Principles of RNA methylation and their implications for biology and medicine.
RNA methylation is a post-transcriptional level of regulation. At present, more than 150 kinds of RNA modifications have been identified. They are widely distributed in messenger RNA (mRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), noncoding small RNA (sncRNA) and long-chain non-coding RNA (lncRNA). In recent years, with the discovery of RNA methylation related proteins and the development of high-throughput sequencing technology, the mystery of RNA methylation has been gradually revealed, and its biological function and application value have gradually emerged. In this review, a large number of research results of RNA methylation in recent years are collected. Through systematic summary and refinement, this review introduced RNA methylation modification-related proteins and RNA methylation sequencing technologies, as well as the biological functions of RNA methylation, expressions and applications of RNA methylation-related genes in physiological or pathological states such as cancer, immunity and virus infection, and discussed the potential therapeutic strategies.


PDK1 upregulates PINK1-mediated pulmonary endothelial cell mitophagy during hypoxia-induced pulmonary vascular remodeling.
Hypoxic pulmonary hypertension (HPH) is a complication of lung diseases with pulmonary vascular remodeling, although the underlying molecular mechanisms have not been fully elucidated. This study investigated the underlying molecular events by using a rat HPH model and primary pulmonary microvascular endothelial cells (PMVECs). This study first established a rat HPH model and cultured PMVECs for transmission electron microscopic analysis and manipulation of 3-phosphoinositide-dependent protein kinase 1 (PDK1) or phosphatase and tensin homolog-induced kinase 1 (PINK1) expression in vitro. After that, the cell viability was assessed and the expression of different proteins was assayed using cell viability and western blot assays, respectively. Reactive oxygen species production, apoptosis, NLR family pyrin domain containing 3 (NLRP3) expression, and the levels of interleukin (IL)-1β, IL-6, and IL-8 were also assessed, while the interaction of PDK1 and PINK1 was determined using co-immunoprecipitation/western blot assays. Hypoxia induced mitophagy in the PMVECs and upregulated PINK1/Parkin expression, whereas knockdown of PINK1 expression under hypoxic conditions inhibited cell proliferation but induced endothelial cell apoptosis in vitro, decreased reactive oxygen species production and NLRP3 expression, and reduced the levels of inflammatory factors in PMVECs. However, hypoxia induced PDK1 expression, whereas knockdown of PDK1 downregulated PINK1 expression. Furthermore, treatment of the model rats with the PDK1 inhibitor dichloroacetate (DCA) was able to decrease PINK1 expression. In addition, the PDK1 and PINK1 proteins could interact with each other in the mitochondria of PMVECs to regulate the cell viability. This study revealed that PDK1 induced PMVEC proliferation but inhibited their apoptosis to participate in pulmonary vascular remodeling, ultimately leading to HPH through regulation of PINK1-mediated mitophagy signaling. Therefore, PINK1 is a novel therapeutic target for the control of HPH.


Loss of orally administered drugs in GI tract.
The aim of this review is to provide a broad perspective on intestinal absorption and the impact of intestinal first-pass metabolism on both clearance and drug-drug interaction prediction along with its historical perspectives. The review also considers abilities to bridge the gap between the increasing amount of intestinal in vitro data and the importance of intestinal first-pass metabolism in vivo. The significance of efflux transporters on the intestinal absorption is also discussed.


The effect of feeding and starvation upon immunoreactive glucagon in the pancreatectomized pig.
The response of 13 pigs to total pancreatectomy was compared with the response of 9 animals to sham operation. Approximately half of each group had free access to food while the remainder received oral water only. Completeness of pancreatectomy was confirmed at autopsy, and the disappearance of insulin and the rise in blood glucose to 14-19 mmol/l. Immunoreactive glucagon (IRG) increased in fed pancreatectomized pigs from the 3rd day postoperatively to a level of 350-400% of basal values by the 8th day. There was no rise in starved pancreatectomized animals, or in either group of sham-operated animals. The rises in glucose, free fatty acids, triglycerides and ketones were similar in the two groups of pigs.


Terminal differentiation of human erythroleukemia cell line K562 induced by aphidicolin.
To analyze the relationship between differentiation and DNA replication, the effect of aphidicolin, a specific inhibitor for DNA polymerase alpha, was measured with respect to erythroid differentiation and activities of DNA polymerases alpha, beta, and gamma. Five micromolar aphidicolin completely blocked the growth of K562 cells and caused 80% of cells to become hemoglobin positive after 5 days exposure. The cessation of K562 cell growth induced by aphidicolin was irreversible, whereas the inhibition of HeLa cell growth was completely reversible. The enzyme activity of DNA polymerase alpha of K562 cells showed a 50-110% increase with aphidicolin treatment as compared to control K562 cells; activities of DNA polymerases beta and gamma were not affected. These features sharply contrasted with the erythroid induction of the same cells by hemin, where cell growth was not suppressed and DNA polymerase alpha was not increased but rather decreased. The enzyme activity of DNA polymerase alpha remained high even after removal of aphidicolin from the culture medium. These results suggest that treatment with aphidicolin might induce an accumulation of protein factors for replication and/or differentiation, causing rapid cell differentiation of cells without cell division.


A neuronal model for syllable representation.
A speculative neuronal template, equivalent to canonical syllable forms and independent of segmental representations, is offered to help account for (1) the inviolate nature of phonotactic constraints in aphasic speech output, and (2) left hemisphere specialization for speech sound access and output. The model, which attempts to relate plausible neuronal systems to linguistic function, is based on cell assemblies that are thought to develop by way of genetic predisposition and ontogenetic language experience, into configurations that can represent canonical slot positions for the consonants and vowel comprising a syllable. The syllable is assumed to be the basic organizational rhythmic unit for serial concatenation of sublexical segments. A scheme for neurological differentiation of vowels and consonants is offered. Phonotactic constraints can become "hard-wired" to help create the automaticity underlying phonological sound organization. Testable predictions are offered to substantiate the claims of the model.


Effect of cystatin C levels on angiographic atherosclerosis progression and events among postmenopausal women with angiographically decompensated coronary artery disease (from the Women's Angiographic Vitamin and Estrogen [WAVE] study).
End-stage renal disease and mild renal insufficiency are associated with increased cardiovascular risk. Cystatin C, a novel marker of kidney function, was found to be associated with a higher frequency of cardiovascular events and mortality independent of glomerular filtration rate. It remained uncertain, however, whether enhanced cardiovascular risk associated with cystatin C is due to accelerated progression of atherosclerosis or to plaque instability. The aim of this study was to examine the effects of baseline cystatin C on annual change in coronary artery narrowing and clinical events in 423 postmenopausal women with angiographically documented coronary artery disease enrolled in the Women's Angiographic Vitamin and Estrogen (WAVE) trial. Baseline and follow-up (mean 2.8 ± 0.9 years) angiography was performed in 320 women. Angiographic progression of disease and clinical events in each cystatin C quartile were compared. Women with cystatin C levels in the highest quartile were older and more likely to have histories of heart failure and stroke. Annualized changes in minimal and average luminal diameters were similar in diseased and nondiseased segments. All-cause death or myocardial infarction (3.6% vs 15.6%, p <0.001), cardiovascular death or myocardial infarction (2.3% vs 13.5%, p <0.001), and cardiovascular events (3.6% vs 13.5%, p <0.001) were significantly higher in women with baseline cystatin C levels in the highest quartile compared with women with cystatin C levels in the lower 3 quartiles. The risk for clinical events associated with cystatin C remained significantly higher in multivariate logistic regression analysis after adjusting for baseline differences and cardiovascular risk factors. The risk for clinical events was also independent of estimated glomerular filtration rate. In conclusion, in postmenopausal women with angiographically documented coronary artery disease, baseline cystatin C levels were associated with worse clinical outcomes without accelerated progression of atherosclerosis.


Developing a New Set of ACGME Milestones for Child Neurology Residency.
The Educational Milestones developed by the Accreditation Council for Graduate Medical Education (ACGME) are a construct used to evaluate the development of core competencies during residency and fellowship training. The milestones were developed to create a framework for professional development during graduate medical education. The first iteration of milestones for the child neurology residency was implemented in 2015. In the years that followed, the ACGME received and reviewed feedback about the milestones and set out to revise them. A committee was assembled to review the original milestones and develop a new set of milestones. The group was also encouraged to not only consider the child neurology residency graduate of today but also the graduate of tomorrow, taking into account growing fields such as genetics and technology. A diverse group of 12 individuals, including 10 child neurologists (all of whom were current or previous program directors or associate program directors), one child neurology resident, and one non-physician program coordinator, were recruited from programs of varying size across the country. The committee developed a revision to the child neurology milestones. All changes made were with a focus on how the milestones can be useful to trainees, program directors, and clinical competency committee members. Implementation and further feedback should help guide future revisions. These changes should help trainees, clinical competency committee members, and program directors find more meaning from their use.


Calcium deposits in the perineurium and their relation to lipid accumulation.
Calcification of the perineurium in sural nerves occurs in human adults. Deposits of calcium salts can be found focally in the middle and outer layers of the perineurium. Evidence indicates that this calcification is related to nerve fibre breakdown and to the local presence of lipids in the perineurium. The origin of the lipids may be senescent cells or degradation myelin.


Growth performance of Epirrita autumnata (Lepidoptera: Geometridae) on mountain birch: trees, broods, and tree x brood interactions.
The outcome of herbivore-host plant interactions is partly a function of variation within the two populations. We partitioned variance in herbivore growth performance into components attributable to differences between trees, differences between (full-sib) insect broods, and tree x brood interactions. Growth performance of Epirrita larvae feeding on a small (0.25 ha) population of mountain birch was greatly influenced by differences between individual trees. Up to 49% of the variation in insect growth rate was due to tree effects; 5th instar growth rates ranged from 0.38 to 0.56 mg·mg-1·day-1 across a sample of 8 trees. About 25% of the variation in pupal weights and larval periods was due to tree effects; on low quality trees larvae required a longer time to attain lower pupal weights. Differences between trees were also evident in physical and chemical characteristics of the leaves. Insect broods differed in the duration of the larval period (15% of the variance) which led to differences in the pupal weight attained (13% of the variance). However, brood-specific differences in growth rate were modest (6% in the 4th instar) or nonexistent (5th instar). There was no evidence for tree x brood interactions, which refutes the possibility of fine scale adaptation to particular tree phenotypes. Hypotheses to explain the existence of this variability, and to predict its evolutionary and ecological consequences, are advanced.


