive Bladder Cancer?
Bladder cancer is the seventh most common cancer and the ninth most common cause of cancer deaths for men worldwide. Cystectomy with pelvic lymph nodes dissection is the standard local treatment of muscle invasive bladder cancer (MIBC) (T2-T4). In the last decade, the management of MIBC had become multidisciplinary involving peri-operative chemotherapy (neo- or adjuvant chemotherapy). Randomized trials and meta-analyses confirmed the survival benefit of neoadjuvant chemotherapy before local treatment (surgery and/or radiotherapy). Consequently, this sequence should be considered as standard treatment of choice, for patients with good performance status (0-1) and good renal function. The benefit of adjuvant chemotherapy is not clear for patients treated with primary surgery.


RFamide-related peptide-3 receptor gene expression in GnRH and kisspeptin neurons and GnRH-dependent mechanism of action.
RFamide-related peptide-3 (RFRP-3) is known to inhibit the activity of GnRH neurons. It is not yet clear whether its G protein-coupled receptors, GPR147 and GPR74, are present on GnRH neurons or on afferent inputs of the GnRH neuronal network or whether RFRP-3 can inhibit gonadotropin secretion independently of GnRH. We tested the following: 1) whether GnRH is essential for the effects of RFRP-3 on LH secretion; 2) whether RFRP-3 neurons project to GnRH and rostral periventricular kisspeptin neurons in mice, and 3) whether Gpr147 and Gpr74 are expressed by these neurons. Intravenous treatment with the GPR147 antagonist RF9 increased plasma LH concentration in castrated male rats but was unable to do so in the presence of the GnRH antagonist cetrorelix. Dual-label immunohistochemistry revealed that approximately 26% of GnRH neurons from male and diestrous female mice were apposed by RFRP-3 fibers, and 19% of kisspeptin neurons from proestrous female mice were apposed by RFRP-3 fibers. Using immunomagnetic purification of GnRH and kisspeptin cells, single-cell nested RT-PCR, and in situ hybridization, we showed that 33% of GnRH neurons and 9-16% of rostral periventricular kisspeptin neurons expressed Gpr147, whereas Gpr74 was not expressed in either population. These data reveal that RFRP-3 can act at two levels of the GnRH neuronal network (i.e. the GnRH neurons and the rostral periventricular kisspeptin neurons) to modulate reproduction but is unable to inhibit gonadotropin secretion independently of GnRH.


Assessment of long-term psychosocial outcomes in anti-NMDA receptor encephalitis.
The purpose of this study was to assess long-term psychosocial outcomes of anti-N-methyl-d-aspartate (NMDA) receptor encephalitis (anti-NMDARE). Adolescents and adults with self-reported anti-NMDARE were invited to complete an online survey distributed by relevant patient organizations. Demographic and clinical information was collected, including the diagnoses initially given for anti-NMDARE symptoms and posthospital care received. Patient-Reported Outcomes Measurement Information System (PROMIS) Psychosocial Impact Illness - Negative short form (Negative PSII) was administered to assess psychosocial outcome of anti-NMDARE. Associations between clinical factors and psychosocial outcomes were evaluated. Sixty-one individuals with anti-NMDARE age 15 years and above participated. Mean age was 33.7 years (standard deviation [SD]: 12.8), and participants were predominantly female (90.2%, n = 55). Mean T-score on PROMIS Negative PSII was 60.7, >1 SD higher (worse psychosocial function) than that of the provided normalized sample enriched for chronic illness (50, SD: 10). Initial misdiagnosis of anti-NMDARE symptoms was associated with decreased odds (odds ratio [OR]: 0.11, p < 0.05), and follow-up with a psychiatrist after hospitalization with increased odds (OR: 8.46, p < 0.05), of return to work/school after illness. Younger age of symptom onset and presence of ongoing neuropsychiatric issues were predictive of worse Negative PSII scores (p < 0.05). Individuals with anti-NMDARE demonstrate poor psychosocial outcomes, yet there are no current standards for long-term assessment or management of such symptoms in this population. These findings highlight the need for use of more comprehensive outcome measures that include assessment of psychosocial function and the importance of developing interventions that address this domain for individuals with anti-NMDARE.


Clinical features and in vivo confocal microscopy assessment in 12 patients with ocular cicatricial pemphigoid.
To describe the clinical features and microstructural characteristics assessed by in vivo confocal microscopy (IVCM) in patients with ocular cicatricial pemphigoid (OCP). A descriptive, uncontrolled case series study. Patients diagnosed with OCP were examined by clinical history, slit-lamp biomicroscopy features and IVCM images. The results of direct immunofluorescence (DIF) biopsies and indirect immunofluorescence (IIF) were also recorded. Local and systemic immunosuppressive therapy were administered and adjusted according to response. A total of 12 consecutive OCP patients (7 male, 5 female; mean age 60.42±10.39y) were recruited. All patients exhibited bilateral progressive conjunctival scarring and recurrent chronic conjunctivitis was the most frequent clinical pattern. The mean duration of symptoms prior to diagnosis of OCP was 2.95±2.85y (range: 5mo to 10y). The Foster classification varied from stage I to IV and 20 eyes (83%) were within or greater than Foster stage III on presentation. Two of the 12 patients (17%) demonstrated positive DIF; 3 of the 12 (25%) patients reported positive IIF. The mean duration of the follow-up period was 20.17±11.88mo (range: 6 to 48mo). IVCM showed variable degrees of abnormality in the conjuctiva-cornea and conjuctival scarring was detected in all the involved eyes. Corneal stromal cell activation and dendritic cell infiltration presented as ocular surface inflammation, ocular surface keratinization along with the destroyed Vogt palisades was noted in eyes with potential limbal stem cell deficiency. After treatment, remission of ocular surface inflammation was achieved in all the patients, 18 eyes (75%) remained stable, 6 eyes (25%) had recurrent conjunctivitis and cicatrization in 2 eyes (8%) was progressing. As an autoimmune disease, OCP manifests as variable degrees of clinical and laboratory abnormalities with both local and systemic immunosuppressive treatment playing important roles in disease therapy. IVCM can be as a valuable non-invasive technique to assess ocular surface changes in a cellular level with a potential value for providing diagnostic evidence and monitoring therapeutic effects during follow-up.


Epidemiology, Clinical Features, and Prescribing Patterns of Aortic Aneurysm in Asian Population From 2005 to 2011.
Aortic aneurysm (AA) is a leading cause of death in Asia and the world. The prevalence in Western countries is around 1.3% to 8%. However, it is still unclear about the incidence, prevalence, and mortality of AA in Asian population. The aim of this study is to investigate the epidemiology of AA for all subtypes in Taiwan, and describe the clinical features and prescribing patterns for AA population.A population-based study was conducted using information from National Health Insurance Research Database (NHIRD) in Taiwan. Patients who were diagnosed with AA and also received computed tomography (CT) were included in this study. Incidence, prevalence, and mortality were calculated in each year during 2005 to 2011. Prevalent comorbidities and prescribing patterns were both evaluated among study population.The average annual incidence of AA in Taiwan was 7.35 per 100,000 population, and the prevalence was 29.04 per 100,000 population. It showed an increased trend of incidence from 2005 to 2011, so as prevalence and mortality. The incidence was associated with age and sex difference. It was much higher in those older than 65 years, especially for male. Hypertension, coronary artery disease (CAD), and chronic obstructive pulmonary disease (COPD) were prevalent comorbidities. Eighty-eight percentages of patients were prescribed antihypertensive agents in acute phase, where 61.4% of calcium channel blocker (CCB) was the most one.Our study found that incidence of AA was lower in Taiwan than in other countries. Nevertheless, it showed an increased trend of AA disease for incidence, prevalence, and also mortality during 2005 to 2011.


Does positive family history of prostate cancer increase the risk of prostate cancer on initial prostate biopsy?
To assess the role of family history (FH) in the risk of a positive prostate biopsy (PBx) in a large North American biopsy population as earlier reports showed increased risk of prostate cancer (PCa) in men with a FH, but the risk has been limited to low grade prostate cancer in smaller studies, and the REDUCE trial found no such risk in North American patients. We evaluated 4360 men undergoing initial extended biopsy (8-14 cores). Indications were elevated prostate-specific antigen (PSA) and/or abnormal digital rectal examination (DRE). Variables including age, FH of PCa, race, PSA, and DRE results were included in our analysis to assess risk factors associated with PCa, high-grade prostate cancer (HGPCa), and low-grade prostate cancer (LGPCa). Two hundred sixty-eight patients had an FH of PCa whereas 4092 had negative FH. Positive biopsy was found in 1976 patients with HGPCa in 1149 and LGPCa in 827. Among 268 patients with an FH, overall PCa was found in 144 of 268 patients (54%); HGPCa in 79 of 144 patients (55%) and LGPCa in 65 of 144 patients (45%). FH was a significant risk factor for PCa, HGPCa, and LGPCa in univariate and multivariate analysis (P = .0001, .02, and .02, respectively). Also, FH was associated with high-risk benign pathology in the form of atypical small acinar cell proliferation (ASAP) or high-grade prostatic intraepithelial neoplasm (HGPIN) (P = .04). Men in North America with an FH of PCa who undergo prostate biopsy are more likely to be diagnosed with both HGPCa and LGPCa.


Does dexamethasone inhibit the growth of human gliomas?
Dexamethasone (10-(4)M) was shown to inhibit the growth of human gliomas in culture. This was indicated by the inhibition of incorporation of radioactively labeled thymidine into the deoxyribonucleic acid (DNA) of the cells, and by the increase in the generation time of cells exposed to the drug in vitro. On the other hand, tumors obtained from patients who had received dexamethasone before craniotomy grew considerably faster in vitro than tumors from patients who had not been given the drug before operation.


