[Effects of insulin on the distribution of actins in vascular smooth muscle cells in the process of proliferation via mitogen-activated protein kinase in vitro].
Proliferation of vascular smooth muscle cells (VSMCs) is often accompanied by changes in intracellular actin distribution. The changes are controlled by the signal transduction pathways of protein kinase C/mitogenic activated protein kinase (PKC-MAPK), but the mechanism is unclear. In order to study the effect of insulin on the intracellular signal transduction (PKC-MAPK) probably involved in the modulation of proliferation and redistribution of actins in the VSMCs, the DNA synthesis, MAPK activities and its gene expression, and the redistribution of intracellular actins were investigated in the isolated VSMCs of SHR pretreated with PKC inhibitor and/or insulin, respectively. We found that insulin treatment resulted in proliferation of the VSMCs and an increase in [(3)H] TdR incorporation. Meanwhile, the activities and expression of MAPK increased significantly compared to the control group. These effects of insulin were blocked by PKC inhibitor. In addition, insulin caused a redistribution of the intracellular actins in VSMCs, which was also inhibited by PKC inhibitor. It is, therefore, suggested that these effects of insulin on VSMCs proliferation and distribution of the intracellular actins may be mediated by the MAPK signal transduction pathway.


Trends in trust of professional groups in Portugal during COVID-19.
We repeated a Portuguese study, 5 months after a first evaluation, to observe any change in the average levels of trust of professional groups during the COVID-19 pandemic. From our sample of the Portuguese population, 1455 completed the questionnaire from June through September 2020, and 1156 during 2 months, February and March 2021. We found a trend toward higher trust of medical doctors, nurses, and researchers. Analysis of changes between the two pandemic periods (first and second lockdown) showed a statistically significant increase in trust in nurses (t = - 2.01; p = 0.04) and researchers (t = - 2.40; p = 0.02), and the opposite trend for journalists and politicians. This study shows a trend in increasing trust of the Portuguese people toward science-based activities during the COVID-19 pandemic.


[Insulin therapy in type 2 diabetes].
Type 2 diabetes is a growing prevalent disease, usually symptomless, with devastating chronic complications for the individual, family and society. Its progressive nature leads to the dose escalating and the association of different drugs which will rapidly become insufficient to achieve the glycemic goals established individually. Insulin is the most effective drug to control diabetes but there is frequently a silent contract to resist to its implementation between the healthcare team and people with type 2 diabetes. This publication aims to share information about this therapeutic option, eliminating old myths and giving an understandable teaching about all the process of insulin therapy centered on the person with type 2 diabetes. Sharing knowledge with different groups of healthcare professionals regarding the glycemic goals and how to reach them with distinct kinds of available insulins, will allow the release by multiprofessional teams of a homogeneous and non contradictory information to the people with type 2 diabetes. Moreover, a good engagement between physicians and nurse educators in the same team is the cornerstone to the initiation and intensification of insulin therapy. The decision of starting insulin therapy is not the end of the process. The progressive nature of the disease compels to empower the patient to adjust its dose of insulin according to the self monitoring blood glucose data and to realize that the decision of a single therapeutic scheme is not definitive, being adapted upon the clinical condition of the person along the diabetes evolution.


Spinal cord injury in an adult patient with thoracic butterfly vertebra: a case report and review of the literature.
Butterfly vertebrae are a rare congenital vertebral anomaly. An overlap of this spinal anomaly with other diseases has been reported. However, to the authors' knowledge, the coexistence of butterfly vertebrae and spinal cord injury has not been reported in the literature. A 42-year-old male was admitted to our emergency department after a motor vehicle accident. His complaint was back pain, and he was unable to move both lower limbs. Upon physical examination, the patient was not ambulatory. Sensory examination revealed the absence of sensation below the T12 level. The strength of the bilateral lower limbs was grade 0. The patient received a radiographic evaluation. The initial diagnosis was T11 fracture with complete paraplegia of the lower limbs. Magnetic resonance imaging (MRI) was then performed. Sagittal MRI demonstrated an isointense lesion on T1-weighted imaging and a high-signal spindle-like lesion on T2-weighted imaging of the spinal cord adjacent to the T11 vertebra. The fat-suppressed sequence also revealed hyperintensities of the cord. There was no evidence of acute injury of the T11 vertebral body except for cuneiform anterior wedging. The patient was ultimately diagnosed with complete paraplegia with a T11 butterfly vertebra. He underwent urgent posterior decompressive and fixation surgery from T10 to T12. His postoperative recovery was uneventful. The coexistence of a butterfly vertebra with spinal cord injury was reported for the first time. Although butterfly vertebrae may be incidentally detected, it is important to be familiar with their radiographic features to distinguish them from fractures.


Prognostic significance of systemic immune-inflammation index in patients with nasopharyngeal carcinoma: a meta-analysis.
Previous studies have investigated the prognostic value of the systemic immune-inflammation index (SII) in nasopharyngeal carcinoma (NPC). However, the results have been inconsistent. Therefore, this study aims to investigate the prognostic significance of SII in NPC through a meta-analysis. The PubMed, Web of Science, Embase, and Cochrane Library databases were thoroughly searched. The pooled hazard ratios (HRs) with 95% confidence intervals (CIs) were calculated to evaluate the prognostic value of the SII for survival outcomes. A total of six studies comprising 2169 patients were included in the meta-analysis. Pooled analyses indicated that a high SII was significantly associated with worse overall survival (OS) (HR = 1.69, 95% CI = 1.36-2.09, P < 0.001) and progression-free survival (PFS) (HR = 1.60, 95% CI = 1.29-1.98, P < 0.001) in patients with NPC. Subgroup analysis showed that SII was a significant prognostic marker for PFS but not for OS in NPC. Our meta-analysis demonstrated that a high SII could be an efficient prognostic indicator of OS and PFS in NPC. In our opinion, SII could be used to predict long-term and short-term outcomes in patients with NPC. Furthermore, we suggest that SII be applied to help individual patients with NPC assess the prognostic risk. PROSPERO CRD42022321570.


