[Presymptomatic detection of Parkinson's disease].
Parkinson's disease (PD) is characterized by a progressive degeneration of mesencephalic dopaminergic neurons. More than half of these neurons are lost in a presymptomatic phase of an estimated 4-6 years duration. It is obvious that any type of treatment aimed at slowing down the disease process should preferably be applied in this presymptomatic phase. Presymptomatic detection of PD has therefore become an important goal. In a recent study in a population of 361 asymptomatic first degree relatives of PD patients, we were able to demonstrate that presymptomatic detection is possible by means of a combination of three olfactory processing tasks and [123l] beta-CIT single photon emission computed tomography (SPECT) scanning of the nigrostriatal dopaminergic system. These results are a first step towards the development of a screening strategy that may be applied in the general population. Impairments of olfactory function, however, are not specific to PD but are also associated with other neurodegenerative disorders (e.g. Alzheimer's disease) and certain lifestyle characteristics (e.g. smoking). In the next few years our research efforts will focus on two different approaches to develop a more specific screening strategy. First, olfactory processing tasks will be combined with tasks aimed at detecting subtle (visuo)motor disturbances and early cognitive impairments. In parallel, an effort will be made to define disease-specific patterns of olfactory dysfunction in neurodegenerative disorders.


Current status of photoprotection by window glass, automobile glass, window films, and sunglasses.
Ultraviolet radiation (UVR) has known adverse effects on the skin and eyes. Practitioners are becoming more aware of the importance of outdoor photoprotection. However, little attention is directed to the exposure of the skin and eyes to UVR through the window glass or sunglasses. The amount of ultraviolet transmission through glass depends mainly on the type of the glass. All types of commercial and automobile glass block the majority of ultraviolet-B; however, the degree of ultraviolet-A transmission depends on the type of glass. Laminated glass offers better UVA protection than tempered glass; new safety regulations for automobiles may result in increased use of laminated glass for side windows. Window films can be applied to glass to increase UVR protection. Sunglasses need to be compliant with one of the national standards; a wraparound style or side shields offer the best protection. Increased understanding by practitioners on the transmission of UVR through glass, window films, and sunglasses would allow them to better educate the public and to better manage photosensitive patients.


Gating-related molecular motions in the extracellular domain of the IKs channel: implications for IKs channelopathy.
Cardiac slow delayed rectifier (I(Ks)) channel complex consists of KCNQ1 channel and KCNE1 auxiliary subunits. The extracellular juxtamembranous region of KCNE1 is an unstructured loop that contacts multiple KCNQ1 positions in a gating-state-dependent manner. Congenital arrhythmia-related mutations have been identified in the extracellular S1-S2 linker of KCNQ1. These mutations manifest abnormal phenotypes only when coexpressed with KCNE1, pointing to the importance of proper KCNQ1/KCNE1 interactions here in I(Ks) channel function. We investigate the interactions between the KCNE1 loop (positions 36-47) and KCNQ1 S1-S2 linker (positions 140-148) by means of disulfide trapping and voltage clamp techniques. During transitions among the resting-state conformations, KCNE1 positions 36-43 make contacts with KCNQ1 positions 144, 145, and 147 in a parallel fashion. During conformational changes in the activated state, KCNE1 position 40 can make contacts with all three KCNQ1 positions, while the neighboring KCNE1 positions (36, 38, 39, and 41) can make contact with KCNQ1 position 147. Furthermore, KCNQ1 positions 143 and 146 are high-impact positions that cannot tolerate cysteine substitution. To maintain the proper I(Ks) channel function, position 143 requires a small side chain with a hydroxyl group, and position 146 requires a negatively charged side chain. These data and the proposed molecular motions provide insights into the mechanisms by which mutations in the extracellular juxtamembranous region of the I(Ks) channel impair its function.


Reduced p120ctn expression correlates with poor survival in patients with adenocarcinoma of the gastroesophageal junction.
P120-catenin (p120ctn) is a member of the E-cadherin-catenin cell-cell adhesion complex. Impairment of one or more of the components of this complex is associated with tumorigenesis. The role of p120ctn in malignancy is not clear yet. We studied the in vivo expression and cellular localization of p120ctn in adenocarcinomas of the gastroesophageal junction. Immunohistochemical staining for p120ctn was performed on 96 tumor samples, 20 cases of Barretts metaplasia and 13 lymph node metastases. The relationship with pathological characteristics and patient survival was also assessed. Loss of normal surface p120ctn expression was found in 4/20 (20%) Barretts metaplasia, in 65/96 (68%) tumors, and 11/13 (85%) lymph node metastases. Nuclear immunoreactivity for p120ctn was seen in five tumors. Loss of normal expression of p120ctn was associated with a higher tumor grade (P < 0.0001) but not with pTNM-stage. Reduced expression of p120ctn was correlated with poor survival (P = 0.0002). Cox regression analysis showed that p120ctn is an independent prognostic marker. Abnormal p120ctn expression is frequently seen in adenocarcinomas of the gastroesophageal junction, and may be a useful as a prognostic marker in these tumors.


Link prediction for tree-like networks.
Link prediction is the problem of predicting the location of either unknown or fake links from uncertain structural information of a network. Link prediction algorithms are useful in gaining insight into different network structures from partial observations of exemplars. However, existing link prediction algorithms only focus on regular complex networks and are overly dependent on either the closed triangular structure of networks or the so-called preferential attachment phenomenon. The performance of these algorithms on highly sparse or treelike networks is poor. In this letter, we proposed a method that is based on the network heterogeneity. We test our algorithms for three real large sparse networks: a metropolitan water distribution network, a Twitter network, and a sexual contact network. We find that our method is effective and performs better than traditional algorithms, especially for the Twitter network. We further argue that heterogeneity is the most obvious defining pattern for complex networks, while other statistical properties failed to be predicted. Moreover, preferential attachment based link prediction performed poorly and hence we infer that preferential attachment is not a plausible model for the genesis of many networks. We also suggest that heterogeneity is an important mechanism for online information propagation.


Case report: Off-label use of low-dose perampanel in a 25-month-old girl with a pathogenic SYNGAP1 variant.
Preclinical studies in a mouse model have shown that SYNGAP1 haploinsufficiency results in an epilepsy phenotype with excessive GluA2-AMPA insertion specifically on the soma of fast-spiking parvalbumin-positive interneurons associated with significant dysfunction of cortical gamma homeostasis that was rescued by perampanel (PER), an AMPA receptor blocker. In this single case, we aimed to investigate the presence of dysregulated cortical gamma in a toddler with a pathogenic SYNGAP1 variant and report on the effect of low-dose PER on electroencephalogram (EEG) and clinical profile. Clinical data from physician's clinic notes; genetic testing reports; developmental scores from occupational therapy, physical therapy, speech and language therapy evaluations; and applied behavioral analysis reports were reviewed. Developmental assessments and EEG analysis were done pre- and post-PER. Clinically, the patient showed improvements in the developmental profile and sleep quality post-PER. EEG spectral power analysis in our patient revealed a loss of gamma power modulation with behavioral-state transitions similar to what was observed in Syngap1+/- mice. Furthermore, the administration of low-dose PER rescued the dysfunctional cortical gamma homeostasis, similar to the preclinical study. However, as in the epileptic mice, PER did not curb epileptiform discharges or clinical seizures. Similar to the Syngap1+/- mice, cortical gamma homeostasis was dysregulated in the patient. This dysfunction was rescued by PER. These encouraging results necessitate further validation of gamma dysregulation as a potential translational EEG biomarker in SYNAP1-DEE. Low-dose PER can be explored as a therapeutic option through clinical trials.


Low contact stress (LCS) complete knee system in revision surgery.
Revision total knee arthroplasty (TKA) should offer the same benefits to patients as primary TKA. As in primary TKA, a main objective of revision TKA is to reduce pain and restore functional range of motion. There are several potential causes of total knee failure, but the principles of repairing each of them is similar. The long-term success of the low contact stress knee system in primary TKA is well established, and clinical evidence for revision TKA with the low contact stress knee is promising.


A first instance of human philophthalmosis in Israel.
Conjunctivitis due to the trematode Philophthalmus in a 13-year-old Israeli girl is described. A single worm, probably a mature Philophthalmus palpebrarum, was detected on the palpebral conjunctiva of the upper eyelid of the right eye. Removal of the worm resulted in rapid abatement of the ocular symptoms. This is the first documentation of human philophthalmosis in Israel.


Tandem autologous stem cell transplantation in multiple myeloma after high-dose chemotherapy with two separate collections: single institution experience.
Presented is a retrospective analysis of multiple myeloma patients transplanted at our institution between November 1993 and August 2007. The objective of this analysis was to assess the feasibility and toxicity of tandem autologous stem cell transplantation (T-ASCT) when stem cells were harvested before first and before second transplantation separately. A total of 90 patients transplanted in our center were analyzed, of whom 43 patients were in tandem transplantation group.The overall response rate (ORR) was 83.7% and 95.1%, estimated five-year overall survival (OS) was 40.1% and 60.0%, probability of five-year event-free survival (EFS) was 18.2% and 25.6%, transplant related mortality (TRM) was 6.3% and 4.6% in the single and tandem transplant group, respectively. In multivariable analysis of all 90 patients, tandem transplantation and ORR attained after induction therapy were favourable prognostic factors for OS (p=0.024 and p=0.002) and EFS (p=0.036 and p=0.008), respectively. In conclusion, tandem transplantation with two separate stem cell harvests, performed separately before the each transplantation, is feasible in majority of patients with acceptable toxicity.


The nipple valve as a sphincter substitute for the ileocecal valve: prevention of bacterial overgrowth in the small bowel.
We evaluated the bacteriologic effect of a nipple valve substitute for the ileocecal valve in a canine model. Resection of the ileocecal valve and the distal 40% of the jejunoileum with end-to-end anastomosis was carried out in 10 dogs. In five of the dogs chosen randomly, a nipple valve was constructed at the anastomosis; in the remainder , a two-layer jejunocolostomy was formed. Microbiologic samples were taken at operation in the ascending colon and at three sites in the small intestine before resection and at reoperation 4 weeks later. These revealed a dramatic increase in anaerobic bacteria in the distal jejunum following jejunocolostomy without valve insertion but no increase following jejunocolostomy with nipple valve. There was a seven-log difference between the two groups in the mean anaerobic bacterial counts (P less than 0.05) measured 60 cm proximal to the anastomosis at the second operation. Results of aerobic cultures were similar but less dramatic. We conclude that construction of a nipple valve jejunocolostomy prevents anaerobic bacterial colonization of the proximal small bowel and may therefore be useful in the treatment or prevention of short bowel syndrome.