Global Discovery and Temporal Changes of Human Albumin Modifications by Pan-Protein Adductomics: Initial Application to Air Pollution Exposure.
Assessing personal exposure to environmental toxicants is a critical challenge for predicting disease risk. Previously, using human serum albumin (HSA)-based biomonitoring, we reported dosimetric relationships between adducts at HSA Cys34 and ambient air pollutant levels (Smith et al., Chem. Res. Toxicol. 2021, 34, 1183). These results provided the foundation to explore modifications at other sites in HSA to reveal novel adducts of complex exposures. Thus, the Pan-Protein Adductomics (PPA) technology reported here is the next step toward an unbiased, comprehensive characterization of the HSA adductome. The PPA workflow requires <2 μL serum/plasma and uses nanoflow-liquid chromatography, gas-phase fractionation, and overlapping-window data-independent acquisition high-resolution tandem mass spectrometry. PPA analysis of albumin from nonsmoking women exposed to high levels of air pollution uncovered 68 unique location-specific modifications (LSMs) across 21 HSA residues. While nearly half were located at Cys34 (33 LSMs), 35 were detected on other residues, including Lys, His, Tyr, Ser, Met, and Arg. HSA adduct relative abundances spanned a ∼400 000-fold range and included putative products of exogenous (SO2, benzene, phycoerythrobilin) and endogenous (oxidation, lipid peroxidation, glycation, carbamylation) origin, as well as 24 modifications without annotations. PPA quantification revealed statistically significant changes in LSM levels across the 84 days of monitoring (∼3 HSA lifetimes) in the following putative adducts: Cys34 trioxidation, β-methylthiolation, benzaldehyde, and benzene diol epoxide; Met329 oxidation; Arg145 dioxidation; and unannotated Cys34 and His146 adducts. Notably, the PPA workflow can be extended to any protein. Pan-Protein Adductomics is a novel and powerful strategy for untargeted global exploration of protein modifications.


Diffuse multifocal chorangiomatosis of the placenta with multiple intestinal stenosis of the fetus: combination of rare causes for nonimmune hydrops fetalis.
Non-immune causes of hydrops fetalis are rare but frequently fatal. Identification of the cause for hydrops fetalis is essential to institute therapy. Chorangiomatosis and intestinal stenosis have not been previously reported as aetiological factors for the development of hydrops. We report a fetus born with hydrops associated with both of these conditions. A 1575 gms preterm neonate was born to a multigravida at 31 weeks of gestation. Emergency caesarean section was performed after detection of hydrops by prenatal ultrasonography. Baby had generalized edema at birth and died after 3 days due to progressive heart failure. At autopsy there was generalized edema and effusions. There were multiple stenotic segments in the intestine. Placenta showed numerous chorangiomas varying from 0.2 to 3 cm in diameter, hence diagnosed as diffuse multifocal chorangiomatosis. Examination of the placenta at neonatal autopsy is an essential part of assessment for hydrops fetalis. Chorangiomatosis is a rare and under reported condition which can cause prematurity, intrauterine growth retardation, pre-eclampsia and rarely hydrops fetalis.


Histo-Blood Group Antigen Presentation Is Critical for Binding of Norovirus VLP to Glycosphingolipids in Model Membranes.
Virus entry depends on biomolecular recognition at the surface of cell membranes. In the case of glycolipid receptors, these events are expected to be influenced by how the glycan epitope close to the membrane is presented to the virus. This presentation of membrane-associated glycans is more restricted than that of glycans in solution, particularly because of orientational constraints imposed on the glycolipid through its lateral interactions with other membrane lipids and proteins. We have developed and employed a total internal reflection fluorescence microscopy-based binding assay and a scheme for molecular dynamics (MD) membrane simulations to investigate the consequences of various glycan presentation effects. The system studied was histo-blood group antigen (HBGA) epitopes of membrane-bound glycosphingolipids (GSLs) derived from small intestinal epithelium of humans (type 1 chain) and dogs (type 2 chain) interacting with GII.4 norovirus-like particles. Our experimental results showed strong binding to all lipid-linked type 1 chain HBGAs but no or only weak binding to the corresponding type 2 chain HBGAs. This is in contrast to results derived from STD experiments with free HBGAs in solution where binding was observed for Lewis x. The MD data suggest that the strong binding to type 1 chain glycolipids was due to the well-exposed (1,2)-linked α-l-Fucp and (1,4)-linked α-l-Fucp residues, while the weaker binding or lack of binding to type 2 chain HBGAs was due to the very restricted accessibility of the (1,3)-linked α-l-Fucp residue when the glycolipid is embedded in a phospholipid membrane. Our results not only contribute to a general understanding of protein-carbohydrate interactions on model membrane surfaces, particularly in the context of virus binding, but also suggest a possible role of human intestinal GSLs as potential receptors for norovirus uptake.


Comparison of blastocyst euploidy rates following luteal versus follicular phase stimulation in a GnRH antagonist protocol: a prospective study with repeated ovarian stimulation cycles.
Is there any difference in the mean number of euploid embryos following luteal phase start (LS) and follicular phase start (FS) of ovarian stimulation? The mean number of euploid blastocysts is equivalent independent of whether the inseminated oocytes are derived from FS or LS. Starting ovarian stimulation at any time of the cycle ('random-start') is commonly used for emergency fertility preservation in cancer patients. A few retrospective studies have been published evaluating LS in women undergoing ovarian stimulation in the context of IVF, but there is a lack of robust data on the comparative efficacy of LS versus FS.Although 'random start' is commonly used in cancer survivors, few retrospective and uncontrolled studies have been published evaluating luteal phase stimulation in women undergoing ovarian stimulation in the context of IVF. Owing to this evident lack of robust data on the efficacy of LS, guidelines typically recommend the LS approach only for medical reasons and not in the context of IVF. This is a prospective, equivalence study, with repeated stimulation cycles, conducted between May 2018 and December 2021. Overall, 44 oocyte donors underwent two identical consecutive ovarian stimulation cycles, one initiated in the FS and the other in the LS. The primary outcome of the study was to evaluate whether FS and LS in the same patient would result in equivalent numbers of euploid embryos following fertilization of oocytes with the same sperm sample. Overall, 44 oocyte donors underwent two consecutive ovarian stimulation protocols with 150 μg corifollitropin alpha followed by 200 IU recombinant FSH (rFSH) in a fixed GnRH antagonist protocol. The only difference between the two cycles was the day of initiation of ovarian stimulation, which was in the early follicular phase (FS) in one cycle, and in the luteal phase (LS) in the other. Forty-four oocyte recipients participated in the study receiving a mean of six metaphase II (MII) oocytes from each stimulation cycle (FS and LS). All MIIs were inseminated with the corresponding recipient's partner sperm (which had been previously frozen) or donor sperm, in order to safeguard the use of the same sample for either the FS or LS. Following fertilization and blastocyst culture, all generated embryos underwent genetic analysis for aneuploidy screening (preimplantation genetic testing for aneuploidy). FS resulted in a significantly shorter duration of ovarian stimulation (difference between means (DBM) -1.05 (95% CI -1.89; -0.20)) and a lower total additional dose of daily rFSH was needed (DBM -196.02 (95% CI -319.92; -72.12)) compared with LS. The donors' hormonal profile on the day of trigger was comparable between the two stimulation cycles, as well as the mean number of oocytes (23.70 ± 10.79 versus 23.70 ± 8.81) (DBM 0.00 (95% CI -3.03; 3.03)) and MII oocytes (20.27 ± 9.60 versus 20.73 ± 8.65) (DBM -0.45 (95% CI -2.82; 1.91)) between FS and LS cycles, respectively. Following fertilization, the overall blastocyst formation rate was 60.70% with a euploid rate of 57.1%. Comparisons between the two stimulation cycles did not reveal any significance differences in terms of fertilization rates (71.9% versus 71.4%), blastocyst formation rates (59.4% versus 62%) and embryo euploidy rates (56.9 versus 57.3%) for the comparison of FS versus LS, respectively. The mean number of euploid blastocysts was equivalent between the FS (1.59 ± 1.30) and the LS (1.61 ± 1.17), (DBM -0.02 (90%CI -0.48; 0.44)). The study was performed in young, potentially fertile oocyte donors who are patients with high blastocyst euploidy rates. Although results may be extrapolated to young infertile women with good ovarian reserve, caution is needed prior to generalizing the results to infertile women of older age. The current study provides evidence that initiation of ovarian stimulation in the luteal phase in young potentially fertile women may result in a comparable number of oocytes and comparable blastocyst euploidy rates compared with follicular phase stimulation. This may imply that in case of a freeze-all protocol in young patients with good ovarian reserve, clinicians may safely consider initiation of ovarian stimulation during the luteal phase. This research was supported by an unrestricted grant from MSD/Organon. N.P.P. has received Research grants and honoraria for lectures from: Merck Serono, MSD/Organon, Ferring Pharmaceuticals, Besins Intenational, Roche Diagnostics, IBSA, Theramex, Gedeon Richter. F.M., E.C., M.R. and S.G. declared no conflict of interests. The study was registered at Clinical Trials Gov (NCT03555942).