Aortic aneurysm leak after streptokinase treatment for myocardial infarction.
A leak from an abdominal aortic aneurysm following the administration of streptokinase treatment for myocardial infarction is reported. It is important to assess candidates for cardiac thrombolytic therapy for aortic pathology and give such treatment with extreme caution in those patients with abdominal aortic aneurysms.


Plasticity of cancer invasion and energy metabolism.
Energy deprivation is a frequent adverse event in tumors that is caused by mutations, malperfusion, hypoxia, and nutrition deficit. The resulting bioenergetic stress leads to signaling and metabolic adaptation responses in tumor cells, secures survival, and adjusts migration activity. The kinetic responses of cancer cells to energy deficit were recently identified, including a switch of invasive cancer cells to energy-conservative amoeboid migration and an enhanced capability for distant metastasis. We review the energy programs employed by different cancer invasion modes including collective, mesenchymal, and amoeboid migration, as well as their interconversion in response to energy deprivation, and we discuss the consequences for metastatic escape. Understanding the energy requirements of amoeboid and other dissemination strategies offers rationales for improving therapeutic targeting of metastatic cancer progression.


Dual modality reflection mode optical coherence and photoacoustic microscopy using an akinetic sensor: publisher's note.
This publisher's note corrects an error in the funding section in Opt. Lett.42, 4319 (2017)OPLEDP0146-959210.1364/OL.42.004319.


Interleukin 1 beta and its association with the periodontal health of pregnant women.
In this study, we measured and compared serum and gingival crevicular fluid (GCF) levels of interleukin 1 beta (IL-1β) in pregnant women with periodontitis and pregnant women with a healthy periodontium. We also determined the prevalence of periodontitis among pregnant women attending Omdurman Midwifery Hospital. This was a hospital-based clinical study and laboratory investigation using ELISA tests of 80 pregnant women in the third trimester conducted at the Omdurman Midwifery Hospital in Khartoum, Sudan. The study group consisted of 50 women while the control group consisted of 30 women. Independent samples t-tests were used to compare serum and GCF levels of IL-1β between the study and control groups. Pearson's correlation analysis was also used to compare gingival parameters and IL-1β levels in the GCF. For each comparison, the p-value was fixed at 0.05. The GCF in the research group showed a considerable increase in IL-1β levels. There was also a strong positive association between high IL-1β levels in the research group's GCF and probing pocket depth (PPD) and clinical attachment level (CAL) levels. Our study provides further evidence that periodontitis, as measured by a PD ≥4 mm and a CAL ≥3 mm, is associated with an increased level of IL-1β in the GCF of pregnant women with active periodontal disease during pregnancy and may include the transient translocation of oral organisms to the utero-placental unit, inciting placental inflammation or oxidative stress early in pregnancy, ultimately resulting in placental damage and clinical manifestations.


Delayed presentation of blunt splenic injury.
Eleven patients with delayed presentation of splenic injury after blunt abdominal trauma treated during a 10 year period have been described. They represented 24 percent of all patients treated for blunt splenic injury in our department in that time period. Ten patients required operative treatment and one was treated nonoperatively. There were no deaths. The value of computerized tomography and ultrasonography in the accurate preoperative assessment of splenic injury has been documented. In addition, the various diagnostic and therapeutic possibilities in blunt splenic trauma have been discussed.


UniFrag and GenomePrimer: selection of primers for genome-wide production of unique amplicons.
The complementary programs UniFrag and GenomePrimer were developed to provide a reliable high-throughput method to select the most unique regions within genomic DNA sequence(s) and design primers therein, involving minimal user intervention and maximum flexibility.


WITHDRAWN: COVID-19, its novel vaccination and fake news - What a brew.
This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at https://www.elsevier.com/about/our-business/policies/article-withdrawal.


Diagnosis of Strongyloides and hookworm infections: comparison of faecal and duodenal fluid microscopy.
Faecal microscopical diagnosis of Strongyloides and hookworm infections is insensitive. We have therefore compared duodenal fluid and faecal microscopy for detection of these parasites in a group of 292 patients being investigated for gastrointestinal symptoms who were examined by both techniques. Thirty-three of these patients (8%) were infected with Strongyloides stercoralis and 88 (30%) had hookworm infections. Microscopical examination of up to 3 faecal specimens detected only 33% and 65% of patients with Strongyloides and hookworm infections, respectively. Microscopical examination of a single specimen of duodenal fluid was more sensitive for detection of strongyloidiasis, identifying 76% of patients; the parasite was found exclusively in duodenal fluid (and not in faeces) in 67% of patients. For hookworm, the diagnostic sensitivity was similar with both techniques but duodenal fluid microscopy detected some patients (35%) who had not been identified by faecal microscopy. This study confirms previous work indicating the insensitivity of faecal microscopy in these infections and emphasizes the need to consider routine examination of duodenal fluid to exclude chronic strongyloidiasis. This may have particular relevance for south-east Asian war veterans and immunocompromised patients.


Unfavorable clinical outcomes in patients with carbamoyl phosphate synthetase 1 deficiency.
Carbamoyl phosphate synthetase 1 (CPS1) deficiency affects the first step of urea cycle and is a severe form of urea cycle disorder (UCD). The severity of hyperammonemic encephalopathy determines the clinical course of UCDs. Here, we describe the genetic and clinical characteristics of CPS1 deficiency in Korea. This study included seven patients with CPS1 deficiency genetically confirmed from January 1992 to September 2020. The peak ammonia level during the first crisis, the half time of peak ammonia level, the initial plasma amino acid levels, and neurological outcomes were compared between CPS1 deficiency and two common UCDs (i.e., 17 patients with argininosuccinate synthetase 1 deficiency and 24 patients with ornithine transcarbamylase deficiency). Eleven CPS1 mutations were identified, including 10 novel mutations. Eight mutations were missense. Six patients with CPS1 deficiency had neonatal type. The peak ammonia level, initial glutamate level, and accompanying rate of irreversible neurological damages were highest in patients with CPS1 deficiency. The patient with late-onset CPS1 deficiency responded dramatically to N-carbamylglutamate treatment. The clinical manifestations of CPS1 deficiency were the most severe among UCDs. Considering the high proportion of missense mutations, responsiveness to N-carbamylglutamate would be evaluated in a future study.


Circ-SMARCA5 suppresses colorectal cancer progression via downregulating miR-39-3p and upregulating ARID4B.
Circular RNAs are crucial in tumorigenesis. However, little is known about their functions in colorectal cancer (CRC). Circ-SMARCA5 was found to be an oncogene or tumor suppresser in different types of cancers, but its exact role in CRC remains unknown. Here, we aim to identify the role of circ-SMARCA5 in CRC development. Circ-SMARCA5 expression was determined by qRT-PCR. CRC cell proliferation, migration, and invasion were detected by CCK-8, wound healing, and Transwell assays, respectively. Bioinformatics analysis was performed to predict target genes. The interaction of microRNA (miR) with circ-SMARCA5 or target genes was detected using luciferase reporter assay. Xenograft model was established to determine the effect of circ-SMARCA5 on CRC tumor growth in vivo. Circ-SMARCA5 expression was dramatically decreased in CRC cell lines and tissues. Circ-SMARCA5 overexpression inhibited CRC cell proliferation, migration and invasion. MiR-93-3p was predicted as a target of circ-SMARCA5 and its overexpression attenuated the anti-tumor effect of circ-SMARCA5 on CRC cells. Furthermore, we predicted AT-rich interaction domain 4B (ARID4B) as the target of miR-39-3p. Functional analysis showed that circ-SMARCA5 upregulated ARID4B expression via miR-39-3p. Additionally, in vivo studies demonstrated that circ-SMARCA5 suppressed CRC tumor progression. Circ-SMARCA5 functions as a tumor suppressor by upregulating ARID4B expression via sponging miR-39-3p, and thereby inhibited CRC progression.


Barasertib impedes chondrocyte senescence and alleviates osteoarthritis by mitigating the destabilization of heterochromatin induced by AURKB.
Osteoarthritis (OA) is a common joint disease characterized by progressive cartilage loss that causes disability worldwide. The accumulation of senescent chondrocytes in aging human cartilage contributes to the high incidence of OA. Heterochromatin instability, the hallmark and driving factor of senescence, regulates the expression of the senescence-associated secretory phenotype that induces inflammation and cartilage destruction. However, the role of heterochromatin instability in OA progression remains unclear. In this work, we identified AURKB as a key senescence-associated chromatin regulator using bioinformatics methods. We found that AURKB was upregulated in OA cartilage and chondrocytes exposed to abnormal mechanical strain. Overexpression of AURKB could cause senescence and heterochromatin instability. Furthermore, the AURKB inhibitor Barasertib reversed senescence and heterochromatin instability in chondrocytes and alleviated OA in a rat model. Mechanistically, abnormal mechanical strain increased AURKB levels through the Piezo1/Ca2+ signaling axis. Blocking Piezo1/Ca2+ signaling by short interfering RNA against Piezo1 and Ca2+ chelator BAPTA could reduce the expression of AURKB and alleviate senescence in chondrocytes exposed to abnormal mechanical strain. In conclusion, our data confirmed that abnormal mechanical strain increases the expression of AURKB by activating the Piezo1/Ca2+ signaling axis, leading to destabilized heterochromatin and senescence in chondrocytes, whereas Barasertib consolidates heterochromatin, counteracts senescence and alleviates OA.


The impact on practice of authentic assessment as part of a university postgraduate taught programme of study for nursing and healthcare professionals: a literature review.
To consider the impact on practice of authentic assessment as part of university postgraduate taught programmes of study for nursing and healthcare professionals. A literature review of the available published articles which include empirical research. An online search for English language quantitative and qualitative literature in the British Education Index, CINAHL Plus and ERIC. No initial limit on publication date was set. Studies were sought to review the impact of authentic assessment strategies on the subsequent practice of postgraduate nursing and healthcare professional learners. Having developed the research question and search strategy, five studies were identified which in some way addressed the question. The data was then synthesised, and themes developed. The studies reviewed hold authentic assessment in high regard, not questioning whether this approach is legitimate, and providing varying degrees of supporting evidence. The studies all scrutinize the learner experience of undertaking a postgraduate healthcare programme of study, although they were different in terms of settings, programmes and assessment strategies, and report positive feedback from learners for authentic assessment. Emergent themes indicate that authentic assessment enhances the contextualisation of learning, provides opportunities to explore roles and supports role development. This review found some evidence to suggest that postgraduate learners respond positively to authentic assessment. The long-term impact in practice remains unproven, and this is where further study is recommended. An authentic assessment impact framework is proposed to evaluate strategies in a consistent and replicable manner. There is a need for university programme directors to take a leadership role in collaborating with stakeholders to ensure assessment strategies align to subsequent role performance, contextualise learning and support role development.