Rapid efflux of Ca2+ from heart mitochondria in the presence of inorganic pyrophosphate.
Inorganic pyrophosphate (PPi) in the intracellular concentration range causes rapid efflux of Ca2+ from rat heart mitochondria oxidizing pyruvate + malate in a low Na+ medium. Half-maximal rates of Ca2+ efflux were given by 20 microM PPi. During and after PPi-stimulated Ca2+ efflux the mitochondria retain their structural integrity and complete respiratory control. Carboxyatractyloside inhibits PPi-stimulated Ca2+ efflux, indicating PPi must enter the matrix in order to promote Ca2+ efflux. Heart mitochondria have a much higher affinity for PPi uptake and PPi-induced Ca2+ efflux than liver mitochondria.


Incidence and impact of cardiac arrhythmias in coronavirus disease 2019 (COVID-19): A systematic review and meta-analysis.
Studies have shown that cardiac arrhythmias may occur in up to 44% of patients with severe coronavirus disease 2019 (COVID-19) and has been associated with an increased risk of death. This systematic review and meta-analysis aimed to evaluate the incidence of cardiac arrhythmias in patients with COVID-19 and their implications on patient prognosis. We performed a systematic literature search from PubMed, SCOPUS, Europe PMC, Cochrane Central Databases, and Google Scholar + Preprint Servers. The primary endpoint of the study was poor outcomes including mortality, severe COVID-19, and the need for ICU care. A total of 4 studies including 784 patients were analyzed. The incidence of arrhythmia in patients with COVID-19 was 19% (9-28%; I2: 91.45). Arrhythmia occurred in 48% (38-57%; I2: 48.08) of patients with poor outcome and 6% (1-12%; I2: 85.33%) of patients without poor outcome. Patients with COVID-19 experiencing arrhythmia had an increased risk of poor outcome (RR 7.96 [3.77, 16.81], p < 0.001; I2: 71.1%). The funnel-plot analysis showed an asymmetrical funnel plot with most of the studies on the right side of the effect estimate. The regression-based Egger's test showed indication of small-study effects (p = 0.001). Cardiac arrhythmias were significantly associated with an increased risk of poor outcome in COVID-19. Arrhythmias were observed in 19% of patients with COVID-19 and in 48% of patients with COVID-19 and poor outcomes.


The Novavax vaccine had 90% efficacy against COVID-19 ≥7 d after the second dose.
Heath PT, Galiza EP, Baxter DN, et al. Safety and efficacy of NVX-CoV2373 Covid-19 vaccine. N Engl J Med. 2021;385:1172-83. 34192426.


Haitian adolescent personality clusters and their problem area correlates.
This study identified personality clusters among a community sample of adolescents of Haitian decent and related cluster subgroup membership to problems in the areas of substance abuse, mental and physical health, family and peer relationships, educational and vocational status, social skills, leisure and recreational pursuits, aggressive behavior-delinquency, and to sexual risk activity. Three cluster subgroups were identified: dependent/conforming (N = 68), high pathology (N = 30); and confident/extroverted/conforming (N = 111). Although the overall sample was relatively healthy based on low average endorsement of problems across areas of expressed concern, significant physical health, mental health, relationship, educational, and HIV risk problems were identified in a MACI identified high psychopathology cluster subgroup. A confident/extraverted/conforming cluster subgroup revealed few problems and appears to reflect a protective style.


Hyperthyroidism impairs pancreatic beta cell adaptations to late pregnancy and maternal liporegulation in the rat.
Hyperthyroidism modifies lipid dynamics (increased oxidation), impairs insulin action and can suppress insulin secretion. We therefore examined the impact of hyperthyroidism on the relationship between glucose-stimulated insulin secretion (GSIS) and insulin action, using late pregnancy as a model of physiological insulin resistance that is associated with compensatory insulin hypersecretion to maintain glucose tolerance. Our aim was to examine whether hyperthyroidism compromises the regulation of insulin secretion and the ability of insulin to modulate circulating lipid concentrations in late pregnancy. Hyperthyroidism was induced by tri-iodothyronine (T(3)) administration from day 17 to 19 of pregnancy. GSIS was assessed during an IVGTT and during hyperglycaemic clamps in vivo and in vitro, using step-up and -down islet perifusions. Hyperthyroidism in pregnancy elevated the glucose threshold for GSIS and impaired GSIS at low and high glucose concentrations in islet perifusions. In the intact animal, insulin secretion (after bolus glucose) was more rapidly curtailed following removal of the glucose stimulus to secretion. In contrast, GSIS was maintained during protracted hyperglycaemia (hyperglycaemic clamps) in the hyperthyroid pregnant state in vivo. Hyperthyroidism in vivo during late pregnancy blunts GSIS in subsequently isolated and perifused islets at low and high glucose concentrations. It also adversely affects GSIS under conditions of an acute glucose challenge in vivo. In contrast, GSIS is maintained during sustained hyperglycaemia in vivo, suggesting that in vivo factors can rescue GSIS. The ability of insulin to suppress systemic lipid levels during hyperglycaemic clamps was impaired. We therefore suggest that higher circulating lipids may preserve GSIS under conditions of sustained hyperglycaemia in the hyperthyroid pregnancy.


Takayasu's arteritis presenting with focal periostitis affecting two limbs.
Takayasu's arteritis (TA) is a vasculitis of large and medium sized arteries. Involvement of bone in TA is very rare. We report a case of young woman who presented with multiple painful bone lesions which were identified as periostitis with new bone formation associated with TA. Our case is unique in that bony involvement in TA could occur independent of vascular stenosis.