Computationally Efficient Implementation of Aperture Domain Model Image Reconstruction.
Aperture domain model image reconstruction (ADMIRE) is a useful tool to mitigate ultrasound imaging artifacts caused by acoustic clutter. However, its lengthy run-time impairs its usefulness. To overcome this drawback, we evaluated the reduced model methods with otherwise similar performance to ADMIRE. We also assessed other approaches to speed up ADMIRE, including the use of different levels of short-time Fourier transform (STFT) window overlap and examining the degrees of freedom of the model fit. In this study, we conducted an analysis of the reduced models, including those using Gram-Schmidt orthonormalization (GSO), singular value decomposition (SVD), and independent component analysis (ICA). We evaluated these reduced models using the data from simulations, experimental phantoms, and in vivo liver scans. We then tested ADMIRE's performance using full, GSO, SVD, and ICA-fourth-order blind identification (ICA-FOBI) models. The results from simulations, experimental phantoms, and in vivo data indicate that a model reduced using the ICA-FOBI method is the most promising for accelerating ADMIRE implementation. In the in vivo liver data, the improvements in contrast relative to delay-and-sum (DAS) using a full model, GSO, SVD, and ICA-FOBI models are 6.29 ± 0.25 dB, 11.88 ± 0.90 dB, 9.01 ± 0.67 dB, and 6.36 ± 0.27 dB, respectively; whereas, the contrast-to-noise ratio (CNR) improvement values in the same order are 0.04 ± 0.06 dB, -1.70 ± 0.17 dB, -1.51 ± 0.19 dB, and 0.12 ± 0.07 dB, respectively. The implementation of ADMIRE using the reduced model methods can decrease ADMIRE's computational complexity over three orders of magnitude. The use of a 50% STFT window overlap reduces ADMIRE's serial run time by more than one order of magnitude without any remarkable loss of image quality, when compared to the use of a 90% window overlap used previously. Based on these findings, a combination of the ICA-FOBI model and the use of a 50% STFT window overlap makes the ADMIRE algorithm more computationally efficient.


Trade-off between Zincophilicity and Zincophobicity: Toward Stable Zn-Based Aqueous Batteries.
Zn-based aqueous batteries (ZABs) hold great promise for large-scale energy storage applications due to the merits of intrinsic safety and low cost. Nevertheless, the thorny issues of metallic Zn anodes, including dendrite growth and parasitic side reactions, have severely limited the application of ZABs. Despite the encouraging improvements for stabilizing Zn anodes through surface modification, electrolyte optimization, and structural design, fundamentally addressing the inherent thermodynamics and kinetics obstacles of Zn anodes remains crucial in realizing reliable ZABs with ultrahigh efficiency, capacity, and cyclability. The target of this perspective is to elucidate the prominent status of Zn metal anode electrochemistry first from the perspective of zincophilicity and zincophobicity. Recent progress in ZABs is critically appraised for addressing the key issues, with special emphasis on the trade-off between zincophilic and zincophobic electrochemistry. Challenges and prospects for further exploration of a reliable Zn anode are presented, which are expected to boost in-depth research and practical applications of advanced ZABs.


Relation of the estrogen receptor and vitamin D receptor polymorphisms with bone mineral density in postmenopausal Mexican-mestizo women.
Since osteoporosis is a complex disease characterized by low bone mineral density (BMD), which is determined by an interaction of genetics with metabolic and environmental factors, the aim of this study was to analyze the possible association among one polymorphism of VDR and two polymorphisms of ESR1; as well as their haplotypes with BMD in postmenopausal Mexican-mestizo women. We studied 742 postmenopausal Mexican-mestizo women. A structured questionnaire for risk factors was applied and BMD was measured in the lumbar spine and total hip by dual-energy X-ray absorptiometry. DNA was obtained from blood leukocytes. One polymorphism of VDR (rs11568820) and two of ESR1 (rs2234693 and rs9340799) were studied. Real-time PCR allelic discrimination was used for genotyping. The differences between the means of the BMDs according to genotype were analyzed with covariance. Hardy-Weinberg equilibrium was tested. Pairwise linkage disequilibrium between single nucleotide polymorphisms was calculated by direct correlation r(2); haplotype analysis was conducted. Rs9340799 of ESR1 and one haplotype formed by the two polymorphisms of the ESR1 were significantly associated with FN-BMD variations. Moreover, analysis of the genotype of rs11568820 of VDR and the rs2234693 of ESR1 showed no significant differences with BMD variations. Our results showed that rs9340799 and one haplotype of ESR1 were significantly associated with BMD only at the femoral neck and this association remained after adjusting for covariates.


Terminal differentiation induction as DNA damage response in hematopoietic stem cells by GADD45A.
Hematopoietic stem cells (HSCs) sustain lifelong blood cell regeneration by balancing their ability for self-renewal with their ability to differentiate into all blood cell types. To prevent organ exhaustion and malignant transformation, long-lived HSCs, in particular, must be protected from exogenous and endogenous stress, which cause severe DNA damage. When DNA is damaged, distinct DNA repair mechanisms and cell fate controls occur in adult HSCs compared with committed cells. Growth arrest and DNA damage-inducible 45 alpha (GADD45A) is known to coordinate a variety of cellular stress responses, indicating the molecule is an important stress mediator. So far, the function of GADD45A in hematopoietic stem and progenitor cells is controversial and appears highly dependent on the cell type and stress stimulus. Recent studies have analyzed its role in cell fate decision control of prospectively isolated HSCs and have revealed unexpected functions of GADD45A, as discussed here. The upregulation of GADD45A by DNA damage-causing conditions results in enhanced HSC differentiation, probably to efficiently eliminate aberrant HSCs from the system. These findings, in concert with a few studies on other stem cell systems, have led us to propose DNA damage-induced differentiation as a novel DNA damage response mechanism in stem cells that circumvents the fatal consequences of cumulative DNA damage in the stem cell compartment.


Magnetic Resonance Spectroscopy in Patients with Insomnia: A Repeated Measurement Study.
Chronic insomnia is one of the most prevalent central nervous system disorders. It is characterized by increased arousal levels, however, the neurobiological causes and correlates of hyperarousal in insomnia remain to be further determined. In the current study, magnetic resonance spectroscopy was used in the morning and evening in a well-characterized sample of 20 primary insomnia patients (12 females; 8 males; 42.7 ± 13.4 years) and 20 healthy good sleepers (12 females; 8 males; 44.1 ± 10.6 years). The most important inhibitory and excitatory neurotransmitters of the central nervous system, γ-aminobutyric acid (GABA) and glutamate/glutamine (Glx), were assessed in the anterior cingulate cortex (ACC) and dorsolateral prefrontal cortex (DLPFC). The primary hypothesis, a diurnal effect on GABA levels in patients with insomnia, could not be confirmed. Moreover, the current results did not support previous findings of altered GABA levels in individuals with insomnia. Exploratory analyses, however, suggested that GABA levels in the ACC may be positively associated with habitual sleep duration, and, thus, reduced GABA levels may be a trait marker of objective sleep disturbances. Moreover, there was a significant GROUP x MEASUREMENT TIME interaction effect on Glx in the DLPFC with increasing Glx levels across the day in the patients but not in the control group. Therefore, Glx levels may reflect hyperarousal at bedtime in those with insomnia. Future confirmatory studies should include larger sample sizes to investigate brain metabolites in different subgroups of insomnia.


Postural orthostatic tachycardia in a teenager with Klinefelter syndrome.
To our knowledge this is the first reported case of postural orthostatic tachycardia syndrome (POTS) in a patient with Klinefelter syndrome. We describe a classic clinical presentation of POTS in an adolescent male with Klinefelter syndrome. Although the etiology of POTS appears to be multifactorial, there is a strong female predominance that suggests a genetic basis. Our patient with Klinefelter syndrome may further support a link with POTS to the X chromosome.


Blocking of SMAD4 expression by shRNA effectively inhibits fibrogenesis of human hepatic stellate cells.
In this study, to clarify the SMAD4 blocking impact on fibrosis process, we investigated its down-regulation by shRNA on activated human LX-2 cell, in vitro. Liver fibrosis is a critical consequence of chronic damage to the liver that can progress toward advanced diseases, liver cirrhosis and hepatocellular carcinoma (HCC). Different SMAD proteins play as major mediators in the fibrogenesis activity of hepatic stellate cells through TGF-β pathways, but the extent of SMAD4 as a co-SMAD protein remained less clear. vector expressing verified shRNA targeting human SMAD4 gene was transfected into LX-2 cells. The GFP expressing plasmid was transfected in the same manner as a control group while leptin treated cells were employed as positive controls. Subsequently, total RNA was extracted and real-time PCR was performed to measure the mRNA levels of SMAD4, COL-1A1, α-SMA, TGF-β and TIMP-1. Furthermore, trypan blue exclusion was performed to test the effect of plasmid transfection and SMAD4 shutting-down on cellular viability. The results indicated that the expression of SMAD4was down-regulated following shRNA transfection intoLX-2 cells (P<0.001). The gene expression analysis of fibrotic genes in LX-2 cells showed that SMAD4 blocking by shRNA significantly reduced the expression level of fibrotic genes when compared to control plasmids (P<0.001). Vector expressing SMAD4-shRNA induced no significant cytotoxic or proliferative effects on LX-2 cells as determined by viability assay (P<0.05). The results of this study suggested that knockdown of SMAD4 expression in stellate cell can control the progression of fibrogenesis through TGF-β pathway blocking.


Modelling the internalization of labelled cells in tumour spheroids.
In this paper a mathematical model is developed to describe the migration of labelled particles within a multicell spheroid. In the model, spatial variations in cell proliferation and death create an internal velocity field which leads to redistribution of the labelled and unlabelled cells. By applying a range of numerical and analytical techniques to the model equations, it is possible to show that, whilst the speed with which the labelled cells migrate through the tumour is independent of the type of cells that are labelled, their limiting distribution depends crucially on whether inert polystyrene microspheres or live tumour cells are labelled. These predictions are shown to be in good qualitative agreement with independent experimental results.