Nanoscale Location Matters: Emerging Principles of Ca2+ Channel Organization at the Presynaptic Active Zone.
How does diversity in the organization of secretory machines determine properties of neurotransmitter release? In this issue of Neuron, Rebola et al. (2019) found that distinct nanoscale assemblies of Ca2+ channels and Munc13, not overall channel abundance, mediate differing release characteristics of two cerebellar synapses.


Transitions of dialysis status and outcomes after the unplanned first dialysis: a nationwide population-based cohort study.
In Taiwan, most first-time dialysis was started without the creation of an arteriovenous shunt. Here, we aimed to elucidate the transitions of dialysis status in the unplanned first dialysis patients and determine factors associated with their outcomes. A total of 50,315 unplanned first dialysis patients aged more than 18 years were identified from the National Health Insurance Dataset in Taiwan between 2001 and 2012. All patients were followed for 5 years for the transitions in dialysis status, including robust (dialysis-free), sporadic dialysis, continued dialysis, and death. Furthermore, factors associated with the development of continued dialysis and death were examined by the Cox proportional hazard models. After 5 years after the first dialysis occurrence, there were 5.39% with robust status, 1.67% with sporadic dialysis, 8.45% with continued dialysis, and 84.48% with death. Notably, we have identified common risk factors for developing maintenance dialysis and deaths, including male gender, older age, diabetes, coronary heart disease, stroke, heart failure, sepsis, and surgery. There was an extremely high mortality rate among the first unplanned dialysis patients in Taiwan. Less than 10% of these patients underwent continued dialysis during the 5-year follow-up period. This study highlighted the urgent need for interventions to improve patient outcomes.


Effects of Korean red ginseng extract on cisplatin-induced nausea and vomiting.
Ginseng, the root of Panax ginseng C.A. Meyer, is well known as a tonic medicine for restoring and enhancing human health. In traditional medicine, ginseng is utilized for the alleviation of emesis, which includes nausea and vomiting. However, it has not yet been demonstrated whether ginseng exhibits in vivo anti-nausea and anti-vomiting properties. In this study, we examined the anti-emetic effect of Korean red ginseng total extract (KRGE) on cisplatin-induced nausea and vomiting using ferrets. Intraperitoneal administration (i.p.) of cisplatin (7.5 mg/kg) induced both nausea and vomiting with one-hour latency. The episodes of nausea and vomiting reached a peak after 1.5 h and persisted for 3 h. Treatment with KRGE via oral route significantly reduced the cisplatin-induced nausea and vomiting in a dose-dependent manner. The anti-emetic effect was 12.7 +/- 8.6, 31.8 +/- 6.9, and 67.6 +/- 4.0% with doses of 0.3, 1.0, and 3.0 g/kg of KRGE, respectively. Pretreatment with KRGE via oral route 1 and 2 h before cisplatin administration also significantly attenuated the cisplatin-induced nausea and vomiting. However this did not occur with a pretreatment 4 h before cisplatin administration. These results are supportive of KRGE being utilized as an anti-emetic agent against nausea and vomiting caused by chemotherapy (i.e. cisplatin).


Compliance of Middle Eastern hospitals with the central line associated bloodstream infection prevention guidelines.
The purposes of this study were to (1) assess compliance with the CLABSI prevention guidelines, (2) assess the predictors of compliance, and (3) investigate the effect of compliance on the rate of CLABSI and related mortality. Implementation of the Central Line Associated Bloodstream Infection (CLABSI) prevention guidelines from the Centers for Disease Control and Prevention (CDC) helps to reduce the rate of CLABSI and related mortality, although the extent to which hospitals implement these guidelines is questionable. A prospective design was used in this study. Observations were conducted over three months in the intensive care units of 58 hospitals in three Middle Eastern countries. An observational checklist, based on the CDC guidelines, was used to assess compliance. The rate of CLABSI and related mortality were obtained from patients' records. The degree of compliance, rate of CLABSI and mortality were highly variable. The multiple regression model showed that the hospitals' characteristics explained 82.0% of the variance of compliance (R2Adj= 0.820, F=29.82, p<0.05). The number of beds in the intensive care unit and patient-nurse ratio were significant predictors of compliance. A lower number of beds and a lower patient-to-nurse ratio were related to higher compliance. Moreover, higher compliance with CLABSI prevention guidelines was associated with lower rate of CLABSI and related mortality. Improvement in the patients' outcome can be achieved through compliance with the CLABSI prevention guidelines. lowering patient-nurse ratio and the number of beds in the ICUs would help to improve compliance.


Evaluation of bisbenzamidines as inhibitors for matriptase-2.
The serine protease matriptase-2 has attracted much attention as a potential target for the treatment of iron overload diseases. In this study, a series of 27 symmetric, achiral bisbenzamidines was evaluated for inhibitory activity against human matriptase-2, against the closely related enzyme human matriptase, as well as against human thrombin, bovine factor Xa and human trypsin. The conformationally restricted piperazine derivative 19 and the oxamide-derived bisbenzamidine 1 were identified as the most potent inhibitors of this series for matriptase-2 and matriptase, respectively.


α-Amino-3-Hydroxy-5-Methyl-4-Isoxazolepropionic Acid Receptor Plasticity Sustains Severe, Fatal Status Epilepticus.
Generalized convulsive status epilepticus is associated with high mortality. We tested whether α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor plasticity plays a role in sustaining seizures, seizure generalization, and mortality observed during focal onset status epilepticus. We also determined whether modified AMPA receptors generated during status epilepticus could be targeted with a drug. Electrically induced status epilepticus was characterized by electroencephalogram and behavior in GluA1 knockout mice and in transgenic mice with selective knockdown of the GluA1 subunit in hippocampal principal neurons. Excitatory and inhibitory synaptic transmission in CA1 neurons was studied using patch clamp electrophysiology. The dose response of N,N,H,-trimethyl-5-([tricyclo(188.8.131.52,7)dec-1-ylmethyl]amino)-1-pentanaminiumbromide hydrobromide (IEM-1460), a calcium-permeable AMPA receptor antagonist, was determined. Global removal of the GluA1 subunit did not affect seizure susceptibility; however, it reduced susceptibility to status epilepticus. GluA1 subunit knockout also reduced mortality, severity, and duration of status epilepticus. Absence of the GluA1 subunit prevented enhancement of glutamatergic synaptic transmission associated with status epilepticus; however, γ-aminobutyric acidergic synaptic inhibition was compromised. Selective removal of the GluA1 subunit from hippocampal principal neurons also reduced mortality, severity, and duration of status epilepticus. IEM-1460 rapidly terminated status epilepticus in a dose-dependent manner. AMPA receptor plasticity mediated by the GluA1 subunit plays a critical role in sustaining and amplifying seizure activity and contributes to mortality. Calcium-permeable AMPA receptors modified during status epilepticus can be inhibited to terminate status epilepticus. ANN NEUROL 2020;87:84-96.


Driving Patients Crazy: Acute Neuropsychiatric Toxicities of Commonly Prescribed Antibiotics in IR.
Neuropsychiatric toxicities are uncommon but well-documented side effects of antibiotics. Society of Interventional Radiology guidelines recommend various antibiotic regimens for patients undergoing interventional radiological procedures. These same classes of drugs are also used to treat infectious complications in patients. Antibiotics have a wide spectrum of affective and cognitive toxicities, the most severe of which can lead to hospitalization or suicide. Fluoroquinolones have the highest incidence of these toxicities.


Spatial distribution of myosin heavy-chain isoforms in mouse masseter.
There is a paucity of information regarding the anatomy and muscle fiber phenotype of the masseter. The objective of this study was to characterize the distribution of each myosin heavy-chain (MyHC) isoform within different anatomical regions of male and female mouse masseters. Masseters from male and female CD-1 mice (2-4 months old) were examined for description of the anatomical partitioning of muscle fibers and endplate distribution. The spatial distribution of MyHC isoforms--embryonic, neonatal, slow, alpha-cardiac, IIa, and IIb--was determined within the defined masseter partitions by means of Western blot analysis and immunofluorescent localization. Types IIa, IIx, and IIb were the predominant MyHC isoforms observed. Distinct differences in the spatial distribution of these MyHC isoforms were found between muscle regions and varied between sexes. The regionalization of muscle fiber types in the mouse masseter is consistent with the functional compartmentalization of the masseter observed in other species.


Cushing's syndrome due to an ACTH-producing primary ovarian carcinoma.
Although Cushing's syndrome has been described in association with various neuroendocrine tumors producing ectopic adrenocorticotropin (ACTH), primary ovarian carcinoma rarely causes this syndrome. We hereby report the case of a 61-year-old woman presented with abdominal distension, facial swelling and skin pigmentation. Clinical and diagnostic evaluation revealed a right ovarian tumor with hypercortisolemia, high plasma ACTH, extremely high plasma ACTH precursors and lack of dexamethasone suppression. Removal of the tumor led to normalization of ACTH, ACTH precursors and cortisol levels. In addition, hypokalemia, hyperglycemia and elevated serum CA 125 were noted, which were also normalized after resection of the tumor. Histopathological examination indicated a primary ovarian carcinoma with a mixed pattern of differentiation. Immunohistochemistry showed immunoreactivity for chromogranin A, synaptophysin and ACTH. Hence, the diagnosis of Cushing's syndrome arising from an ovarian carcinoma secreting ectopic ACTH was made. The case was managed successfully, with subsequent good recovery and no evidence of recurrence at 4 years of follow-up.


A search for allelic recombination in Chinese hamster cell hybrids.
Mutants resistant to 6-thioguanine were selected from CHO cells which were either temperature sensitive or proline requiring. These mutants were stable and had low levels of hypoxanthine guanine phosphoribosyl transferase (HGPRT). Hybrids were selected which were heteroallelic at the hgprt locus and complementation between the mutants used was not observed. Interallelic recombination at this locus would generate hgprt+ cells which could be selected in Littlefield's HAT medium. Selection experiments with hybrids containing three different pairs of mutants yielded no recombinants among populations of 4 x 106 - 2 x 107 cells. After treatment with the recombinagen mitomycin C, 3 putative recombinants were detected amongst 1.4 x 107 surviving cells from one hybrid. One of these strains was examined and shown to have a normal level of HGPRT and its heterozygosity at this locus was demonstrated by the segregation of colonies resistant to 6-thioguanine. It cannot be excluded that the rare hgprt+ colonies seen arose by mutation rather than by recombination. Mitotic allelic recombination therefore appears to be a much less frequent event in CHO cells than it is in lower eukaryotes. It is possible that mitotic recombination is effectively suppressed in mammalian cells to prevent the expression of deleterious recessive mutants.