Improvement of large copy number variant detection by whole genome nanopore sequencing.
Whole-genome sequencing using nanopore technologies can uncover structural variants, which are DNA rearrangements larger than 50 base pairs. Nanopore technologies can also characterize their boundaries with single-base accuracy, owing to the kilobase-long reads that encompass either full variants or their junctions. Other methods, such as next-generation short read sequencing or PCR assays, are limited in their capabilities to detect or characterize structural variants. However, the existing software for nanopore sequencing data analysis still reports incomplete variant sets, which also contain erroneous calls, a considerable obstacle for the molecular diagnosis or accurate genotyping of populations. We compared multiple factors affecting variant calling, such as reference genome version, aligner (minimap2, NGMLR, and lra) choice, and variant caller combinations (Sniffles, CuteSV, SVIM, and NanoVar), to find the optimal group of tools for calling large (>50 kb) deletions and duplications, using data from seven patients exhibiting gross gene defects on SERPINC1 and from a reference variant set as the control. The goal was to obtain the most complete, yet reasonably specific group of large variants using a single cell of PromethION sequencing, which yielded lower depth coverage than short-read sequencing. We also used a custom method for the statistical analysis of the coverage value to refine the resulting datasets. We found that for large deletions and duplications (>50 kb), the existing software performed worse than for smaller ones, in terms of both sensitivity and specificity, and newer tools had not improved this. Our novel software, disCoverage, could polish variant callers' results, improving specificity by up to 62% and sensitivity by 15%, the latter requiring other data or samples. We analyzed the current situation of >50-kb copy number variants with nanopore sequencing, which could be improved. The methods presented in this work could help to identify the known deletions and duplications in a set of patients, while also helping to filter out erroneous calls for these variants, which might aid the efforts to characterize a not-yet well-known fraction of genetic variability in the human genome.


Angina Bullosa Hemorrhagica: Frightening Experience-A Case Series.
Angina bullosa haemorrhagica (ABH) is characterized by the recurrent appearance of haemorrhagic bullae on the oropharyngeal mucosa which rupture spontaneously leading to complete recovery within a weeks' time without any scarring. We report the clinical features of six cases of ABH. A cross-sectional observational study was performed. A total of six cases of ABH fulfilling the Ordioni et. al. criteria for diagnosis of ABH were enrolled.The age of our patients were 65, 25, 20, 35, 55 and 48 years. Four of them were females (67%), whereas two were males (33%).Retromolar trigone involvement was most common.The chief complaint in all was reddish bulla(e) in the oral cavity of 1 day duration. Five of the patients had solitary lesions, while one had multiple lesions. The lesions measured from 1 to 3 cm in diameter. Complete blood counts and clotting factors were normal in all patients. All cases healed within a week's time. ABH is not a very common disorder encountered by ENT surgeons, dermatologists, general practitioners, and the lack of knowledge of its normal presentation can lead to unnecessary anxiety and incorrect treatment. The typical hemorrhagic bulla(e) usually erupt after eating hard, hot, or spicy food. These lesions heal spontaneously and treatment is not necessary except for reassurance and mild anxiololytics.


Kidneys contribute to the extrahepatic clearance of propofol in humans, but not lungs and brain.
The principal site for the metabolism of propofol is the liver. However, the total body clearance of propofol is greater than the generally accepted hepatic blood flow. In this study, we determined the elimination of propofol in the liver, lungs, brain and kidneys by measuring the arterial-venous blood concentration at steady state in patients undergoing cardiac surgery. After induction of anaesthesia, propofol was infused continuously during surgery. For measurement of propofol concentration, blood samples were collected from the radial and pulmonary artery at predetermined intervals. In addition, blood samples from hepatic and internal jugular vein were collected at the same times in 19 patients in whom a hepatic venous catheter was fitted and the other six in whom an internal jugular venous catheter was fitted, respectively. In six out of 19 patients fitted with a hepatic venous catheter, blood samples from the radial artery and the renal vein were also collected at the same time, when the catheter was inserted into the right renal vein before insertion into the hepatic vein. Hepatic clearance of propofol was approximately 60% of total body clearance. The hepatic extraction ratio of propofol was 0.87 +/- 0.09. There was no significant difference in the concentration of propofol between the radial, pulmonary arteries and internal jugular vein. However, a high level of propofol extraction in the kidneys was observed--the renal extraction ratio being 0.70 +/- 0.13. We have demonstrated substantial renal extraction of propofol in human. Metabolic clearance of propofol by the kidneys accounts for almost one-third of total body clearance and may be the major contributor to the extrahepatic elimination of this drug.


Cerebellar atrophy: pneumoencephalography and computerized tomography correlation.
Ten consecutive patients with clinical diagnosis of cerebellar atrophy were studied by both CT and PEG. CT and PEG findings on the fourth ventricle, cerebellar cortical sulci and cisterns around the brain stem were evaluated independently and compared according to the criteria based on actual measurements. In the diagnosis of cerebellar atrophy, CT and PEG were complementary, but PEG provided better diagnostic information in detecting minimal changes than CT at this stage of development.


Relationships between Isometric Muscle Strength, Gait Parameters, and Gross Motor Function Measure in Patients with Cerebral Palsy.
This study investigated the correlation between isometric muscle strength, gross motor function, and gait parameters in patients with spastic cerebral palsy and to find which muscle groups play an important role for gait pattern in a flexed knee gait. Twenty-four ambulatory patients (mean age, 10.0 years) with spastic cerebral palsy who were scheduled for single event multilevel surgery, including distal hamstring lengthening, were included. Preoperatively, peak isometric muscle strength was measured for the hip flexor, hip extensor, knee flexor, and knee extensor muscle groups using a handheld dynamometer, and three-dimensional (3D) gait analysis and gross motor function measure (GMFM) scoring were also performed. Correlations between peak isometric strength and GMFM, gait kinematics, and gait kinetics were analyzed. Peak isometric muscle strength of all muscle groups was not related to the GMFM score and the gross motor function classification system level. Peak isometric strength of the hip extensor and knee extensor was significantly correlated with the mean pelvic tilt (r=-0.588, p=0.003 and r=-0.436, p=0.033) and maximum pelvic obliquity (r=-0.450, p=0.031 and r=-0.419, p=0.041). There were significant correlations between peak isometric strength of the knee extensor and peak knee extensor moment in early stance (r=0.467, p=0.021) and in terminal stance (r=0.416, p=0.043). There is no correlation between muscle strength and gross motor function. However, this study showed that muscle strength, especially of the extensor muscle group of the hip and knee joints, might play a critical role in gait by stabilizing pelvic motion and decreasing energy consumption in a flexed knee gait.