Structural basis of Mos1 transposase inhibition by the anti-retroviral drug Raltegravir.
DNA transposases catalyze the movement of transposons around genomes by a cut-and-paste mechanism related to retroviral integration. Transposases and retroviral integrases share a common RNaseH-like domain with a catalytic DDE/D triad that coordinates the divalent cations required for DNA cleavage and integration. The anti-retroviral drugs Raltegravir and Elvitegravir inhibit integrases by displacing viral DNA ends from the catalytic metal ions. We demonstrate that Raltegravir, but not Elvitegravir, binds to Mos1 transposase in the presence of Mg(2+) or Mn(2+), without the requirement for transposon DNA, and inhibits transposon cleavage and DNA integration in biochemical assays. Crystal structures at 1.7 Å resolution show Raltegravir, in common with integrases, coordinating two Mg(2+) or Mn(2+) ions in the Mos1 active site. However, in the absence of transposon ends, the drug adopts an unusual, compact binding mode distinct from that observed in the active site of the prototype foamy virus integrase.


Enhanced Production of Adenosine Triphosphate by Pharmacological Activation of Adenosine Monophosphate-Activated Protein Kinase Ameliorates Acetaminophen-Induced Liver Injury.
The hepatic cell death induced by acetaminophen (APAP) is closely related to cellular adenosine triphosphate (ATP) depletion, which is mainly caused by mitochondrial dysfunction. Adenosine monophosphate (AMP)-activated protein kinase (AMPK) is a key sensor of low energy status. AMPK regulates metabolic homeostasis by stimulating catabolic metabolism and suppressing anabolic pathways to increase cellular energy levels. We found that the decrease in active phosphorylation of AMPK in response to APAP correlates with decreased ATP levels, in vivo. Therefore, we hypothesized that the enhanced production of ATP via AMPK stimulation can lead to amelioration of APAP-induced liver failure. A769662, an allosteric activator of AMPK, produced a strong synergistic effect on AMPK Thr172 phosphorylation with APAP in primary hepatocytes and liver tissue. Interestingly, activation of AMPK by A769662 ameliorated the APAP-induced hepatotoxicity in C57BL/6N mice treated with APAP at a dose of 400 mg/kg intraperitoneally. However, mice treated with APAP alone developed massive centrilobular necrosis, and APAP increased their serum alanine aminotransferase and aspartate aminotransferase levels. Furthermore, A769662 administration prevented the loss of intracellular ATP without interfering with the APAP-mediated reduction of mitochondrial dysfunction. In contrast, inhibition of glycolysis by 2-deoxy-glucose eliminated the beneficial effects of A769662 on APAP-mediated liver injury. In conclusion, A769662 can effectively protect mice against APAP-induced liver injury through ATP synthesis by anaerobic glycolysis. Furthermore, stimulation of AMPK may have potential therapeutic application for APAP overdose.


Identification and characterization of ADAM41, a novel ADAM metalloproteinase in Xenopus.
The ADAM family of transmembrane metalloproteinases has important functions in fertilization, development and disease, and is widely distributed throughout the Metazoa. In this study, we identified a novel ADAM protein in Xenopus tropicalis (X. tropicalis) with closest overall sequence similarity to the Xenopus ADAM10 protein. Based on comparisons of available sequence information, putative orthologs of this ADAM (which we designate ADAM41) are identified in several other vertebrate species including non-placental mammals, but absent from placental mammals and aves. ADAM41 mRNA is maternally deposited in X. tropicalis with subsequent zygotic expression detected in the neural plate at neurula stages. Antisense morpholino knockdown of ADAM41 results in a delay in early neuronal marker expression, which can be rescued by a non-targeted ADAM41 transcript. Thus, ADAM41 is likely required for maintaining proper timing of neurogenesis in X. tropicalis.


Cap Assisted Upper Endoscopy for Examination of the Major Duodenal Papilla: A Randomized, Blinded, Controlled Crossover Study (CAPPA Study).
Examination of major duodenal papilla (MDP) by standard forward-viewing esophagogastroduodenoscopy (S-EGD) is limited. Cap assisted esophagogastroduodenoscopy (CA-EGD) utilizes a cap fitted to the tip of the endoscope that can depress the mucosal folds and thus might improve visualization of MDP. The aim of this study was to compare CA-EGD to S-EGD for complete examination of the MDP. Prospective, randomized, blinded, controlled crossover study. Subjects scheduled for elective EGD were randomized to undergo S-EGD (group A) or CA-EGD (group B) before undergoing a second examination by the alternate method. Images of the MDP were evaluated by three blinded multicenter-experts. Our primary outcome measure was complete examination of the papilla. Secondary outcome measures were duration and overall diagnostic yield. A total of 101 patients were randomized and completed the study. Complete examination of MDP was achieved in 98 patients using CA-EGD compared to 24 patients using S-EGD (97 vs. 24%, P<0.001). Median duration from intubation of the esophagus until localization of the MDP was shorter with CA-EGD (46. vs. 96 s., P<0.001). In group A, 11 extra lesions and 12 additional incidental findings were detected by secondary CA-EGD, whereas neither were detected by secondary S-EGD in group B (22 vs. 0% and 24 vs. 0%, P<0.001 and P<0.001). CA-EGD enabled complete examination of MDP in almost all cases compared to a low success rate of S-EGD. CA-EGD detected a significant amount of lesions and incidental findings when added to S-EGD. CA-EGD is a safe and effective method for examination of MDP.


Intrauterine insemination with recombinant or urinary human chorionic gonadotropin: A prospective randomized trial.
Our aim was to compare the efficacy and safety of recombinant and urinary human chorionic gonadotropin (rhCG and uhCG, respectively) for the induction of follicle maturation in women undergoing intrauterine insemination (IUI). Patients were randomized to receive rhCG or uhCG. IUI was carried out 24 h (day 1) and 48 h (day 2) after hCG administration, except for all cases in which ovulation occurred after 24 h. The two treatments were comparable in terms of progesterone levels on day 7 and day 12. Pregnancy rates were comparable between the treatment groups. Of the 64 women who received rhCG, 29.7% became pregnant; there were 16.7% clinical pregnancies and 3.1% biochemical pregnancies per started cycle, and an ongoing pregnancy rate of 93.7% was reported. Of the 61 patients who received uhCG, 24.6% became pregnant; there were 15.9% clinical pregnancies and 1.1% biochemical pregnancies per started cycle, and ongoing pregnancy rate was 92.9%. No adverse effects were noted in either group. The recombinant products can be effectively used instead of urinary products; moreover, apart from the equivalent efficacy in ovulation induction and safety described in this study, it is necessary to consider the advantages provided by the recombinant form.


Lysophosphatidylcholine-induced gastric injury and ulceration in the guinea pig.
The effect of lysophosphatidylcholine (lysoPC) on the guinea pig stomach was studied. At concentrations observed in gastric secretions of gastric ulcer patients (1 to 2 mM), lysoPC induced both functional and morphologic changes in the gastric mucosa. Two millimolar lysoPC caused back diffusion of H+ and forward diffusion of Na+, indicating impairment of the gastric mucosal barrier. In the lysoPC-treated stomachs, an increase was observed in 1) the number of mucosal erosions, 2) the intensity of inflammation, and 3) the degree of vascular congestion. Microvascular plugging by platelets, vascular stasis, and polymorphonuclear leukocyte margination were dose-responsive to lysoPC. Pretreatment of animals with aspirin (20 mg/kg) did not alter injury. These results indicate a role of lysoPC reflux in gastric mucosal injury, including mucosal erosions and ulcerations.


Let's work together to spread good practice.
Hundreds of the country's most senior nurses and midwives met in Birmingham last month at the annual summit of England's chief nursing officer (CNO). It proved to be a great opportunity to hear some of the most influential thinkers and speakers in healthcare, including the chief nurse herself, Jane Cummings.


Thermal response of a microgel system.
Recent experimental studies [Z. Wu, B. Zhou, Z.B. Hu, Phys. Rev. Lett. 90 (2003) 048304] on an uncharged aqueous poly-N-isopropylacrylamide (PNIPAM) dispersion have shown that this microgel system is sensitive to temperature. This system was also experimentally found to be modeled quite well by microgel particles interacting via a hard-sphere repulsive plus an inverse power (temperature-dependent) attractive potential. To understand theoretically this thermally responsive PNIPAM dispersion, we apply a novel approach [G.F. Wang, S.K. Lai, Phys. Rev. E 70 (2004) 051402] to calculate its thermodynamic phase diagram. Differing from the conventional method in which the boundaries of the coexisting phases are the ultimate target, the present work places emphasis on crosshatching colloidal domains which include the homogeneous phase (gas, liquid or solid), two coexisting phases and perhaps also multi-phases in coexistence. Strategically, this was done by treating the coexisting phases as one composite system whose Helmholtz free energy density is written as the sum of constituent free energy densities each of which is weighed by its respective volume proportion. We show here that by minimizing the composite system's free energy density the phase-diagram domains can all be determined in addition to the phase boundaries customarily obtained by imposing the conditions of equal pressure and equal chemical potential. Also, we present the theoretically predicted phase diagram of PNIPAM dispersion and compare it with the one observed experimentally.


Monitoring of changes in lipid profiles during PLK1 knockdown in cancer cells using DESI MS.
The importance of the polo-like kinase 1 (PLK1) gene is increasing substantially both as a biomarker and as a target for highly specific cancer therapy. This is due to its involvement in multiple points of cell progression and carcinogenesis. PLK1 inhibitors' efficacy in treating human cancers has been limited due to the lack of a specific targeting strategy. Here, we describe a method of targeted downregulation of PLK1 in cancer cells and the concomitant rapid detection of surface lipidomic perturbations using desorption electrospray ionization mass spectrometry (DESI MS). The efficient delivery of siRNA targeting PLK1 gene selectively to the cancer cells is achieved by targeting overexpressed cell surface epithelial cell adhesion molecule (EpCAM) by the EpDT3 aptamer. The chimeric aptamer (EpDT3-siPLK1) showed the knockdown of PLK1 gene expression and PLK1 protein levels by quantitative PCR and western blotting, respectively. The abundant surface lipids, phosphatidylcholines (PCs), such as PC(32:1) (m/z 754.6), PC(34:1) (m/z 782.6), and PC(36:2) (m/z 808.6), were highly expressed in MCF-7 and WERI-RB1 cancer cells compared to normal MIO-M1 cells and they were observed using DESI MS. These overexpressed cell surface lipids in the cancer cells were downregulated upon the treatment of EpDT3-siPLK1 chimera indicating a novel role of PLK1 to regulate surface lipid expression in addition to the efficient selective cancer targeting ability. Our results indicate that DESI MS has a potential ability to rapidly monitor aptamer-mediated cancer therapy and accelerate the drug discovery process. Graphical abstract Binding of aptamer chimera to the cells and changes in lipid profile.