Advance Care Planning Prior to Death in Older Adults with Hip Fracture.
Although hip fractures in older adults are associated with a high degree of mortality and disability, the use of advance care planning (ACP) in this population is unknown. To determine the prevalence of ACP and need for surrogate decision-making prior to death in older adults with hip fracture and to identify factors associated with ACP. Retrospective cohort study using Health and Retirement Study (HRS) interviews linked to Medicare fee-for-service claims data. Six hundred six decedent participants aged 65 or older who sustained a hip fracture during HRS enrollment and had a proxy participate in the exit HRS survey. Survey responses by proxies were used to determine ACP, defined by either advance directive completion or surrogate designation, and to assess decision-making at the end of life. Multivariate logistic regression was used to analyze correlates of ACP. Prior to death, 54.9% of all participants had an advance directive and 68.9% had designated a surrogate decision-maker; however, 24.5% had no ACP. Of the total cohort, 32.5% required decisions to be made about treatment at the end of life and lacked capacity to make these decisions themselves. In this subset, 19.9% had no ACP. In all participants, ACP was less likely in non-white individuals (adjusted odds ratio (aOR) 0.14, 95% CI 0.06-0.31), those with less than a high school education (aOR 0.58, 95% CI 0.35-0.97), and those with a net worth below the median of the cohort (aOR 0.49, 95% CI 0.26-0.72). No clinical factors were found to be associated with ACP completion prior to death. A considerable number of older adults with hip fracture required surrogate decision-making at the end of life, of whom one fifth had no ACP prior to death. Clinicians providing care for these patients are uniquely poised to address ACP.


Enabling cultural safety in Indigenous primary healthcare.
The Aboriginal Health Access Centre (AHAC) and Aboriginal Community Health Centre Model of Wholistic Health and Wellbeing is critical to addressing inequities and barriers that limit access to comprehensive primary healthcare for Indigenous people. Even with this model in place, there are multiple points of intersection with mainstream healthcare service providers across health sectors. Further, there is considerable cultural diversity among Indigenous healthcare staff and professional groups. These factors place Indigenous people at risk of culturally unsafe experiences causing harm. Given this, it is essential that leaders focus on cultural safety education to address both intercultural frictions within the Indigenous centres and systemic and structural racism widespread within the broader healthcare system. This article explores how one AHAC has undertaken to examine these complex challenges, while offering some direction on leadership within the sector.


Outcomes and Complications After Bone Marrow Aspirate Harvest From the Calcaneus.
Bone marrow aspirate (BMA) is used with the putative goal of enhancing healing of injured tissue. The most common sites to harvest BMA are the iliac crest, the tibia, and the calcaneus. Investigators have found that the tibia and calcaneus have fewer progenitor cells than the iliac crest. This retrospective review evaluates the efficacy and safety profile of harvesting BMA from the calcaneus bone. We hypothesized that harvesting BMA from the calcaneus will have high efficacy and safety profile with low complication rates. A retrospective chart review of patients undergoing bone marrow aspiration from the calcaneus bone from January 2019 to October 2022 was performed. The main data points evaluated were patient satisfaction and pain level, follow-up times, quantity of BMA harvested, fusion rates, and complications including nerve damage, infections, and nonunions. There were 45 (34 female and 11 male) patients who underwent 45 procedures. The average age, BMI, and amount of BMA were 45.1 years (range 23-79), 33.1 (range 19.2-61.3), and 10.3 mL (range 2-40), respectively. There was no pain reported at the final follow-up in 32 patients (71.1%), and there was minimal to no pain in the area of surgery in the remaining 13 patients (28.9%). The 10 cases of fusion successfully healed with combined use of allograft and BMA. The average follow-up time was 12.3 months (range 2.4-33.7). There were no infections, wound complications, or nerve injuries. Harvesting BMA from the calcaneus bone is a safe procedure. In this heterogenous series where BMA augmented other surgical strategies, patients had little to no pain and there were no cases of wound complication or iatrogenic neuropraxia after undergoing bone marrow harvest from the calcaneus. Level IV, case series.


Semi-quantitative measurement of a specific glycoform using a DNA-tagged antibody and lectin affinity chromatography for glyco-biomarker development.
Aberrant glycosylation-targeted disease biomarker development is based on cumulative evidence that certain glycoforms are mass-produced in a disease-specific manner. However, the development process has been hampered by the absence of an efficient validation method based on a sensitive and multiplexed platform. In particular, ELISA-based analytical tools are not adequate for this purpose, mainly because of the presence of a pair of N-glycans of IgG-type antibodies. To overcome the associated hurdles in this study, antibodies were tagged with oligonucleotides with T7 promoter and then allowed to form a complex with corresponding antigens. An antibody-bound specific glycoform was isolated by lectin chromatography and quantitatively measured on a DNA microarray chip following production of fluorescent RNA by T7-trascription. This tool ensured measurement of targeted glycoforms of multiple biomarkers with high sensitivity and multiplexity. This analytical method was applied to an in vitro diagnostic multivariate index assay where a panel of hepatocellular carcinoma (HCC) biomarkers comprising alpha-fetoprotein, hemopexin, and alpha-2-macroglobulin (A2M) was examined in terms of the serum level and their fuco-fractions. The results indicated that the tests using the multiplexed fuco-biomarkers provided improved discriminatory power between non- hepatocellular carcinoma and hepatocellular carcinoma subjects compared with the alpha-fetoprotein level or fuco-alpha-fetoprotein test alone. The developed method is expected to facilitate the validation of disease-specific glycan biomarker candidates.


[Drugs that aggravate the course of COVID-19 : really ?]
The safety of NSAIDs, corticosteroids and renin-angiotensin inhibitors in COVID-19 is challenged. NSAIDs may interfere with the defense process against viral infection and are best avoided. Systemic corticosteroids have not shown benefit in viral infection, including other coronavirus; thus they should be avoided, unless prescribed for another indication. The benefit-risk ratio is however clearly in favor of continuing inhaled corticosteroids in patients with asthma or COPD. ACE inhibitors and sartans upregulate the expression of angiotensin-converting enzyme 2 (ACE2), the pulmonary receptor for SARS-CoV-2. Any possible clinical impact of these treatments on COVID-19 infection remains to be clarified; in the meantime, they should be continued.


Correlates of suicidality: investigation of a representative sample of Manitoba First Nations adolescents.
We examined individual, friend or family, and community or tribe correlates of suicidality in a representative on-reserve sample of First Nations adolescents. Data came from the 2002-2003 Manitoba First Nations Regional Longitudinal Health Survey of Youth. Interviews were conducted with adolescents aged 12 to 17 years (n=1125) from 23 First Nations communities in Manitoba. We used bivariate logistic regression analyses to examine the relationships between a range of factors and lifetime suicidality. We conducted sex-by-correlate interactions for each significant correlate at the bivariate level. A multivariate logistic regression analysis identified those correlates most strongly related to suicidality. We found several variables to be associated with an increased likelihood of suicidality in the multivariate model, including being female, depressed mood, abuse or fear of abuse, a hospital stay, and substance use (adjusted odds ratio range=2.43-11.73). Perceived community caring was protective against suicidality (adjusted odds ratio=0.93; 95% confidence interval=0.88, 0.97) in the same model. Results of this study may be important in informing First Nations and government policy related to the implementation of suicide prevention strategies in First Nations communities.


Hypophysectomy and hemivasectomy can inhibit the testicular hemicastration response of the mature rat.
Three questions were asked in an attempt to understand how testosterone (T) concentration in the veins of the remaining testis can double within 24 h after hemicastration in the mature rat without a change in plasma luteinizing hormone (LH) levels. These three questions (and their answers) were: 1) Can the testicular hemicastration response occur in hypophysectomized rats? Answer, No. 2) Does LH binding to the testis increase after hemicastration? Answer, No. 3) Is there a neural route to the testis alternate to the superior spermatic plexi? Answer, Yes, apparently there is, since hemivasectomy contralateral to the excised testis partially suppressed the testicular hemicastration response (150.4 +/- 13.2 ng/ml in hemicastrated, sham- hemivasectomized rats [n = 18] vs. 109.4 +/- 11.6 ng/ml in hemicastrated, hemivasectomized rats [n = 18], P less than 0.026). It was concluded that LH was probably necessary to the testicular hemicastration response but that its presence did not provide a mechanism. The response was mediated at least partly through the inferior spermatic nerves associated with the vas deferens. A possible reason, although highly speculative, for failure to previously block the testicular hemicastration response by bilateral denervation of the superior spermatic plexi (Mock and Frankel , 1982) was that during the 12-wk interval between denervation and hemicastration, testicular innervation functionally transferred from the superior spermatic to the inferior spermatic nerves.


[Identification of a new antigenic variant of simian adenovirus (SV30N)].
A new variant of simian adenovirus SV30-N is described. The variant is antigenically close to SV30 virus in the neutralization test but has some antigenic relationship to SA7 virus. The properties of SV30-N virus were retained after 5 clonings by the plaque method, after passage and cloning of the virus treated with immune sera to SV30 or SA7. The heteroduplex analysis showed DNA of the virus under study to be by 80% and 40% homologous to SV30 DNA and SA7 DNA, respectively, whereas no heteroduplex molecules between SV30 and SA7 DNA were found. By the set of polypeptides the antigenic variant SV30-N is close to SV30 but differs from SV30 and, particularly from SA7. Unlike SV30 and SA7 viruses, the SV30-N virus showed no oncogenic activity for hamsters.


Suppression of polymethyl methacrylate dust explosion by ultrafine water mist/additives.
The suppressions of ultrafine water mists containing additives (NaCl and NaHCO3) on 100 nm, 5 μm, and 30 μm polymethyl methacrylate (PMMA) dust explosions were experimentally studied in a dust-explosion apparatus. High-speed photography showed that maximum vertical positions and flame propagation velocities were significantly decreased by suppression with ultrafine water mist/additives. Flame propagation velocities in 100 nm, 5 μm, and 30 μm dust explosions suppressed by the ultrafine pure water mist were reduced by 48.2%, 27.7%, and 15.3%, respectively. Maximum temperatures and temperature rising rates measured by a fine thermocouple in nano- and micro-PMMA dust explosions were also significantly decreased. It was proved that the addition of NaCl and NaHCO3 improved the suppression effects of the ultrafine pure water mist. The improvement of explosion suppression by an 8% NaHCO3 mist was superior to that of a 16% NaCl mist. The suppression mechanisms of ultrafine water mist/additives are further discussed by analyzing the physical and chemical effects.