Impaired, spared, and enhanced ACh efflux across the hippocampus and striatum in diencephalic amnesia is dependent on task demands.
Diencephalic amnesia manifests itself through a host of neurological and memory impairments. A commonly employed animal model of diencephalic amnesia, pyrithiamine-induced thiamine deficiency (PTD), results in brain lesions and impairments similar in nature and distribution to those observed in humans with Wernicke-Korsakoff syndrome (WKS). In the current investigation, 2 separate experiments were conducted in which acetylcholine (ACh) efflux was assessed in the hippocampus and striatum of PTD-treated and pair-fed (PF) control male Sprague-Dawley rats. The goal was to determine under what behavioral conditions and in which brain structures ACh efflux was spared, impaired, or adaptively enhanced. In Experiment 1, rats were assessed on a spontaneous alternation task; in Experiment 2, rats were tested on a T-maze discrimination task that could be learned via a hippocampal- or striatal-based strategy. In Experiment 1, PTD-treated rats were impaired on the spontaneous alternation task and ACh efflux in the hippocampus during testing was significantly reduced, but spared in the striatum. In Experiment 2, PTD- and PF-treated rats did not differ in the number of trials to criterion, but PTD-treated rats demonstrated greater reliance upon egocentric cues to solve the task. Furthermore, ACh efflux in the striatum was greater during maze learning in the PTD-treated animals when compared to the PF animals. These results suggest that there is behavioral and systems level plasticity that can facilitate the use of alternative strategies to solve a task following diencephalic damage and WKS.


A Polytherapy Strategy Using Vincristine and ALK Inhibitors to Sensitise EML4-ALK-Positive NSCLC.
The oncogenic fusion of EML4-ALK is present in about 4-6% of non-small cell lung cancer (NSCLC). A targeted approach with ALK tyrosine kinase inhibitors (TKIs) has been proven highly effective in ALK-positive NSCLC patients. However, despite the initial responses, the outcome of the treatment is variable. Previous studies have shown that the differential response depends in part on the type of EML4-ALK variant. Here, we examined the combination of ALK inhibitors and microtubule poison, vincristine, in cells expressing EML4-ALK V1 and V3, the two most common variants in NSCLC. We showed that combination therapy of ALK-TKIs with vincristine had anti-proliferative effects and blocked RAS/MAPK, PI3K/AKT and JAK/STAT3 signalling pathways in EML4-ALK V1 but not V3 cells. Our results demonstrate that high levels of tubulin acetylation are associated with poor response to vincristine in EML4-ALK V3 cells. Additionally, we demonstrated differences in microtubule stability between the two EML4-ALK fusions. EML4-ALK V3 cells exhibited dynamic microtubules that confer poor response to vincristine compared to V1 cells. Hence, we suggested that the portion of EML4 in the fusion has an important role for the outcome of the combination treatment.


Calf implants: a 25-year experience and an anatomical review.
Body contouring in the calf region is becoming a more frequently requested procedure. There are several techniques for calf enhancement, including implants, liposuction, and free flaps. Alloplastic augmentation can be performed with several implant types and several layers of pocket dissection. We present our 25 years of experience using the subfascial technique for calf implantation along with an anatomical study to illustrate all the important steps and relevant anatomy of this augmentation technique. The subfascial technique was performed in 25 cadavers, in which the important layers were dissected for high-resolution photos of the surgery to learn about the relevant anatomy of the region. Also, we did a retrospective study of our experience with calf implants, studying the aesthetic outcome, the presence of early complications, and the presence of late complications. We performed dissections in 25 cadavers and surgery in 63 patients (126 implants). In our series of patients the final aesthetic index was of 9.8. The early complications were severe postoperative pain (11.11%), infection (0.79%), seroma (21.42%), hematoma (0%), and wound dehiscence (7.14%). The late complications were capsular contracture (Baker grades III and IV) (3.17%), implant rupture (1.58%), implant leaking (0%), implant displacement (3.96%), numbness at the ankle (2.38%), and palpability of the implant (0%). The subfascial technique for calf augmentation has complication rates low enough and surgical outcomes good enough to recommend it as the gold standard for alloplastic calf augmentation.


Assessing disease activity and damage in antiphospholipid syndrome.
Antiphospholipid syndrome (APS) has been characterized by a variety of vascular and pregnancy manifestations related to an interplay between thrombotic and inflammatory mechanisms, a progressive accrual of irreversible organ damage and increased morbidity and mortality rates, supporting a high need of optimal treatment approach. The lack of standardized outcome measures is a significant barrier in the design of clinical studies in APS. Disease activity (in principle reversible) and its distinction from disease damage (in principle irreversible) needs to be evaluated by validated scores for use in clinical trials but also in daily clinical practice in APS. A disease damage score in APS, the DIAPS score, has been developed and validated in external cohorts. The development of a disease activity score that will provide an accurate and reproducible rating of each disease domain, can help clinicians and researchers to comprehensively assess the activity of disease and the response to treatment, in an attempt to prevent future damage.


Intramedullary fixation of unstable metacarpal fractures.
Intramedullary stabilization of metacarpal shaft and neck fractures is a relatively simple, cost-effective, and safe technique with good published outcomes (Fig. 9); however definite advantages over other techniques of fracture stabilization, or indeed simple early mobilization in some instances, have not been clearly demonstrated. A recent publication does suggest that the technique is comparable to percutaneous transverse fixation in the context of fifth metacarpal neck fractures.


Phorbol ester-induced protein kinase C translocation and lysosomal enzyme release in normal and cystic fibrosis fibroblasts.
The ability of the tumor-promoting phorbol ester 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA) to induce protein kinase C (PKC) translocation and lysosomal enzyme release was examined in skin fibroblasts from normal subjects and from patients with cystic fibrosis (CF). As compared to normal fibroblasts, those CF exhibited: (i) an increased sensitivity to the effect of PMA on the disappearance of PKC from cytosolic fractions as well as a greater and earlier recovery, in the membrane fraction, of the PKC activity lost in the cytosolic fraction; (ii) an earlier response to PMA for its effect on beta-N-acetylglucosaminidase release. In contrast, the inactive phorbol ester 4 alpha-phorbol 12,13-didecanoate (4 alpha PDD) proved ineffective in inducing PKC translocation and beta-N-acetylglucosaminidase release in both normal and CF fibroblasts. The data suggest a defect in the regulation of PKC activity in CF fibroblasts, which may lead to altered secretion.


Immunofluorescent localization of four serum proteinase inhibitors in normal rat liver.
Four serum proteinase inhibitors, alpha 1-macroglobulin, alpha 1-proteinase inhibitor, alpha 1-inhibitor3 and alpha 2-acute phase macroglobulin, were localized in rat liver by immunofluorescent techniques. alpha 1-Macroglobulin was observed predominantly in the sinusoids and alpha 1-inhibitor3 in hepatocytes. In contrast, alpha 1-proteinase inhibitor was localized in two sites, sinusoids and parenchymal cells. The fourth inhibitor tested, alpha 2-acute phase macroglobulin, was barely detectable in normal liver. However, some appeared to be present in the extrahepatocyte space.


Perceptions and practices of family physicians regarding diet and cancer.
This study was undertaken to determine the beliefs of family physicians about the role of diet and cancer, as well as to determine how often family physicians assess dietary risks in patient encounters and recommend preventive dietary practices to reduce cancer risk. This study consisted of a survey of a national random sample of 486 family physicians from the American Academy of Family Physicians (AAFP), and 237 responded (49%). The demographic characteristics of the respondents were determined to be similar to those of the AAFP membership. Thirty-five percent of the physicians surveyed responded that diet is important in the development of cancer. While 31% responded they recommend dietary changes to reduce cancer risk to their patients, only 21% agreed that they find counseling patients about dietary issues professionally gratifying. Respondents agreed with most of the American Cancer Society, National Cancer Institute, and surgeon general's dietary recommendations to reduce cancer risk. The level of agreement to broad general recommendations was higher than the level of agreement to specific dietary recommendations that stated a specific level or amount of consumption.


Oxidative DNA damage induced by simultaneous generation of nitric oxide and superoxide.
Incubation of calf thymus DNA with 3-morpholinosydnonimine (SIN-1), which simultaneously generates nitric oxide (NO) and superoxide (O2-), induced a significant increase of 8-hydroxydeoxyguanosine (8-OH-dG). Peroxynitrite also increased 8-OH-dG in calf thymus DNA. Addition of free hydroxyl radical (.OH) scavengers inhibited the increase of 8-OH-dG by SIN-1 or peroxynitrite. Incubation of 32P-labeled DNA fragment with SIN-1 or peroxynitrite caused DNA cleavage at every nucleotide with a little dominance at guanine residues. The results suggest that NO reacts with O2- to form peroxynitrite and the peroxynitrite induces oxidative DNA damage through an active intermediate of which reactivity is similar to .OH.


Beneficial effects of hydralazine in severe mitral regurgitation.
The severity of mitral regurgitation is, in part, determined by aortic impedance to left ventricular outflow. Sodium nitroprusside acutely decreases regurgitant flow, but the importance of its dual vasodilating effects, the lowering of peripheral vascular resistance and increasing of venous capacitance, is unclear. We studied the hemodynamic response to intravenous hydralazine, which selectively acts on the arteriolar resistance bed, in 10 patients with severe mitral regurgitation. Hydralazine produced a 50% increase in forward stroke volume (22 +/- 2 to 33 +/- 3 ml/m2, P less than 0.001) and a 33% reduction in regurgitant stroke volume (40 +/- 6 to 27 +/- 6 ml/m2, P less than 0.001), with a resultant fall in pulmonary capillary wedge v wave and mean pressures. Unlike nitroprusside, it did not alter left ventricular end-diastolic volume or pressure. Oral hydralazine maintained this hemodynamic improvement for at least 48 hours and, in three patients, provided more sustained clinical improvement. We conclude that hydralazine, by virtue of its selective lowering of aortic impedance, reduces the amount of mitral regurgitation and thus may be a useful mode of interim or chronic therapy in selected patients.