Quantitative estimation of hemorrhage in chronic subdural hematoma using the 51Cr erythrocyte labeling method.
Red cell survival studies using an infusion of chromium-51-labeled erythrocytes were performed to quantitatively estimate hemorrhage in the chronic subdural hematoma cavity of 50 patients. The amount of hemorrhage was determined during craniotomy. Between 6 and 24 hours after infusion of the labeled red cells, hemorrhage accounted for a mean of 6.7% of the hematoma content, indicating continuous or intermittent hemorrhage into the cavity. The clinical state of the patients and the density of the chronic subdural hematoma on computerized tomography scans were related to the amount of hemorrhage. Chronic subdural hematomas with a greater amount of hemorrhage frequently consisted of clots rather than fluid.


The thyroidal production of reverse triiodothyronine in autonomous adenoma.
The ratios of reverse triiodothyronine (rT3) to triiodothyronine (T3) concentrations were similar (0.89 +/- 0.09) in twenty-three autonomous thyroid nodules (fourteen decompensated, nine compensated) and paranodular tissues (0.82 +/- 0.08), whereas the serum ratio was significantly lower (0.098 +/- 0.014). This is compatible with the hypothesis that thyroidal rT3 and T3 production from mono- and diiodotyrosine may be a random process in contrast to non-random extrathyroidal rT3 and T3 formation.


Functional analysis of three Nav1.6 mutations causing early infantile epileptic encephalopathy.
The voltage-gated sodium channel Nav1.6 is associated with more than 300 cases of epileptic encephalopathy. Nav1.6 epilepsy-causing mutations are spread over the entire channel's structure and only 10% of mutations have been characterized at the molecular level, with most of them being gain of function mutations. In this study, we analyzed three previously uncharacterized Nav1.6 epilepsy-causing mutations: G214D, N215D and V216D, located within a mutation hot-spot at the S3-S4 extracellular loop of Domain1. Voltage clamp experiments showed a 6-16 mV hyperpolarizing shift in the activation mid-point for all three mutants. V216D presented the largest shift along with decreased current amplitude, enhanced inactivation and a lack of persistent current. Recordings at hyperpolarized potentials indicated that all three mutants presented gating pore currents. Furthermore, trafficking experiments performed in cultured hippocampal neurons demonstrated that the mutants trafficked properly to the cell surface, with no significant differences regarding surface expression within the axon initial segment or soma compared to wild-type. These trafficking data suggest that the disease-causing consequences are due to only changes in the biophysical properties of the channel. Interestingly, the patient carrying the V216D mutation, which is the mutant with the greatest electrophysiological changes as compared to wild-type, exhibited the most severe phenotype. These results emphasize that these mutations will mandate unique treatment approaches, for normal sodium channel blockers may not work given that the studied mutations present gating pore currents. This study emphasizes the importance of molecular characterization of disease-causing mutations in order to improve the pharmacological treatment of patients.


Clinical relevance of health-related quality of life outcomes with darifenacin.
To determine the clinical relevance of changes in health-related quality of life (HRQoL) in patients with overactive bladder (OAB) treated with darifenacin. Data were pooled from three randomized, placebo-controlled, parallel-group, fixed-dose, 12-week studies. After 2-week washout, treatment-free or placebo run-in periods, patients with OAB (n = 1059; 85% women; age 19-88 years) were randomized to 12 weeks' treatment with darifenacin controlled-release 7.5 mg (n = 337) or 15 mg once daily (n = 334) or placebo (n = 388). The King's Health Questionnaire (KHQ) was used to assess HRQoL at baseline and Week 12. The clinical significance of changes in KHQ domain scores was assessed using the concept of minimum important difference (MID), using two different methods. Darifenacin treatment was associated with significantly greater improvements than placebo in six primary KHQ domain scores known to be of importance to patients with OAB. In addition, a significantly greater proportion of darifenacin-treated patients met or exceeded reference MID vs placebo in these domains (Incontinence Impact, Severity Measures, Role Limitations, Social Limitations, Emotions and Physical Limitations; P = 0.01). In darifenacin-treated patients, there were significant correlations between the reductions in incontinence episodes per week and improvements in KHQ scores (P < 0.001). The strongest correlations were in the Incontinence Impact, Social Limitations, Role Limitations, Severity Measures and Emotions domains. Darifenacin treatment was associated with significant, clinically relevant improvements in HRQoL in patients with OAB, shown using the concept of MID to interpret change in KHQ scores.


Evaluation of the SPECTRUM training programme for real-time continuous glucose monitoring: A real-world multicentre prospective study in 120 adults with type 1 diabetes.
Comprehensive knowledge, specific skills and data-analysis competences are prerequisites for the successful use of continuous glucose monitoring (CGM) systems. SPECTRUM is a structured training programme for real-time CGM (rtCGM) consisting of a web-based introduction and six group sessions of 90 min each. The 'CGM-TRAIN study' evaluated the efficacy and acceptance of SPECTRUM and rtCGM systems among adults with insulin therapy. Participants (n = 120) were recruited from 10 German diabetes centres in which they were treated under usual care conditions. Outcome measures were rtCGM knowledge, practical skills, satisfaction with the training programme, satisfaction and acceptance of rtCGM system and glycaemic control. Data were collected at study entry, after training completion and at 6-month follow-up. All participants were diagnosed with type 1 diabetes (56% women, mean age 42.4 ± 13.4 years, diabetes duration 21.6 ± 11.6 years), 110 participants completed the course. After training completion, rtCGM-specific knowledge had improved by 43% (scale: 0-40 points) from 21.2 ± 7.6 to 30.4 ± 4.5 points; p < 0.001. The knowledge-level persisted until follow-up (29.4 ± 4.5). Participants were able to master nearly all the practical requirements of the technology. In addition, rtCGM was highly accepted, and participants were motivated to use their systems continuously. HbA1c improved slightly from 61 ± 14 mmol/mol (7.7 ± 1.3%) before training to 60 ± 14 mmol/mol (7.6 ± 1.3%) at follow-up (p = 0.04). The training programme itself was favourably rated by participants. Under usual out-patient daily care conditions, the training programme SPECTRUM improved knowledge and skills about rtCGM in adults with type 1 diabetes. This was associated with a reduced HbA1c , high satisfaction and acceptance of rtCGM (Clinical Trials Registry no.: DRKS00014380).