Cognitive functioning in patients with familial bipolar I disorder and their unaffected relatives.
Impairments in verbal learning and memory, executive functions and attention are manifest in some euthymic patients with bipolar disorder (BPD). However, evidence is sparse on their putative role as aetiologically important genetic vulnerability markers for the disorder. This population-based study examined the cognitive functions of affected and unaffected individuals in families with BPD. The aim was to discover whether any cognitive function would indicate genetic liability to the disorder and could thus be regarded as endophenotypes of BPD. A diagnostic interview and a neuropsychological test battery were administered to 32 familial bipolar I disorder patients, 40 of their unaffected first-degree relatives and 55 controls, all representing population-based samples. Unaffected first-degree relatives showed impairment in psychomotor performance speed and slight impairment in executive function. Bipolar patients were impaired in verbal learning and memory compared with unaffected relatives and controls. They also differed from controls in tasks of executive functions. There were no difference between the groups in simple attention and working memory tasks. Impaired psychomotor performance speed and executive function may represent endophenotypes of BPD, reflecting possible underlying vulnerability to the disorder. Verbal memory impairments appear to be more related to the fully developed disorder.


Survivorship of immature stages of Anopheles gambiae s.l. (Diptera: Culicidae) in natural habitats in western Kenya highlands.
We examined the survivorship of Anopheles gambiae s.l. Giles (Diptera: Culicidae) larvae and habitat productivity in three major habitat types in the western Kenya highlands. The age-specific distribution was determined for larvae and pupae, and survivorship curves were constructed. Larval-to-pupal survivorship was 6.8% in drainage ditches, 4.3% in cow hoofprints, and 1.8% in disused goldmines, respectively. High mortality rates were observed in all developmental stages. The estimated daily survival rate was highest in drainage ditches (0.74), followed by cow hoofprints (0.71), and it was lowest in disused goldmines (0.62). Productivity of emerging An. gambiae adults was generally low in these larval habitats (1.35, 1.55, and 1.84 mosquitoes per m2 per wk in drainage ditches, disused goldmines, and cow hoofprints, respectively). In total, seven families of larval mosquito predators were identified from the larval habitats, including Hydrophilidae, Dytiscidae, Corixidae, Nepidae, Notonectidae, Belostomatidae, and Cordulidae. Predator density in disused goldmines was significantly higher than that of other habitat types. Determination of the relative importance of predation, habitat stability and food contents on natural mosquito habitat productivity would help to design cost-effective vector control methods specifically targeted at the productive habitats.


Does Cardiovascular Comorbidity Influence the Prescribing of Bronchodilators in Chronic Obstructive Pulmonary Disease?
Cardiovascular disease (CVD) is the most prevalent comorbidity for chronic obstructive pulmonary disease (COPD). Potential adverse cardiovascular events of bronchodilators warrant their cautionary use in the comorbid COPD-CVD population, yet little is known about the prescribing of bronchodilators in this high-risk patient group. To determine whether comorbid CVD is associated with reduced bronchodilator prescribing in patients with COPD. In addition, we explored how the association was modified by gender, concurrent asthma, and concomitant β-blocker (BB) use. A cross-sectional study was conducted using the 2010 National Ambulatory Medical Care Survey. All visits among diagnosed COPD patients 40 years and older were examined. Logistic regression on survey-weighted data was used to predict treatment with bronchodilators and determine the influence of gender, asthma, and BBs on bronchodilator prescribing. Among 11 627 061 ambulatory COPD visits, we found a significantly lower bronchodilator treatment rate among patients with comorbid CVD (32.3%) than among patients without CVD (57.6%). The observed effect was modified by gender, asthma, and BBs. Deprescribing was more pronounced for females than males, for nonusers of BBs than users of BBs, and for asthma patients than nonasthma patients. CVD did not affect bronchodilator prescribing in either asthmatic or nonasthmatic males taking BBs. Most COPD patients with concurrent CVD were less likely to be prescribed bronchodilators, with the exception of males who were also prescribed BBs. Thus, this study highlights a specific patient subgroup for whom the guidelines are less likely to be observed.


Large coronary intramural hematomas: a case series and focused literature review.
Isolated spontaneous coronary intramural hematoma is a unique subset of spontaneous coronary artery dissection that is characterized by a hemorrhage limited to the medial-adventitial layers, causing subsequent hematoma formation without visible intimal flaps. It is an infrequent and serious coronary vessel wall pathology, with poorly understood underlying pathogenic mechanisms. Affected individuals may present with a broad spectrum of symptoms ranging from acute coronary syndromes (ACS) to cardiogenic shock or even sudden cardiac death. The disease entity causes challenges in terms of both diagnostics and treatment strategy. Coronary intramural hematomas can also occur iatrogenically, as a complication to percutaneous coronary intervention (PCI). Coronary angiography (CAG) has limited diagnostic value in the absence of intimal dissections, and lesions are often angiographically ambiguous. Intravascular ultrasound (IVUS) is an important diagnostic tool in establishing the correct diagnosis, as it provides a complete vessel wall assessment, and enables morphometric information regarding the magnitude and severity of the underlying hematoma. Due to the rarity of this clinical scenario, no randomized, controlled trials exist to guide treatment, and no consensus regarding management is available. Currently, treatment strategies are based on a case-by-case clinical assessment, and experiences described in previous, limited retrospective studies and case reports.


The effect of delay on flap survival in an irradiated field.
Chronic radiation skin injury without ulceration was induced in rats by administering either 3000 or 5000 rads in staged doses. Either 4 or 8 months later, McFarlane et al. dorsal flaps were elevated, with half being delayed and half non-delayed. Measurements showed that flap survival in irradiated skin was significantly increased by delay [approximately two of four experimental groups (4 months, 5000 rads; and 8 months, 3000 rads)]; flap survival was increased with borderline significance in a third experimental group (8 months, 5000 rads). These data indicate that flap survival can be increased by delay in an irradiated field. The presence of the vascular delay phenomenon suggests that microvascular occlusion alone cannot account for radiation-induced complications in skin.


Non-invasive identification of stable rotors and focal sources for human atrial fibrillation: mechanistic classification of atrial fibrillation from the electrocardiogram.
To develop electrocardiogram (ECG) tools to quantify the number of sources for atrial fibrillation (AF), i.e. spatially stable rotors and focal impulses, and whether they lie in right or left atrium. Intracardiac mapping has recently shown that paroxysmal and persistent AF is sustained by rotors or focal sources that are stable in location and thus targets for limited ablation [focal impulse and rotor modulation (FIRM)] to eliminate AF. Importantly, the numbers and locations of concurrent sources determine both the complexity of AF and the approach for ablation. In 36 AF patients (n = 29 persistent, 63 ± 9 years) in the CONventional ablation with or without Focal Impulse and Rotor Modulation (CONFIRM) trial, we developed phase lock (PL) to quantify spatial repeatability of ECG 'F-waves' between leads over time. Phase lock spectrally quantifies the angle θ between F-wave voltages in planes formed by ECG leads I, aVF, and V1 at successive points in time. We compared PL with ECG spectral dominant frequency (DF) and organizational index (OI) to characterize stable rotors and focal sources validated by intracardiac FIRM mapping. Focal impulse and rotor modulation ablation alone at ≤3 sources acutely terminated and rendered AF non-inducible or substantially slowed AF in 31 of 36 patients. Receiver operating characteristics of PL for this endpoint had area under the curve (AUC) = 0.72, and the optimum cut-point (PL = 0.09) had 74% sensitivity, 92% positive predictive value (PPV). Receiver operating characteristics areas for OI and DF were 0.50 and 0.58, respectively. Left (n = 28) or right (n = 3) atrial sources were localized by PL with AUC = 0.85, sensitivity 100%, PPV 30%, and negative predictive value 100%. Spectral DF provided AUC = 0.79. Notably, PL did not comigrate with diagnosis of paroxysmal or persistent AF (P = NS), unlike ECG DF. The novel metric of ECG PL identifies patients with fewer (≤3) or greater numbers of stable rotors/focal sources for AF, validated by intracardiac FIRM mapping, and localized them to right or left atria. These data open the possibility of using 12-lead ECG analyses to classify AF mechanistically and plan procedures for right- or left-sided FIRM ablation.


Diet in the Pathogenesis and Management of Ulcerative Colitis; A Review of Randomized Controlled Dietary Interventions.
Epidemiological and experimental studies have suggested that diet is one of the environmental factors that contributes to the onset and pathophysiology of ulcerative colitis. Although many patients suffering from ulcerative colitis attribute their symptoms or disease relapse to dietary factors, only a few well-designed randomized controlled trials have been done to investigate the role of diet in the management of ulcerative colitis. Here, we review the potential mechanisms of the relationship between diet and pathogenesis of ulcerative colitis and summarize randomized controlled dietary interventions that have been conducted in ulcerative colitis patients.


The role of chemokines in hypertension and consequent target organ damage.
Immune cells infiltrate the kidney, vasculature, and central nervous system during hypertension, consequently amplifying tissue damage and/or blood pressure elevation. Mononuclear cell motility depends partly on chemokines, which are small cytokines that guide cells through an increasing concentration gradient via ligation of their receptors. Tissue expression of several chemokines is elevated in clinical and experimental hypertension. Likewise, immune cells have enhanced chemokine receptor expression during hypertension, driving immune cell infiltration and inappropriate inflammation in cardiovascular control centers. T lymphocytes and monocytes/macrophages are pivotal mediators of hypertensive inflammation, and these cells migrate in response to several chemokines. As powerful drivers of diapedesis, the chemokines CCL2 and CCL5 have long been implicated in hypertension, but experimental data highlight divergent, context-specific effects of these chemokines on blood pressure and tissue injury. Several other chemokines, particularly those of the CXC family, contribute to blood pressure elevation and target organ damage. Given the significant interplay and chemotactic redundancy among chemokines during disease, future work must not only describe the actions of individual chemokines in hypertension, but also characterize how manipulating a single chemokine modulates the expression and/or function of other chemokines and their cognate receptors. This information will facilitate the design of precise chemotactic immunotherapies to limit cardiovascular and renal morbidity in hypertensive patients.


Erbium:YAG laser skin resurfacing.
The Erbium:YAG laser is a useful flexible tool for the surgeon interested in providing overall skin rejuvenation. With its unique wavelength, greater absorption, and less thermal injury, it can offer more precise ablation with greater safety and shorter healing times. All skin types as well as all body parts can safely be treated, from simple lentigines to deeper rhytides. The Erbium:YAG laser is an effective tool for complete skin rejuvenation. System specifications and parameters are reviewed along with pathology,uses, and techniques including postoperative care, risks, and safety profile.


