Deglycosylation of isoflavones in isoflavone-rich soy germ flour by Aspergillus oryzae KACC 40247.
Aspergillus oryzae KACC 40247 was selected as an efficient daidzein-producing fungus from strains of the genus Aspergillus by using 5% (w/v) soy germ flour (SGF) as an isoflavone-glycoside-rich medium. The culture conditions, including SGF concentration, agitation speed, initial pH, temperature, and time, were optimized as follows: 7% (w/v) SGF, initial pH 6.0, 33 °C, 300 rpm, and 24 h in a 100 mL baffled flask. The determined amount of isoflavone aglycons in SGF using 50% ethyl acetate was the highest among the solvent systems tested and it was 3.7-fold higher than that using 70% methanol. Under the optimized conditions, the content and concentration of daidzein were 134 mg/g of SGF and 9.4 g/L, respectively, with a productivity of 391 ± 2.8 mg/L/h, and those of isoflavone aglycons were 165 mg/g of SGF and 11.5 g/L, respectively, with a productivity of 479 mg/L/h. Optimization of culture conditions increased the content, concentration, and productivity of isoflavone aglycons by 3.1-, 3.0-, and 3.7-fold, respectively. To our knowledge, this is the highest production of isoflavone aglycons reported to date.


Current practices, needs, and expectations of discussing work with a medical specialist from a patient's perspective: a qualitative study.
Attention to paid work in clinical health care-clinical work-integrating care (CWIC)-might be beneficial for patients of working age. However, the perceptions and expectations of patients about CWIC are unknown. The aim of this study was to develop an understanding of current practices, needs, and expectations among patients for discussing work with a medical specialist. A qualitative study was undertaken involving patients with diverse medical conditions (n = 33). Eight online synchronous focus groups were held. A thematic analysis was then performed. Three themes emerged from the data: (1) the process of becoming a patient while wanting to work again, (2) different needs for different patients, (3) patients' expectations of CWIC. We identified three different overarching categories of work-concerns: (a) the impact of work on disease, (b) the impact of disease or treatment on work ability, and (c) concerns when work ability remained decreased. For each category of concerns, patients expected medical specialists to perform differing roles. Patients indicated that they need support for work-related concerns from their medical specialists and/or other professionals. Currently, not all work concerns received the requested attention, leaving a portion of the patients with unmet needs regarding CWIC.


Sorafenib and radiation therapy for the treatment of advanced hepatocellular carcinoma.
Treatment options for advanced hepatocellular cancer (HCC) are limited. Sorafenib, an orally active multi-kinase inhibitor, is the only systemic agent with a survival benefit in randomized studies. However, the gain in survival is modest and new treatment strategies are needed. We report two cases of advanced HCC treated with a combination of sorafenib and radiation therapy. Impressive and sustained clinical and radiological responses were seen. Treatment was well-tolerated in both cases with no unexpected toxicities reported. There is sound rationale for the combination of sorafenib and radiation therapy for treatment of HCC. These two cases suggest both feasibility and efficacy of this combination, in selected patients. Prospective studies addressing this combination are ongoing.


[Cardiovascular risk in HIV infected patients].
The increased cardiovascular risk has been implicated in both HIV infection and antirretroviral therapy. The endothelial dysfunction triggered by the infection and the metabolic abnormalities associated with therapy predispose to premature atherosclerosis. The recognition of the increased cardiovascular risk in these patients will allow more appropriate prevention strategies, as well as therapy optimization with lipid-lowering agents and antirretroviral therapy modification.


Home ventilator dependence after high cervical cord injury.
Home ventilator support, though somewhat unorthodox, is economically desirable and is now technically feasible. Close cooperation between public health providers, ventilator suppliers, and the family physician, as well as strong patient and family compliance, is essential for success in maintaining a ventilator-dependent patient at home.


Significance of chromosome 9p status in renal cell carcinoma: a systematic review and quality of the reported studies.
Defining the prognosis of renal cell carcinoma (RCC) using genetic tests is an evolving area. The prognostic significance of 9p status in RCC, although described in the literature, remains underutilised in clinical practice. The study explored the causes of this translational gap. A systematic review on the significance of 9p status in RCC was performed to assess its clinical applicability and impact on clinical decision-making. Medline, Embase, and other electronic searches were made for studies reporting on 9p status in RCC. We collected data on: genetic techniques, pathological parameters, clinical outcomes, and completeness of follow-up assessment. Eleven studies reporting on 1,431 patients using different genetic techniques were included. The most commonly used genetic technique for the assessment of 9p status in RCC was fluorescence in situ hybridization. Combined genomic hybridisation (CGH), microsatellite analysis, karyotyping, and sequencing were other reported techniques. Various thresholds and cut-off values were used for the diagnosis of 9p deletion in different studies. Standardization, interobserver agreement, and consensus on the interpretation of test remained poor. The studies lacked validation and had high risk of bias and poor clinical applicability as assessed by two independent reviewers using a modified quality assessment tool. Further protocol driven studies with standardised methodology including use of appropriate positive and negative controls, assessment of interobserver variations, and evidenced based follow-up protocols are needed to clarify the role of 9p status in predicting oncological outcomes in renal cell cancer.


Circular RNAs: biology and clinical significance of breast cancer.
Circular RNAs (circRNAs) are novel noncoding RNAs with covalently closed-loop structures that can regulate eukaryotic gene expression. Due to their stable structure, circRNAs are widely distributed in the cytoplasm and have important biological functions, including as microRNA sponges, RNA-binding protein conjugates, transcription regulators, and translation templates. Breast cancer is among the most common malignant cancers diagnosed in women worldwide. Despite the development of comprehensive treatments, breast cancer still has high mortality rates. Recent studies have unmasked critical roles for circRNAs in breast cancer as regulators of tumour initiation, progression, and metastasis. Further, research has revealed that some circRNAs have the potential for use as diagnostic and prognostic biomarkers in clinical practice. Herein, we review the biogenesis and biological functions of circRNAs, as well as their roles in different breast cancer subtypes. Moreover, we provide a comprehensive summary of the clinical significance of circRNAs in breast cancer. CircRNAs are believed to be a hot focus in basic and clinical research of breast cancer, and innovative future research directions of circRNAs could be used as biomarkers, therapeutic targets, or novel drugs.Abbreviations: CeRNA: Competitive endogenous RNA; ciRNA: Circular intronic RNA; circRNA: Circular RNA; EIciRNA: Exon-intron circRNA; EMT: Epithelial-mesenchymal transition; IRES: Internal ribosome entry site; lncRNA: Long non-coding RNA; miRNA: MicroRNA; MRE: MiRNA response element; ncRNA: Non-coding RNA; RBP: RNA-binding protein; RNA-seq: RNA sequencing; RT-PCR: Reverse transcription-polymerase chain reaction.


Hamstring grafts are tenogenic constructs for ACL reconstruction and Pulsed Electromagnetic Fields improve tendon specific markers expression. An in-vitro study.
Hamstring tendons represent one of the commonest autologous graft used during ACL reconstruction. The harvest of the tendon and the time of tendon processing on the operating table, together with the pretensioning maneuvers and the permanence out of the joint during the time of surgery, might impair tendon derived cells (TCs) viability. The aim of the study was: i) to assess the effective viability of the TCs at the end of the surgical procedure; ii) to investigate if TCs viability and the expression of tendon specific markers may be improved through exposure to prolonged pulsed electromagnetic fields (PEMF) similar to that of clinical practice. Remnants of semitendinosus and gracilis tendons (discarded at the end of the ACL reconstruction) were collected from 13 healthy donors. To isolate TCs, the tendon tissue was minced and digested enzymatically with 0.3% type I collagenase in DMEM with continuous agitation for 15 h at 37°C. The isolated nucleated cells were then plated at 5x103 cells/cm2 in a complete medium composed of DMEM, 10% fetal bovine serum, 50 U/ml Penicillin, 50 mg/ml Streptomycin, 2 mM L-glutamine, and supplemented with 5 ng/ml basic fibroblast growth factor (b-FGF). They were maintained at 37 °C in a humidified atmosphere with 5% CO2, changing culture medium every 3 days. When they reached 80-90% of confluence, the cells were detached by incubation with trypsin/EDTA and then cultured at a density of 5x103 cells/cm2. TCs were cultured in complete medium for 7, 14, 21 days (in chamber slides, to optimize the final immunofluorescence analysis). The following cell cultures were set up: i) TCs cultured with differentiation medium + exposure to PEMF 8 h/day; ii) TCs cultured with differentiation medium without exposure to PEMF. The stimulation with PEMF was generated by a pair of electrical coils, connected with the generator of pulsed electromagnetic fields (PEMF generator system IGEA, Carpi, Italy, intensity of magnetic field = 1.5 mT, frequency = 75 Hz). At day 0, day 7, day 14 and day 21 immunofluorescence analysis was performed to evaluate the expression of tendon specific markers (collagen type I, collagen type VI, scleraxis) and proliferative markers (PCNA, beta-catenin). The TCs from the hamstring tendon fragments at the end of the ACL reconstruction were alive and they expressed markers of proliferation and tendon phenotype at the end of the culture period. The TCs in the presence of PEMF 8h/day showed a greater production of collagen type I, collagen type VI and scleraxis than TCs cultured without PEMF (p<0.05). The expression of these markers increased from 7 to 21 days of culture. The expression of proliferative markers in the presence of PEMF stimulus was significantly lower (p<0.05) than that of TCs cultured without PEMF. Hamstring tendons are not simple "tenoconductive" scaffolds but biologic alive tenogenic constructs rich in cells that can sustain tenogenic behavior and tendon matrix synthesis. Prolonged exposure to PEMF improves their phenotype. Thus, from a clinical perspective, the use of PEMF may represent a possible future strategy to positively influence the early phase of graft remodeling and, ultimately, improve the ligamentization process. Following these concepts, further studies might also exploit the anabolic role of PEMF as an adjunctive postoperative strategy in different tendon pathologies.


Gross anatomy videos: student satisfaction, usage, and effect on student performance in a condensed curriculum.
Anatomy educators are being tasked with delivering the same quantity and quality of material in the face of fewer classroom and laboratory hours. As a result they have turned to computer-aided instruction (CAI) to supplement and augment curriculum delivery. Research on the satisfaction and use of anatomy videos, a form of CAI, on examination performance continues to grow. The purpose of this study was to describe the usage and effect on examination scores of a series of locally produced anatomy videos after an 11% curriculum reduction. First-year medical students (n = 40) were given access to the videos and the prior year's students (n = 40) were used as historical controls. There was no significant difference in demographics between the two groups. The survey response rate was 85% (n = 34) in the experimental group. The students found the videos to be highly satisfying (median = 5 on a five-point Likert scale, interquartile range = 1) and used them on average 1.55 times/week (SD ± 0.77). Availability of the videos did have a statistically significant effect (4% improvement) on the final laboratory examination (p = 0.039). This suggests that the videos were a well-received form of CAI that may be useful in bridging the gap created by a reduction in gross anatomy course contact hours.