Immunochemical differentiation of glucose phosphate isomerase (GPI) allozymes of the mouse.
Polyclonal xenoantisera against mouse GPI-1B and GPI-1C were produced in rabbits and analyzed for their ability to recognize allozyme-specific determinants. These studies showed a high degree of serological similarity among the three allozymes of mouse glucose phosphate isomerase (GPI). However, GPI-1B and GPI-1C could be differentiated from GPI-1A as well as GPI-1A and GPI-1B from GPI-1C using quantitative solid-phase immunobinding assays. In addition, polyclonal and monoclonal alloantibodies specific for GPI-1C were produced in BALB/c (Gpi-1a/Gpi-1a) mice. As indicated by immunoblotting data, the allozyme specificity of rabbit antisera and monoclonal alloantibodies against GPI-1C is dependent on the native structure of that allozyme.


Phoenix atherectomy for patients with peripheral artery disease.
Endovascular atherectomy enables minimally invasive plaque removal in peripheral artery disease (PAD). We aimed to evaluate the safety and the long-term effectiveness of the Phoenix atherectomy for the treatment of complex and calcified lesions in PAD patients. Consecutive all-comer patients with PAD underwent the Phoenix atherectomy. Device safety in terms of perforation and distal embolisation were evaluated. Lesion calcifications were categorised by the Peripheral Arterial Calcium Scoring System (PACSS) and lesion complexity was assessed by the Transatlantic Inter-Society Consensus (TASC). Clinically driven target lesion revascularisation (TLR) was assessed. A total of 558 lesions were treated in 402 consecutive patients. Clinical follow-up was available at 15.7±10.2 months for 365 (91%) patients. Of 402 patients, 135 (33.6%) had claudication, 37 (9.2%) had ischaemic rest pain and 230 (57%) exhibited ischaemic ulcerations. Lesions were mostly identified in the femoropopliteal segments (55%), followed by below-the-knee (BTK) segments (32%). Complex TASC C/D lesions and moderate to severe calcifications (PACSS score ≥2) were present in 331 (82%) and 323 (80%) patients, respectively. The mean lesion length was 20.6±14.3 cm. Five (1%) perforations and 10 (2%) asymptomatic embolisations occurred. Bail-out stenting was performed in 4%, 16% and 3% of patients with common femoral artery, femoropopliteal and BTK lesions, respectively. During follow-up, 5 (3.9%) patients with claudication and 52 (21.9%) patients with critical limb-threatening ischaemia (CLTI) died (hazard ratio [HR] 3.7; p<0.001). Freedom from TLR was 87.5% (112 of 128) in patients with claudication and 82.3% (195 of 237) in patients with CLTI, respectively (HR 1.8; p=0.03). The Phoenix atherectomy can be safely performed in patients with complex lesions with a relatively low rate of bail-out stenting and clinically acceptable TLR rates. DRKS00016708.


Regulation of Carcinogenesis by Sensory Neurons and Neuromediators.
Interactions between the immune system and the nervous system are crucial in maintaining homeostasis, and disturbances of these neuro-immune interactions may participate in carcinogenesis and metastasis. Nerve endings have been identified within solid tumors in humans and experimental animals. Although the involvement of the efferent sympathetic and parasympathetic innervation in carcinogenesis has been extensively investigated, the role of the afferent sensory neurons and the neuropeptides in tumor development, growth, and progression is recently appreciated. Similarly, current findings point to the significant role of Schwann cells as part of neuro-immune interactions. Hence, in this review, we mainly focus on local and systemic effects of sensory nerve activity as well as Schwann cells in carcinogenesis and metastasis. Specific denervation of vagal sensory nerve fibers, or vagotomy, in animal models, has been reported to markedly increase lung metastases of breast carcinoma as well as pancreatic and gastric tumor growth, with the formation of liver metastases demonstrating the protective role of vagal sensory fibers against cancer. Clinical studies have revealed that patients with gastric ulcers who have undergone a vagotomy have a greater risk of stomach, colorectal, biliary tract, and lung cancers. Protective effects of vagal activity have also been documented by epidemiological studies demonstrating that high vagal activity predicts longer survival rates in patients with colon, non-small cell lung, prostate, and breast cancers. However, several studies have reported that inhibition of sensory neuronal activity reduces the development of solid tumors, including prostate, gastric, pancreatic, head and neck, cervical, ovarian, and skin cancers. These contradictory findings are likely to be due to the post-nerve injury-induced activation of systemic sensory fibers, the level of aggressiveness of the tumor model used, and the local heterogeneity of sensory fibers. As the aggressiveness of the tumor model and the level of the inflammatory response increase, the protective role of sensory nerve fibers is apparent and might be mostly due to systemic alterations in the neuro-immune response. Hence, more insights into inductive and permissive mechanisms, such as systemic, cellular neuro-immunological mechanisms of carcinogenesis and metastasis formation, are needed to understand the role of sensory neurons in tumor growth and spread.


Third molar treatment outcome: a comparison of patients' preferences in Sweden and Wales.
To elucidate and compare patients' outcome preferences for removal and retention of mandibular third molars in Sweden and Wales. The subjects comprised patients referred and scheduled for removal of one or both mandibular third molars in Sweden and Wales. The multi-attribute utility (MAU) methodology was applied to study patients' preferences for outcomes of removal and retention of the mandibular third molar. Relative weighting of domains was similar in the two countries. "Home and social life" received the highest relative weighting in Sweden and "general health and wellbeing" in Wales. "Your appearance" received the lowest relative weighting in both countries. In both Sweden and Wales operative jaw fracture was considered to be the outcome with most impact, and dentigerous cyst and imbricated incisors the least impact. Outcome ranking was similar in both countries and operative outcomes were considered by patients to be more detrimental to health than retention outcomes. This comparison showed that patients' preferences in Sweden and Wales were similar and that the outcomes of surgery were considered worse after third molar removal than retention. Patient-orientated treatment decisions were less subject to variation than clinician-orientated decisions.