Gas chromatography-chemical ionization-mass spectrometric fatty acid analysis of a commercial supercritical carbon dioxide lipid extract from New Zealand green-lipped mussel (Perna canaliculus).
Supercritical fluid extracts of New Zealand green-lipped mussels (NZGLM) have been suggested to have therapeutic properties related to their oil components. The large number of minor FA in NZGLM extract was characterized by a GC-CIMS/MS method that excels at identification of double-bond positions in FAME. The extract contained five major lipid classes: sterol esters, TAG, FFA, sterols, and polar lipids. The total FA content of the lipid extract was 0.664 g/mL. Fifty-three unsaturated FA (UFA) were fully identified, of which 37 were PUFA, and a further 21 UFA were detected for which concentrations were too low for assignment of double-bond positions. There were 17 saturated FA, with 14:0, 16:0, and 18:0 present in the greatest concentration. The 10 n-3 PUFA detected included 20:5n-3 and 22:6n-3, the two main n-3 FA; n-3 PUFA at low concentrations were 18:3, 18:4, 20:3, 20:4, 21:5, 22:5, 24:6, and 28:8. There were 43 UFA from the n-4, n-5, n-6, n-7, n-8, n-9, n-10, n-1 families, with 16:2n-4, 16:1n-5, 18:1n-5, 18:2n-6, 20:4n-6, 16:1n-7, 20:1n-7, 16:1n-9, 18:1 n-9, and 20:1 n-9 being the most abundant. In general, we estimated that FAME concentrations greater than 0.05% (w/w) were sufficient to assign double-bond positions. In total, 91 FA were detected in an extract of the NZGLM, whereas previous studies of fresh flesh from the NZGLM had reported identification of 42 FA. These data demonstrate a remarkable diversity of NZGLM FA.


Disease course and efficacy of medical therapy in stricturing paediatric Crohn's disease.
Stricturing is the most common complicated phenotype in paediatric Crohn's disease, but only few studies have described its course, while data on the outcome of medical treatment are scanty. To retrospectively describes the course of paediatric stricturing Crohn's disease and assess clinical and imaging response to medical therapy. Thirty-six patients with stricturing Crohn's disease were identified by our department database. Paediatric Crohn's disease activity index, need of surgery and magnetic resonance were evaluated as outcomes at 6, 12, 18 and 24 months after detection of stenosis. Strictures were ileal, ileocolonic and colonic in 61%, 28% and 11% of patients. Thirteen (36%) had stricturing disease at the diagnosis of Crohn's disease, while 64% developed it at the follow-up. At baseline, 89% had medical treatment, while 11% surgery. At 6, 12, 18, and 24 months, 53%, 50%, 42%, and 35% had complete response to medical treatment, respectively. Overall, 44% were unresponsive to medical therapy and required surgery at the follow-up. Responders and non-responders significantly differed for inflammatory imaging findings at the stenosis detection. A stricturing phenotype is not uncommon at the diagnosis of Crohn's disease in children. Medical therapy seems poorly effective in avoiding intestinal resection. Magnetic resonance imaging is valuable in identifying patients who will benefit from medical therapy.


Estimating upper-extremity function from kinematics in stroke patients following goal-oriented computer-based training.
After a stroke, a wide range of deficits can occur with varying onset latencies. As a result, assessing impairment and recovery are enormous challenges in neurorehabilitation. Although several clinical scales are generally accepted, they are time-consuming, show high inter-rater variability, have low ecological validity, and are vulnerable to biases introduced by compensatory movements and action modifications. Alternative methods need to be developed for efficient and objective assessment. In this study, we explore the potential of computer-based body tracking systems and classification tools to estimate the motor impairment of the more affected arm in stroke patients. We present a method for estimating clinical scores from movement parameters that are extracted from kinematic data recorded during unsupervised computer-based rehabilitation sessions. We identify a number of kinematic descriptors that characterise the patients' hemiparesis (e.g., movement smoothness, work area), we implement a double-noise model and perform a multivariate regression using clinical data from 98 stroke patients who completed a total of 191 sessions with RGS. Our results reveal a new digital biomarker of arm function, the Total Goal-Directed Movement (TGDM), which relates to the patients work area during the execution of goal-oriented reaching movements. The model's performance to estimate FM-UE scores reaches an accuracy of [Formula: see text]: 0.38 with an error ([Formula: see text]: 12.8). Next, we evaluate its reliability ([Formula: see text] for test-retest), longitudinal external validity ([Formula: see text] true positive rate), sensitivity, and generalisation to other tasks that involve planar reaching movements ([Formula: see text]: 0.39). The model achieves comparable accuracy also for the Chedoke Arm and Hand Activity Inventory ([Formula: see text]: 0.40) and Barthel Index ([Formula: see text]: 0.35). Our results highlight the clinical value of kinematic data collected during unsupervised goal-oriented motor training with the RGS combined with data science techniques, and provide new insight into factors underlying recovery and its biomarkers.


Attenuation of cadmium-induced testicular injury in metallothionein-III null mice.
In order to evaluate the role of metallothionein (MT)-III in cadmium (Cd)-induced testicular toxicity, we examined the sensitivity of MT-III null mice to severe testicular injury caused by Cd. Male MT-III null mice, MT-I/II null mice and wild-type mice were given a subcutaneous injection of CdCl(2) (15μmol/kg). The testis was collected from each mouse at 6, 12 and 24h after Cd administration. Testicular hemorrhages by evaluating morphology, hemoglobin content and histological parameters in the 3 types of mice were elevated by Cd injection in a time-dependent manner. The degree of hemorrhage in Cd-injected MT-I/II null mice was similar to that in the wild-type mice. In contrast, hemorrhage in the MT-III null mice was attenuated compared with that in wild-type mice and MT-I/II null mice. Cd levels, MT-I and MT-II mRNA levels and Cd-binding molecules in the testis were similar between MT-III null mice and wild-type mice. In microarray analysis, high expression of purine-nucleoside phosphorylase 2 (Pnp2), retinal degeneration 3 (Rd3), and cadherin-like 24 (Cdh24) was revealed in the testis of MT-III null mice under normal or Cd-treated conditions. MT-III null mice were found to show attenuation of Cd-induced severe testicular toxicity. These results suggest the lack of MT-III contributes to protection of testis from Cd. In addition, regulation of Pnp2, Rd3, and Cdh24 mRNA levels may involve the sensitivity of MT-III null mice to Cd.


Viability of a FRET dual binding technique to detect calpastatin.
We have been investigating a fluorescence dual binding biosensor to detect calpastatin. Calpastatin is a protein found in meat and it is a regulator of meat tenderness. The ability to accurately predict the calpastatin concentration of beef with a biological sensor at the time of grading would lead to a more accurate assessment of the overall palatability of beef when it reaches the consumer. Meat can then be labeled as tender or tough, which would greatly enhance meat processors' ability to grade meat, allowing them to recover lost revenue. The biosensor technique utilized the chemical transduction principle of fluorescence resonance energy transfer (FRET). FRET requires the use of two fluorophores, termed a donor and acceptor. In this study, the donor fluorophore was conjugated to the protein, mu-calpain, while the acceptor fluorophore was conjugated to a monoclonal antibody. The results showed that in the presence of calpastatin, the labeled mu-calpain and antibody would bind to calpastatin, reducing the distance between the two proteins and eliciting a measurable change in fluorescence. The FRET dual binding technique was tested in heated and unheated meat extract, and a limit of detection for calpastatin was 120 ng/ml in diluted heated meat extract with no significant response in the unheated meat extract. Stable response times were achieved within 5 min. The proof-of-principle of utilizing a FRET dual binding technique to detect calpastatin in heated meat extract has been established.


Improved two-point frequency shift power method for measurement of shear wave attenuation.
Quantitative assessment of mechanical properties of biological soft tissues is frequently evaluated using a noninvasive modality, called ultrasound shear wave elastography (SWE). SWE typically exerts an acoustic radiation force (ARF) to produce shear waves propagating in the lateral direction for which velocities and attenuations are measured. The tissue viscoelasticity is commonly studied by investigating the shear wave phase velocity curves. Viscoelastic tissue properties can also be characterized through utilizing various shear wave attenuation techniques. In this study, we propose an improved method for measuring the shear wave attenuation, called two-point frequency shift power (2P-FSP), which is an improved version of the two-point frequency shift (2P-FS) method. The technique is fully data driven and does not use a rheological model for mathematical modeling. The 2P-FSP method utilizes the power spectra frequency shift of shear waves measured at two spatial positions, which provides robustness to noise. The conceptual basis for the 2P-FSP is provided and tested with numerical and experimental data. We investigated how the location of the first signal and the distance interval between the two locations influence the shear wave attenuation measurement in the 2P-FSP technique. We utilized the 2P-FSP method on numerical phantom data generated using a finite-difference-based method in tissue-mimicking viscoelastic media. Moreover, we tested the 2P-FSP method with data from custom-made tissue-mimicking viscoelastic phantom experiments, and ex vivo porcine liver. We compared results from the proposed technique with results from 2P-FS and analytical values in the case of simulations. The results showed that the 2P-FSP method provides improved results over the 2P-FS technique for lower signal-to-noise ratio (SNR) and locations farther from the push location considered, and can be used to measure attenuation of viscoelastic soft tissues.


[Blood vessel access, skill and tricks].
From a personal viewpoint, the author explains diverse "tricks" and skills which can be used when facing various emergency situation. In this case, the author deals with means of applying blood vessel access.


Identification of the novel HLA-B*58:138 allele in a Korean individual.
HLA-B*58:138 differs from HLA-B*58:01:01:01 by one nucleotide substitution in exon 3 at codon 145.


Measure quantity of mitochondrial DNA in aged bones or calculate it from nuclear DNA quantitative PCR results?
Mitochondrial DNA (mtDNA) is of great value in forensics to procure information about a person when a next of kin, personal belongings, or other sources of nuclear DNA (nDNA) are unavailable, or nDNA is lacking in quality and quantity. The quality and reliability of the results depend greatly on ensuring optimal conditions for the given method, for instance, the optimal input of the copy number (CN) in next-generation sequencing (NGS) methods. The unavailability of commercial quantitative PCR (qPCR) methods to determine mtDNA CN creates the necessity to rely on recommendations to infer mtDNA CN from nDNA yield. Because nDNA yield varies between individuals, tissues, parts of the same tissue, and because mtDNA CN varies between tissues, such assumptions must be examined for a specific context, rather than be generalized. This study compares mtDNA CN calculated from nDNA yield and qPCR measured mtDNA CN. Seventy-five femurs from the Second World War victims were used as samples; they were cut below the greater trochanter, surface contaminants were removed by mechanical and chemical cleaning, samples were fully demineralized, and DNA was isolated. PowerQuant® Kit (Promega) was used to analyze DNA yield. An in-house method was used to determine mtDNA CN. Comparison of mtDNA CN from nDNA derived calculations and measured mtDNA CN highlighted vast differences. The results emphasize the need to perform qPCR to assess mtDNA CN before NGS analyses of aged bones' mitogenomes rather than estimating mtDNA CN from nDNA yield to ensure the quality and reliability of the results of NGS analysis.