Rac2 GTPase activation by angiotensin II is modulated by Ca2+/calcineurin and mitogen-activated protein kinases in human neutrophils.
Angiotensin II (Ang II) highly stimulates superoxide anion production by neutrophils. The G-protein Rac2 modulates the activity of NADPH oxidase in response to various stimuli. Here, we describe that Ang II induced both Rac2 translocation from the cytosol to the plasma membrane and Rac2 GTP-binding activity. Furthermore, Clostridium difficile toxin A, an inhibitor of the Rho-GTPases family Rho, Rac and Cdc42, prevented Ang II-elicited O2-/ROS production, phosphorylation of the mitogen-activated protein kinases (MAPKs) p38, extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase 1/2, and Rac2 activation. Rac2 GTPase inhibition by C. difficile toxin A was accompanied by a robust reduction of the cytosolic Ca(2)(+) elevation induced by Ang II in human neutrophils. Furthermore, SB203580 and PD098059 act as inhibitors of p38MAPK and ERK1/2 respectively, wortmannin, an inhibitor of phosphatidylinositol-3-kinase, and cyclosporin A, a calcineurin inhibitor, hindered both translocation of Rac2 from the cytosol to the plasma membrane and enhancement of Rac2 GTP-binding elicited by Ang II. These results provide evidence that the activation of Rac2 by Ang II is exerted through multiple signalling pathways, involving Ca(2)(+)/calcineurin and protein kinases, the elucidation of which should be insightful in the design of new therapies aimed at reversing the inflammation of vessel walls found in a number of cardiovascular diseases.


Recovery of complete genome sequences of Crimean-Congo haemorrhagic fever virus (CCHFV) directly from clinical samples: A comparative study between targeted enrichment and metagenomic approaches.
Crimean-Congo haemorrhagic fever (CCHF) is the most prevalent human tick-borne viral disease, endemic to the Balkans, Africa, Middle East and Asia. There are currently no licensed vaccines or effective antivirals against CCHF. CCHF virus (CCHFV) has a negative sense segmented tripartite RNA genome consisting of the small (S), medium (M) and large (L) segments. Depending on the segment utilised for genetic affiliation, there are up to 7 circulating lineages of CCHFV. The current lack of geographical representation of CCHFV sequences in various repositories highlights a requirement for increased CCHFV sequencing capabilities in endemic regions. We have optimised and established a multiplex PCR tiling methodology for the targeted enrichment of complete genomes of Europe 1 CCHFV lineage directly from clinical samples and compared its performance to a non-targeted enrichment approach on both short-read and long-read sequencing platforms. We have found a statistically significant increase in mapped viral sequencing reads produced with our targeted enrichment approach. This has allowed us to recover near complete S segment sequences and above 90% of the M and L segment sequences for samples with Ct values as high as 31.3. This study demonstrates the superiority of a targeted enrichment approach for recovery of CCHFV genomic sequences from samples with low virus titre. CCHFV is an important vector-borne human pathogen with wide geographical distribution. The validated methodology reported here adds value to front-line public health laboratories employing genomic sequencing for CCHFV Europe 1 lineage surveillance, particularly in the Balkan and Middle Eastern territories currently monitoring the spread of the pathogen. Tracking the genomic evolution of the virus across regions improves risk assessment and directly informs the development of diagnostics, therapeutics, and vaccines.


Spina bifida and birth outcome before and after fortification of flour with iron and folic acid in Oman.
This paper presents the trend of spina bifida and other neural tube defects in Oman after the nationwide implementation of folate supplementation of pregnant women in 1990 and the fortification of wheat flour with iron and folate in 1996. The annual incidence of spina bifida fluctuated from 2.34 to 4.03 per 1000 deliveries between 1991 and 1996, but fell sharply to 2.11 per 1000 deliveries in 1997, after which the downward trend continued, reaching 0.29 per 1000 deliveries by 2006. The rate of other neural tube defects remained almost constant. The reduction in spina bifida rates in Oman could be linked to the start of flour fortification but not the supplementation programme.


[Fibroma of the small intestine with heterotopia of the pyloric glands in the mucous membrane covering the tumor].
Small bowel tumour, 5 X 3 X X 3 cm in size, in a woman of 52 provoked intestinal invagination. Small bowel was resected with the tumour which was diagnosed histologically as a soft fibroma. Focal areas of the pyloric glands with single argyrophil endocrine cells as well as "stomach metaplasia" of the epithelium of one crypt were detected in the intestinal mucous membrane covering tumour. In the author's opinion, this heterotopia is related to the developmental malformation.


Enhanced recovery after thoracic surgery reduces discharge on highly dependent narcotics.
There is large prescription drug epidemic in United States. We want to determine if ERATS (enhanced recovery after thoracic surgery) program can reduce discharge on highly dependent narcotics. We performed a retrospective analysis of prospectively collected data on patients who underwent lung resection and foregut procedures on thoracic surgery service over an 8-month time period. Patients underwent preoperative conditioning instructions, multimodal non-narcotic pharmaceutical usage, total intravenous anesthesia (TIVA) and minimizing highly addictive narcotics during the post-operative period. We gathered information on demographics, indication and type of surgery, morbidity, mortality and length of stay. We also recorded the type of pain medication patients were given as a prescription based on the Drug Enforcement Agency's classification schedule. Fifty-two patients underwent lung resection and 54 patients underwent foregut surgery. There were no mortalities in either group. Ten percent of patients after lung surgery and 6% after foregut surgery had a greater than grade II complication. The median length of stay after lung resection was 2 days and foregut surgery was 1 day. Only 10% of patients went home after lung resection and 2% after foregut surgery with a prescription for schedule II narcotics. We found that patients who were on schedule II narcotics prior to surgery all went home with schedule II narcotics. We found that ERATS program for thoracic surgical cases can reduce the number of patients going home with highly dependent narcotics. This strategy will decrease the availability of highly addictive prescription drugs in the community.


Utilization and costs of health care after geriatric traumatic brain injury.
Despite the growing number of older adults experiencing traumatic brain injury (TBI), little information exists regarding their utilization and cost of health care services. Identifying patterns in the type of care received and determining their costs is an important first step toward understanding the return on investment and potential areas for improvement. We performed a health care utilization and cost analysis using the National Study on the Costs and Outcomes of Trauma (NSCOT) dataset. Subjects were persons 55-84 years of age with TBI treated in 69 U.S. hospitals located in 14 states (n=414, weighted n=1038). Health outcomes, health care utilization, and 1-year costs of care following TBI in 2005 U.S. dollars were estimated from hospital bills, patient surveys, medical records, and Medicare claims data. The subjects were further analyzed in three subgroups (55-64, 65-74, and 75-84 years of age). Unadjusted cost models were built, followed by a second set of models adjusting for demographic and pre-injury health status. Those in the oldest category (75-84 years) had significantly higher numbers of re-hospitalizations, home health care visits, and hours per week of unpaid care, and significantly lower numbers of physician and mental health professional visits than younger age groups (age 55-64 and 65-74 years). Significant age-related differences were seen in all health outcomes tested at 12 months post-injury except for incidence of depressive symptoms. One-year total treatment costs did not differ significantly across age categories for brain-injured older adults in either the unadjusted or adjusted models. The unadjusted total mean 1-year cost of care was $77,872 in persons aged 55-64 years, $76,903 in persons aged 65-74 years, and $72,733 in persons aged 75-84 years. There were significant differences in cost drivers among the age groups. In the unadjusted model index hospitalization costs and inpatient rehabilitation costs were significantly lower in the oldest age category, while outpatient care costs and nursing home stays were lower in the younger age categories. In the adjusted model, in addition to these cost drivers, re-hospitalization costs were significantly higher among those 75-84 years of age, and receipt of informal care from friends and family was significantly different, being lowest among those aged 65-74 years, and highest among those aged 75-84 years. Identifying variations in care that these patients are receiving and determining the costs versus benefits is an important next step in understanding potential areas for improvement.


Evidence-based support for autistic people across the lifespan: maximising potential, minimising barriers, and optimising the person-environment fit.
Autism is both a medical condition that gives rise to disability and an example of human variation that is characterised by neurological and cognitive differences. The goal of evidence-based intervention and support is to alleviate distress, improve adaptation, and promote wellbeing. Support should be collaborative, with autistic individuals, families, and service providers taking a shared decision-making approach to maximise the individual's potential, minimise barriers, and optimise the person-environment fit. Comprehensive, naturalistic early intervention with active caregiver involvement can facilitate early social communication, adaptive functioning, and cognitive development; targeted intervention can help to enhance social skills and aspects of cognition. Augmentative and alternative communication interventions show preliminary evidence of benefit in minimising communication barriers. Co-occurring health issues, such as epilepsy and other neurodevelopmental disorders, sleep problems, and mental health challenges, should be treated in a timely fashion. The creation of autism-friendly contexts is best achieved by supporting families, reducing stigma, enhancing peer understanding, promoting inclusion in education, the community, and at work, and through advocacy.


An empirical study of choosing efficient discriminative seeds for oligonucleotide design.
Oligonucleotide design is known as a time-consuming work in bioinformatics. In order to accelerate and be efficient the oligonucleotide design process, one of widely used approach is the prescreening unreliable regions using a hashing (or seeding) algorithm. Since the seeding algorithm is originally proposed to increase sensitivity for local alignment, the specificity should be considered as well as the sensitivity for the oligonucleotide design problem. However, a measure of evaluating the seeds regarding how adequate and efficient they are in the oligo design is not yet proposed. Here, we propose novel measures of evaluating the seeding algorithms based on the discriminability and the efficiency. To evaluate the proposed measures, we examine five seeding algorithms in oligonucleotide design. We carried out a series of experiments to compare the seeding algorithms. As the result, the spaced seed is recorded as the most efficient discriminative seed for oligo design. The performance of transition-constrained seed is slightly lower than the spaced seed. Because BLAT seeding algorithm and Vector seeding algorithm give poor scores in specificity and efficiency, we conclude that these algorithms are not adequate to design oligos. Consequently, we recommend spaced seeds or transition-constrained seeds with 15 approximately 18 weight in order to design oligos with the length of 50 mer. The empirical experiments in real biological data reveal that the recommended seeds show consequently good performance. We also propose a software package which enables the users to get the adequate seeds under their own experimental conditions. Our study is valuable to the two points. One is that our study can be applied to the oligo design programs in order to improve the performance by suggesting the experiment-specific seeds. The other is that our study is useful to improve the performance of the mapping assembly in the field of Next-Generation Sequencing. Our proposed measures are originally designed to be used for oligo design but we expect that our study will be helpful to the other genomic tasks.