Predictive Performance for Hospital Mortality of SAPS 3, SOFA, ISS, and New ISS in Critically Ill Trauma Patients: A Validation Cohort Study.
Background: It is not known whether anatomical scores perform better than general critical care scores for trauma patients admitted to the intensive care unit (ICU). We compare the predictive performance for hospital mortality of general critical care scores (SAPS 3 and SOFA) with anatomical injury-based scores (Injury Severity Score [ISS] and New ISS [NISS]). Methods: Retrospective cohort study of patients admitted to a specialized trauma ICU from a tertiary hospital in São Paulo, Brazil between May, 2012 and January, 2016. We retrieved data from the ICU database for critical care scores and calculated ISS and NISS from chart data and whole body computed tomography results. We compared the predictive performance for hospital mortality of each model through discrimination, calibration, and decision-curve analysis. Results: The sample comprised 1053 victims of trauma admitted to the ICU, with 84.2% male patients and mean age of 40 (±18) years. Main injury mechanism was blunt trauma (90.7%). Traumatic brain injury was present in 67.8% of patients; 43.3% with severe TBI. At the time of ICU admission, 846 patients (80.3%) were on mechanical ventilation and 644 (64.3%) on vasoactive drugs. Hospital mortality was 23.8% (251). Median SAPS 3 was 41; median maximum SOFA within 24 h of admission, 7; ISS, 29; and NISS, 41. AUROCs (95% CI) were: SAPS 3 = 0.786 (0.756-0.817), SOFA = 0.807 (0.778-0.837), ISS = 0.616 (0.577-0.656), and NISS = 0.689 (0.649-0.729). In pairwise comparisons, SAPS 3 and SOFA did not differ, while both outperformed the anatomical scores (p < .001). Maximum SOFA within 24 h of admission presented the best calibration and net benefit in decision-curve analysis. Conclusions: Trauma-specific anatomical scores have fair performance in critically ill trauma patients and are outperformed by SAPS 3 and SOFA. Illness severity is best characterized by organ dysfunction and physiological variables than anatomical injuries.


Multiple testing issues in discriminating compound-related peaks and chromatograms from high frequency noise, spikes and solvent-based noise in LC-MS data sets.
Liquid Chromatography--Mass Spectrometry (LC-MS) is a powerful method for sensitive detection and quantification of proteins and peptides in complex biological fluids like serum. LC-MS produces complex data sets, consisting of some hundreds of millions of data points per sample at a resolution of 0.1 amu in the m/z domain and 7000 data points in the time domain. However, the detection of the lower abundance proteins from this data is hampered by the presence of artefacts, such as high frequency noise and spikes. Moreover, not all of the tens of thousands of the chromatograms produced per sample are relevant for the pursuit of the biomarkers. Thus in analysing the LC-MS data, two critical pre-processing issues arise. Which of the thousands of the: 1. chromatograms per sample are relevant for the detection of the biomarkers?, and 2. signals per chromatogram are truly compound-related? Each of these issues involves assessing the significance (deviation from noise) of multiple observations and the issue of multiple comparisons arises. Current methods disregard the multiplicity and provide no concrete threshold for significance. However, with such procedures, the probability of one or more false-positives is high as the number of tests to be performed is large, and must be controlled. Realizing that the cut-offs for declaring a chromatogram (or a signal) to be compound-related can hugely influence which proteins are detected, it seems natural to define thresholds that are neither arbitrary nor subjective. We suggest the choice of thresholds guided by the critical aim of controlling the False Discovery Rate (FDR) in multiple hypotheses testing for significance over a large set of features produced per sample. This involves the use of the regression diagnostics to characterize the signals of a chromatogram (e.g. as outliers or influential) and to suggest suitable tests statistics for the multiple testing procedures (MTP) for discriminating noise and spikes from true signals. The role of the Generalized Linear Models (GLM) in this MTP is investigated. The method is applied to LC-MS datasets from trypsin-digested serum spiked with varying levels of horse heart cytochrome C (cytoc).


Identification of STN7/STN8 kinase targets reveals connections between electron transport, metabolism and gene expression.
The thylakoid-associated kinases STN7 and STN8 are involved in short- and long-term acclimation of photosynthetic electron transport to changing light conditions. Here we report the identification of STN7/STN8 in vivo targets that connect photosynthetic electron transport with metabolism and gene expression. Comparative phosphoproteomics with the stn7 and stn8 single and double mutants identified two proteases, one RNA-binding protein, a ribosomal protein, the large subunit of Rubisco and a ferredoxin-NADP reductase as targets for the thylakoid-associated kinases. Phosphorylation of three of the above proteins can be partially complemented by STN8 in the stn7 single mutant, albeit at lower efficiency, while phosphorylation of the remaining three proteins strictly depends on STN7. The properties of the STN7-dependent phosphorylation site are similar to those of phosphorylated light-harvesting complex proteins entailing glycine or another small hydrophobic amino acid in the -1 position. Our analysis uncovers the STN7/STN8 kinases as mediators between photosynthetic electron transport, its immediate downstream sinks and long-term adaptation processes affecting metabolite accumulation and gene expression.


Role of Molecular Singlet Oxygen in Photochemical Degradation of NTO: DFT Study.
NTO (5-nitro-1,2,4-triazol-3-one), an energetic material used in military applications, may be released to the environment and dissolved in surface water and groundwater due to its good water solubility. Singlet oxygen is an important reactive oxygen species produced in the aquatic environment under sunlight irradiation. A detailed investigation of the possible mechanism for NTO decomposition in water induced by singlet oxygen as one of the pathways for NTO environmental degradation was performed by a computational study at PCM(Pauling)/M06-2X/6-311++G(d,p) level. Decomposition of NTO was found to be a multistep process that may begin with singlet oxygen attachment to the carbon atom of the C═N double bond. The formed intermediate undergoes cycle opening, and nitrogen gas, nitrous acid, and carbon (IV) oxide elimination. Isocyanic acid, arisen transiently, hydrolyzes into ammonia and carbon (IV) oxide. The obtained results show a significant increase in reactivity of the anionic form of NTO as compared to its neutral form. The calculated activation energies and high exothermicity of the studied processes support the contribution of singlet oxygen to NTO degradation into low-weight inorganic compounds in the environment.


Renal Endometriosis Mimicking Cystic Renal Tumor: Case Report and Literature Review.
Background: Endometriosis mainly affects female pelvic tissues and organs, and the presence of endometriosis in the kidney is extremely rare. Case Presentation: We report a case of a 48-year-old woman who presented with intermittent hematuria. She was found to have a cystic mass on renal ultrasonography, and contrast-enhanced computed tomography (CT) showed slight enhancement of the cystic wall and septa. These findings were indicative of cystic renal tumor. The patient subsequently underwent partial right nephrectomy. Histopathology revealed endometriosis of the right renal parenchyma. The patient recovered well and had no evidence of a recurrent renal mass at the 3 months' follow up. Conclusion: The possibility of renal endometriosis should be considered in a female patient with a cystic renal mass and clinical symptoms related to the menstrual cycle.


New simple dye-uptake assay for interferon.
Using the spectrophotometer that the authors developed, the amounts of human leukocyte and mouse L cell interferons on FL cells and L929 cells were measured and values were compared with those measured by the cytopathogenic effect (CPE) reduction method (CPE method). The spectrophotometric method, which was simpler than the original dye-uptake method, was found to be more sensitive than the latter. When Sindbis virus was used instead of vesicular stomatitis virus (VSV), there were no significant differences in the sensitivities of the two methods or the interferon titers estimated. When FL cells or L929 cells were treated with interferon at the time of their dispersion, their interferon titers were almost the same as those of cells treated with interferon 2 days after dispersion. It is concluded that this new dye-uptake method is useful for assay of human and mouse interferons.


Purification and characterization of proteins that bind to yeast ARSs.
Two proteins that bind to yeast ARS DNA have been purified using conventional and oligonucleotide affinity chromatography. One protein has been purified to homogeneity and has a mass of 135 kDa. Competitive binding studies and DNase I footprinting show that the protein binds to a sequence about 80 base pairs away from the core consensus in the region known as domain B. This region has previously been shown to be required for efficient replication of plasmids carrying ARS1 elements. To investigate further whether the protein might have a function related to the ability of ARSs to act as replicators, binding to another ARS was tested. The protein binds to the functional ARS adjacent to the silent mating type locus HMR, called the HMR-E ARS, about 60 base pairs from the core consensus sequence. Surprisingly, there is little homology between the binding site at the HMR-E ARS and the binding site at ARS1. The 135-kDa protein is probably the same as ABF-I (SBF I) (Shore, D., Stillman, D. J. Brand, A. H., and Nasmyth, K. A. (1987) EMBO J. 6, 461-467; Buchman, A. R., Kimmerly, W. J., Rine, J., and Kornberg, R. D. (1988) Mol. Cell. Biol. 8, 210-225). A second DNA-binding protein was separated from ABF-I during later stages of the purification. This protein, which we designate ABF-III, also binds specifically to the ARS1 sequence, as shown by DNase I footprinting, at a site adjacent to the ABF-I recognition site. Purification of these two ARS binding proteins should aid in our understanding of the complex mechanisms that regulate eukaryotic DNA replication.


Chemical transformation of gilvocarcin V. Modification of the side-chain.
Gilvocarcin V was chemically transformed to alter its biological activities as well as its solubility by mainly focusing on the vinyl side chain. The oxirane and oxime derivatives showed slightly decreased in vivo antitumor activity, while the aminoethylmorpholine derivative turned out to be soluble in some organic solvents.


Immunochemical characterization of six monoclonal antibodies to human apolipoprotein A-I: epitope mapping and expression.
We have produced and characterized six murine monoclonal antibodies to human apolipoprotein A-I named A-I-9, A-I-12, A-I-15, A-I-16, A-I-19, and A-I-57. All monoclonal antibodies were specific for apolipoprotein A-I and bound between 55% and 100% of 125I-labeled high density lipoproteins (HDL) in a fluid phase radioimmunoassay. All antibodies possessed a higher affinity to apoA-I in HDL than to free, delipidated apoA-I. Two of them, particularly A-I-12 and A-I-15, which were directed to the same or very close epitopes on the molecule, recognized very poorly the delipidated protein. Binding of apoA-I to phospholipid restored the immunoreactivity of the monoclonal antibodies to the protein suggesting that lipids play an important role in determining the immunochemical structure of apoA-I. Using CNBr fragments and synthetic peptides, the epitopes for the antibodies were mapped as follows: A-I-19, CNBr fragment 1; A-I-12 and 15, CNBr fragment 2; A-I-9 and A-I-16, CNBr fragment 3; A-I-57, CNBr fragment 4. Antibody A-I-57 failed to recognized a mutant form of apoA-I, A-IMilano (Arg173----Cys) by immunoblotting and by competitive radioimmunoassay demonstrating that substitution of a single amino acid in human apoA-I may cause the loss of an antigenic determinant.