Integrative, multifunctional plasmids for hypha-specific or constitutive expression of green fluorescent protein in Candida albicans.
The authors have engineered plasmid constructs for developmental and constitutive expression of yeast-enhanced green fluorescent protein (yEGFP3) in Candida albicans. The promoter for the hyphae-specific gene Hyphal Wall Protein 1 (HWP1) conferred developmental expression of yEGFP3 in germ tubes and hyphae but not in yeasts or pseudohyphae when targeted to the ENO1 (enolase) locus in single copy. The pHWP1GFP3 construct allows for the easy visualization of HWP1 promoter activity in individual cells expressing true hyphae without having to prepare RNA for analysis. Constitutive expression of yEGFP was seen in all cell morphologies when the HWP1 promoter was replaced with the ENO1 promoter region. The use of the plasmids for expression of genes other than yEGFP3 was examined by substituting the putative C. albicans BCY1 (SRA1) gene, a component of the cAMP signalling pathway involved in yeast to hyphae transitions, for yEGFP3. Strains overexpressing BCY1 from the ENO1 promoter were inhibited in germ tube formation and filamentation in both liquid and solid media, a phenotype consistent with keeping protein kinase A in its inactive form by association with Bcy1p. The plasmids are suitable for studies of germ tube induction or assessing germ tube formation by measuring yEGFP3 expression, for inducible expression of genes concomitant with germ tube formation by the HWP1 promoter, for constitutive expression of genes by the ENO1 promoter, and for expressing yEGFP3 using a promoter of choice.


Cu-Catalyzed C-H Allylation of Benzimidazoles with Allenes.
CuH-catalyzed intramolecular cyclization and intermolecular allylation of benzimidazoles with allenes have been described. The reaction proceeded smoothly with the catalytic system of Cu(OAc)2/Xantphos and catalytic amount of (MeO)2MeSiH. This protocol features mild reaction conditions and a good tolerance of substrates bearing electron-withdrawing, electron-donating, or electron-neutral groups. A new catalytic mechanism was proposed for this copper hydride catalytic system.


Identification of substrates of palmitoyl protein thioesterase 1 highlights roles of depalmitoylation in disulfide bond formation and synaptic function.
Loss-of-function mutations in the depalmitoylating enzyme palmitoyl protein thioesterase 1 (PPT1) cause neuronal ceroid lipofuscinosis (NCL), a devastating neurodegenerative disease. The substrates of PPT1 are largely undescribed, posing a limitation on molecular dissection of disease mechanisms and therapeutic development. Here, we provide a resource identifying >100 novel PPT1 substrates. We utilized Acyl Resin-Assisted Capture (Acyl RAC) and mass spectrometry to identify proteins with increased in vivo palmitoylation in PPT1 knockout (KO) mouse brains. We then validated putative substrates through direct depalmitoylation with recombinant PPT1. This stringent screen elucidated diverse PPT1 substrates at the synapse, including channels and transporters, G-protein-associated molecules, endo/exocytic components, synaptic adhesion molecules, and mitochondrial proteins. Cysteine depalmitoylation sites in transmembrane PPT1 substrates frequently participate in disulfide bonds in the mature protein. We confirmed that depalmitoylation plays a role in disulfide bond formation in a tertiary screen analyzing posttranslational modifications (PTMs). Collectively, these data highlight the role of PPT1 in mediating synapse functions, implicate molecular pathways in the etiology of NCL and other neurodegenerative diseases, and advance our basic understanding of the purpose of depalmitoylation.


Alkaline phosphatase of cancerous larynx tissue in comparison with the placental enzyme. Biochemical and histochemical studies.
Tissue specimens of squamous cell carcinoma of the larynx from twenty patients were processed for histological and histopathological characterization. A histochemical study of alkaline phosphatase (ALP) was carried out using the simultaneous azo coupling method, and biochemical studies were performed using disodium phenylphosphate as substrate. Full-term, normal placentae were used for comparison. The specific activity of ALP from cancerous laryngeal tissue was 8.9 mKAU/mg protein compared with 154.7 mKAU/mg protein in the placenta. The ALP was localized histochemically in tumor cells (tumor-specific), blood vessels (vascular) and fibrous tissue (interstitial). The tumor-specific phosphatase was sensitive to inhibition by L-phenylalanine, L-leucine and to a lesser degree by L-tryptophan and levamisole. Placental ALP, on the other hand, was completely inhibited by levamisole, more resistant to leucine and more sensitive to phenylalanine and tryptophan. Biochemical estimation of ALP in cancerous laryngeal tissue combined with inhibition studies revealed that the tumor-specific activity of ALP constitutes 15% of the total ALP activity while the major isoenzyme was the vascular ALP, and around one-third of ALP activity was attributed to the interstitial enzyme. The characterization and localization of these isoenzymes are described and compared with that of the placenta. The significance and implications of the above findings are presented.


Proteomic profile in glomeruli of type-2 diabetic KKAy mice using 2-dimensional differential gel electrophoresis.
Diabetic nephropathy (DN) is a leading cause of end-stage renal disease. To search for glomerular proteins associated with early-stage DN, glomeruli of spontaneous type 2 diabetic KKAy mice were analyzed by 2-dimensional differential gel electrophoresis (2D-DIGE). Glomeruli of 20-week spontaneous type 2 diabetic KKAy mice and age-matched C57BL/6 mice were isolated by kidney perfusion with magnetic beads. Proteomic profiles of glomeruli were investigated by using 2D-DIGE and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Western blot analysis was used to confirm the results of proteomics. Immunohistochemical and semi-quantitative analysis were used to confirm the differential expression of prohibitin and annexin A2 in glomeruli. We identified 19 differentially expressed proteins - 17 proteins were significantly up-regulated and 2 proteins were significantly down-regulated in glomeruli of diabetic KKAy mice. Among them, prohibitin and annexin A2 were up-regulated and Western blot analysis validated the same result in proteomics. Immunohistochemical analysis also revealed up-regulation of prohibitin and annexin A2 in glomeruli of KKAy mice. Our findings suggest that prohibitin and annexin A2 may be associated with early-stage DN. Further functional research might help to reveal the pathogenesis of DN.


Protective effect of capsaicin against methyl methanesulphonate induced toxicity in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg9.
Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide) is the main component in hot peppers, including red chili peppers, jalapenos, and habanero, belonging to the genus Capsicum. Capsaicin is a potent antioxidant that interferes with free radical activities. In the present study, the possible protective effect of capsaicin was studied against methyl methanesulphonate (MMS) induced toxicity in third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg9. The third instar was allowed to feed on the diet having different doses of capsaicin and MMS separately and in combination. The results suggested that the exposure of third instar larvae to the diet having MMS alone showed significant hsp70 expression as well as tissue DNA and oxidative damage, whereas the larvae feed on the diet having MMS and capsaicin showed a decrease in the toxic effects for 48-h of exposure. In conclusion, capsaicin showed a dose-dependent decrease in the toxic effects induced by MMS in the third instar larvae of transgenic Drosophila melanogaster.


The effect of ultraviolet radiation on the cornea - experimental study.
Ultraviolet (UV) radiation in high doses may have harmful effects on the eye. The sources of UV radiation are the sun, as well as some artificial sources such as UV lamps or voltaic arcs. Chronic exposure to UV can cause damage to the anterior pole of the eye, ranging from minor (pterygium) to serious photokeratitis. In our study, we applied a UV dose of 6.5 J/cm(2) in the wavelength range of 290-400 nm, for five consecutive days per rat anterior pole of the eye. Seven days after the last dose of radiation, the animals were sacrificed, harvesting both the irradiated and the non-irradiated eye. Histological and immunohistochemical examination of the lesions revealed that the greatest damage to the epithelium was recorded prior to and 2/3 of the remaining corneal stroma. The epithelial lesions we found varied from pseudokeratosis and detachment of the Bowman epithelium membrane to deep epithelial necrosis. Within the corneal stroma, we observed the formation of interstitial edema with disruption of the collagen structure. We also noticed the presence of an inflammatory infiltrate composed mainly of lymphocytes and CD68+ and CD163+ macrophages, as well as the occurrence of vascular devices. These consisted of angiogenesis capillaries with structured wall composed mainly of endothelial CD34+ precursor cells and a basal membrane rich in collagen IV fibers.