Clonal composition of T cells in lymphomatoid papulosis.
A cDNA of the C beta 2 gene of the T-cell receptor was used as a probe to investigate the clonal composition of T cells in skin lesions of 5 patients with lymphomatoid papulosis (LyP), a chronic recurrent eruption characterized by morphologically abnormal activated T cells in the cutaneous infiltrate. Clonal T-cell populations, as evidenced by rearranged DNA bands, were demonstrated in the skin lesions of four patients, one of whom has shown clinical progression toward lymphoma. Three of these patients had lesions of type A histology, a type previously shown to be associated with aneuploidy. The remaining patient with clonal lesions appeared to have the same gene rearrangement pattern in DNA obtained from separate lesions taken 11 months apart, providing evidence that the T cells in both sites were derived from the same clone. This patient had lesions of type B histology, which is not associated with aneuploidy. Absence of a rearranged band and deletion or near absence of the 10.8 kb band in Eco RI digests was interpreted as evidence of polyclonal T-cell hyperplasia, accounting for the skin infiltrate of a fifth patient who had a prolonged clinical course without progression to lymphoma. This patient had lesions of type A histology with frequent Ki-1-positive Reed-Sternberg-like cells. Our results show that gene rearrangement analysis provides information that is independent of histology in LyP and may in part explain the variable progression of LyP to lymphoma in 10-20% of patients.


[Alpha-methylglucoside transmembrane phosphorylation and regulation of the beta-galactoside permease activity in E. coli K12].
The interaction between alpha-methylglucoside (MeGlc) and beta-galactoside transport systems in E. coli K12 was studied. It was shown that an addition of MeGlc to bacterial cells leads to repression of [14C] lactose accumulation and o-nitrophenyl-beta-D-galactopyranoside (NO2PheGal) hydrolysis. The mutational damage of one of the components of the MeGlc transport system is accompanied by elimination of the glucoside inhibitory action. Intracellular MeGlcP and GlcP lower the efficiency of the transmembraneous transfer of beta-galactosides. The data obtained suggest that repression of the beta-galactoside permease activity during transport of the MeGlc is a result of two processes: i.e. phosphorylation-coupled translocation of MeGlc and intracellular accumulation of MeGlcP. An assumption on the intramembraneous interaction of enzyme IIGlc with beta-galactoside permease is made.


EEG Channel Correlation Based Model for Emotion Recognition.
Emotion recognition using Artificial Intelligence (AI) is a fundamental prerequisite to improve Human-Computer Interaction (HCI). Recognizing emotion from Electroencephalogram (EEG) has been globally accepted in many applications such as intelligent thinking, decision-making, social communication, feeling detection, affective computing, etc. Nevertheless, due to having too low amplitude variation related to time on EEG signal, the proper recognition of emotion from this signal has become too challenging. Usually, considerable effort is required to identify the proper feature or feature set for an effective feature-based emotion recognition system. To extenuate the manual human effort of feature extraction, we proposed a deep machine-learning-based model with Convolutional Neural Network (CNN). At first, the one-dimensional EEG data were converted to Pearson's Correlation Coefficient (PCC) featured images of channel correlation of EEG sub-bands. Then the images were fed into the CNN model to recognize emotion. Two protocols were conducted, namely, protocol-1 to identify two levels and protocol-2 to recognize three levels of valence and arousal that demonstrate emotion. We investigated that only the upper triangular portion of the PCC featured images reduced the computational complexity and size of memory without hampering the model accuracy. The maximum accuracy of 78.22% on valence and 74.92% on arousal were obtained using the internationally authorized DEAP dataset.


A resistance determinant to nucleic acid-binding compounds in methicillin-resistant Staphylococcus aureus.
Recent isolates of methicillin-resistant Staphylococcus aureus from Australia and several other countries carry a plasmid coding for high levels of resistance to propamidine isethionate and low levels of resistance to cetyltrimethyl-ammonium bromide. The reasons for the acquisition and retention of such a determinant are not known. To define the properties of this resistance determinant in more detail, the minimum inhibitory concentrations of a series of cationic agents, including ethidium bromide, were determined and a cationic-resistance profile prepared for several strains of S. aureus and their isogenic, sensitive derivatives. The compounds for which resistance is coded by this determinant all bind to nucleic acids. Hence the determinant has been designated the NAB (nucleic acid-binding compounds)-resistance determinant.


Immunological regulation of synaptic transmission in isolated guinea pig autonomic ganglia.
We have attempted to identify a role for mast cells in autonomic ganglia by examining the effects of antigen challenge on mast cell-associated mediator release and synaptic transmission through the superior cervical ganglion isolated from ovalbumin-sensitized guinea pigs. Ovalbumin induced the release of 7.9 ng of histamine, 40 pg of immunoreactive sulfidopeptide-leukotriene, and 140 pg of immunoreactive-PgD2 per ganglion. Ovalbumin produced long-lasting potentiation (51 +/- 4%, mean +/- SEM, n = 66) of synaptic transmission, the protracted nature of which could not be mimicked by exogenous histamine (10(-5) M). Selective histamine H1 antagonists inhibited the antigen-induced potentiation, but did not reverse it when added any time after antigen exposure. These results indicate that immunologic activation of mast cells can directly potentiate neurotransmission in sympathetic ganglia. Histamine appears to be a mediator involved in the induction of antigen-induced potentiation of synaptic transmission, but alone cannot account for the long term nature of this phenomenon.


A DNA nick at Ku-blocked double-strand break ends serves as an entry site for exonuclease 1 (Exo1) or Sgs1-Dna2 in long-range DNA end resection.
The repair of DNA double-strand breaks (DSBs) by homologous recombination (HR) is initiated by nucleolytic resection of the DNA break ends. The current model, being based primarily on genetic analyses in Saccharomyces cerevisiae and companion biochemical reconstitution studies, posits that end resection proceeds in two distinct stages. Specifically, the initiation of resection is mediated by the nuclease activity of the Mre11-Rad50-Xrs2 (MRX) complex in conjunction with its cofactor Sae2, and long-range resection is carried out by exonuclease 1 (Exo1) or the Sgs1-Top3-Rmi1-Dna2 ensemble. Using fully reconstituted systems, we show here that DNA with ends occluded by the DNA end-joining factor Ku70-Ku80 becomes a suitable substrate for long-range 5'-3' resection when a nick is introduced at a locale proximal to one of the Ku-bound DNA ends. We also show that Sgs1 can unwind duplex DNA harboring a nick, in a manner dependent on a species-specific interaction with the ssDNA-binding factor replication protein A (RPA). These biochemical systems and results will be valuable for guiding future endeavors directed at delineating the mechanistic intricacy of DNA end resection in eukaryotes.


Experiences of Family Caregivers of Patients with Borderline Personality Disorder of Social Stigma.
Social stigma is the most common and challenging burden of care on the family of people with Borderline Personality Disorder (BPD) In Iran, despite the cultural and social influences, this issue has been less studied. Therefore, present study was conducted to determine the lived experiences of caregivers of patients with BPD of social stigma. This qualitative study was performed at Ibn Sina hospital in Mashhad, Iran from 2017 to 2019. Participants were selected by purposive and snowball sampling method. Data were collected through semi-structured interviews. Data saturation was achieved after 16 interviews. Finally, the data were analyzed by the method proposed by Diekelmann (1989). In data analysis, one main theme and two sub-themes emerged. The main themes include Black shadow. Two sub-themes consisted of society dagger and secrecy. The sub-theme of society dagger included the two common meanings (inner turmoil in response to the stigma of others and weakening of family status among relatives and acquaintances). The sub-theme of secrecy comprised of the three common meanings (concealment of disease, hide hospitalization, and seclusion). An understanding of the experience of family stigma can lead to the development of supportive strategies to manage this problem among caregivers of patients with BPD. Nurses can support caregivers by offering them opportunities to discuss how stigma is disrupting their caregiving roles. They can also support the caregivers in negotiating the experienced social and emotional distress and when necessary, refer them to the other members of healthcare teams.


Within-visit SBP variability from childhood to adulthood and markers of cardiovascular end-organ damage in mid-life.
Within-visit SBP variability is associated with age and SBP, but its long-term clinical significance is unknown. We examined the association between child, adult, and life-time within-visit SBP variability with markers of end-organ damage using data from a 31-year longitudinal study. Within-visit SBP variability was calculated as the standard deviation of three sitting SBP readings among up to 3010 participants aged 6-18 years (childhood) who were re-measured up to seven times to mid-adulthood. Markers of cardiovascular end-organ damage in adulthood were carotid intima--media thickness, brachial flow-mediated dilatation, carotid distensibility, pulse wave velocity, left ventricular mass index, carotid plaque, and coronary artery calcification. The mean (standard deviation) cumulative within-visit SBP variability was 2.7 (1.5) mmHg in childhood, 3.9 (1.9) mmHg in adulthood and 3.7 (1.5) mmHg across the observed life-time. Childhood within-visit SBP variability was not correlated with its subsequent values measured from 3 to 31 years later. With adjustment for age, sex, cumulative SBP, BMI and serum lipids, neither child, adult, or life-time cumulative within-visit SBP variability associated with markers of cardiovascular end-organ damage. However, higher child, adult, and life-time cumulative SBP significantly associated with higher carotid intima--media thickness, higher pulse wave velocity, lower brachial flow-mediated dilatation, lower carotid distensibility in adulthood. Within-visit SBP variability from childhood to adulthood does not provide additional predictive utility over SBP over the same period of the life course.


Premature ovarian failure in nobox-deficient mice is caused by defects in somatic cell invasion and germ cell cyst breakdown.
To understand the mechanism of premature ovarian failure (POF). The ultrastructural (electron microscopy) analysis of primordial ovarian follicles in Nobox deficient mice. We studied, for the first time, the fate of oogonia in embryonic (prenatal) mouse ovaries and showed that the abolishment of the transition from germ cell cysts to primordial follicles in the ovaries of Nobox deficient mice is caused by defects in germ cell cyst breakdown, leading to the formation of syncytial follicles instead of primordial follicles. These results indicate that POF syndrome in Nobox deficient mice results from the faulty signaling between somatic and germ line components during embryonic development. In addition, the extremely unusual and abnormal presence of adherens junctions between unseparated oocytes within syncytial follicles indicates that faulty communication between somatic and germ cells is involved in, or leads to, abnormalities in the cell adhesion program.