Dynamic Changes in the Quantitative Electroencephalographic Spectrum During Attention Tasks in Patients With Prader-Willi Syndrome.
Introduction: Attention problems are frequently observed in patients with Prader-Willi syndrome (PWS); however, only few studies have investigated the severity and mechanisms of attention problems in them. In this study, we aim to evaluate dynamic changes in the quantitative electroencephalographic (EEG) spectrum during attention tasks in patients with PWS. Method: From January to June 2019, 10 patients with PWS and 10 age-matched neurotypical control participants were recruited at Taipei Tzu Chi Hospital. Each participant completed Conners' continuous performance test, third edition (CPT-3), tasks with simultaneous EEG monitoring. The dynamic changes in the quantitative EEG spectrum between the resting state and during CPT-3 tasks were compared. Results: Behaviorally, patients with PWS experienced significant attention problems, indicated by the high scores for several CPT-3 variables. The theta/beta ratio of the resting-state EEG spectrum revealed no significant differences between the control participants and patients with PWS. During CPT-3 tasks, a significant decrease in the alpha power was noted in controls compared with that in patients with PWS. The attention-to-resting alpha power ratio was positively correlated with many CPT-3 variables. After adjusting for genotype, age, intelligence, and body mass index, the attention-to-resting alpha power ratio was still significantly correlated with participants' commission errors. Conclusion: This study provides evidence that attention problems are frequently observed in patients with PWS, while attention impairment can be demonstrated by dynamic changes in the quantitative EEG spectrum.


Effects of menopause on brain structural changes in schizophrenia.
The aim of this study was to investigate the influences of menopause on brain morphological changes in schizophrenia using magnetic resonance imaging (MRI). Forty female schizophrenia patients, 20 premenopausal and 20 postmenopausal, and 50 female controls underwent cerebral MRI. Optimized voxel-based morphometry was performed with Statistical Parametric Mapping version 5. Compared with controls, regional gray matter reductions in schizophrenia patients were observed in the insula, superior temporal gyrus, anterior cingulate, parahippocampal gyrus, and thalamus. Direct comparison between the patient groups showed that the gray matter of postmenopausal patients was significantly smaller when compared with premenopausal patients in the left middle frontal gyrus, and no region had significantly lower gray matter volume in premenopausal patients relative to postmenopausal patients. Significant negative correlation between gray matter volume and the interval after menopause was found in the right superior frontal gyrus in the postmenopause patient group. Differential morphological alterations between postmenopausal and premenopausal schizophrenia patients were observed, suggesting that the female hormone plays a protective role against schizophrenia.


[Lysis therapy in aortic dissection: by hypertrophic cardiomyopathy simulated myocardial infarct].
A 52-year-old patient presenting with severe thoracic pain of more than 30 minutes duration and ST-segment elevation > 1 mV in leads V1-3 was given 100 mg rt-PA. With persisting thoracic pain and ST-segment elevation, coronary angiography was performed 90 minutes after administration of thrombolytic therapy. Surprisingly, angiography revealed normal coronary arteries in presence of marked hypertrophic cardiomyopathy. New onset of thoracic pain after four hours led to a chest radiograph demonstrating increased mediastinal widening and new pleural effusion as compared to a baseline chest film prior to rt-PA. Spiral computed tomography revealed overt Type B aortic dissection with an intimal flap and communication distal to the origin of the left subclavian artery with signs of a mediastinal hematoma. Immediate cardiac surgery with replacement of the dissected aortic segment was successfully performed. A literature review revealed 4% of aortic dissection to present with electrocardiographic signs of acute myocardial infarction and 13% to have concomitant coronary artery disease; moreover 34 cases of aortic dissection were misdiagnosed as infarction and subjected to thrombolytic agents with a mortality of 64% underlining the importance of exclusion of dissection prior to thrombolytic therapy.


Moderate-intensity exercise: for our patients, for ourselves.
Even more than the general population, orthopaedic patients suffer from multiple consequences of immobility. Fortunately, most of these patients are physically able to participate in some degree of moderately intense exercise. Helping the orthopaedic patient initiate and adhere to a moderately intense exercise routine is a nursing challenge. This challenge is even more difficult because as nurses, we are often expecting our patients to make a lifestyle change that we have been unable or unwilling to make ourselves. Understanding the health-related benefits that can be derived from participation in moderately intense exercise routines and the recommended guidelines for exercise frequency and intensity is a first step toward initiating a fitness routine. Using information acquired from research, nurses can assess for specific facilitators and barriers to exercise participation. This assessment data can then be used to individualize plans for exercise that meet the fitness needs of our patients and ourselves.


African American Young Adults' Pain and Pain Reduction Strategies.
Effective acute pain management strategies are important for young adults in order to reduce risk for transition to chronic pain. To describe pain and pain self-management strategies used by African American young adults. A national online cross-sectional survey design was used. Ninety-four African Americans Qualtrics panelists ages 18-25 who reported previous experience with acute pain responded. Methods: Respondents completed the Brief Pain Inventory Short Form to describe their pain intensity, pain interference with function, pain self-management, and percent of relief obtained from their self-management. African American young adults reported pain primarily in the back (n = 22, 23.4%) and head (n = 19, 20.2%), with moderate pain intensity M = 4.5 (standard deviation [SD] = 1.79) and pain interference with function M = 4.6 (SD = 2.36). African American young adults described their worst pain in the last 24 hours as M = 5.7 (SD = 2.01), least pain as M = 3.4 (SD = 2.41), and average pain as M = 5.1 (SD = 2.09). They reported 61.3% pain relief from self-treatment. A total of 45 (47.9%) reported no pain self-management strategies. African American young adults report moderate levels of pain intensity and pain interference with function. A significant number report no pain self-management strategies. Focused pain assessment and education about efficacious pain self-management strategies, both pharmacological and complementary, could assist young African Americans to reduce their pain and risk of chronic pain in the future.


Intraoperative Detection of Efferent Lymphatic Vessels Emerging from Lymph Node during Lymphatic Microsurgery.
Although the usefulness of efferent lymphaticovenular anastomosis (ELVA) in lymphatic microsurgery has been reported, the optimal method to distinguish efferent from afferent lymphatics is not yet established. We propose a novel technique to detect efferent lymphatics appropriate for anastomosis. In total, 62 groin lymph nodes (LNs) of 46 limbs were divided into four groups based on the findings of indocyanine green lymphography: n = 15 in normal, 15 in dermal backflow stage 0, 18 in stage I, and 14 in stage II groups. The target LN and afferent lymphatic connecting it were preoperatively detected using ultrasonography. Intraoperatively, 0.05 mL patent blue dye was slowly manually injected from the afferent lymphatic; the lymphatic(s) subsequently stained was diagnosed as the efferent lymphatic(s) emerging from the node. The success rates of efferent lymphatic detection, sizes of LN, and diameter of efferent lymphatics were compared among the groups. Both LN size and diameter of efferent lymphatic were significantly larger in the stage 0 and I groups than the other groups (p < 0.01). Efferent lymphatic could be stained in 13, 13, 18, and 9 LNs in the normal and stage 0, I, and II groups, respectively. A significant difference was observed between the stage I and II groups regarding the success rate of efferent lymphatic detection (p = 0.04). Efferent lymphatics could be detected using patent blue dye in 85.5% of the cases. The patients in early-stage lymphostasis might be the most appropriate candidates for ELVA.


Secretion properties, clearance, and therapy in airway disease.
Chronic airway diseases like cystic fibrosis, chronic bronchitis, asthma, diffuse panbronchiolitis, and bronchiectasis are all associated with chronic inflammation. The airway mucosa responds to infection and inflammation in part by surface mucous (goblet) cell and submucosal gland hyperplasia and hypertrophy with mucus hypersecretion. Products of inflammation including neutrophil derived DNA and filamentous actin, effete cells, bacteria, and cell debris all contribute to mucus purulence and, when this is expectorated it is called sputum. Mucus is usually cleared by ciliary movement, and sputum is cleared by cough. These airway diseases each are associated with the production of mucus and sputum with characteristic composition, polymer structure, and biophysical properties. These properties change with the progress of the disease making it possible to use sputum analysis to identify the potential cause and severity of airway diseases. This information has also been important for the development of effective mucoactive therapy to promote airway hygiene.


Data quality of general practice electronic health records: the impact of a program of assessments, feedback, and training.
The aim of this study was to investigate the impact of a program of repeated assessments, feedback, and training on the quality of coded clinical data in general practice. A prospective uncontrolled intervention study was conducted in a general practice research network. Percentage of recorded consultations with a coded problem title and percentage of patients receiving a specific drug (e.g., tamoxifen) who had the relevant morbidity code (e.g., breast cancer) were calculated. Annual period prevalence of 12 selected morbidities was compared with parallel data derived from the fourth National Study of Morbidity Statistics from General Practice (MSGP4). The first two measures showed variation between practices at baseline, but on repeat assessments all practices improved or maintained their levels of coding. The period prevalence figures also were variable, but over time rates increased to levels comparable with, or above, MSGP4 rates. Practices were able to provide time and resources for feedback and training sessions. A program of repeated assessments, feedback, and training appears to improve data quality in a range of practices. The program is likely to be generalizable to other practices but needs a trained support team to implement it that has implications for cost and resources.


Disrupted knee - disrupted me: a strenuous process of regaining balance in the aftermath of an anterior cruciate ligament injury.
Individuals describe both short and long term consequences after an anterior cruciate ligament (ACL) injury. Functional impairments are well documented while psychological, social and contextual factors need to be further investigated. By the use of a qualitative method incorporating a biopsychosocial lens, we aimed to explore individuals' experiences of living and coping with an ACL rupture with a specific focus on experiences significant to overall life, activity in daily living and physical activity more than one year after injury. Twelve participants were chosen strategically by a purposive sampling. Four men and eight women (19-41 years) with an ACL rupture 2-25 years ago, were included. Semi-structured interviews were used and analysed with qualitative content analysis. The results consisted of one overarching theme: 'A strenuous process towards regaining balance' which built on three categories 'Disrupted knee', 'Disrupted me' and 'Moving forward with new insights'. The overarching theme captures the participants' experiences of a strenuous process towards regaining both physical and mental balance in the aftermath of an ACL injury. The results illuminate how participants were forced to cope with a physically 'disrupted knee', as well as facing mental challenges, identity challenges and a 'disrupted me'. By gradual acceptance and re-orientation they were moving forward with new insights - although still struggling with the consequences of the injury. Individuals with an ACL injury experience both physical, psychological, and social challenges several years after injury. In addition to the functional impairments, diverse psychological, social and contextual 'disruptions' and struggles may also be present and influence the rehabilitation process. It is important that physiotherapists identify individuals who face such challenges and individually tailor the rehabilitation and support. A biopsychosocial approach is recommended in the clinical practice and future studies focusing on psychosocial processes in the context of ACL rehabilitation are warranted.