Persistence of parasite antigenemia following diethylcarbamazine therapy of bancroftian filariasis.
This study was designed to reexamine the efficacy of diethylcarbamazine for bancroftian filariasis with special reference to changes in serum parasite antigen levels and antifilarial antibody titers after treatment. Patients with asymptomatic microfilaremia were treated with 6 mg/kg diethylcarbamazine daily for 12 days. Microfilaria counts fell dramatically after treatment, as expected. IgG antibody titers to adult and microfilarial antigens of B. malayi were increased 1 month after treatment in most patients. Titers fell slowly to or below pretreatment levels, but remained positive during subsequent months. Parasite antigen levels, measured by monoclonal antibody-based enzyme immunoassay, decreased to 72%, 58%, 53%, and 48% of pretreatment values 1, 3, 6, and 12 months after diethylcarbamazine treatment, respectively. Parasite antigen levels decreased similarly in subjects with and without residual microfilaremia after treatment. These results suggest that diethylcarbamazine has only partial macrofilaricidal activity against W. bancrofti with this dosage schedule. The sustained, impressive reductions in microfilaria counts after treatment despite significant persistence of parasite antigenemia may be explained by sublethal effects of the drug on adult worms. We believe that parasite antigen detection represents a valuable new approach for monitoring the efficacy of antifilarial drug therapy which we hope will lead to improved use of existing drugs and aid in the evaluation of new drugs for filariasis.


Moderately Hypofractionated Intensity-modulated Radiotherapy With a Simultaneous Integrated Boost for Locally Advanced Head and Neck Cancer - Do Modern Techniques Fulfil Their Promise?
Intensity-modulated radiotherapy (IMRT) with simultaneous integrated boost (SIB) and moderate hypofractionation offers an opportunity for defining individual doses and a reduction in overall treatment time in locally advanced head and neck cancer (HNSCC). We present retrospective data on toxicity and locoregional control of a patient cohort treated with an IMRT-SIB concept in comparison to normo-fractionated 3D-conformal radiotherapy (3D-RT). Between 2012 and 2014, 67 patients with HNSCC (stages III-IVB) were treated with IMRT-SIB either definitively or in the postoperative setting. These patients were matched with those of patients treated with normo-fractionated 3D-RT before mid-2012 and their clinical courses were compared. Chemotherapy or cetuximab was given concomitantly in both groups in the definitive situation (postoperatively, dependent on risk factors). Significantly less toxicity was found in favor of IMRT-SIB concerning dysphagia, dermatitis, xerostomia, fibrosis, and lymphedema. After a median follow-up of 31 months (range=2-104 months), 3-year locoregional control was 73% for those treated with IMRT-SIB versus 78% for those treated with 3D-RT. This moderately hypofractionated IMRT-SIB concept was shown to be feasible, incurring less toxicity than conventional 3D-RT.


Altered expression of inhibitory guanine nucleotide regulatory proteins (Gi-proteins) in experimental hepatocellular carcinoma.
Guanine nucleotide regulatory proteins (G-proteins) play an important role in the onset and progression of malignancy. We hypothesized that alterations in inhibitory G-protein (Gi) expression and/or function may contribute to cellular invasion and formation of hepatocellular carcinoma (HCC). H4IIE hepatoma cells were inoculated directly into the liver parenchyma of ACI strain rats, and membranes were prepared from HCC livers and adjacent nonneoplastic livers 12 days following the initial inoculation. Expression of inhibitory Gialpha proteins was determined by Western blot analysis and changes in the functional activity of these proteins confirmed by pertussis toxin catalyzed ADP ribosylation and adenylyl cyclase activity. Inhibitory Gialpha1, Gialpha1/2, and Gialpha3 protein expression was significantly elevated in HCC when compared to adjacent nonneoplastic liver and sham-operated hepatic tissue. Pertussis toxin catalyzed ADP ribosylation of Gialpha substrates was significantly enhanced in HCC concomitant with increased basal and stimulated adenylyl cyclase activity following uncoupling of Gi-proteins with manganese ions. The role of Gi-proteins in cellular proliferation was confirmed using cultured H4IIE cells and normal hepatocytes. In quiescent H4IIE cells, mastoparan (Gialpha activator) increased [3H] thymidine incorporation and cell growth in a dose-dependent manner, whereas both pertussis toxin (a Gi-protein inhibitor) and 8-bromo-cAMP inhibited mitogenesis. In contrast, in isolated cultured hepatocytes, mastoparan inhibited [3H] thymidine incorporation, while pertussis toxin and 8-bromo-cAMP were mitogenic. We conclude that HCC is associated with marked changes in Gialpha-protein expression in vivo and in vitro, direct activation of which leads to increased mitogenesis in H4IIE cells in vitro.


The association of gender concordance and primary care physicians' perceptions of their patients.
In this article, we examined the effect of gender concordance on physicians' perception of their patients and of their medical condition, analyzing a data set of 8,258 visit questionnaires from the New Zealand National Primary Care Medical Care Survey conducted in 2001 2002. Multivariate analysis indicated that the concordant female patient/female physician dyad had a positive independent association with physicians' reporting high rapport and a negative independent association with reporting uncertainty about the diagnosis. The discordant female patient/male physician dyad had a positive independent association with physicians' perceptions of uncertainty of diagnosis and hidden agenda, and a negative independent association with rating the patient's condition of high severity. The findings suggest a need to raise male physicians' awareness to possible biases when treating female patients. The findings also suggest the need to empower female patients to take an active partnership role to improve their communication with male physicians.


Current and potential roles of immuno-PET/-SPECT in CAR T-cell therapy.
Chimeric antigen receptor (CAR) T-cell therapies have evolved as breakthrough treatment options for the management of hematological malignancies and are also being developed as therapeutics for solid tumors. However, despite the impressive patient responses from CD19-directed CAR T-cell therapies, ~ 40%-60% of these patients' cancers eventually relapse, with variable prognosis. Such relapses may occur due to a combination of molecular resistance mechanisms, including antigen loss or mutations, T-cell exhaustion, and progression of the immunosuppressive tumor microenvironment. This class of therapeutics is also associated with certain unique toxicities, such as cytokine release syndrome, immune effector cell-associated neurotoxicity syndrome, and other "on-target, off-tumor" toxicities, as well as anaphylactic effects. Furthermore, manufacturing limitations and challenges associated with solid tumor infiltration have delayed extensive applications. The molecular imaging modalities of immunological positron emission tomography and single-photon emission computed tomography (immuno-PET/-SPECT) offer a target-specific and highly sensitive, quantitative, non-invasive platform for longitudinal detection of dynamic variations in target antigen expression in the body. Leveraging these imaging strategies as guidance tools for use with CAR T-cell therapies may enable the timely identification of resistance mechanisms and/or toxic events when they occur, permitting effective therapeutic interventions. In addition, the utilization of these approaches in tracking the CAR T-cell pharmacokinetics during product development and optimization may help to assess their efficacy and accordingly to predict treatment outcomes. In this review, we focus on current challenges and potential opportunities in the application of immuno-PET/-SPECT imaging strategies to address the challenges encountered with CAR T-cell therapies.


Wavelength dependence of laser-induced damage: determining the damage initiation mechanisms.
A novel experimental approach is employed to understand the mechanisms of laser induced damage. Using an OPO (optical parametric oscillator) laser, we have measured the damage thresholds of deuterated potassium dihydrogen phosphate (DKDP) from the near ultraviolet into the visible. Distinct steps, whose width is of the order of k(B)T, are observed in the damage threshold at photon energies associated with the number of photons (3-->2 or 4-->3) needed to promote a ground state electron across the energy gap. The wavelength dependence of the damage threshold suggests that a primary mechanism for damage initiation in DKDP is a multiphoton process in which the order is reduced through excited defect state absorption.


Expression and characterization of three new glutathione transferases, an epsilon (AcGSTE2-2), omega (AcGSTO1-1), and theta (AcGSTT1-1) from Anopheles cracens (Diptera: Culicidae), a major Thai malaria vector.
Glutathione transferases (GSTs) (E.C.188.8.131.52) are multifunctional enzymes involved in the detoxification of many exogenous and endogenous compounds. This study aimed to characterize several new GSTs from Anopheles cracens, a major Thai malaria vector formerly known as Anopheles dirus. The three recombinant enzymes obtained were from the epsilon, theta and omega classes. They showed 80-93% identity to orthologous An. gambiae GSTs. AcGSTE2-2 possessed peroxidase activity that cannot be detected for the An. gambiae AgGSTE2-2. AcGSTT1-1 had high activity toward several substrates that are specific for mammalian theta class. The AcGSTO1-1 can use 1-chloro-2,4-dinitrobenzene, dichloroacetic acid, and hydroxyethyl disulfide substrates. The enzymes bound but did not metabolize the organophosphate temephos. The epsilon AcGSTE2-2 functioned as a peroxidase and DDT metabolizing enzyme. The theta AcGSTT1-1 functioned not only as peroxidase but also acted as a binding protein for organophosphates. The omega GST had thiol transferase activity suggesting a role in oxidative stress response.


[Chronic occupational mercury exposure in renal damage in workers in the chlorine-alkali electrolysis industry].
The authors investigated renal damage in 46 chlorine-alkaly plant workers (mean age was 38.8 +/- 5.7 years) under conditions of continued occupational exposure to metallic mercury vapour. The mercury unexposed control group consisted of 32 workers who works in the plant area. Significantly low of serum globulin level was found in exposed evaluated group compared with control subjects (P < 0.001). The serum globulin level was in correlation with urine mercury level (P < 0.001). Analyses of urine chemistry indicated that exposed workers had cell death produces in sediment urine as the most common signs (P < 0.001). The proteinuria was found in 4 out 32 and high level of gamma-glutamyl-transpeptidase in 8 out 32 exposed workers to high mercury level workers. Additionally, disuria and ejaculatory pain as symptoms occurred without evidence of urological disease. Mercury induced nephropathy usually associated with proteinuria, but is not with renal insufficiency.