Left atrial distension and antiorthostatic decrease in arterial pressure and heart rate in humans.
It was investigated to what degree left atrial distension augments the hypotensive effects of a 15-min moderate antiorthostatic maneuver in humans. Ten healthy males underwent a posture change from upright seated (Seat, legs horizontal) to supine (Sup) or to supine with simultaneous lower body negative pressure (Sup + LBNP) to keep left atrial diameter (LAD) unchanged. After 2.5 min of Sup, mean arterial pressure (MAP) decreased from 94 +/- 3 to 86 +/- 3 mmHg (P < 0.05), whereas a similar decrease was delayed 7.5 min into Sup + LBNP. Heart rate (HR) decreased within 2.5 min of Sup from 68 +/- 2 to 60 +/- 3 beats/min (P < 0.05) and remained significantly decreased for at least 2.5 min longer than during Sup + LBNP. Aortic systolic distension (ASD) increased by 59 +/- 17% during Sup (P < 0.05) but was unchanged during Sup + LBNP. The 29 +/- 4% decrease in plasma norepinephrine (NE) during Sup (P < 0.05) was abolished during Sup + LBNP. In conclusion, the increases in LAD and ASD seem important stimuli for the prompt decrease in MAP, the 2.5-min longer-lasting decrease in HR, and the sustained decrease in NE during a 15-min moderate antiorthostatic posture change in humans.


Stent-grafts placement for treatment of massive hemorrhage from ruptured hepatic artery after pancreaticoduodenectomy.
To present a series of cases with life-threatening hemorrhage from ruptured hepatic artery pseudoaneurysm after pancreaticoduodenectomy (PD) treated with placement of stent-grafts. Massive hemorrhage from ruptured hepatic artery pseudoaneurysm after PD in 9 patients (6 men, 3 women) at the age of 23-75 years (mean 48 years), were treated with placement of percutaneous endovascular balloon-expandable coronary stent-grafts. All patients were not suitable for embolization because of a non-patent portal vein. One or more stent-grafts, ranging 3-6 mm in diameter and 16-55 mm in length, were placed to exclude ruptured pseudoaneurysm. Follow-up data, including clinical condition, liver function tests, and Doppler ultrasound examination, were recorded at the outpatient clinic. Immediate technical success was achieved in all the 9 patients. All stent-grafts were deployed in the intended position for immediate cessation of bleeding and preservation of satisfactory hepatic arterial blood flow. No significant procedure-related complications occurred. Recurrent bleeding occurred in 2 patients at 16 and 24 h, respectively, after placement of stent-grafts and treated with surgical revision. One patient died of sepsis 12 d after the interventional procedure. The remaining 6 patients were survived when they were discharged. The mean follow-up time was 10.5 mo (range 4-16 mo). No patient had recurrent bleeding after discharge. Doppler ultrasound examination verified the patency of hepatic artery and stent-grafts during the follow-up. Placement of stent-grafts is an effective and safe procedure for acute life-threatening hemorrhage from ruptured hepatic artery pseudoaneurysm.


Induction of monocyte-macrophage procoagulant activity by transformed cell lines.
Human peripheral blood monocytes and mature macrophages were found to produce significant procoagulant activity (PCA), identified as tissue factor, on exposure to a variety of human (K562, HL60, Raji) and murine (TU5, NS-1) transformed cell lines. The monocyte procoagulant response was vigorous, generating PCA to a level nearly comparable to the response to endotoxin, a known stimulant for monocyte PCA. The response was rapid and could be fully elicited, in a dose-dependent fashion, within 4 hr with HL60 and Raji cell lines and within 14 hr with K562, TU5, and NS-1 cells. The monocyte PCA-inducing activity was found to reside in the membrane fraction of transformed cells. Other transformed human (Laz 509, Laz 221, Laz 156, U937, CEM) and murine (L1210, P815, TLX9, WEHI 164) cell lines had little, if any, activity. The induction of monocyte PCA by transformed cells most probably was not due to an allogeneic signal, as 1) the K562 and HL60 cell lines were potent PCA inducers despite the lack of class II histocompatibility antigen expression, whereas Laz 156, which did express HLA antigens, was ineffective; 2) mouse peritoneal macrophages responded with the production of strong PCA to the syngeneic transformed cell lines TU5 and NS-1. The monocyte-macrophage procoagulant response to transformed cell lines appeared to be independent of T lymphocytes. Indeed, monocytes purified on the basis of reactivity with monoclonal antibody Mo2 and sorting or depleted of contaminating T cells by anti-T3 antibody and complement responded similarly to conventional monocyte preparations. The production of tissue factor by monocyte-macrophages in response to exposure to some tumor cells may represent a mechanism whereby blood coagulation is activated in malignancy.


Electrospray ionization quadrupole time-of-flight and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometric analyses to solve micro-heterogeneity in post-translationally modified peptides from Phoneutria nigriventer (Aranea, Ctenidae) venom.
Previous studies of the fractionated venom of the Brazilian armed spider Phoneutria nigriventer, obtained by gel filtration, have demonstrated the presence of a fraction PhM, a pool of small peptides (up to 2000 Da) that provoke contractions in smooth muscle of guinea pig ileum. Initial attempts to sequence these peptides were largely unsuccessful because of the low purification yield and the fact that the majority seemed to be blocked at their N-termini. In the present work, analysis of this venom fraction by mass spectrometry has revealed the existence of a highly complex mixture of peptides with molecular weights corresponding to those observed for the muscle-active peptides previously described (800-1800 Da). These peptides appear to be a family of isoforms with some particular features. The amino acid sequences of 15 isoforms have been determined by tandem mass spectrometry (MS/MS) using both electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q/ToFMS) and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-ToF/ToFMS). These molecules contain post-translational modifications such as proteolysis and C-terminal amidation, which combine to generate additional isoforms. All the isoforms sequenced in this study possess an N-terminal pyroglutamic acid residue. A search for sequence similarities with other peptides in databanks revealed that these peptides are structurally related to the tachykinins, a family of neuro-hormone peptides. The data obtained in this study will be essential for the subsequent steps of this research, the synthesis of these peptides and pharmacological characterization of their biological activity.


Investigating the Chemolithoautotrophic and Formate Metabolism of Nitrospira moscoviensis by Constraint-Based Metabolic Modeling and 13C-Tracer Analysis.
Nitrite-oxidizing bacteria belonging to the genus Nitrospira mediate a key step in nitrification and play important roles in the biogeochemical nitrogen cycle and wastewater treatment. While these organisms have recently been shown to exhibit metabolic flexibility beyond their chemolithoautotrophic lifestyle, including the use of simple organic compounds to fuel their energy metabolism, the metabolic networks controlling their autotrophic and mixotrophic growth remain poorly understood. Here, we reconstructed a genome-scale metabolic model for Nitrospira moscoviensis (iNmo686) and used flux balance analysis to evaluate the metabolic networks controlling autotrophic and formatotrophic growth on nitrite and formate, respectively. Subsequently, proteomic analysis and [13C]bicarbonate and [13C]formate tracer experiments coupled to metabolomic analysis were performed to experimentally validate model predictions. Our findings corroborate that N. moscoviensis uses the reductive tricarboxylic acid cycle for CO2 fixation, and we also show that N. moscoviensis can indirectly use formate as a carbon source by oxidizing it first to CO2 followed by reassimilation, rather than direct incorporation via the reductive glycine pathway. Our study offers the first measurements of Nitrospira's in vivo central carbon metabolism and provides a quantitative tool that can be used for understanding and predicting their metabolic processes. IMPORTANCE Nitrospira spp. are globally abundant nitrifying bacteria in soil and aquatic ecosystems and in wastewater treatment plants, where they control the oxidation of nitrite to nitrate. Despite their critical contribution to nitrogen cycling across diverse environments, detailed understanding of their metabolic network and prediction of their function under different environmental conditions remains a major challenge. Here, we provide the first constraint-based metabolic model of Nitrospira moscoviensis representing the ubiquitous Nitrospira lineage II and subsequently validate this model using proteomics and 13C-tracers combined with intracellular metabolomic analysis. The resulting genome-scale model will serve as a knowledge base of Nitrospira metabolism and lays the foundation for quantitative systems biology studies of these globally important nitrite-oxidizing bacteria.


The effects of elevated glucose on Na+/K(+)-ATPase of cultured bovine retinal pigment epithelial cells measured by a new nonradioactive rubidium uptake assay.
The effects of stimulated hyperglycemia on the Na+/K(+)-ATPase activity of cultured bovine retinal pigment epithelial (RPE) cells were investigated. Total Rb+ uptake, measured by a chromatographic method, was decreased 20-30% by 55.5 mM glucose relative to 5.55 mM glucose for culture periods of 2 to 28 days. An acute hyperglycemic stress (< 1 week) had no effect on ouabain-inhibition of Rb+ uptake or ouabain binding to RPE cells (IC50 = 55 nM for both processes) and did not alter the IC50 value (near 10 nM) for binding of strophanthidin, another selective Na+/K(+)-ATPase inhibitor. A small increase in the apparent K(m) of Rb+ for Na+/K(+)-ATPase accompanied the decrease in maximal Rb+ uptake at 55.5 mM glucose. The continuous presence of AL-1576, an aldose reductase inhibitor (ARI), normalized the effect of severe hyperglycemia on Rb+ uptake in the chronic (28 days) but not the acute exposure protocols. Thus, decreased efficiency of Na+/K(+)-ATPase caused by chronic accumulation of intracellular sorbitol can account for previously reported functional and structural alterations in the RPE cell layer of diabetic rodents. The results of the present study suggest that hyperglycemia-induced loss of Na+/K(+)-ATPase function in RPE cells, which responds to aldose reductase inhibitor treatment, contributes to the pathogenesis of diabetic retinopathy.


Beta-defensin 22 is a major component of the mouse sperm glycocalyx.
Surface components of sperm isolated from the cauda epididymides were stabilized by whole sperm fixation for immunization of rabbits. The resulting immunoglobulins (Igs) recognized a single protein of 130 kDa (non-reduced) or 54-57 kDa (reduced) on western blots of cauda sperm. Igs recognized the same 54-57 kDa protein band on whole tissue blots of the corpus and cauda epididymidis and vas deferens. No immunoreactive bands were detected on blots of the prostate, seminal vesicles, testes, caput epididymis, or any of various non-reproductive tissues. Removal of sperm from the vas deferens prior to blotting eliminated the detection of the sperm antigen. Antibodies raised to synthetic peptides, identical in amino acid sequence to two unique spans of DEFB22, recognized the same 130/54-57 kDa antigen on western blots of both caudal sperm and the purified antigen isolated with the anti-sperm Ig. From indirect immunofluorescence, both the anti-sperm and anti-peptide Igs appeared to localize to the entire sperm surface, a pattern confirmed at the ultrastructural level. Real-time PCR identified the corpus epididymides as the major site of expression of DEFB22, with negligible expression in the testes, caput epididymides, and vas deferens. Immunostaining of epididymal sections showed DEFB22 being released into the lumen at the distal caput/proximal corpus, with sperm becoming intensely coated with DEFB22 as they reached the distal corpus. Most uterine sperm recovered from mice 4 h following copulation exhibited DEFB22 coating the entire sperm surface. By contrast, some sperm recovered from the oviduct and cumulus extracellular matrix showed loss of DEFB22 from the sperm head.