[Late results of mesocaval and distal splenorenal shunts].
Shunt operations were carried out in 63 patients. Child A and B patients received alternatingly a distal splenorenal or mesocaval shunt, Child C and emergency patients exclusively mesocaval shunts. The postoperative mortality in the emergency group was 29%, in the elective group with mesocaval shunts 13% and with distal splenorenal shunts 17%. After 3 years the cumulative survival rates were similar in all patient groups and amounted to 54-64%. Episodes of acute encephalopathy in the late phase were observed only after mesocaval shunts (7%).


Non-alcoholic fatty liver disease proteomics.
Non-alcoholic fatty liver disease (NAFLD) is an important cause of chronic liver injury that has gained concern in clinical hepatology. The principal aim of this study was to find differences in protein expression between patients with NAFLD and healthy controls. Changes in protein expression of liver samples from each of the three groups of subjects, controls, non-alcoholic steatosis, and non-alcoholic steatohepatitis (NASH), were analyzed by DIGE combined with MALDI TOF/TOF analysis, a proteomic approach that allows to compare hundreds of proteins simultaneously. Forty-three proteins exhibiting significant changes (ratio ≥1.5, p<0.05) were characterized, 22 comparing steatosis samples versus control samples and 21 comparing NASH versus control samples. Ten of these proteins were further analyzed by Western blot in tissue samples to confirm the observed changes of protein expression using DIGE. The proteins validated were further tested in serum samples of different cohorts of patients. Following this approach we identified two candidate markers, carbamoyl phosphate synthase 1 and 78 kDa glucose-regulated protein, differentially expressed between control and NASH. This proteomics approach demonstrates that DIGE combined with MALDI TOF/TOF and Western blot analysis of tissue and serum samples is a useful approach to identify candidate markers associated with NAFLD, resulting in proteins whose level of expression can be correlated to a disease state.


Rare cause of absence of femoral arterial pulse: bilateral common iliac artery hypoplasia.
Congenital anomaly of the iliac arteries is rare, and, given that patients are generally asymptomatic, diagnosis in the newborn period is difficult. Herein is presented the case of a newborn with bilateral hypoplasia of the common iliac arteries, seen on multi-slice computed tomography while investigating the absence of femoral pulse. This anomaly is a rare cause of absence of femoral arterial pulse.


Reconstructing network topology and coupling strengths in directed networks of discrete-time dynamics.
Reconstructing network connection topology and interaction strengths solely from measurement of the dynamics of the nodes is a challenging inverse problem of broad applicability in various areas of science and engineering. For a discrete-time step network under noises whose noise-free dynamics is stationary, we derive general analytic results relating the weighted connection matrix of the network to the correlation functions obtained from time-series measurements of the nodes for networks with one-dimensional intrinsic node dynamics. Information about the intrinsic node dynamics and the noise strengths acting on the nodes can also be obtained. Based on these results, we develop a scheme that can reconstruct the above information of the network using only the time-series measurements of node dynamics as input. Reconstruction formulas for higher-dimensional node dynamics are also derived and illustrated with a two-dimensional node dynamics network system. Furthermore, we extend our results and obtain a reconstruction scheme even for the cases when the noise-free dynamics is periodic. We demonstrate that our method can give accurate reconstruction results for weighted directed networks with linear or nonlinear node dynamics of various connection topologies, and with linear or nonlinear couplings.


L-dopa modulates motor cortex excitability in Alzheimer's disease patients.
In Alzheimer's disease (AD), transcranial magnetic stimulation (TMS) studies have shown abnormalities of motor cortical excitability, such as a decreased intra-cortical inhibition (ICI) and changes in resting motor threshold (rMT). We studied the effects of L-dopa on rMT and ICI in a cohort of moderate AD patients after paired-pulse TMS. Results were compared with a control group of healthy subjects. As expected, AD patients showed a significant reduction in ICI and a lower rMT. L-dopa administration (soluble form, melevodopa 200 mg) promptly reversed the ICI impairment up to normalization. This effect was specific, since it was not mimicked in control subjects. These results indicate a possible role of dopamine in modulating AD cortical excitability, thus suggesting an interaction between dopaminergic ascending pathways and endogenous intracortical transmitters. In addition, considering that L-dopa showed a pharmacological profile similar to the one of cholinomimetics, L-dopa might represent a reliable tool to study new therapeutic perspective and strategies for AD.


New class of RNA biomarker for endometriosis diagnosis: The potential of salivary piRNA expression.
In contrast to miRNA expression, little attention has been given to piwiRNA (piRNA) expression among endometriosis patients. The aim of the present study was to explore the human piRNAome and to investigate a potential piRNA saliva-based diagnostic signature for endometriosis. Data from the prospective "ENDOmiRNA" study (ClinicalTrials.gov Identifier: NCT04728152) were used. Saliva samples from 200 patients were analyzed in order to evaluate human piRNA expression using the piRNA bank. Next Generation Sequencing (NGS), barcoding of unique molecular identifiers and both Artificial Intelligence (AI) and machine learning (ML) were used. For each piRNA, sensitivity, specificity, and ROC AUC values were calculated for the diagnosis of endometriosis. 201 piRNAs were identified, none had an AUC ≥ 0.70, and only three piRNAs (piR-004153, piR001918, piR-020401) had an AUC between ≥ 0.6 and < 0.70. Seven were differentially expressed: piR-004153, piR-001918, piR-020401, piR-012864, piR-017716, piR-020326 and piR-016904. The respective correlation and accuracy to diagnose endometriosis according to the F1-score, sensitivity, specificity, and AUC ranged from 0 to 0.862 %, 0-0.961 %, 0.085-1, and 0.425-0.618. A correlation was observed between the patients' age (≥35 years) and piR-004153 (p = 0.002) and piR-017716 (p = 0.030). Among the 201 piRNAs, four were differentially expressed in patients with and without hormonal treatment: piR-004153 (p = 0.015), piR-020401 (p = 0.001), piR-012864 (p = 0.036) and piR-017716 (p = 0.009). Our results support the link between piRNAs and endometriosis physiopathology and establish its utility as a potential diagnostic biomarker using saliva samples. Per se, piRNA expression should be analyzed along with the clinical status of a patient.


Small Bowel Perforation during Suprapubic Trocar Cystotomy: A Case Report.
A 35 years old man presented with retention of urine secondary to meatal stenosis with bulbar urethral stricture. He had a distended, palpable, tender urinary bladder. Urethral catheterization and dilatation was tried but failed. A trocar cystostomy was performed under local anaesthesia, which led to the injury to the small bowel when least expected. This is a rare but well recognized complication of small bowel injury following blind trocar suprapubic cystostomy when it was least expected and as such had a significant bearing on its management. We discuss its subsequent management and possible mechanism underlying this unexpected and unfortunate complication in the given circumstances.


Primary Localized Cutaneous Amyloidosis Affecting Female Individuals of a Pakistani Pedigree.
Primary localized cutaneous amyloidosis is a group of rare conditions where amyloid deposition is limited to the skin without systemic manifestations. Most cases are sporadic; however, mutations in the oncostatin M receptor (OSMR) and interleukin-31 receptor A (IL31RA) genes can cause a familial form of the condition in up to 10% of cases. Here, we describe a family in which 8 female individuals are affected by either macular amyloidosis or amyloidosis cutis dyschromica. To the best of our knowledge, a sex-specific expression or the coexistence of 2 different phenotypes of primary localized cutaneous amyloidosis in 1 pedigree has not yet been reported.


Effects of habitual alcohol intake on ambulatory blood pressure, heart rate, and its variability among Japanese men.
We sought to examine effects of habitual alcohol intake on ambulatory blood pressure (BP), heart rate (HR), and HR variability among Japanese men. Subjects were 539 men aged 35 to 65 years from rural and urban communities. Ambulatory BP and HR were monitored with an automated, portable, noninvasive multibiomedical recorder. Power spectral analysis of the RR intervals on the ECG was performed every 5 minutes. Compared with nondrinkers, moderate drinkers (alcohol intake 23 to 45 g/d) and heavy drinkers (alcohol intake >or=46 g/d) showed higher age- and field-adjusted mean values of systolic and diastolic BPs during the morning and while awake, but there were no differences in BPs over 24-hour periods and while asleep among the alcohol intake categories. Alcohol intake was positively associated with mean values of sleep-morning differences and daytime variability in BPs, HRs while awake and asleep, and low frequency:high frequency ratio while asleep. The results were virtually unchanged after adjustment for body mass index, smoking, and diabetes mellitus. Compared with the nondrinkers, age- and field-adjusted odds ratios of the morning BP surge (excess elevation of BP in the morning: morning systolic BP minus sleep systolic BP >or=37 mm Hg) for light (alcohol intake 0 to 22 g/d), moderate, and heavy drinkers were 0.96 (95% CI: 0.34 to 2.78), 1.68 (95% CI: 0.64 to 4.38), and 2.73 (95% CI: 1.12 to 6.67), respectively. Habitual alcohol intake was associated with increased BP in the morning, HR while awake and asleep, and sympathetic activity while asleep, which may explain some of the mechanisms of the relationship between heavy alcohol intake and risk of cardiovascular diseases.