Impact of the barium enema on patient management.
The medical records of 214 consecutive inpatients who had a barium enema examination were reviewed, and the clinical indications, efficacy of the barium enema, and patient outcome were correlated to determine the impact of the barium enema on patient management. The most frequent indications were rectal bleeding (33%), abdominal pain (31%), anemia (17%), weight loss (12%), and previous lesions needing reevaluation (12%). Diverticular disease (30%), colonic polyps (10%), and primary or secondary malignancies (12%) were the most common abnormalities detected radiographically. The sensitivity of the barium enema for colonic neoplasms was 89%, with only 1 small cecal polyp being undetected. The effects of barium enema on patient management were serious pathology excluded (64%), diagnosis made that changed therapy (24%), existing therapy continued (10%), and a further study ordered (2%). No serious or life-threatening lesion was missed by barium enema.


Characterization of speed adaptation while walking on an omnidirectional treadmill.
Conventional treadmills are widely used for gait retraining in rehabilitation setting. Their usefulness for training more complex locomotor tasks, however, remains limited given that they do not allow changing the speed nor the direction of walking which are essential walking adaptations for efficient and safe community ambulation. These drawbacks can be addressed by using a self-pace omnidirectional treadmill, as those recently developed by the gaming industry, which allows speed changes and locomotor movements in any direction. The extent to which these treadmills yield a walking pattern that is similar to overground walking, however, is yet to be determined. The objective of this study was to compare spatiotemporal parameters, body kinematics and lower limb muscle activation of healthy young individuals walking at different speeds (slow, comfortable, fast) on a low-cost non-motorized omnidirectional treadmill with and without virtual reality (VR) vs. overground. Results obtained from 12 young healthy individuals (18-29 years) showed that participants achieved slower speed on the treadmill compared to overground. On the treadmill, faster walking speeds were achieved by a mere increase in cadence, as opposed to a combined increase in cadence and step length when walking overground. At matched speed, enhanced stance phase knee flexion, reduced late stance ankle plantarflexion, as well as enhanced activation amplitudes of hip extensors in late stance and hip extensors in early swing were observed. The addition of VR to treadmill walking had little or no effect of walking outcomes. Collectively, results show that the omnidirectional treadmill yields a different walking pattern and lead to different adaptations to speed compared to overground walking. We suggest that these alterations are mainly driven by the reduced shear forces between the weight bearing foot and supporting surface and a perceived threat to balance on the omnidirectional treadmill. Since such treadmills are likely to be used for prolonged periods of time by gamers or patients undergoing physical rehabilitation, further research should aim at determining the impact of repeated exposure on gait biomechanics and lower limb musculoskeletal integrity.


[Reconciling temporality in the caregiver-user-family trilogy].
Multiple factors are at play in a patient's care. One such factor, time, is a unique notion which establishes itself as a third party in the caregiver-patient relationship. Understanding the challenges involved makes it easier to appreciate how it can interfere in care and how to use it as a real resource in the patient's treatment.


Epidemiology, Clinical Characteristics and Risk Factors for Severity of Chronic Disseminated Candidiasis in Jerusalem, Israel.
Chronic disseminated candidiasis (CDC) occurs mostly in patients with acute hematologic malignancy and its clinical manifestations derive from immune reconstitution following neutrophil recovery. The aim of this study was to describe epidemiological and clinical characteristics of CDC and define risk factors for disease severity. Demographic and clinical data were collected from medical files of patients with CDC hospitalized in two tertiary medical centers in Jerusalem between 2005 and 2020. Associations between different variables and disease severity were evaluated, as well as characterization of Candida species. The study included 35 patients. CDC incidence slightly increased during study years and the average number of involved organs and disease duration was 3 ± 1.26 and 178 ± 123 days, respectively. Candida grew in blood in less than third of cases and the most common isolated pathogen was Candida tropicalis (50%). Histopathological or microbiological workup in patients who underwent an organ biopsy demonstrated Candida in about half of the patients. Nine months after starting antifungals, 43% of the patients still didn't have resolution of organ lesions in imaging modalities. Factors associated with protracted and extensive disease were prolonged fever prior to CDC and absence of candidemia. A C- Reactive Protein (CRP) cutoff level of 7.18 mg/dL was found to predict extensive disease. In conclusion, CDC incidence is increasing and the number of involved organs is higher than previously described. Clinical factors such as fever duration prior to CDC and absence of candidemia can predict severe course of disease and assist in treatment decisions and follow-up planning.


[A new technique for harvesting of internal thoracic artery].
A new, unique harvesting technique of internal thoracic artery is described. We have found that the technique can be applied easily and quickly and gives excellent exposure of the pedicle from origin to bifurcation.


An ultrastructural study of the development of afferent and efferent synapses on outer hair cells of the guinea pig organ of Corti.
The guinea pig organ of Corti was studied using transmission electron microscopy, the second turn of the cochlea being examined at various ages between 20 days before birth and 30 days postnatal. Outer hair cells were examined at each of these ages. At all ages studied, the efferent (presynaptic) terminals are large and are packed with synaptic vesicles, whereas the afferent (postsynaptic) terminals are generally smaller, with a relatively small number of vesicles. During development, the subsynaptic cistern changes from a fragmented, diffuse profile extending over 50-70% of the length of the efferent contact zones, to a continuous, compact structure spanning neighbouring synapses. Synaptic vesicles in the efferent terminals are predominantly rounded in early development, flattened vesicles appearing postnatally. The synaptic bodies at afferent synapses do not change noticeably during development. Quantitative analysis revealed that the area of efferent terminals and the length of their active zone increase with increasing age, the same parameters decreasing in afferent terminals. Synaptic vesicles in the efferent terminals decrease in diameter, but remain constant in afferent terminals, with increasing age. The number of hair cell membrane invaginations decreases as development proceeds.


S-100 protein in normal, osteoarthrotic, and arthritic cartilage.
The occurrence of the S-100 protein was studied in normal and pathological cartilage from patients with osteoarthrosis and rheumatoid arthritis. In normal cartilage all cells of the different zones exhibit immunoreactivity for this protein. An identical distribution is observed in cases of osteoarthrosis and rheumatoid arthritis. Cellular pannus tissue usually is devoid of the S-100 protein. However, if chondroid metaplasia occurs in this tissue the cells become positive for the S-100 protein. From these results it is concluded that a chondroid metaplasia not only leads to a matrix comparable to cartilage but also contains cells that gain further characteristics of original chondrocytes.


Co-variates associated with outcomes of tyrosine kinase-inhibitor therapy in persons with chronic myeloid leukaemia initially presenting in accelerated phase.
We interrogated data from 278 consecutive subjects with chronic myeloid leukaemia (CML) presenting in accelerated phase diagnosed by European LeukemiaNet (ELN) criteria receiving initial imatinib (n = 187) or a 2nd-generation tyrosine kinase-inhibitor (2G-TKI; n = 91). In multi-variable analyses, blood and/or bone marrow blasts ≥15% (Hazard ratio [HR] = 3.7 [1.6, 8.5], p = 0.003) and blood basophils <3% (HR = 4.6 [2.0, 10.7], p < 0.001) were significantly-associated with worse transformation-free survival (TFS). Age ≥60 years (HR = 4.3 [1.7, 11.4], p = 0.003), platelet concentration <230 × 10E + 9/L (HR = 4.7 [2.0, 10.7], p < 0.001) and blood and/or bone marrow blasts ≥9% (HR = 3.9 [1.7, 8.7], p = 0.001) were significantly-associated with worse survival. Based on number of adverse prognostic co-variates of TFS and survival, respectively, subjects were classified into the low- (none), intermediate- (one) and high-risk (≥2) cohorts with significant difference in TFS and survival (all p < 0.001). In propensity-score matching analysis subjects initially receiving a 2G-TKI had higher cumulative incidences of cytogenetic and molecular responses but similar TFS and survival to those receiving imatinib. Our data should help inform physicians treating person with CML initially presenting in accelerated phase.


The treatment of stage III carcinoma of the uterine cervix with telecobalt irradiation.
This is a retrospective analysis of 148 patients with histologically proven carcinoma of the cervix, stage III, treated with irradiation. All patients received external irradiation with cobalt 60 followed by intracavitary radium application. The median age was 55 years. Squamous cell carcinoma was found in 96.5% of the cases. The 8-year actuarial survival rate was 41%. Bilateral parametrial invasion proved to be a strong prognostic factor. Patients with unilateral disease had a survival rate of 43% whereas in those with bilateral involvement it was only 15% (P less than 0.005). The total pelvic failure rate was 29.5%. The overall incidence of distant metastasis was 11%. The complication rate (minor and major complications) was high, with vaginal stenosis (22.5%), proctitis (21.5%), cystitis (13.5%), and fistulae (4%) occurring in 33, 32, 20, and 6 patients, respectively. New treatment modalities are urgently needed for advanced carcinoma of the cervix. Bilateral parametrial involvement is an unfavorable prognostic factor and this should be kept in mind when designing new protocols.