Coflex interspinous process dynamic stabilization for lumbar spinal stenosis: Long-term follow-up.
To evaluate the long-term efficacy of Coflex dynamic stabilization device in the treatment of lumbar spinal stenosis. The clinical and imaging data of 73 patients undergoing Coflex dynamic stabilization surgery from July 2008 to June 2012 were retrospectively analyzed. All patients had a minimum of 8 years of follow-up. Clinical data were used to assess the clinical efficacy, and radiographic parameters were measured for evaluation of ASD. 56 Patients were followed up for 107.6 ± 13.3 months. The visual analogue scale of pain (VAS), Owestry disability index (ODI) and Japanese Orthopedic Association Scores (JOA) improved significantly after surgery. At 6 months after surgery and the last follow-up, lumbar range of motion (ROM) was significantly lower than that before surgery (P < 0.001). ROM was slightly increased at the last follow-up compared with that 6 months after operation (P > 0.05). ROM of adjacent segments increased at 6 months and at the last follow-up compared with that before surgery (P > 0.05). At 6 months after surgery, intervertebral space height (ISH) and intervertebral foramen height (IFH) of implanted segment was significantly higher than that before surgery (P < 0.05). At the last follow-up, there was a decrease in ISH and IFH (P > 0.05). During the follow-up period, a total of 11 patients (19.6%) experienced complications and 6 patients (10.7%) underwent secondary surgery. Coflex interspinous process dynamic stabilization is effective in the long-term treatment of lumbar spinal stenosis, the ISH and IFH of implanted segment could be increased in a short period of time.


Preferable timing of intraductal ultrasonography during endoscopic retrograde cholangiopancreatography lithotomy: A prospective cohort study.
Intraductal ultrasonography (IDUS) is a highly sensitive and non-invasive detective method that can be used to detect complete calculus clearance during endoscopic retrograde cholangiopancreatography (ERCP). In this study, we examined the preferable timing of IDUS during ERCP lithotomy. From 2017 to 2020, patients with choledocholithiasis were randomized into IDUS-BL (IDUS performed before lithotomy) group, IDUS-ALC (cholangiography and IDUS performed after lithotomy) group, and IDUS-AL group (IDUS performed after lithotomy) group. The influence of IDUS on the accuracy of prejudgment, the incidence of residual stones, the need for repeated lithotomy (RL), and fluoroscopy time were analyzed. A total of 184 patients were enrolled. No residual stones were found during follow-up in any of the three groups. There was no difference in prejudgment accuracy rate on size and number of stones between different groups (all P > 0.05). RL were performed in 5, 9, and 9 cases of IDUS-BL, IDUS-ALC, and IDUS-AL group, respectively (P > 0.05). IDUS-AL group had a shorter fluoroscopy time than the other two groups (1.5 ± 0.6 vs. 2.8 ± 1.2, 2.5 ± 1.0 min, P < 0.05). Incidence of RL was related to the location of calculus [middle or lower part of common bile duct (CBD)], lithotripsy, dilated CBD (2.12 ± 0.46 vs. 1.78 ± 0.40 cm, P < 0.01), and inaccuracy prejudgment. IDUS performed after lithotomy is preferable for shorten fluoroscopy time during ERCP. IDUS is a reliable solution for the stone omission, which may be more valuable for patients with high-risk factors of RL.


The effects of major depression disorder on neurogenic thoracic outlet syndrome surgery outcomes.
Thoracic outlet syndrome (TOS) is a group of disorders caused by impingement of the neurovascular structures at the thoracic outlet. Neurogenic TOS (nTOS), which is thought to be caused by a compression of the brachial plexus, accounts for more than 90% of the cases. Although treatment for nTOS is successful through physiotherapy and/or surgical decompression, little is known about the impact of psychosocial factors, namely, major depressive disorder (MDD), on postoperative outcomes such as non-routine discharge (NRD). Here, we assess whether MDD predicts the type of discharge following nTOS surgical intervention. A retrospective analysis of the National Inpatient Sample database from the years 2005-2018 was performed. Using the International Classification of Diseases Clinical Modification, Ninth and Tenth revisions, patients who underwent a surgical intervention for nTOS were identified. Our primary outcome was to investigate the effects of MDD on nTOS patient disposition status after surgical management; secondary outcomes included analysis of total hospital charges and length of stay. NRD was defined as anything beyond discharge home without healthcare services. Univariate and multivariable logistic regression analyses were conducted to assess MDD and other potential independent predictors of NRD and prolonged hospital stay (> 2 days) following surgical intervention. A total of 6099 patients were identified: 596 (9.77%) patients with MDD and 5503 (90.23%) without MDD. On average, patients with MDD were older (39.6 ± 12.0 years vs. 36.0 ± 13.0 years; p < 0.001), female (80.7% vs. 63.5%; p < 0.001), white (89.6% vs. 85.6%; p = 0.030), and on Medicare (9.6% vs 5.2%; p < 0.001). Univariate and multivariable logistic regression models identified MDD as an independent risk factor associated with a higher risk of NRD (adjusted odds ratio [aOR], 1.50; 95% confidence interval [CI], 1.0-2.2). Additionally, chronic kidney disease (aOR, 2.60; 95% CI, 1.2-5.4), postoperative complications (aOR, 1.87; 95% CI, 1.2-2.9), and Medicare (aOR, 2.95; 95% CI, 1.9-4.7) were statistically significant predictors for higher risk of NRD. However, MDD was not associated with prolonged hospital stay (aOR, 1.00; 95% CI, 0.8-1.2) or higher median of total charges (MDD group: $27,867 vs. non-MDD group: $28,123; p = 0.799). Comorbid MDD was strongly associated with higher NRD rates following nTOS surgical intervention. MDD had no significant impact on length of hospital stay or total hospital charges. Additional prospective research is necessary in order to better evaluate the impact of MDD in patients with nTOS.