Evaluation of radial and ulnar blood flow after radial artery cannulation with 20- and 22-gauge cannulae using duplex Doppler ultrasound.
This study evaluated ulnar and radial artery blood flow after radial artery cannulation during general anaesthesia using Doppler ultrasound. A total of 80 patients were randomly assigned to receive radial artery cannulation with either a 20-G or 22-G cannula. Arterial diameter, peak systolic velocity, end-diastolic velocity, resistance index and mean volume flow were measured at four time points in both arteries: before anaesthesia; 5 min after intubation; immediately after cannulation; and 5 min after cannulation. After radial artery cannulation, ulnar diameters and blood flow were significantly increased, and persisted until 5 min after cannulation. Radial blood flow was decreased immediately after cannulation and recovered to pre-cannulation values 5 min after cannulation. There were no statistical differences between groups at each time point. Radial artery cannulation causes compensatory increase in ulnar artery blood flow, and the difference in cannula size has minimal effect on this change.


Amphotericin B resistance of Aspergillus terreus in a murine model of disseminated aspergillosis.
The in-vivo activity of amphotericin B and itraconazole against a clinical isolate of Aspergillus terreus was determined in a murine model of disseminated aspergillosis. MICs of amphotericin B and itraconazole for the strain, determined by an NCCLS-based technique, were 2 microg/ml and 1 microg/ml, respectively. Mice infected intravenously were treated with either itraconazole (50 or 100 mg/kg/day) or amphotericin B 4.5 mg/kg/day for 10 days. Treatment with both doses of itraconazole significantly prolonged the survival rates compared with those for untreated mice. In comparison, mortality rate and median survival time were identical for mice treated with amphotericin B and for mice given no therapy, indicating that the strain was highly resistant to amphotericin B in this model. Analysis of sterol composition showed that the major sterol was ergosterol. This suggests that amphotericin B resistance was not related to a modified sterol profile.


Vagus nerve stimulation: 2-year outcomes for bipolar versus unipolar treatment-resistant depression.
The outcome of vagus nerve stimulation (VNS) for patients with bipolar treatment-resistant depression (TRD) has not been well characterized. This study describes the outcome of VNS for bipolar TRD patients participating in the acute and longitudinal pivotal trials and compares their outcome with unipolar TRD patients in the same trials. Of 235 participants enrolled in the acute study, 25 (11%) were diagnosed with DSM-IV bipolar I or II disorder. A sham-controlled 12-week trial of VNS preceded 2 years of open treatment. Bipolar and unipolar subjects were compared on baseline characteristics as well as acute and long-term outcomes. At baseline, bipolar TRD was as severe as unipolar TRD but with depressive episodes of shorter duration and more failed antidepressant trials/year. Acute, 1-year, and 2-year outcomes were similar for both groups, even when the definition of response for bipolar TRD was expanded to include lack of manic symptoms. Bipolar TRD is a serious condition. In this hypothesis-generating analysis, VNS short- and long-term effects on bipolar and unipolar TRD were similar. Because these analyses were post hoc, these findings should not be interpreted as warranting clinical inference regarding effectiveness of VNS in patients with bipolar depression.


Everolimus and basiliximab permit suppression by human CD4+CD25+ cells in vitro.
Immunosuppressive drugs are essential for the prevention of acute transplant rejection but some may not promote long-term tolerance. Tolerance is dependent on the presence and regulatory function of CD4(+)CD25(+) T cells in a number of animal models. The direct effects of immunosuppressive drugs on CD4(+)CD25(+) cells, particularly those that interfere with IL-2 signaling are uncertain. We studied the effects of the rapamycin derivative everolimus and the anti-CD25 monoclonal antibody basiliximab on the regulatory capacity of human CD4(+)CD25(+) cells in vitro. Both drugs permitted the suppression of proliferation and IFN-gamma secretion by CD4(+)CD25(-) cells responding to allogeneic and other polyclonal stimuli; CTLA-4 expression was abolished on CD4(+)CD25(+) cells without compromising their suppressive ability. Everolimus reduced IFN-gamma secretion by CD4(+)CD25(-) cells before the anti-proliferative effect: this is a novel finding. Exogenous IL-2 and IL-15 could prevent the suppression of proliferation by CD4(+)CD25(+) cells and the drugs could not restore suppression. By contrast, suppression of IFN-gamma secretion was only slightly impeded with the exogenous cytokines. Finally, CD4(+)CD25(+) cells were more resistant than CD4(+)CD25(-) cells to the pro-apoptotic action of the drugs. Together these data suggest that CD4(+)CD25(+) cells may still exert their effects in transplant patients taking immunosuppression that interferes with IL-2 signaling.


Paternal sex chromosome aneuploidy as a possible origin of Turner syndrome in monozygotic twins: case report.
The meiotic or mitotic origin of most cases of Turner syndrome remains unknown, due to the difficulty in detecting hidden mosaicisms and to the lack of meiotic segregation studies. We have had the opportunity to study one pair of monozygotic twins concordant for Turner syndrome of paternal origin. The paternal origin of the single X chromosome was determined by polymerase chain reaction (PCR) amplification. No mosaicism was detected for the X or Y chromosome. In this case, a meiotic error during gametogenesis would be a likely origin of X monosomy. To determine if meiotic errors are more frequent in the father of these monozygotic twins concordant for Turner syndrome of paternal origin, molecular studies in spermatozoa were conducted to analyse sex chromosome numerical abnormalities. A total of 12520 sperm nuclei from the twins' father and 85338 sperm nuclei from eight normal donors were analysed using three-colour fluorescent in-situ hybridization. There were significant differences between the twins' father and control donors for XY disomy (0.22 versus 0.11%, P < 0.001) and total sex chromosome disomy (0.38 versus 0.21%, P < 0.001). These results could indicate an increased tendency to meiotic sex chromosome non-disjunction in the father of the Turner twins.


Sex Differences in the Phosphoproteomic Profiles of APP/PS1 Mice after Chronic Unpredictable Mild Stress.
Approximately two-thirds of those suffering with Alzheimer's disease (AD) are women, however, the biological mechanisms underlying this sex divergence of AD prevalence remain unknown. Previous research has shown sex-specific biochemical differences that bias female mice toward pro-AD signaling on the phosphoproteomic level via corticotropin releasing factor (CRF) receptor 1 activation after CRF overexpression. Here we aimed to determine if chronic stress would induce a similar response in AD mouse models. We stressed 4-month-old APP/PS1 mice using a chronic unpredictable mild stress (CUMS) paradigm for up to 1 month. Following CUMS and behavioral assessments, we quantified whole protein and phosphoprotein levels in the cortex of stressed and non-stressed APP/PS1 mice using mass spectrometry-based proteomics. While there were no statistically significant differences at the total protein and peptide abundance levels, we found 909 and 841 statistically significant phosphopeptides between stressed and unstressed females and males, respectively, using a false discovery rate of 5%. Of these significant phosphopeptides, only 301 were the same in males and females. These results indicate that while both males and females undergo protein phosphorylation changes following stress, the peptides that are phosphorylated differ between sexes. We then used Metacore analysis to determine which biological pathways were affected. We found that several pathways were changed differently between male and female mice including NMDA receptor trafficking, cytoskeleton organization, and tau pathology. The differing biological pathways affected between males and females in response to chronic stress may help us to better understand why women are at a higher risk of AD.


Fetal sheep endocrine responses to sustained hypoxemic stress after chronic fetal placental embolization.
The purpose of this study was to determine the endocrine and circulatory responses of the ovine fetus, near term, to sustained hypoxemic stress superimposed on chronic hypoxemia. Fetal sheep were chronically embolized (n = 7) for 10 days between 0.84 and 0.91 of gestation via the descending aorta until arterial oxygen content was decreased by approximately 30%. Control animals (n = 8) received saline only. On experimental day 10, both groups were embolized over a 6-h period until fetal arterial pH decreased to approximately 7.00. Regional distribution of lower body blood flows was measured on day 10, before and at the end of acute embolization. On day 10, the chronically embolized group had lower arterial oxygen content (P < 0.05), Po2 (P < 0.01), and placental blood flow (P < 0.05) than controls and higher prostaglandin E2 (PGE2) and norepinephrine plasma concentrations (both P < 0.05). In response to a superimposed sustained hypoxemic stress, there was a twofold greater increase in PGE2 in the chronically embolized group than in the control group (P < 0.05). However, the increase in fetal plasma cortisol in response to superimposed hypoxemic stress was similar in both groups, despite significantly lower adrenocorticotropic hormone and adrenal cortex blood flow responses in the chronically hypoxemic group (both P < 0.05). We conclude that PGE2 response to a sustained superimposed reduction in placental blood flow, leading to metabolic acidosis, is enhanced under conditions of chronic hypoxemia and may play an important role for the maintenance of the fetal cortisol response to an episode of superimposed acute stress.


Effects of atorvastatin on the clearance of triglyceride-rich lipoproteins in familial combined hyperlipidemia.
Familial combined hyperlipidemia (FCHL) patients have an impaired catabolism of postprandial triglyceride (TG)-rich lipoproteins (TRLs). We investigated whether atorvastatin corrects the delayed clearance of large TRLs in FCHL by evaluating the acute clearance of Intralipid (10%) and TRLs after oral fat-loading tests. Sixteen matched controls were included. Atorvastatin reduced fasting plasma TG (from 3.6 +/- 0.4 to 2.5 +/- 0.3 mM; mean +/- SEM) without major effects on fasting apolipoprotein B48 (apoB48) and apoB100 in large TRLs. Atorvastatin significantly reduced fasting intermediate density lipoprotein (Svedberg flotation, 12-20)-apoB100 concentrations. After Intralipid, TG in plasma and TRL showed similar kinetics in FCHL before and after atorvastatin treatment, although compared with controls, the clearance of large TRLs was only significantly slower in untreated FCHL, suggesting an improvement by atorvastatin. Investigated with oral fat-loading tests, the clearance of very low density lipoprotein (Sf20-60)-apoB100 improved by 24%, without major changes in the other fractions. The most striking effects of atorvastatin on postprandial lipemia in FCHL were on hepatic TRL, without major improvements on intestinal TRLs. Fasting plasma TG should be reduced more aggressively in FCHL to overcome the lipolytic disturbance causing delayed clearance of postprandial TRLs.