Beneficial effects of resveratrol on atherosclerosis.
Atherosclerosis, a progressive disease characterized by the accumulation of lipids and fibrous elements in the arteries, is a most important contributor to cardiovascular diseases. Resveratrol is a naturally occurring phytopolyphenol compound and shows the ability to reduce the risk of cardiovascular diseases. In this review, beneficial effects of resveratrol on the initiation and progression of atherosclerosis, including regulation of vasodilator and vasoconstrictor production, inhibition of oxidative stress/reactive oxygen species generation, anti-inflammation, inhibition of modification of low-density lipoproteins, anti-platelet aggregation, and its abilities to impede progression and modulate complications of atherosclerosis, are discussed.


Immune Checkpoint Blockade Enhances Shared Neoantigen-Induced T-cell Immunity Directed against Mutated Calreticulin in Myeloproliferative Neoplasms.
Somatic frameshift mutations in the calreticulin (CALR) gene are key drivers of cellular transformation in myeloproliferative neoplasms (MPN). All patients carrying these mutations (CALR + MPN) share an identical sequence in the C-terminus of the mutated CALR protein (mut-CALR), with the potential for utility as a shared neoantigen. Here, we demonstrate that although a subset of patients with CALR + MPN develop specific T-cell responses against the mut-CALR C-terminus, PD-1 or CTLA4 expression abrogates the full complement of responses. Significantly, blockade of PD-1 and CLTA4 ex vivo by mAbs and of PD-1 in vivo by pembrolizumab administration restores mut-CALR-specific T-cell immunity in some patients with CALR + MPN. Moreover, mut-CALR elicits antigen-specific responses from both CD4+ and CD8+ T cells, confirming its broad applicability as an immunogen. Collectively, these results establish mut-CALR as a shared, MPN-specific neoantigen and inform the design of novel immunotherapies targeting mut-CALR. SIGNIFICANCE: Current treatment modalities for MPN are not effective in eliminating malignant cells. Here, we show that mutations in the CALR gene, which drive transformation in MPN, elicit T-cell responses that can be further enhanced by checkpoint blockade, suggesting immunotherapies could be employed to eliminate CALR + malignant cells in MPN.This article is highlighted in the In This Issue feature, p. 1143.


[25 years without reactions to a grown-in laparitimy gauze].
We report the remarkable history of a case with intraperitoneal retention of a gauze sponge, left in place for 24 years without any symptoms. The foreign body was removed in a selective operation after being detected by routine gynecologic examination. The rare and unusually fortunate course is discussed.


Rehabilitation in patients with dementia following hip fracture: a systematic review.
Best rehabilitation practices after hip fracture for people with dementia have not been established. A systematic review was conducted to determine current evidence for rehabilitation in this population, including residents in continuing care. Standardized review methodology was used to search eight databases for literature on hip-fracture rehabilitation for people with dementia. Eligible studies included participants with dementia who had a hip fracture; performed a rehabilitation intervention; and evaluated one or more of function, ambulation, discharge location, or falls. The Newcastle-Ottawa Scale was used to assess validity. A total of 13 studies were included: five randomized controlled trials (RCTs), seven prospective cohort series, and one retrospective cohort study. Average quality ratings for RCTs and cohort studies were good and fair respectively. Participants with mild to moderate dementia receiving rehabilitation showed similar relative gains in function to those without dementia. Only one study examined the effect of rehabilitation among residents in continuing care. People with mild or moderate dementia may show improved function and ambulation and decreased fall risk after rehabilitation post hip fracture, similar to gains achieved by those without dementia. More research is required to ascertain the effect of rehabilitation in people with moderate to severe dementia, including those residing in continuing-care settings. Purpose: Best rehabilitation practices after hip fracture for people with dementia have not been established. A systematic review was conducted to determine current evidence for rehabilitation in this population, including residents in continuing care. Methods: Standardized review methodology was used to search eight databases for literature on hip-fracture rehabilitation for people with dementia. Eligible studies included participants with dementia who had a hip fracture; performed a rehabilitation intervention; and evaluated one or more of function, ambulation, discharge location, or falls. The Newcastle–Ottawa Scale was used to assess validity. Results: A total of 13 studies were included: five randomized controlled trials (RCTs), seven prospective cohort series, and one retrospective cohort study. Average quality ratings for RCTs and cohort studies were good and fair respectively. Participants with mild to moderate dementia receiving rehabilitation showed similar relative gains in function to those without dementia. Only one study examined the effect of rehabilitation among residents in continuing care. Conclusions: People with mild or moderate dementia may show improved function and ambulation and decreased fall risk after rehabilitation post hip fracture, similar to gains achieved by those without dementia. More research is required to ascertain the effect of rehabilitation in people with moderate to severe dementia, including those residing in continuing-care settings.


ZmGluTR1 is involved in chlorophyll biosynthesis and is essential for maize development.
Chlorophyll is the most important carrier of photosynthesis in plants and is therefore vital for plant growth and development. Synthesis of 5-aminolevulinic acid (ALA) is initiated and catalyzed by glutamyl-tRNA reductase (GluTR) and is the rate-limiting step in chlorophyll biosynthesis. GluTR is controlled by several regulating factors. Although many studies have investigated the structure and function of GluTR in plants, the maize (Zea mays L.) GluTR has not yet been reported. Here, we isolated and identified the first loss-of-function mutant of GluTR in plants from a maize mutagenic population. The stop-gain mutation in ZmGluTR1 resulted in leaf etiolation throughout the growing season. The level of intermediates of chlorophyll biosynthesis and photosynthetic pigments decreased markedly and abnormal chloroplast structure was also observed in the mutants. Further analysis revealed that the deletion of carboxyl terminal (C-terminal) led to premature transcription termination and this hindered the interaction with FLUORESCENT (FLU), thereby influencing the stability of mutated ZmGluTR1 and leading to abolish interaction with GluTR-binding protein (GluBP). Moreover, mutations in the catalytic domain or nicotinamide adenine dinucleotide phosphate (NADPH) binding domain were lethal under normal growth conditions. These results indicate that ZmGluTR1 plays a fundamental role in chlorophyll biosynthesis and maize development.


[Results of managing prostate adenomyomatosis over a period of 5 years].
It is reported on the treatment of 382 patients with adenomas in the period from 1974 to 1978. The operative process is briefly described. The results and the complications are shown. The shorter duration of stay, the smaller number of the postoperative complications and the smaller postoperative lethality prove the advantage of the TUR in the treatment of the adenomyomatosis of the prostate compared with the adenomectomy.


Forward models for extending the mechanical damage evaluation capability of resonant ultrasound spectroscopy.
Finite element (FE) modeling has been coupled with resonant ultrasound spectroscopy (RUS) for nondestructive evaluation (NDE) of high temperature damage induced by mechanical loading. Forward FE models predict mode-specific changes in resonance frequencies (ΔfR), inform RUS measurements of mode-type, and identify diagnostic resonance modes sensitive to individual or multiple concurrent damage mechanisms. The magnitude of modeled ΔfR correlate very well with the magnitude of measured ΔfR from RUS, affording quantitative assessments of damage. This approach was employed to study creep damage in a polycrystalline Ni-based superalloy (Mar-M247) at 950°C. After iterative applications of creep strains up to 8.8%, RUS measurements recorded ΔfR that correspond to the accumulation of plastic deformation and cracks in the gauge section of a cylindrical dog-bone specimen. Of the first 50 resonance modes that occur, ranging from 3 to 220kHz, modes classified as longitudinal bending were most sensitive to creep damage while transverse bending modes were found to be largely unaffected. Measure to model comparisons of ΔfR show that the deformation experienced by the specimen during creep, specifically uniform elongation of the gauge section, is responsible for a majority of the measured ΔfR until at least 6.1% creep strain. After 8.8% strain considerable surface cracking along the gauge section of the dog-bone was observed, for which FE models indicate low-frequency longitudinal bending modes are significantly affected. Key differences between historical implementations of RUS for NDE and the FE model-based framework developed herein are discussed, with attention to general implementation of a FE model-based framework for NDE of damage.


Interaction of structure-specific and promiscuous G-protein-coupled receptors mediates small-molecule signaling in Caenorhabditis elegans.
A chemically diverse family of small-molecule signals, the ascarosides, control developmental diapause (dauer), olfactory learning, and social behaviors of the nematode model organism, Caenorhabditis elegans. The ascarosides act upstream of conserved signaling pathways, including the insulin, TGF-β, serotonin, and guanylyl cyclase pathways; however, the sensory processes underlying ascaroside function are poorly understood. Because ascarosides often are multifunctional and show strongly synergistic effects, characterization of their receptors will be essential for understanding ascaroside biology and may provide insight into molecular mechanisms that produce synergistic outcomes in small-molecule sensing. Based on DAF-8 immunoprecipitation, we here identify two G-protein-coupled receptors, DAF-37 and DAF-38, which cooperatively mediate ascaroside perception. daf-37 mutants are defective in all responses to ascr#2, one of the most potent dauer-inducing ascarosides, although this mutant responds normally to other ascarosides. In contrast, daf-38 mutants are partially defective in responses to several different ascarosides. Through cell-specific overexpression, we show that DAF-37 regulates dauer when expressed in ASI neurons and adult behavior when expressed in ASK neurons. Using a photoaffinity-labeled ascr#2 probe and amplified luminescence assays (AlphaScreen), we demonstrate that ascr#2 binds to DAF-37. Photobleaching fluorescent energy transfer assays revealed that DAF-37 and DAF-38 form heterodimers, and we show that heterodimerization strongly increases cAMP inhibition in response to ascr#2. These results suggest that that the ascarosides' intricate signaling properties result in part from the interaction of highly structure-specific G-protein-coupled receptors such as DAF-37 with more promiscuous G-protein-coupled receptors such as DAF-38.