Diverse clinical and genetic characteristics of six cases of inherited epidermolysis bullosa.
Inherited epidermolysis bullosa (IEB) represents a group of rare genetic dermatoses comprising various phenotypes ranging from severe cutaneous and extracutaneous involvement to mild cutaneous fragility. Pathogenic variants have been identified in at least 20 genes responsible for IEB. In the present study, six cases of epidermolysis bullosa were recruited and subjected to a combination of clinical and genetic analysis. The family history of each case was surveyed. Whole exome sequencing was performed to identify the causative variation. The six patients showed typical EB symptoms. In all cases, WES detected the diagnostic variations of the COL7A1 or DST gene. A total of 10 variants were identified and verified. The findings of the present study further expanded the mutation spectrum of IEB, provided evidence for genetic counseling to the affected families, as well as highlighted the complexity of the pathogenesis of IEB.


A Simple One-Parameter Percent Dissolved Versus Time Dissolution Equation that Accommodates Sink and Non-sink Conditions via Drug Solubility and Dissolution Volume.
In vitro dissolution generally involves sink conditions, so dissolution equations generally do not need to accommodate non-sink conditions. Greater use of biorelevant media, which are typically less able to provide sink conditions than pharmaceutical surfactants, necessitates equations that accommodate non-sink conditions. One objective was to derive an integrated, one-parameter dissolution equation for percent dissolved versus time that accommodates non-sink effects via drug solubility and dissolution volume parameters, including incomplete solubility. A second objective was to characterize the novel equation by fitting it to biorelevant dissolution profiles of tablets of two poorly water-soluble drugs, as well as by conducting simulations of the effect of dose on dissolution profile. The Polli dissolution equation was derived, [Formula: see text], where M0 is the drug dose (mg), cs is drug solubility (mg/ml), V is dissolution volume (ml), and kd is dissolution rate coefficient (ml/mg per min). Maximum allowable percent dissolved was determined by drug solubility and not a fitted extent of dissolution parameter. The equation fit tablet profiles in the presence and absence of sink conditions, using a single fitted parameter, kd, and where solubility ranged over a 1000-fold range. kd was generally smaller when cs was larger. FeSSGF provided relatively small kd values, reflecting FeSSGF colloids are large and slowly diffusing. Simulations showed impact of non-sink conditions, as well as plausible kd values for various cs scenarios, in agreement with observed kd values. The equation has advantages over first-order and z-factor dissolution rate equations. An Excel file for regression is provided.


Histopathological and immunohistochemical evaluation of malignant potential in canine aortic body tumours.
In order to verify the malignant potential of aortic body tumours (ABTs) in dogs, 13 cases of canine ABT were studied histopathologically and immunohistochemically. The cases were divided into two groups according to the presence or absence of metastases to other organs at necropsy examination (metastasis group [n = 9] and non-metastasis group [n = 4]). The mean tumour weight:body weight ratio (TW:BW; g/kg) in the metastasis group (9.3 ± 6.7) was significantly higher than that in the non-metastasis group (1.5 ± 1.7) (P <0.05). In both groups, the neoplastic cells had malignant features including pleomorphism, anisocytosis and anisokaryosis, and mononuclear giant cells were present, showing invasion through the capsule and into the vascular lumen and other adjacent tissues. The mitotic index (MI), mean nuclear area (NA) for size value and coefficient of variation of the nuclear area (CVNA) for anisonucleosis did not differ significantly between the two groups. These findings show that anaplastic characteristics are present regardless of the tumour size or the presence or absence of metastases, suggesting that these tumours are generally malignant or potentially malignant. Immunohistochemical analysis using neuroendocrine markers including neuron-specific enolase, chromogranin A and S100 revealed no obvious differences in labelling intensity of neoplastic cells related to the presence or absence of metastases or associated with the mean TW:BW, MI, NA or CVNA value, indicating that immunohistochemistry has no practical value for determining the tumour grade of canine ABTs.


Stereoscopic Display Is Superior to Conventional Display for Three-Dimensional Echocardiography of Congenital Heart Anatomy.
Three-dimensional echocardiography (3DE) improves visualization of cardiac lesions. Current viewing of 3DE studies on a conventional display diminishes the encoded stereoscopic (stereo) information for depth perception. This study aims to evaluate clinician subjective and objective experience of stereo display compared with nonstereo display of 3DE in congenital heart disease. In this prospective study, 22 cardiologists, advanced cardiology trainees, and cardiothoracic surgeons used a commercially available stereo display system with proprietary software to view 10 3DE data sets, alternating between simple and complex lesions. In part A, participants viewed each data set, randomized to 1 minute of stereo display followed by 1 minute of nonstereo display, or vice versa. In part B, participants could freely toggle between stereo and nonstereo display for an additional 90 seconds per data set. Participants answered a series of questions and rated their subjective experience using stereo versus nonstereo display mode on a Likert scale. Objective data on time spent in each display mode during part B and duration of interaction and degree of movement of the 3DE data set in parts A and B were also collected. All clinician groups found stereo display preferable to nonstereo display of 3DE (P < .0001). Viewing complex lesions was rated lower than simple lesions when using nonstereo display (P < .01). Simple and complex lesions were equally well rated when using stereo display (P = .14). When given a choice of display modes in part B, participants spent more time in stereo display (P < .0001) and interacted more with the 3DE data sets in stereo display (P < .0001). Interactive stereoscopic display of 3DE was preferred over conventional nonstereo display by all clinician groups for viewing both simple and complex lesions. This preference is especially true for viewing complex lesions.