Nutrition and lung cancer: a case control study in Iran.
Despite many prospective and retrospective studies about the association of dietary habit and lung cancer, the topic still remains controversial. So, this study aims to investigate the association of lung cancer with dietary factors. In this study 242 lung cancer patients and their 484 matched controls on age, sex, and place of residence were enrolled between October 2002 to 2005. Trained physicians interviewed all participants with standardized questionnaires. The middle and upper third consumer groups were compared to the lower third according to the distribution in controls unless the linear trend was significant across exposure groups. Conditional logistic regression was used to evaluate the association with lung cancer. In a multivariate analysis fruit (Ptrend < 0.0001), vegetable (P = 0.001) and sunflower oil (P = 0.006) remained as protective factors and rice (P = 0.008), bread (Ptrend = 0.04), liver (P = 0.004), butter (Ptrend = 0.04), white cheese (Ptrend < 0.0001), beef (Ptrend = 0.005), vegetable ghee (P < 0.0001) and, animal ghee (P = 0.015) remained as risk factors of lung cancer. Generally, we found positive trend between consumption of beef (P = 0.002), bread (P < 0.0001), and dairy products (P < 0.0001) with lung cancer. In contrast, only fruits were inversely related to lung cancer (P < 0.0001). It seems that vegetables, fruits, and sunflower oil could be protective factors and bread, rice, beef, liver, dairy products, vegetable ghee, and animal ghee found to be possible risk factors for the development of lung cancer in Iran.


Inferior vena cava obstruction due to cecum adenocarcinoma.
In this paper, we report the radiologic findings of cecal adenocarcinoma with caval invasion in a 60-year-old man. Caval invasion was confirmed by ultrasonographically guided fine-needle aspiration biopsy and by surgery. To our knowledge, this is the first case report demonstrating malignant caval thrombus due to colon cancer. In addition, we suggest that fine-needle aspiration biopsy may be helpful for prompt diagnosis and treatment in such a case.


In vitro Susceptibility of Human Cell Lines Infection by Bovine Leukemia Virus.
Evidence of the presence of bovine leukemia virus (BLV) in human beings and its association with breast cancer has been published in the literature, proposing it as a zoonotic infection. However, not enough evidence exists about transmission pathways nor biological mechanisms in human beings. This study was aimed at gathering experimental evidence about susceptibility of human cell lines to BLV infection. Malignant and non-malignant human cell lines were co-cultured with BLV-infected FLK cells using a cell-to-cell model of infection. Infected human cell lines were harvested and cultured for 3 to 6 months to determine stability of infection. BLV detection was performed through liquid-phase PCR and visualized through in situ PCR. Seven out of nine cell lines were susceptible to BLV infection as determined by at least one positive liquid-phase PCR result in the 3-month culture period. iSLK and MCF7 cell lines were able to produce a stable infection throughout the 3-month period, with both cytoplasmic and/or nuclear BLV-DNA visualized by IS-PCR. Our results support experimental evidence of BLV infection in humans by demonstrating the susceptibility of human cells to BLV infection, supporting the hypothesis of a natural transmission from cattle to humans.


Partial Purification and Characterization of a Heat Stable α-Amylase from a Thermophilic Actinobacteria, Streptomyces sp. MSC702.
A partial purification and biochemical characterization of the α-amylase from Streptomyces sp. MSC702 were carried out in this study. The optimum operational conditions for enzyme substrate reaction for amylolytic enzyme activity from the strain were evaluated. The optimum pH, temperature, and incubation period for assaying the enzyme were observed to be 5.0, 55°C, and 30 min, respectively. The extracellular extract was concentrated using ammonium sulfate precipitation. It was stable in the presence of metal ions (5 mM) such as K(+), Co(2+), and Mo(2+), whereas Pb(2+), Mn(2+), Mg(2+), Cu(2+), Zn(2+), Ba(2+), Ca(2+), Hg(2+), Sn(2+), Cr(3+), Al(3+), Ag(+), and Fe(2+) were found to have inhibitory effects. The enzyme activity was also unstable in the presence of 1% Triton X-100, 1% Tween 80, 5 mM sodium lauryl sulphate, 1% glycerol, 5 mM EDTA, and 5 mM denaturant urea. At temperature 60°C and pH 5.0, the enzyme stability was maximum. α-amylase retained 100% and 34.18% stability for 1 h and 4 h, respectively, at 60°C (pH 7.0). The enzyme exhibited a half-life of 195 min at 60°C temperature. The analysis of kinetic showed that the enzyme has K m of 2.4 mg/mL and V max of 21853.0 μmol/min/mg for soluble potato starch. The results indicate that the enzyme reflects their potentiality towards industrial utilization.


Functional immunoglobulin transgenes guide ordered B-cell differentiation in Rag-1-deficient mice.
We have examined the regulatory role of the individual components of the immunoglobulin antigen receptor in B-cell development by transgenic complementation of Rag-1 deficient (Rag-1-) mice. Complementation with a membrane mu heavy chain (mu HC) gene allows progression of developmentally arrested Rag-1- pro-B-cells to the small pre-B cell stage, whereas the introduction of independently integrated mu HC and kappa light chain (kappa LC) transgenes promotes the appearance of peripheral lymphocytes which, however, remain unresponsive to external stimuli. Complete reconstitution of the B-cell lineage and the emergence of functionally nature Rag-1- peripheral B cells is achieved by the introduction of cointegrated heavy and light chain transgenes encoding an anti-H-2k antibody. This experimental system demonstrates the competence of the mu HC and kappa LC to direct and regulate the sequential stages of B-cell differentiation, defines the time at which negative selection of self-reactive B cells occurs, and shows that elimination of these cells occurs equally well in the absence of Rag-1 as in its presence. These data also support the hypothesis that Rag-1 directly participates in the V(D)J recombination process.


A novel assay for factor XIII based on cross-linking of synthetic peptides: analysis of different substrates.
A novel assay for factor XIII is described that utilizes exclusively small synthetic peptides as substrates for the cross-linking reaction catalyzed by activated factor XIII (FXIIIa). The acyl donor substrate (selection peptide) is immobilized on a microplate via biotin while the acyl acceptor substrate (detection peptide) is labeled with the fluorochrome Oregon green to allow sensitive detection without the need for secondary enzyme systems for signal amplification. Starting with an amino acid sequence from the fibrin gamma-chain (GQQHHLGGAKQAGDV) as a prototype peptide, the influence of amino acid exchanges were investigated with respect to their impact on the FXIIIa-catalyzed reaction. It was found that FXIIIa readily accepts a broad range of substrate peptides, with a proline neighboring the essential lysine having the most detrimental effect. The assay appears to be valuable for the molecular characterization of factor XIII and may be used for a deeper investigation into the substrate requirements of this final enzyme of wound repair, and eventually also for the characterization of other transglutaminases.


In situ peeling of one-dimensional nanostructures using a dual-probe nanotweezer.
We reported a method for in situ peeling force measurement of one-dimensional nanostructures using a dual-probe nanotweezer, which is developed on the principle of force microscopy. Benefiting from capabilities of image scanning and accurate force sensing, the nanotweezer is capable of positioning one-dimensional nanostructures deposited on a surface and then performing in situ peeling tests with pick-and-place operations at different peeling locations of interest along a selected nanostructure. In experiments, nanoscale peeling of silicon nanowires (SiNWs) on a silicon substrate has been studied. Peeling locations at the end and in the middle of the SiNW were tested and the results indicate that approximate peeling energies are needed.


Weight management in community pharmacy: what do the experts think?
The increasing prevalence of obesity and overweight adults creates a significant public health burden and there is great potential for pharmacists to be involved in the provision of weight management services, other than the mundane supply of commercial products. In order to provide optimal services that can be integrated into the healthcare system, a best practice model for weight management services in community pharmacy should be in place. We sought experts' and key stakeholders' opinions on this matter. (1) To identify components of a best practice model of a weight management service feasible in Australian community pharmacy. (2) To identify the role of pharmacists and the training requirements to up-skill pharmacists to competently provide weight management services. (3) To elicit any practical suggestions that would contribute to successful implementation of weight management services in pharmacy. Australian primary care sector. Semi-structured interviews were conducted with a purposive sample of 12 participants including Australian experts in obesity and representatives of main Australian professional organisations in pharmacy. Interviews were digitally recorded, transcribed verbatim and thematically analysed using the framework approach. Recommended components of pharmacy-based weight management services and training requirements. Participants perceived two potential roles for pharmacists involved in weight management: health promotion and individualised service. Multi-component interventions targeting all three areas: diet, physical activity and behaviour change were emphasised. Physical assessment (e.g. weight, waist circumference measurements), goal setting, referral to allied healthcare professionals and on-going support for weight maintenance were also proposed. Participants suggested pharmacists should undergo formal training and identified various training topics to improve pharmacists' knowledge, attributes and skills to acquire competencies necessary for delivery of this service. Some physical and financial barriers in providing these services were also identified including infrastructure, pharmacists' time and cost-effectiveness. Pharmacists are well-positioned to promote healthy weight and/or implement weight management interventions. Furthering pharmacists' role would involve training and up-skilling; and addressing key practice change facilitators such as pharmacy layout and remuneration. This study provides some insight into the design and implementation of a best practice model for pharmacy-based weight management services in Australia.


The Role of Patent Foramen Ovale in Cryptogenic Stroke.
Almost one-third of ischemic strokes has an unknown etiology and are classified as cryptogenic stroke. Paradoxical embolism because of a patent foramen ovale (PFO) is detected in 40%-50% of these patients. Recently, PFO has been reported as a risk factor for patients of all age groups. In this study, 1080 ischemic stroke patients admitted to our clinic (2011-2013) were retrospectively evaluated. Age, sex, risk factors, complete blood count, vasculitis, biochemical and hypercoagulability tests, magnetic resonance imaging, magnetic resonance angiography, transthoracic echocardiography, transeosophageal echocardiography (TEE) findings, and therapeutic approaches were evaluated. The age range of the participants (seven male and four female patients) was 20-60 years (mean=43.09±11.13 years). Hemiparesis (n=10), diplopia (n=2), hemianopsia (n=2), and dysarthria (n=2) were the main findings of the neurological examinations. Patient medical history revealed hypertension (n=3), asthma (n=1), deep venous thrombosis (n=1), and smoking (n=4). Diffusion-weighted imaging showed middle cerebral artery (n=8) and posterior cerebral artery (n=3) infarctions. In one case, symptomatic severe carotid stenosis was detected. In eight cases, TEE showed PFO without any other abnormalities, but PFO was associated with atrial septal aneurysm in two cases, and in one case it was associated with ventricular hypokinesia and pulmonary arterial hypertension. Antiplatelet therapy was applied to nine patients and percutaneous PFO closure operation to two patients. In a 2-year follow-up, no recurrent ischemic stroke was recorded. PFO, especially in terms of the etiology of cryptogenic stroke in young patients, should not be underestimated. We want to emphasize the importance of TEE in identifying potential cardioembolic sources not only in young but also in all ischemic stroke patients with unknown etiology; we also discuss the controversial management options of PFO.