Making pro formas perform.
Regulators are trying to clear up the muddle created by earnings-report adjustments called "pro formas" that companies issue. Constraining such reporting, as the regulators seem bent on doing, isn't the solution. Firms should increase alternative reporting--and fully account for their accounting.


[Evaluation of a number of commercial diets for iguana (Iguana iguana), bearded dragons (Pogona vitticeps), and land and marsh tortoises].
Fifteen commercially available, complete diets for iguanas, bearded dragons, and chelonians were analysed and compared with the ideal composition of a diet formulated by the authors. The clinical risks of feeding with a too high or a too low content of specific nutrients are described. Nutrient deficiencies are not expected if these diets are the sole source of nutrients. However, one diet had only a marginally adequate protein content, five diets for herbivorous reptiles contained a too low percentage of crude fibre, three diets contained an undesirably high percentage of calcium, and three diets had an extremely high iron content. As a rule, the calcium and iron contents of diets are not declared on the food label, which complicates evaluation of the diets.


Phylogenetic Relationships among Whiteflies in the Bemisia tabaci (Gennadius) Species Complex from Major Cassava Growing Areas in Kenya.
Whiteflies, Bemisia tabaci (Gennadius) are major insect pests that affect many crops such as cassava, tomato, beans, cotton, cucurbits, potato, sweet potato, and ornamental crops. Bemisia tabaci transmits viral diseases, namely cassava mosaic and cassava brown streak diseases, which are the main constraints to cassava production, causing huge losses to many small-scale farmers. The aim of this work was to determine the phylogenetic relationships among Bemisia tabaci species in major cassava growing areas of Kenya. Surveys were carried out between 2013 and 2015 in major cassava growing areas (Western, Nyanza, Eastern, and Coast regions), for cassava mosaic disease (CMD) and cassava brown streak disease (CBSD). Mitochondrial cytochrome oxidase I (mtCOI-DNA) was used to determine the genetic diversity of B. tabaci. Phylogenetic trees were constructed using Bayesian methods to understand the genetic diversity across the study regions. Phylogenetic analysis revealed two B. tabaci species present in Kenya, sub-Saharan Africa 1 and 2 comprising five distinct clades (A-E) with percent sequence similarity ranging from 97.7 % to 99.5%. Clades B, C, D, and E are predominantly distributed in the Western and Nyanza regions of Kenya whereas clade B is dominantly found along the coast, the eastern region, and parts of Nyanza. Our B. tabaci clade A groups with sub-Saharan Africa 2-(SSA2) recorded a percent sequence similarity of 99.5%. In this study, we also report the identification of SSA2 after a 15 year absence in Kenya. The SSA2 species associated with CMD has been found in the Western region of Kenya bordering Uganda. More information is needed to determine if these species are differentially involved in the epidemiology of the cassava viruses.


Protein intake in cancer: Does it improve nutritional status and/or modify tumour response to chemotherapy?
Combating malnutrition and cachexia is a core challenge in oncology. To limit muscle mass loss, the use of proteins in cancer is encouraged by experts in the field, but it is still debated due to their antagonist effects. Indeed, a high protein intake could preserve lean body mass but may promote tumour growth, whereas a low-protein diet could reduce tumour size but without addressing cachexia. Here we used a realistic rodent model of cancer and chemotherapy to evaluate the influence of different protein intakes on cachexia, tumour response to chemotherapy and immune system response. The goal is to gain a closer understanding of the effect of protein intake in cancer patients undergoing chemotherapy. Female Fischer 344 rats were divided into six groups: five groups (n = 14 per group) with cancer (Ward colon tumour) and chemotherapy were fed with isocaloric diets with 8%, 12%, 16%, 24% or 32% of caloric intake from protein and one healthy control group (n = 8) fed a 16% protein diet, considered as a standard diet. Chemotherapy included two cycles, 1 week apart, each consisting of an injection of CPT-11 (50 mg/kg) followed by 5-fluorouracil (50 mg/kg) the day after. Food intake, body weight, and tumour size were measured daily. On day 9, the rats were euthanized and organs were weighed. Body composition was determined and protein content and protein synthesis (SUnSET method) were measured in the muscle, liver, intestine, and tumour. Immune function was explored by flow cytometry. Cancer and chemotherapy led to a decrease in body weight characterized by a decrease of both fat mass (-56 ± 3%, P < 0.05) and fat-free mass (-8 ± 1%, P < 0.05). Surprisingly, there was no effect of protein diet on body composition, muscle or tumour parameters (weight, protein content, or protein synthesis) but a high cumulative protein intake was positively associated with a high relative body weight and high fat-free mass. The immune system was impacted by cancer and chemotherapy but not by the different amount of protein intake. Using a realistic model of cancer and chemotherapy, we demonstrated for the first time that protein intake did not positively or negatively modulate tumour growth. Moreover, our results suggested that a high cumulative protein intake was able to improve moderately nutritional status in chemotherapy treated cancer rodents. Although this work cannot be evaluated clinically for ethical reasons, it nevertheless brings an essential contribution to nutrition management for cancer patients.


Plasma amino-acid concentrations during development in the rat.
The plasma individual amino-acid concentrations in the rat during late foetal development and from the birth to the end of the weaning have been studied. Both the very low foetal amino-acid degrading capabilities in addition to the high amino-acid concentration ability of the placenta help to sustain considerably high plasma foetal amino-acid levels that favour considerably protein synthesis. There is a very important decrease in most amino-acid concentrations with birth, due to the cessation of placental transfer, relative immaturity of intake and assimilation processes and to the highly anabolic environment found in the pup. The changes in postnatal development plasma amino-acid patterns reflect directly the availability of amino acids in the diet, the maturation of the amino-acid metabolism pathways and transport systems and their use for protein synthesis (and other nitrogenous compounds). The main situation that affects individual amino-acid levels is the shift from high biological value milk protein to low biological value plant proteins in the rat chow pellets. The postnatal development is characterized by a high combined total amino-acids homeostasis with ample changes in individual amino-acid concentrations, that is maintained in spite of the deep changes in diet, size, metabolism, hormone environment, maturation, etc. observed in the rat from birth to weaning and adulthood.


Emerging trends and new developments in monoclonal antibodies: A scientometric analysis (1980-2016).
This article aims to explore the intellectual landscape of the study of monoclonal antibody (mAb), mainly to identify thematic trends, landmark articles and emerging trends involving mAb. This work is based on 4 sets of bibliographic records retrieved from the Web of Science. The final data set, consisting of 7,385 bibliographic records, was combined from the 4 individual data sets. This study explores the document co-citation clusters of 7,385 bibliographic records to identify the origin of mAb and the hot research specialty of this domain by applying CiteSpace software. We examined the mAb evolution from 4 perspectives: (1) Clusters of cited references regarding mAb; (2) Cited authors as contributors to mAb research; (3) Institutions participating in mAb research; and (4) Cited journals regarding mAb. The technical development, drug development and clinical applications of mAbs were analyzed. Through data analysis, we have identified the new directions for the exploration of mAbs, interactions between mAb technologies and diseases, and evolving global collaboration among institutions.


Effect of reoxygenation on hypofractionated radiotherapy of prostate cancer.
To assess the role of reoxygenation of hypoxic tumor cells in hypofractionated radiotherapy of prostate cancer. The considered radiobiological model is based on the assumption of two populations (compartments) of cells: oxygenated (aerobic) cells and hypoxic cells. After each fraction of radiation, some of the hypoxic cells reoxygenate while a fraction of initially aerobic cells becomes hypoxic. The kinetics of this process between successive treatments is described by coupled, first-order differential equations. To determine the effect of reoxygenation on cell kill in the treatment target, we utilize the linear-quadratic (LQ) model assuming different radiosensitivities for the aerobic and hypoxic cells. Analytical solutions for the number of surviving malignant cells are obtained for special cases of slow and fast reoxygenation. The radiobiological effect of reoxygenation for different fractionation regimens is also evaluated numerically. In this study, a radiobiological model for kinetics of reoxygenation in tumors is used to evaluate different fractionation schedules in radiotherapy of prostate cancer. The obtained results indicate that in the case of low alpha/beta ratio for malignant cells (e.g., α / β = 1.5 Gy), treatment schedule with 4-10 fractions and dose per fraction >4-5 Gy can result in increased cell kill in the treatment target at the same level of rectal toxicity as compared to conventional fractionation. The findings of this study also suggest that radiotherapy of the prostate with 1-3 fractions can be radiobiologically inferior to treatments with greater number of fractions.


It is not just muscle mass: a review of muscle quality, composition and metabolism during ageing as determinants of muscle function and mobility in later life.
Worldwide estimates predict 2 billion people will be aged over 65 years by 2050. A major current challenge is maintaining mobility and quality of life into old age. Impaired mobility is often a precursor of functional decline, disability and loss of independence. Sarcopenia which represents the age-related decline in muscle mass is a well-established factor associated with mobility limitations in older adults. However, there is now evidence that not only changes in muscle mass but other factors underpinning muscle quality including composition, metabolism, aerobic capacity, insulin resistance, fat infiltration, fibrosis and neural activation may also play a role in the decline in muscle function and impaired mobility associated with ageing. Importantly, changes in muscle quality may precede loss of muscle mass and therefore provide new opportunities for the assessment of muscle quality particularly in middle-aged adults who could benefit from interventions to improve muscle function. This review will discuss the accumulating evidence that in addition to muscle mass, factors underpinning muscle quality influence muscle function and mobility with age. Further development of tools to assess muscle quality in community settings is needed. Preventative diet, exercise or treatment interventions particularly in middle-aged adults at the low end of the spectrum of muscle function may help preserve mobility in later years and improve healthspan.