Expression of Active Fluorophore Proteins in the Milk of Transgenic Pigs Bypassing the Secretory Pathway.
We describe the expression of recombinant fluorescent proteins in the milk of two lines of transgenic pigs generated by Sleeping Beauty transposon-mediated genetic engineering. The Sleeping Beauty transposon consisted of an ubiquitously active CAGGS promoter driving a fluorophore cDNA, encoding either Venus or mCherry. Importantly, the fluorophore cDNAs did not encode for a signal peptide for the secretory pathway, and in previous studies of the transgenic animals a cytoplasmic localization of the fluorophore proteins was found. Unexpectedly, milk samples from lactating sows contained high levels of bioactive Venus or mCherry fluorophores. A detailed analysis suggested that exfoliated cells of the mammary epithelium carried the recombinant proteins passively into the milk. This is the first description of reporter fluorophore expression in the milk of livestock, and the findings may contribute to the development of an alternative concept for the production of bioactive recombinant proteins in the udder.


Cleaning symbiosis in coral reefs of Jardines de la Reina National Park.
Cleaning symbiotic interactions are an important component of coral reef biodiversity and the study of the characteristics of these interacting species networks allows to assess the health of communities. The coral reefs of Jardines de la Reina National Park (JRNP) are subject to a protection gradient and there is a lack of knowledge about the effect of different levels of protection on the cleaning mutualistic networks in the area. The present study aims to characterize the mutualistic cleaning networks in the reefs of JRNP and to assess the potential effect of the protection gradient on their characteristics. We visited 26 reef sites distributed along the National Park and performed 96 band transects (50 m × 4 m). Low, medium and highly protected regions were compared according to the number of cleaning stations and the abundance and number of species of clients and cleaners associated with them. Additionally, we built interaction networks for the three regions and the entire archipelago based on a total of 150 minutes' video records of active cleaning stations. We assessed ecological networks characteristics (specialization, nestedness) using network topological metrics. We found a high diversity and complex cleaning interaction network with 6 cleaner species and 39 client species, among them, the threatened grouper Epinephelus striatus was one of the most common clients. No clear effect of the protection level on the density, abundance or diversity of cleaners and clients was detected during this study. However, we found that the network structure varied among regions, with the highly protected region being more specialized and less nested than the other regions. Our research reveals some patterns that suggest the effect of fishing pressure on cleaning symbiosis, as fishing may reduce the abundance and composition of client species, especially those that are targeted by fishers. However, fishing pressure may not be the main factor influencing cleaning symbiosis inside of the National Park, as other factors, such as habitat quality or environmental conditions may have stronger effects on the demand for cleaning services and the interactions between cleaners and clients. Our research provides insights into the factors that influence cleaning symbiosis and its implications for coral reef conservation and management.


Allelopathic interactions between the brown algal genus Lobophora (Dictyotales, Phaeophyceae) and scleractinian corals.
Allelopathy has been recently suggested as a mechanism by which macroalgae may outcompete corals in damaged reefs. Members of the brown algal genus Lobophora are commonly observed in close contact with scleractinian corals and have been considered responsible for negative effects of macroalgae to scleractinian corals. Recent field assays have suggested the potential role of chemical mediators in this interaction. We performed in situ bioassays testing the allelopathy of crude extracts and isolated compounds of several Lobophora species, naturally associated or not with corals, against four corals in New Caledonia. Our results showed that, regardless of their natural association with corals, organic extracts from species of the genus Lobophora are intrinsically capable of bleaching some coral species upon direct contact. Additionally, three new C21 polyunsaturated alcohols named lobophorenols A-C (1-3) were isolated and identified. Significant allelopathic effects against Acropora muricata were identified for these compounds. In situ observations in New Caledonia, however, indicated that while allelopathic interactions are likely to occur at the macroalgal-coral interface, Lobophora spp. rarely bleached their coral hosts. These findings are important toward our understanding of the importance of allelopathy versus other processes such as herbivory in the interaction between macroalgae and corals in reef ecosystems.


Acquired defects of glycogen synthase activity in cultured human skeletal muscle cells: influence of high glucose and insulin levels.
To determine whether defects of muscle glycogen synthase (GS) activity can be acquired by exposure to elevated glucose or insulin levels, human skeletal muscle cells obtained by needle biopsy from normal control subjects were grown in culture for 4-6 weeks followed by 4 days of fusion and differentiation in media containing either normal (5.5 mmol/l glucose and 22 pmol/l insulin) or increased concentrations of glucose (20 mmol/l), insulin (30 micromol/l), or both. After fusion in normal media, acute stimulation by 33 nmol/l insulin for 1 h increased GS fractional velocity (FV) approximately twofold (from 9.01 +/- 1.26 to 16.31 +/- 2.40, P < 0.05). Increasing the media glucose concentration alone to 20 mmol/l during fusion had no effect on basal FV but caused a marginal impairment of the insulin-stimulated GS response (from 8.51 +/- 1.33 to 12.99 +/- 1.90, P = 0.08). Increasing the media insulin concentration to 30 micromol/l during fusion at 5.5 mmol/l glucose also did not alter basal GS FV (10.61 +/- 1.69%) but completely abolished the normal insulin-stimulated increase in GS activity (to 11.63 +/- 1.55%, NS). The combination of high insulin (30 micromol/l) and high glucose (20 mmol/l) during fusion had no greater effect on the FV of either basal (11.66 +/- 2.16%, NS) or insulin-stimulated (9.20 +/- 1.80%, NS) GS activity than high insulin alone. Fusion in hyperinsulinemic media altered the kinetic parameters of GS with a near doubling of the basal Km0.1 and Vmax0.1 for uridinediphospho-glucose. Hyperinsulinemia also totally prevented the normal insulin-stimulated threefold increase in the Vmax0.1 and the 65% decrease in the A0.5 for glucose-6-phosphate. GS mRNA and protein expression, determined by RNase protection assay and immunoblotting, respectively, were unaffected by changes in media conditions. We conclude that exposure of human skeletal muscle cells primarily to high insulin induces severe insulin resistance through multiple acquired posttranslational defects, which affect both the kinetic characteristics and absolute activity of the GS enzyme.


Quasiparticle Breakdown and Spin Hamiltonian of the Frustrated Quantum Pyrochlore Yb_{2}Ti_{2}O_{7} in a Magnetic Field.
The frustrated pyrochlore magnet Yb_{2}Ti_{2}O_{7} has the remarkable property that it orders magnetically but has no propagating magnons over wide regions of the Brillouin zone. Here we use inelastic neutron scattering to follow how the spectrum evolves in cubic-axis magnetic fields. At high fields we observe, in addition to dispersive magnons, a two-magnon continuum, which grows in intensity upon reducing the field and overlaps with the one-magnon states at intermediate fields leading to strong renormalization of the dispersion relations, and magnon decays. Using heat capacity measurements we find that the low- and high-field regions are smoothly connected with no sharp phase transition, with the spin gap increasing monotonically in field. Through fits to an extensive data set of dispersion relations combined with magnetization measurements, we reevaluate the spin Hamiltonian, finding dominant quantum exchange terms, which we propose are responsible for the anomalously strong fluctuations and quasiparticle breakdown effects observed at low fields.


Antibiotic-loaded spacer arthroplasty in a terrible triad injury with unreconstructable radial head fracture: a case report.
Radial head arthroplasty is a viable option in cases with terrible triad injuries of elbow, wherein the radial head sustains significant comminution that precludes reconstruction. Nevertheless, this alternative is not recommended for individuals with poor elbow skin conditions, accompanied neuropsychiatric disorders, or low patient compliance. This case report presents a patient with bilateral terrible triad injury, along with the aforementioned conditions. The report outlines the treatment challenges of such a case and proposes potential solutions. A 37-year-old Persian male patient presenting with a bilateral terrible triad fracture-dislocation and a history of psychoactive substance abuse, was admitted to our emergency department. The patient underwent radial head replacement using a cement spacer containing antibiotics, due to the comminuted radial head in the presence of a contaminated wound on the left elbow. The fracture of the right side was successfully fixed. Subsequent to discharge, the patient did not attend any follow-up appointments. After a period of 6 months, he was admitted to the psychiatric ward and orthopedic consultation was requested to evaluate the patient. In acute terrible triad injuries with unreconstructable radial head fractures where arthroplasty with metallic prostheses may not be suitable due to contaminated wounds, unstable psychiatric condition, and low patient cooperation, temporary orthopedic cement spacers can maintain elbow biomechanics, stability, and sterility.


Response of myocardial cellular energy metabolism to variation of buffer composition during open-chest experimental cardiopulmonary resuscitation in the pig.
The aim of the present study was to investigate possible relationships in piglets between myocardial energy-related metabolites and intracellular electrolytes during open-chest cardiopulmonary resuscitation (OCCPR) supplemented by the administration of alkaline buffers with varying sodium content. Our hypothesis was that an increasing myocardial intracellular sodium content would decrease the intracellular energy stores. In addition to haemodynamics, acid-base and blood gas variables were analysed, and myocardial biopsies were collected before and during OCCPR as well as after the return of spontaneous circulation. After a period of 4 min of untreated ventricular fibrillation (VF). 25 piglets were randomly allocated to one of four groups: OCCPR with normal saline (n = 5); OCCPR with sodium bicarbonate (SB) (n = 7); OCCPR with Tris buffer mixture (TBM) (n = 7); and a totally untreated control group (n = 6). The results showed that 4 min of untreated VF almost eradicated creatine phosphate (CrP) and that the ATP/ADP ratio decreased to 1.5-2.0. During OCCPR with normal saline, the myocardial content of CrP increased, whereas lactate, ATP and ADP levelled off and AMP decreased, causing an increased ATP/ADP ratio. The adenosine and inosine contents increased, whereas inosine monophosphate was unchanged at a low level, the adenosine and inosine contents being inversely correlated with the total content of adenine nucleotides. In both buffered groups, the increase in most energy-related metabolites (CrP, ATP, ADP, AMP and the ATP/ADP quotient) was less and in lactate more pronounced than in the group not being buffered, with no difference between the groups receiving SB or TBM. Although the intracellular potassium content was unaltered, the sodium, chloride and calcium concentration increased, more so in the group receiving SB. The intracellular content of sodium was correlated with that of calcium. Thus, buffering increased the myocardial AMP degradation during OCCPR by increasing the flux via the 5'-nucleotidase reaction, and SB increased the intracellular contents of sodium and calcium to a greater extent than did TBM.