Prevalence of syphilis and associated factors among female sex workers in Ethiopia: findings from a multilevel analysis of a national bio-behavioral survey.
Syphilis is a highly contagious sexually transmitted infection posing a significant public health challenge, especially in developing countries, including sub-Saharan Africa. Female sex workers are exposed to sexually transmitted infections, including syphilis, because of their sexual behavior and limited access to health services. However, data on national syphilis prevalence estimates and the associated factors are scarce in Ethiopia. This, as well as our limited knowledge about the extent of clustering among female sex workers in the country, is a critical gap in information we aimed to fill through this analysis. The study was a cross-sectional, bio-behavioral survey conducted among female sex workers in six cities and ten major towns in Ethiopia. Participants were selected using a respondent-driven sampling method. Survey participants provided blood samples for syphilis, HIV, and hepatitis serological testing. Survey data were collected via an interviewer-administered questionnaire. In this analysis, we employed descriptive statistics to summarize data on the study variables. In addition, we used multilevel bivariable and multivariable logistic regression models to examine the association between independent variables and the dependent variable (syphilis prevalence) while accounting for the clustering effect. A total of 6085 female sex workers participated in the survey. Their median age [Interquartile Range (IQR) was 25 (8)] years, and a majority (96.1%) were in the 20-24-year-old age group. The prevalence of syphilis among female sex workers in Ethiopia's six cities and ten major towns was 6.2%. Being in the age group of 30-34 (AOR = 2.64; 95% CI = 1.40, 4.98) and 35-59 (AOR = 4.7; 95% CI = 2.5, 8.86), being divorced/widowed (AOR = 1.37; 95% CI = 1.03, 1.82), having no formal education (AOR = 3.38; 95% CI = 2.34, 5.11), primary 1st cycle (grades 1-4) education (AOR = 2.77; 95% CI = 1.79, 4.30), and having primary 2nd cycle (grades 5-8) education (AOR = 1.80; 95% CI = 1.21, 2.69) were significantly associated with syphilis among female sex workers. The prevalence of syphilis among female sex workers was high. Being divorced/widowed or in the older age group and having a low level of education were significantly associated with an increased risk of syphilis. The high prevalence and associated factors identified need to be considered in planning comprehensive interventions to control syphilis among female sex workers in Ethiopia.


Disturbances of menstrual cycle after immunization against SARS-CoV-2 and their risk factors: Cross-sectional clinical study.
Abnormalities of the menstrual cycle were reported after infection with SARS-CoV-2 and vaccination against it, but the available data are very heterogeneous, do not reflect intermenstrual variations or regional differences, and their risk factors are missing. We performed a survey-based study among 6383 employees and students of Ulm University Hospital in Germany between March 1 and 31, 2021. Attributes of menstrual cycles such as cycle length (CL), menses duration (MD), and bleeding volume (BV) were reported as categorical variables before and after immunization against SARS-CoV-2 (first, second, third vaccination or infection). Additionally, the potential risk factors for cycle changes were evaluated and all participants reported the subjective perception of changes, their duration, and time of occurrence. The final analysis included 1726 participants. CL and BV significantly changed after vaccination, but not MD. The subjective perception showed only slight levels of agreement with the objective changes, with the highest Cohen's kappa for CL. The risk factors for the variations in CL were previous cycle irregularities, and risk factors for the changes in BV were age and body mass index. The combination of vaccines (homogenous or heterogeneous) and different types of immunization (infection and vaccination) had no significant effect on cycle irregularities. In summary, immunization against SARS-CoV-2 causes changes in the characteristics of the menstrual cycle, which are mostly temporary. The individual risk factors, but not the type of immunization, can affect the mentioned changes.


Effect of bicarbonate and other buffers on choroid plexus Na+/K+pump.
The Na+/K+ pump on the apical membrane of the choroid plexus epithelium was found to be sensitive to bicarbonate. Glycodiazine, a non-volatile, lipid soluble buffer with a pK of 5.7, mimicked the effect of bicarbonate, and was transported across the epithelium in the same direction as sodium. These results are explained in terms of a Na+/H+ exchange mechanism on the basal-lateral membrane.


Atrial fibrillatory rate as predictor of recurrence of atrial fibrillation in horses treated medically or with electrical cardioversion.
The recurrence rate of atrial fibrillation (AF) in horses after cardioversion to sinus rhythm (SR) is relatively high. Atrial fibrillatory rate (AFR) derived from surface ECG is considered a biomarker for electrical remodelling and could potentially be used for the prediction of successful AF cardioversion and AF recurrence. Evaluate if AFR was associated with successful treatment and could predict AF recurrence in horses. Retrospective multicentre study. Electrocardiograms (ECG) from horses with persistent AF admitted for cardioversion with either medical treatment (quinidine) or transvenous electrical cardioversion (TVEC) were included. Bipolar surface ECG recordings were analysed by spatiotemporal cancellation of QRST complexes and calculation of AFR from the remaining atrial signal. Kaplan-Meier survival curve and Cox regression analyses were performed to assess the relationship between AFR and the risk of AF recurrence. Of the 195 horses included, 74 received quinidine treatment and 121 were treated with TVEC. Ten horses did not cardiovert to SR after quinidine treatment and AFR was higher in these, compared with the horses that successfully cardioverted to SR (median [interquartile range]), (383 [367-422] vs 351 [332-389] fibrillations per minute (fpm), P < .01). Within the first 180 days following AF cardioversion, 12% of the quinidine and 34% of TVEC horses had AF recurrence. For the horses successfully cardioverted with TVEC, AFR above 380 fpm was significantly associated with AF recurrence (hazard ratio = 2.4, 95% confidence interval 1.2-4.8, P = .01). The treatment groups were different and not randomly allocated, therefore the two treatments cannot be compared. Medical records and the follow-up strategy varied between the centres. High AFR is associated with failure of quinidine cardioversion and AF recurrence after successful TVEC. As a noninvasive marker that can be retrieved from surface ECG, AFR can be clinically useful in predicting the probability of responding to quinidine treatment as well as maintaining SR after electrical cardioversion.


Expression of a recombinant branched chain alpha-oxo acid dehydrogenase complex E2 (BCOADC-E2) in insect cells and its immunoreactivity to autoimmune sera.
Preparation of a pure autoantigen by way of recombinant DNA technology has an important value in an accurate diagnosis or prognosis of an autoimmune disease. BCOADC-E2 subunit, a mitochondrial protein, has been known to be the autoantigen of primary biliary cirrhosis (PBC), a chronic autoimmune liver disease, as well as idiopathic dilated cardiomypathy (IDCM), a chronic autoimmune heart disease. Recombinant form of this molecule had been expressed in E. coli but with low yield and severe degradation. Furthermore, sera from IDCM patients failed to recognized BCOADC-E2 molecule produced in prokaryotic expression system. In this study, a recombinant bovine BCOADC-E2 fusion protein has been expressed in insect cells using baculovirus expression system and analyzed anti-BCOADC-E2 reactivity in sera from patients with PBC or with IDCM. Optimal production of the recombinant fusion protein has been achieved at 20 multiplicity of infection (MOI), and the protein was affinity-purified using metal-binding resins. The affinity-purified BCOADC-E2 protein was successfully recognized by sera from PBC patients, but not by sera from IDCM patients suggesting that the different auto-immune response against BCOADC-E2 is needed to be elucidated in terms of epitope recognition.


Role of acetylcholine receptors in proliferation and differentiation of P19 embryonal carcinoma cells.
Coordinated proliferation and differentiation of progenitor cells is the base for production of appropriate numbers of neurons and glia during neuronal development in order to establish normal brain functions. We have used murine embryonal carcinoma P19 cells as an in vitro model for early differentiation to study participation of nicotinic (nAChR) and muscarinic acetylcholine (mAChR) receptors in the proliferation of neural progenitor cells and their differentiation to neurons. We have previously shown that functional nicotinic acetylcholine receptors (nAChRs) already expressed in embryonic cells mediate elevations in cytosolic free calcium concentration ([Ca2+]i) via calcium influx through nAChR channels whereas intracellular stores contribute to nAChR- and mAChR-mediated calcium fluxes in differentiated cells [Resende et al., Cell Calcium 43 (2008) 107-121]. In the present study, we have demonstrated that nicotine provoked inhibition of proliferation in embryonic cells as determined by BrdU labeling. However, in neural progenitor cells nicotine stimulated proliferation which was reversed in the presence of inhibitors of calcium mobilization from intracellular stores, indicating that liberation of intracellular calcium contributed to this proliferation induction. Muscarine induced proliferation stimulation in progenitor cells by activation of Galphaq/11-coupled M1, M3 and M5 receptors and intracellular calcium stores, whereas Galphai/o-protein coupled M2 receptor activity mediated neuronal differentiation.


Conventional kinesin KIF5B mediates adiponectin secretion in 3T3-L1 adipocytes.
Insulin stimulates adiponectin secretion and glucose transporter type 4 (GLUT4) translocation in adipocyte to regulate metabolism homeostasis. Similar to GLUT4 translocation, intracellular trafficking and release of adiponectin in adipocytes relies on the trans-Golgi network and endosomal system. Recent studies show that the heavy chain of conventional kinesin (KIF5B) mediates GLUT4 translocation in murine 3T3-L1 adipocytes, however, the motor machinery involved in mediating intracellular trafficking and release of adiponectin is unknown. Here, we examined the role of KIF5B in the regulation of adiponectin secretion. The KIF5B level was up-regulated during 3T3-L1 adipogenesis. This increase in cytosolic KIF5B was synchronized with the induction of adiponectin. Endogenous KIF5B and adiponectin were partially colocalized at the peri-nuclear and cytosolic regions. In addition, adiponectin-containing vesicles were co-immunoprecipitated with KIF5B. Knockdown of KIF5B resulted in a marked inhibition of adiponectin secretion and overexpression of KIF5B enhanced adiponectin release, whereas leptin secretion was not affected by changes in KIF5B expression. These data suggest that the secretion of adiponectin, but not leptin, is dependent on functional KIF5B.


Motility response of levamisole/benzimidazole-resistant Haemonchus contortus larvae.
Electronic measurements of the motility of third stage larvae of a susceptible and a levamisole/benzimidazole-resistant strain of Haemonchus contortus were made after incubation in solutions of anthelmintics for 24 h. Results confirmed the resistance to benzimidazoles, but failed to show differences in response to levamisole. Visual observations of paralysis of first and third stage larvae in solutions of levamisole also failed to show significant differences between the two strains. The tests as conducted failed to demonstrate levamisole resistance, suggesting that nematode strains may vary in their mechanisms of resistance to levamisole.


In vivo and in vitro studies with agents that cause quasi-morphine withdrawal syndromes.
Compounds that elicit a "quasi-morphine withdrawal syndrome" (QMWS) after acute administration to rats constitute a new class of behavior-modifying agent. In studying the pharmacological bases of quasi-opiate withdrawal syndromes, we found that the dihydrocodeinone, RX 336-M, a standard QMWS-inducing agent, caused a naloxone-insensitive increase in twitch tension on the field-stimulated rat was deferens preparation. In contrast, IBMX (3-isobutyl-1-methylxanthine), the historical inducer of quasi-abstinence in rats, gave a naloxone-insensitive decrease in twitch tension. Tolerance developed to the "wet-dog" shakes elicited by twice-daily s.c. injections of RX 336-M or IBMX in rats for 5 or 15 days, respectively. The parallel development of tolerance to the in vitro effects of these compounds could not be demonstrated.