Shutoff pulse illusion.
Visual signals produced by sharp illumination decrements are commonly misinterpreted because of the presence of an illusory sharp increment at the moment of switching. Conditions for occurrence of the illusion are outlined, as well as conditions under which it is not reported.


Reduction of infarct size and infiltration of polymorphonuclear leukocytes by a thromboxane synthetase inhibitor. Studies in a rabbit ischemic heart model.
The effect of sodium 6-(2-(1-(1H)-imidazolyl)methyl-4,5-dihydrobenzo(b) thiophene)carboxylate (RS-5186), a potent and long acting thromboxane synthetase inhibitor in vitro and in vivo, on infarct size and on the infiltration of polymorphonuclear leukocytes (PMNs), was studied in a rabbit coronary artery occlusion (1 h)--reperfusion (0.5 h or 3 h) model. The infarcted region was stained with triphenyltetrazolium, and the ratio of infarcted area/left ventricular area was calculated. The infiltration of PMNs into the infarcted region was determined by measuring the PMNs specific enzyme, myeloperoxidase (MPO) activity. In the vehicle treated group, infarct size and MPO activity were increased with increased reperfusion time from 0.5 h to 3 h (infarct size: 15.3 +/- 2.7 to 25.2 +/- 3.2%; MPO activity: 255 +/- 51 to 825.3 +/- 169.4 units/g wet weight). There was also a significant correlation (r = 0.90, p less than 0.01) between the infarct size and MPO activity. In contrast, in the RS-5186 treated group (2 mg/kg i.v.), both infarct size and MPO activity did not increase with prolongation of the reperfusion period (infarct size: 12.8 +/- 5.5 to 10.3 +/- 3.6%; MPO activity: 318.8 +/- 36.7 to 381.2 +/- 72.6 units/g wet weight). In 0.5 h reperfused samples, there was no significant difference in infarct size or in MPO activity between the vehicle treated group and RS-5186 treated group.(ABSTRACT TRUNCATED AT 250 WORDS)


Analysis of a New Delayed HBV Model with Exposed State and Immune Response to Infected Cells and Viruses.
We propose a comprehensive delayed HBV model, which not only considers the immune response to both infected cells and viruses and a time delay for the immune system to clear viruses but also incorporates an exposed state and the proliferation of hepatocytes. We prove the positivity and boundedness of solutions and analyze the global stability of two boundary equilibria and then study the local asymptotic stability and Hopf bifurcation of the positive (infection) equilibrium and also the stability of the bifurcating periodic solutions. Moreover, we illustrate how the factors such as the time delay, the immune response to infected cells and viruses, and the proliferation of hepatocytes affect the dynamics of the model by numerical simulation.


Accuracy assessment of the Dinamap ProCare 400 in pregnancy and preeclampsia.
To assess the accuracy of the Dinamap ProCare 400 in pregnancy and preeclampsia, according to the British Hypertension Society protocol. Ethical approval was obtained. Forty-five women were recruited at two large teaching hospitals. Nine sequential same arm measurements were taken from each woman alternating between the Dinamap and mercury sphygmomanometry. The device achieved an A/A grade (pregnancy) and an A/B grade (preeclampsia). Mean difference (SD) were 1.1(5.8)/0.1(5.7) mmHg (pregnancy) and -0.5(5.4)/0.9(9.5) mmHg (preeclampsia) for systolic/diastolic. The Dinamap ProCare 400 is recommended for use in pregnancy and has good accuracy in preeclampsia, although caution is still advised.


Effects of exercise training on the distribution of metallic mercury in mice.
1. The purpose of this study was to correlate exercise induced changes of antioxidant enzymes with the distribution of mercury after mercury vapour exposure in mice. 2. Exercise training consisted of swimming (1 h/day for 5 days/week) for 9 weeks. After 9 weeks of training, swim-trained mice showed significantly elevated levels of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSHpx) in their red blood cells, CAT and GSHpx in their kidneys and SOD in the liver. 3. Exercised mice (Ex) and non-exercised mice (N.Ex) were exposed to mercury vapour (3.5 mg m-3) for 1 h. Mercury concentrations were assayed in the blood, brain, heart, lungs, liver and kidneys along with the mercury content of the entire body. The whole body mercury content showed no significant difference in any measurement (immediately, 24 h and 48 h after mercury exposure) between the Ex and N.Ex groups. Mercury concentrations in the Ex group were significantly higher than the N.Ex group in the heart, whole blood, red blood cells and the brain at 24 and 48 h; and in the plasma and kidneys at 24 h. 4. It was concluded that exercise training is a factor in distribution changes of mercury after exposure to mercury vapour, though it is not a factor in the total absorption and excretion of mercury.


Imprinted status of pleomorphic adenoma gene-like I and paternal expression gene 10 genes in pigs.
Genomic imprinting is theorized to exist in all placental mammals and some marsupials. Imprinted genes play important roles in the regulation of fetal growth, development, and postnatal behavior, but the study of imprinted genes has been limited in livestock. In this study, the polymorphism-based approach was used to detect the expression patterns of the porcine pleomorphic adenoma gene-like I (PLAGL1) and paternal expression gene 10 (PEG10) genes. Single nucleotide polymorphisms in the exons were detected between the Meishan and Large White breeds in the PLAGL1 and PEG10 genes. The polymorphisms were used to determine the monoallelic or biallelic expression with reverse transcription-PCR-RFLP in 44 tissues from 4 heterozygous pigs (based on SNP). Imprinting analysis indicated that the PLAGL1 and PEG10 genes were both paternally expressed in all tissues tested (heart, liver, spleen, lung, kidney, stomach, small intestine, skeletal muscle, fat, uterus, and ovary). Our study showed that the method of identifying polymorphic transcripts with reverse transcription-PCR-RFLP may be beneficial for detecting the imprinting status of some candidate imprinted genes.


[Multiple analysis of the difference in intestinal absorption between the main components and the extract of Glycyrrhiza uralensis].
The aim of this study is to investigate the rat intestinal absorption behavior of two main active components, liquiritin, glycyrrhizin and the extract of Glycyrrhiza uralensis. The rat intestinal perfusion model was employed. Concentrations of the compounds of the interest in the intestinal perfusate, bile and plasma samples were determined by HPLC and UPLC. At the same time, the intestinal enzymes incubation test and the partition coefficient determination, the absorption of liquiritin and glycyrrhizin alone and the extract were multiple analyzed. The results showed that the P(eff) (effective permeability) of liquiritin or glycyrrhizin alone or the extract was less than 0.3, which suggested their poor absorption in the intestine. The P(eff) of the two main active components or the extract was not significantly different in duodenum, jejunum, colon and ileum segment. The P(eff) of the glycyrrhizin in the extract had no significant difference in the four intestinal segments compared with the glycyrrhizin alone. The absorption of the liquiritin displayed significant difference (P < 0.05) at ileum segment compared with the liquiritin alone, while it had no markedly change in the other three segments. This phenomenon indicated that some ingredients in the extract might improve the absorption of liquiritin. Moreover, no parent compounds and their metabolites were found in the intestinal perfusate, bile and the plasma samples. The results demonstrated that the influence of the other ingredients in the extract on the two components might not increase the amount of liquiritin and glycyrrhizin in the bile and plasma within the duration of the test.


Mucosal biotransformation.
About 4 million compounds have been described by chemists, and some 60,000 are presently on the market. The search for new chemicals with better properties and less toxicity continues, and future life quality will depend on our ability to find the safest compounds in each field of application. During development of new drugs and chemicals, studies on biotransformation should be done very early, and with adequate analytical tools, in order to get an early understanding of data on bioavailability, metabolic pattern, and toxicity. Though the liver is generally the organ with the highest drug metabolizing activity, it becomes increasingly evident that some extrahepatic organs, such as intestine, kidney, skin, and lung also participate in drug metabolism. The peculiar property of intestinal metabolism is the fact that it modifies chemicals before they enter the circulation. Therefore, an understanding of intestinal metabolism is important for proper interpretation of all pharmacological and toxicological data during development of a new compound. Mucosal biotransformation has recently been reviewed. The present work gives a schematic survey on the topic, shows new trends, and discusses the consequences for toxicological testing of new chemicals.


Increased striatal mRNA and protein levels of the immunophilin FKBP-12 in experimental Parkinson's disease and identification of FKBP-12-binding proteins.
FKBP-12, a 12 kDa FK506-binding protein (neuroimmunophilin), acts as a receptor for the immunosuppressant drug FK506. Neuroimmunophilins, including FKBP-12, are abundant in the brain and have been shown to be involved in reversing neuronal degeneration and preventing cell death. In this report, we have utilized several analytical techniques, such as in situ hybridization, Western blotting, two-dimensional gel electrophoresis, and liquid chromatography electrospray tandem mass spectrometry to study the transcriptional expression as well as protein levels of FKBP-12 in the unilateral 6-hydroxydopamine (6-OHDA) rat model of Parkinson's disease. The FKBP-12 protein was also detected directly on brain tissue sections using mass spectrometry profiling. We found increased levels of FKBP-12 mRNA and protein in the dorsal and middle part of the 6-OHDA lesioned striatum. Thus, these studies clearly demonstrate that FKBP-12 is increased in the brain of a common animal model of Parkinson's disease (PD). Additionally, we have identified potential binding partners to FKBP-12 that may be implicated in the pathophysiology of Parkinson's disease, such as alpha-enolase, 14-3-3 zeta/delta, pyruvate kinase isozymes, and heat shock protein 70, using surface plasmon resonance sensor technology in combination with mass spectrometry. In conclusion, these data strongly suggests that FKBP-12 is altered in an experimental model of PD.