A Microethnographic and Ethnobotanical Approach to Llayta Consumption Among Andes Feeding Practices.
Llayta is a dietary supplement that has been used by rural communities in Perú and northern Chile since pre-Columbian days. Llayta is the biomass of colonies of a Nostoc cyanobacterium grown in wetlands of the Andean highlands, harvested, sun-dried and sold as an ingredient for human consumption. The biomass has a substantial content of essential amino acids (58% of total amino acids) and polyunsaturated fatty acids (33% total fatty acids). This ancestral practice is being lost and the causes were investigated by an ethnographic approach to register the social representations of Llayta, to document how this Andean feeding practice is perceived and how much the community knows about Llayta. Only 37% of the participants (mostly adults) have had a direct experience with Llayta; other participants (mostly children) did not have any knowledge about it. These social responses reflect anthropological and cultural tensions associated with a lack of knowledge on Andean algae, sites where to find Llayta, where it is commercialized, how it is cooked and on its nutritional benefits. The loss of this ancestral feeding practice, mostly in northern Chile, is probably associated with cultural changes, migration of the rural communities, and very limited access to the available information. We propose that Llayta consumption can be revitalized by developing appropriate educational strategies and investigating potential new food derivatives based on the biomass from the isolated Llayta cyanobacterium.


New lipid-lowering agents acting on LDL receptors.
The treatment of dyslipoproteinemia has proven a successful strategy in the prevention of cardiovascular diseases. The major target of hypolipidemic drugs is the reduction of low density lipoprotein cholesterol (LDL-C). HMG-CoA reductase inhibitors (HMGRI) effectively lower LDL-C by inhibiting the mevalonate pathway and enhancing the activity of the LDL receptor (LDL-R). Numerous clinical studies demonstrated convincingly, that the reduction of LDL-C lowers the incidence of cardiovascular events in primary and secondary prevention. Two new HMGRI, rosuvastatin and pitavastatin, have been evaluated in clinical trials. Both drugs demonstrated efficacy in lowering atherogenic lipoproteins. In addition to the reduction of LDL-C, they may have a higher potency to lower triacylglycerides (TG) and to increase HDL cholesterol (HDL-C) compared to currently available HMGRI. Other therapeutic strategies examined in experimental animals are the inhibition of squalene synthase, the first enzyme of the mevalonate pathway, which is specifically committed to cholesterol biosynthesis, and the direct up-regulation of LDL receptor activity. The latter compounds, the SCAP ligands, are the first members of a new class of hypolipidemic agents affecting the transcriptional regulation of genes involved in lipid metabolism. Recent treatment guidelines emphasise the importance of modifying lipid metabolism beyond lowering LDL-C, mainly by lowering TG and raising HDL-C. Although these actions are not primary targets of the compounds discussed here, it is interesting that drugs inducing the LDL-R usually also lower TG and, in the case of HMGRI, increase HDL-C.


Benzo[a]pyrene differentially alters mitochondrial and nuclear DNA synthesis in primary hepatocyte cultures.
The incubation of non-proliferating primary cultures of rat hepatocytes with benzo[a]Pyrene for 24 or 48 hours produced a concentration-dependent stimulation of nuclear DNA synthesis (measured as [3H]thymidine incorporation into isolated, purified DNA) whereas the normal processes of D-loop formation and expansion synthesis on mitochondrial DNA were inhibited by as much as 62% of that in control cells. Incubation of cultures with [3H]benzo[a]pyrene showed that highly purified mitochondrial DNA retained 4-15 times more covalently bound adducts per unit length of DNA than did nuclear DNA. We suggest, therefore, that this carcinogenic, environmental pollutant may exert its most profound effects on the mitochondrial, rather than the nuclear, genetic apparatus.


Evaluation of DNA Methyltransferase Activity and Inhibition via Isothermal Enzyme-Free Concatenated Hybridization Chain Reaction.
Methyltransferase (MTase)-catalyzed DNA methylation plays a vital role in the biological epigenetic processes of key diseases and has attracted increasing attention, making the amplified detection of MTase activity of great significance in clinical disease diagnosis and treatment. Herein, we developed an isothermal, enzyme-free, and autonomous strategy for analyzing MTase activity based on concatenated hybridization chain reaction (C-HCR)-mediated Förster resonance energy transfer (FRET). In a typical C-HCR procedure without MTase (Dam), Y-shaped initiator DNA activates upstream HCR-1 to assemble a double-stranded DNA (dsDNA) copolymeric nanowire consisting of multiple tandem DNA trigger units that motivate downstream HCR-2 to successively bring a fluorophore donor/acceptor (FAM/TAMRA) pair into close proximity, leading to the generation of an amplified FRET readout signal. The target Dam MTase and auxiliary DpnI endonuclease can sequentially and specifically recognize/methylate and cleave the Y-shaped initiator oligonucleotide, respectively, and thus prohibit the C-HCR process and FRET signal generation, resulting in the construction of a signal-on sensing platform for MTase assay. Our proposed isothermal enzyme-free C-HCR amplification approach was further utilized for screening MTase inhibitors. Furthermore, the proposed C-HCR approach can be easily adapted for probing other different MTases and for screening the corresponding inhibitors just by changing the recognition sequence of Y-shaped initiator DNA through a "plug-and-play" format. It provides a versatile and robust tool for highly sensitive detection of various biotransformations and thus holds great promise in clinical assessment and diagnosis.


Bacterial bioluminescence response to long-term exposure to reverse osmosis treated effluents from dye industries.
The bacterial bioluminescence assay is one of the novel means for toxicity detection. The bioluminescence response of 2 marine bioluminescent bacteria was tested upon their long-term exposure to 9 different reverse osmosis (RO) rejects with varying chemical composition sampled from various dye industries. Bioluminescent bacteria were cultured in the RO reject samples, at different concentrations, and their growth rate and luminescence was measured for 24 h. The RO reject samples caused sublethal effects upon exposure and retarded the growth of bacteria, confirming their toxic nature. Further, continuation of the exposure showed that the initial luminescence, though reduced, recovered and increased beyond the control cultures irrespective of cell density, and finally decreased once again. The present study emphasizes the need of evolving a long-term exposure assay and shows that the method followed in this study is suitable to evaluate the toxicants that exert delayed toxicity, using lower concentrations of toxicants as well as coloured samples.


Mössbauer and EPR study of recombinant acetyl-CoA synthase from Moorella thermoacetica.
Mössbauer and EPR spectroscopies were used to study the electronic structure of the A-cluster from recombinant acetyl-CoA synthase (the alpha subunit of the alpha2beta2 acetyl-CoA synthase/CO dehydrogenase). Once activated with Ni, these subunits have properties mimicking those associated with the alpha2beta2 tetramer, including structural heterogeneities. The Fe4S4 portion of the A-cluster in oxidized, methylated, and acetylated states was in the 2+ core oxidation state. Upon reduction with dithionite or Ti3+ citrate, samples of Ni-activated alpha developed the ability to accept a methyl group. Corresponding Mössbauer spectra exhibited two populations of A-clusters; roughly, 70% contained [Fe4S4]1+ cubanes, while approximately 30% contained [Fe4S4]2+ cubanes, suggesting an extremely low [Fe4S4](1+/2+) reduction potential for the 30% portion (perhaps <-800 mV vs NHE). The same population ratio was observed when Ni-free unactivated alpha was used. The 70% fraction exhibited paramagnetic hyperfine structure in the absence of an applied magnetic field, excluding the possibility that it represents an [Fe4S4]1+ cluster coupled to a (proximal) Ni(p)1+. EPR spectra of dithionite-reduced, Ni-activated alpha exhibited features at g = 5.8 and g(ave) approximately 1.93, consistent with a physical mixture of {S = 3/2; S = 1/2} spin-states for A-clusters containing [Fe4S4]1+ clusters. Incubation of Ni-activated alpha with dithionite and CO converted 25% of alpha subunits into the S = 1/2 A(red)-CO state. Previous correlation of this state to functional A-clusters suggests that only the 30% fraction not reduced by dithionite or Ti3+ citrate represents functional A-clusters. Comparison of spin states in oxidized and methylated states suggests that two electrons are required for reductive activation, starting from the oxidized state containing Ni(p)2+. Refitting published activity-vs-potential data supports an n = 2 reductive activation. Enzyme starting in the methylated state exhibited catalytic activity in the absence of an external reductant, suggesting that the two electrons used in reductive activation are retained by the enzyme after each catalytic cycle and that the enzyme does not have to pass through the A(red)-CO state during catalysis. Taken together, our results suggest that a Ni(p)0 state may form upon reductive activation and reform after each catalytic cycle.


Gemfibrozil-Induced Polyuria.
Gemfibrozil is a lipid-regulating agent used mainly to treat patients with hypertriglyceridaemia, especially those at risk for acute pancreatitis. Like any other pharmacological agent, gemfibrozil has known adverse effects, mainly gastrointestinal, such as cholelithiasis, gallstones, elevated transaminase, and other non-specific symptoms including dyspepsia, nausea and vomiting. Other reported adverse reactions are dizziness and vertigo, myopathy and rhabdomyolysis, angioedema, urticaria and rash. As far as we knew, gemfibrozil does not have urinary tract adverse reactions. In this report, we present a case of polyuria secondary to gemfibrozil with a score of 9 on the Naranjo scale, and a literature review. Gemfibrozil has known, mainly gastrointestinal, adverse effects.We aim to increase awareness of the urinary side effects of gemfibrozil so unnecessary investigations can be avoided.


Chromosomal Micro-aberration in a Saudi Family with Juvenile Myoclonic Epilepsy.
Epilepsy is etiologically and genetically complex neurological disorder affecting millions of people worldwide. Juvenile myoclonic epilepsy (JME) is the most common epilepsy syndrome that starts in the teen age group commonly between ages 12, 18, and lasts till adulthood. One out of fourteen people with epilepsy suffers with JME. Myoclonic seizures and muscle twitching or uncontrolled jerking are the most common type of seizures in the people suffering with JME. To observe the novel CNVs involved in JME, we investigated a Saudi family with nine siblings including one male and one female affected members. In this study we used high density whole genome Agilent sure print G3 Hmn CGH 2x 400K array-CGH chips. Our results showed CNVs including the amplifications and deletions in different chromosomal regions in the patients as compared to the normal members of the family. Amplifications were observed in the chromosome 22 cytoband 22q11.23 with LDL receptor related protein 5 like (LRP5L), Immunoglobulin Lambda-Like Polypeptide 3 (IGLL3) and crystallin beta B2 pseudogene (CRYBB2P) genes respectively whereas the deletions were observed in the chromosomal regions 4q22.2 with Glutamate receptor, ionotropic, delta 2 (GRID2) as potential gene cytoband 1p31.1 with potential Neuronal Growth Regulator 1 gene (NEGR1) gene in this region and NME/NM23 family member (NME7) gene cytoband 1q24. Moreover, the array CGH resulting in deletions and duplication were also validated by using primer for simple PCR or also by using quantitative real time PCR analysis. We found deletions and duplication in JME patients in our study for the first time in Saudi population. The findings in this study suggest that the array-CGH may be considered as a first line of genetic testing for diagnosis of epilepsy unless strong evidence is presented for a monogenic syndrome. The use of high throughput technique in this study will help to identify novel mechanisms underlying epileptic disorder in order to lower the burden of epilepsy in Saudi Arabia.