High concentration powdered activated carbon-membrane bioreactor (PAC-MBR) for slightly polluted surface water treatment at low temperature.
In this study, different concentrations of PAC combined with MBR were carried out to treat slightly polluted surface water (SPSW) at low temperature (10°C). Effects of PAC on the efficiencies of operation, treatment, and the performance of the process were investigated. It was found that the effluent quality, performance efficiency, resistance of shock load were all enhanced and chemical irreversible membrane fouling was reduced with increasing dosage of PAC in MBR. Only when the concentration of PAC which acted as biological carriers was high enough (i.g., 50 g/L), nitrification without initial inoculation in the filtration tank could start within 19 days and be completed within 35 days at 10°C. Fifty grams per liter PAC was the optimal dosage in MBR for stable and extended operation. Under this condition, mean removal efficiencies of ammonia nitrogen (NH(3)-N), dissolved organic carbon (DOC) and UV(254) were 93%, 75%, and 85%, respectively.


Conditions for self-collimation in three-dimensional photonic crystals.
We introduce the theoretical criterion for achieving three-dimensional self-collimation of light in a photonic crystal. Based on this criterion, we numerically demonstrate a body-center-cubic structure that supports wide-angle self-collimation and is directly compatible with the recently developed holographic fabrication technique. We further show that both bends and beam splitters can be introduced into this structure by the use of interfaces.


Anti-inflammatory effect of taurocholate on TNBS-induced ulcerative colitis in mice.
Taurocholate is a natural conjugated bile acid. The aim of this study was to evaluate the anti-inflammatory effect of taurocholate in TNBS-induced ulcerative colitis in mice. The colitis were induced by rectal administration of TNBS. After 24h, the experimental animals were treated with sulfasalazine (SASP, 500mg/kg/day) and taurocholate (20, 40 and 60mg/kg) for 7 consecutive days. The anti-inflammatory effects of taurocholate for colitis were assessed by body weight, colonic weight and length, macroscopic scores, and histopathological examinations. In addition, the colonic tissue levels of myeloperoxidase (MPO) activity, interleukin (IL)-1β, interferon (IFN-γ) and tumour necrosis factor-α (TNF-α) were also determined to assess the effect of taurocholate. Compared with the model group, treatment with taurocholate (20, 40 and 60mg/kg) significantly inhibited the body weight loss, improved colonic weight and length, and decreased macroscopic and histopathological scores. Furthermore, the activity accumulation of MPO and the colonic tissue levels of IL-1β, IFN-γ and TNF-α were also decreased by administration of taurocholate. All the findings of this study suggested that taurocholate has the anti-inflammatory effect in ulcerative colitis in mice and indicated it as a good candidate to treat inflammatory bowel disease.


Molecular characterization of hypothetical scaffolding-like protein S1 in multienzyme complex produced by Paenibacillus curdlanolyticus B-6.
Paenibacillus curdlanolyticus B-6 produces an extracellular multienzyme complex containing a hypothetical scaffolding-like protein and several xylanases and cellulases. The largest (280-kDa) component protein, called S1, has cellulose-binding ability and xylanase activity, thus was considered to function like the scaffolding proteins found in cellulosomes. S1 consists of 863 amino acid residues with predicted molecular mass 91,029 Da and includes two N-terminal surface layer homology (SLH) domains, but most of its sequence shows no homology with proteins of known function. Native S1 (nS1) was highly glycosylated. Purified nS1 and recombinant Xyn11A (rXyn11A) as a major xylanase subunit could assemble in a complex, but recombinant S1 (rS1) could not interact with rXyn11A, indicating that S1 glycosylation is necessary for assembly of the multienzyme complex. nS1 and rS1 showed weak, typical endo-xylanase activity, even though they have no homology with known glycosyl hydrolase family enzymes. S1 and its SLH domains bound tightly to the peptide-glycan layer of P. curdlanolyticus B-6, microcrystalline cellulose, and insoluble xylan, indicating that the SLHs of S1 bind to carbohydrate polymers and the cell surface. When nS1 and rXyn11A were co-incubated with birchwood xylan, the degradation ability was synergistically increased compared with that for each protein; however synergy was not observed for rS1 and rXynA. These results indicate that S1 may have a scaffolding protein-like function by interaction with enzyme subunits and polysaccharides through its glycosylated sites and SLH domains.


Characterization of Ffh of Mycobacterium tuberculosis and its interaction with 4.5S RNA.
Signal recognition particle (SRP) mediates targeting of proteins to appropriate cellular compartments, which is an important process in all living organisms. In prokaryotes, SRP consists of Ffh, a protein, and 4.5S RNA that recognizes signal peptide emerging from ribosomes. The SRP (Ffh) of one the most successful intracellular pathogen, Mycobacterium tuberculosis, has been investigated with respect to biochemical properties. In the present study, Ffh of M. tuberculosis was overexpressed and was confirmed to be a GTPase using thin layer chromatography and malachite green assay. The GTP binding ability was confirmed by GTP overlay assay. The 4.5S RNA sequence of M. tuberculosis was synthesized by in vitro transcription assay. The interaction between Ffh and 4.5S RNA was confirmed by overlay assay and RNA gel shift assay. The results show that the biochemical properties of M. tuberculosis Ffh have been conserved, and this is the first report that shows the interaction of components of SRP in M. tuberculosis, namely Ffh protein and 4.5S RNA.


Antimicrobial resistance acquisition via natural transformation: context is everything.
Natural transformation is a process where bacterial cells actively take up free DNA from the environment and recombine it into their genome or reconvert it into extra-chromosomal genetic elements. Although this mechanism is known to mediate the uptake of antibiotic resistance determinants in a range of human pathogens, its importance in the spread of antimicrobial resistance is not always appreciated. This review highlights the context in which transformation takes place: in diverse microbiomes, in interaction with other forms of horizontal gene transfer and in increasingly polluted environments. This examination of the abiotic and biotic drivers of transformation reveals that it could be more important in the dissemination of resistance genes than is often recognised.


Underweight, stunting and wasting in black and white South African schoolchildren: malnutrition or adaptation?
Prevalences of underweight for age in South African schoolchildren rose from seven years to reach high levels during puberty. In Black children at the height of the pubertal growth spurt, half to three-quarters were underweight for age and hence classifiable as "malnourished". However, percentages fell rapidly to one-fifth or less at 17 years. High prevalences of stunting were common in Blacks. Wasting occurred in similar proportions of urban Black compared with White children, i.e., the large majority of both Blacks and Whites were normally proportioned; younger children were more wasted than older pupils. Factors additional to nutrition appear to share in the regulation of growth. In Whites on adequate diets, there were (i) similar patterns, but smaller rises in underweight prevalences over the growth spurt, also (ii) small but not insignificant prevalences of stunting: (iii) closely similar "normal" patterns of weight for height occurred in Whites as well as Blacks, and (iv) in Blacks the sharp fall in prevalences of underweight and wasting, after the rapid growth phase, occurred without any dietary intervention. A plea is made to use not only weight, but height, particularly relative to weight, at all ages, in assessing nutritional status and "at risk" groups. The stunted and wasted child is likely to be at greater risk than a similarly stunted but normally proportioned or overweight child--both could be underweight for age. Further definitive studies are required.


Silver-ion redox sensing based on colloid formation by gallate ester derivatives.
Plasmon absorption of silver colloid, which was formed by reduction of silver ion with compounds bearing gallate ester groups in ethanol, was observed by absorption spectroscopy. The time-dependent absorption of silver colloid was found, which shows that the silver colloid is unstable and easy to aggregate. It seemed that a calixarene derivative bearing two gallate ester groups was the most suitable for determining silver ion. In order to prevent the silver colloid aggregation, poly(N-vinylpyrrolidone) (PVP) was added into the solution at a constant time after the preparation of sample solutions. In the calibration graph, the absorbance increased with increasing silver ion concentration, especially in the concentration range of 2.00 x 10(-6) to 1.00 x 10(-5) M. It is possible to determine silver ion concentrations using this method.


Does circadian rhythm disruption induced by light-at-night has beneficial effect of melatonin on sciatic nerve injury?
Melatonin stimulates peripheral nerve regeneration. However, the precise effect of Melatonin on nerve repair in dark period have not been clarified. The aim of the present study was to investigate the effect of melatonin on sciatic nerve injury after melatonin was given to rats in the morning or evening by means of combined analysis. This is the first study to investigate the influence of melatonin on sciatic nerve in cut injury two different times of the day. 60 adult female Wistar rats were divided into 4 groups: control (Group 1), sham-operated (Group 2), sciatic nerve cut+melatonin treatment in light (Group 3), sciatic nerve cut+melatonin treatment in dark (Group 4). Melatonin was administered intraperitoneally at dose of 50 mg/kg/day for six weeks. Recovery of function was analyzed by structural (biochemical properties of the antioxidant levels and ultrastructural analysis) and functional analyses (Sciatic function index, pinch test). The data demonstrated beneficial effect of melatonin in light period. However significant beneficial effect of melatonin was detected on the recovery of the cut sciatic nerve in dark period. Melatonin treatment was unable to influence on the recovery of the cut sciatic nerve in dark period. This means that the effect of melatonin the recovery of the cut injured sciatic nerve depends on the time of treatment may be attributed to its circadian rhythm.


Integrated pretreatment of poplar biomass employing p-toluenesulfonic acid catalyzed liquid hot water and short-time ball milling for complete conversion to xylooligosaccharides, glucose, and native-like lignin.
This work aimed to study an integrated pretreatment technology employing p-toluenesulfonic acid (TsOH)-catalyzed liquid hot water (LHW) and short-time ball milling for the complete conversion of poplar biomass to xylooligosaccharides (XOS), glucose, and native-like lignin. The optimized TsOH-catalyzed LHW pretreatment solubilized 98.5% of hemicellulose at 160 °C for 40 min, releasing 49.8% XOS. Moreover, subsequent ball milling (20 min) maximized the enzymatic hydrolysis of cellulose from 65.8% to 96.5%, owing to the reduced particle sizes and cellulose crystallinity index. The combined pretreatment reduced the crystallinity by 70.9% while enlarging the average pore size and pore volume of the substrate by 29.5% and 52.4%, respectively. The residual lignin after enzymatic hydrolysis was rich in β-O-4 linkages (55.7/100 Ar) with less condensed structures. This lignin exhibited excellent antioxidant activity (RSI of 66.22) and ultraviolet absorbance. Thus, this research suggested a sustainable waste-free biorefinery for the holistic valorization of biomass through two-step biomass fractionation.