No Evidence for Improved Associative Memory Performance Following Process-Based Associative Memory Training in Older Adults.
Studies attempting to improve episodic memory performance with strategy instructions and training have had limited success in older adults: their training gains are limited in comparison to those of younger adults and do not generalize to untrained tasks and contexts. This limited success has been partly attributed to age-related impairments in associative binding of information into coherent episodes. We therefore investigated potential training and transfer effects of process-based associative memory training (i.e., repeated practice). Thirty-nine older adults (Mage = 68.8) underwent 6 weeks of either adaptive associative memory training or item recognition training. Both groups improved performance in item memory, spatial memory (object-context binding) and reasoning. A disproportionate effect of associative memory training was only observed for item memory, whereas no training-related performance changes were observed for associative memory. Self-reported strategies showed no signs of spontaneous development of memory-enhancing associative memory strategies. Hence, the results do not support the hypothesis that process-based associative memory training leads to higher associative memory performance in older adults.


The effects of drying technique and surface pre-treatment on the cytotoxicity and dissolution rate of luminescent porous silicon quantum dots in model fluids and living cells.
Tailoring of the biodegradation of photoluminescent silicon quantum dots (Si QDs) is important for their future applications in diagnostics and therapy. Here, the effect of drying and surface pretreatment on the dissolution rate of Si QDs in model liquids and living cells was studied in vitro using a combination of photoluminescence and Raman micro-spectroscopy. Porous silicon particles were obtained by mechanical milling of electrochemically etched mesoporous silicon films, and consist of interlinked silicon nanocrystals (QDs) and pores. The samples were subjected to super-critical drying with CO2 solvent (SCD) or air drying (AD) and then annealed at 600 °C for 16 hours in 1% oxygen to obtain nano-sized Si QDs. The obtained samples were characterized by a core-shell structure with a crystalline silicon core and a SiO2 layer on the surface. The sizes of the crystalline silicon cores, calculated from Raman scattering spectra, were about 4.5 nm for the initial AD-SiQDs, and about 2 nm for the initial SCD-SiQDs. Both the AD-Si QDs and the SCD-Si QDs exhibited visible photoluminescence (PL) properties due to quantum confinement effects. The dissolution of the nanocrystals was evaluated through their PL quenching, as well as by the presence of a low-frequency shift, broadening, and a decrease in the intensity of the Raman signal. The stability of the AD-Si QDs and the complete dissolution of the SCD-Si QDs during 24 hours of incubation with cells have been demonstrated. This might explain the apparent lower cytotoxicity observed for SCD-Si QDs.


Proteomic analysis of testis biopsies in men treated with transient scrotal hyperthermia reveals the potential targets for contraceptive development.
Mild testicular heating safely and reversibly suppresses spermatogenesis. In this study, we attempted to clarify the underlying molecular mechanism(s) involved in heat-induced spermatogenesis suppression in human testis. We conducted global proteomic analyses of human testicular biopsies before, and at 2 and 9 wk after heat treatment. Thirty-one and Twenty-six known proteins were identified with significant differential expression at 2 and 9 wk after heat treatment, respectively. These were used to characterize the cellular and molecular events in the testes when seminiferous epithelia became damaged (2 wk) and recovered (9 wk). At 2 wk post-treatment, the changed expression of a series of proteins could promote apoptosis or suppress proliferation and cell survival. At 9 wk post-treatment, the changed expression of proteins mainly promoted cell proliferation, differentiation and survival, but resisted cell apoptosis. Among those heat-regulated proteins, HNRNPH1 was selected for the further functional study. We found that HNRNPH1 was an anti-apoptosis protein that could regulate the expression of other heat-induced proteins. In conclusion, heat-induced reversible suppression of spermatogenesis occurred by modulating the expression of proteins related to proliferation, differentiation, apoptosis and cell survival pathways. These differentially expressed proteins were found to be key molecular targets affecting spermatogenesis after heat treatment.


Estimating the validity of the guilty knowledge test from simulated experiments: the external validity of mock crime studies.
This experiment was designed to examine the external validity of the standard mock-crime procedure used extensively to evaluate the validity of polygraph tests. The authors manipulated the type of mock-crime procedure (standard vs. a more realistic version) and the time of test (immediate vs. delayed) and examined their effects on the validity of the Guilty Knowledge Test (GKT) and the recall rate of the relevant items. The results indicated that only the type of mock-crime affected the 2 outcome variables. The realistic procedure was associated with a lower recall rate and weaker detection efficiency than the standard procedure. However, these effects were mediated by the type of GKT questions used. Practical implications of these results are discussed.


TRPM2 as a conserved gatekeeper determines the vulnerability of DA neurons by mediating ROS sensing and calcium dyshomeostasis.
Different dopaminergic (DA) neuronal subgroups exhibit distinct vulnerability to stress, while the underlying mechanisms are elusive. Here we report that the transient receptor potential melastatin 2 (TRPM2) channel is preferentially expressed in vulnerable DA neuronal subgroups, which correlates positively with aging in Parkinson's Disease (PD) patients. Overexpression of human TRPM2 in the DA neurons of C. elegans resulted in selective death of ADE but not CEP neurons in aged worms. Mechanistically, TRPM2 activation mediates FZO-1/CED-9-dependent mitochondrial hyperfusion and mitochondrial permeability transition (MPT), leading to ADE death. In mice, TRPM2 knockout reduced vulnerable substantia nigra pars compacta (SNc) DA neuronal death induced by stress. Moreover, the TRPM2-mediated vulnerable DA neuronal death pathway is conserved from C. elegans to toxin-treated mice model and PD patient iPSC-derived DA neurons. The vulnerable SNc DA neuronal loss is the major symptom and cause of PD, and therefore the TRPM2-mediated pathway serves as a promising therapeutic target against PD.


Photoluminescent sensor for acetylcholinesterase inhibitor determination.
An acetylcholinesterase immobilized p-type porous silicon material for sensing acetylcholinesterase inhibitors was demonstrated. The enzyme immobilized photoluminescence surface upon incubation with standard acetylcholinesterase inhibitors (neostigmine methyl sulfate and tacrine) for 30 minutes at 37 °C triggers the photoluminescence with up to a 3 fold enhancement in the intensity with a hypsochromic shift of 3 nm. The dramatic change in photoluminescence of the enzyme immobilized surface upon reaction with the enzyme inhibitor can be used as a tool for the detection of acetylcholinesterase inhibitors by utilizing the subsequent photoluminescence response after the incubation with the corresponding inhibitor drug.


The use of polymorphic state modifiers in solid lipid microparticles: The role of structural modifications on drug release performance.
This study investigates the correlation between the structural and release properties of solid lipid microparticles (MPs) of tristearin containing 5 % w/w of four different liquid additives used as crystal modifiers: isopropyl myristate (IM), ethyl oleate (EO), oleic acid (OA) and medium chain triglycerides (MCT). All additives accelerated the conversion of the unstable α-form of tristearin, formed after the MPs manufacturing, to the stable β-polymorph and the transformation was completed within 24 h (for IM and EO) or 48 h (for OA and MCT). The kinetic of polymorphic transition at 25 °C was investigated by simultaneous synchrotron SAXS/WAXS and DSC analysis after melting and subsequent cooling of the lipid mixture. After crystallization in the α-phase, additives accelerate the solid-solid phase transformation to β-tristearin. SAXS data showed that two types of structural modifications occurred on MPs during storage: compaction of the crystal packing (slight decrease in lamellar thickness) and crystal growth (increased number of stacked lipid lamellae). The release behavior of a model hydrophilic drug (caffeine) at two different amounts (15 % and 30 %) from MPs was studied in water and biorelevant media simulated the gastric and intestinal environment. It was particularly significant that the introduction of IM, EO and MCT were able to prolong the drug release in water, passing from a diffusion-based Higuchi kinetics to a perfect zero-order kinetic. Moreover, the overall release profiles were higher in biorelevant media, where erosion/digestion of MPs was observed. After 6 months, a moderate but statistically significant change in release profile was observed for the MPs with IM and EO, which can be correlated with the time-dependent structural alterations (i.e. larger average crystallite size) of these formulations; while MPs with OA or MCT displayed stable release profiles. These findings help to understand the correlation between release behavior, polymorphism and supramolecular-level structural modification of lipid formulations containing crystal modifiers.


Time to treatment in rheumatoid arthritis: factors associated with time to treatment initiation and urgent triage assessment of general practitioner referrals.
The importance of early diagnosis and treatment of rheumatoid arthritis (RA) is increasingly being recognized. This requires accurate triaging of suspected RA referrals from primary care and expedient assessment of these patients in secondary care. To assess the factors associated with urgent triage for first specialist appointment and early disease modifying agents in rheumatic disease treatment. The general practitioner (GP) referrals to a single rheumatology service from 128 new RA patients were assessed for their information content and triage allocation by the Rheumatologists. Information on symptoms, signs, and investigation results were collected. Factors associated with urgent triage allocation, time to treatment, and a GP requesting urgency were assessed. Median time from symptom onset to treatment was 6.1 months. Triage allocation to urgent was associated with earlier treatment (difference of 97 days, P = 0.003). GP perception of urgency (odds ratio = 13.34, 95% confidence interval: 2.20-81.02) was independently associated with an urgent triage allocation by the triaging rheumatologist. Swollen joints and a raised C-reactive protein predicted GP request for urgency. Triage is important to facilitate early treatment; however, rheumatologists in this service are not currently triaging suspected RA referrals with reference to known poor prognostic indicators. Several interventions could improve both informative referrals and triaging of referrals to decrease time to diagnosis and treatment. These interventions could include public education, GP education sessions with associated distribution of referral guidelines, and reminding triaging rheumatology clinicians about the available prognostic factors often present in GP referrals that assist with correct triage.