Molecular epidemiology of the attachment glycoprotein (G) gene in respiratory syncytial virus in children with acute respiratory infection in Japan in 2009/2010.
This study performed a detailed genetic analysis of the glycoprotein (G) gene of respiratory syncytial virus (RSV) detected in 50 Japanese children with acute respiratory infection (ARI) in the 2009/2010 season. A phylogenetic tree constructed by the neighbour-joining method showed that 34 and 16 of the RSV strains could be classified into subgroups A and B, respectively. Strains belonging to subgroups A and B were further subdivided into GA2 and BA, respectively. The nucleotide and deduced amino acid sequence identities were relatively high among these strains (>90%). The deduced amino acid sequences implied that a relatively high frequency of amino acid substitutions occurred in the C-terminal 3rd hypervariable region of the G protein in these strains. In addition, some positively selected sites were estimated. The results suggest that RSV with genotypes GA2 and BA was associated with ARI in Japanese children in 2009/2010.


Monolayer MoS2 field effect transistor with low Schottky barrier height with ferromagnetic metal contacts.
Two-dimensional MoS2 has emerged as promising material for nanoelectronics and spintronics due to its exotic properties. However, high contact resistance at metal semiconductor MoS2 interface still remains an open issue. Here, we report electronic properties of field effect transistor devices using monolayer MoS2 channels and permalloy (Py) as ferromagnetic (FM) metal contacts. Monolayer MoS2 channels were directly grown on SiO2/Si substrate via chemical vapor deposition technique. The increase in current with back gate voltage (Vg) shows the tunability of FET characteristics. The Schottky barrier height (SBH) estimated for Py/MoS2 contacts is found to be +28.8 meV (at Vg = 0V), which is the smallest value reported so-far for any direct metal (magnetic or non-magnetic)/monolayer MoS2 contact. With the application of positive gate voltage, SBH shows a reduction, which reveals ohmic behavior of Py/MoS2 contacts. Low SBH with controlled ohmic nature of FM contacts is a primary requirement for MoS2 based spintronics and therefore using directly grown MoS2 channels in the present study can pave a path towards high performance devices for large scale applications.


Dose- and time-related effects of acute diisopropylfluorophosphate intoxication on forced swim behavior and sucrose preference in rats.
Acute intoxication by organophosphorus anticholinesterases (OPs) has been associated with depression and other neuropsychiatric disorders. We previously reported that adult male rats treated with diisopropylfluorophosphate (2.5 mg/kg, sc) showed acute cholinergic signs followed by changes (increased immobility/decreased swimming) in the forced swim test (a measure of behavioral despair) for at least one month. This study was conducted to evaluate the further persistence of changes in the forced swim test out to 4 months and to compare responses in a sucrose preference test, a measure of anhedonia. Adult male rats were treated with vehicle (peanut oil, 1 mL/kg, sc) or DFP (2.0, 2.25 or 2.5 mg/kg) followed by sacrifice 4 h later for measurement of OP-sensitive serine hydrolases (cholinesterase [ChE], fatty acid amide hydrolase [FAAH], and monoacylglycerol lipase [MAGL]) in hippocampus. Additional rats were treated similarly and evaluated for functional signs of acute toxicity from 30 min to 6 days, and then motor activity, forced swim behavior and sucrose preference at approximately 1 week, 1 month and 4 months after dosing. All dosages of DFP elicited serine hydrolase inhibition (ChE, 92-96 %; FAAH, 46-63 %; MAGL, 26-33 %). Body weight was reduced in all DFP-treated groups during the first two weeks, and lethality was noted with the higher dosages. Involuntary movements were elicited in all DFP treatment groups during the first week, but both time of onset and rate of recovery were dose-related. There was a significant reduction in ambulation at one week after the highest dosage (2.5 mg/kg), but no other significant locomotor changes were noted. Immobility was increased and swimming was decreased in the forced swim test at all three time-points by 2.25 mg/kg DFP, and at 2 of 3 time-points by the other dosages. While length of water deprivation and time after DFP dosing affected sucrose preference, DFP treatment had no main effect. We conclude that the forced swim test (a measure of behavioral despair/coping mechanism for inescapable stress) is a robust and persistent neurobehavioral consequence of acute DFP intoxication while sucrose preference, a measure of anhedonia and a common symptom of major clinical depression, is not affected.


Reliability of the Kinematic Steadiness Index during one-leg standing in subjects with recurrent low back pain.
To assess the reliability of standing time and the Kinematic Steadiness Index (KSI) in one-leg standing compared with the Timed Up and Go (TUG) test while considering anthropometric factors in subjects with recurrent low back pain (LBP). Sixty-six individuals participated in the study. The data were collected on two different days, 1 week apart. The KSI of the core spine, using video motion-capture techniques, was based on the relative standing time and relative standstill time. The intraclass correlation coefficient (ICC2,1) was compared for the reliability between measures. The covariates, such as age, Body Mass Index, and the Oswestry Disability Index (ODI), were analyzed for any interactions based on these measures. The standing time (t = - 1.01, p = 0.32) and the KSI (t = - 1.70, p = 0.09) were not significantly different between measures. The TUG results were not different between measures (t = 1.01, p = 0.32). The Cronbach's alpha for the standing time was 0.84, for KSI was 0.89, and for TUG was 0.76. The standing time and KSI demonstrated an interaction with age, while the TUG demonstrated an interaction with the ODI score. The KSI during one-leg standing could help to develop a practical tool to justify quantity and quality of balance outcome measures, which identify balance deficits and core spine rehabilitation strategies in subjects with recurrent LBP.


Method development for the characterization of biofuel intermediate products using gas chromatography with simultaneous mass spectrometric and flame ionization detections.
Accurate analytical methods are required to develop and evaluate the quality of new renewable transportation fuels and intermediate organic liquid products (OLPs). Unfortunately, existing methods developed for the detailed characterization of petroleum products, are not accurate for many of the OLPs generated from non-petroleum feedstocks. In this study, a method was developed and applied to the detailed characterization of complex OLPs formed during triacylglyceride (TG) pyrolysis which is the basis for generating one class of emerging biofuels. This method uses gas chromatography coupled simultaneously with flame ionization and mass spectrometry detectors (GC-FID/MS). The FID provided accurate quantification of carbonaceous species while MS enabled identification of unknown compounds. A programed temperature vaporizer using a 25 °C, 0.1 min, 720 °C min(-1), 350 °C, 5 min temperature program is employed which minimizes compound discrimination better than the more commonly utilized split/splitless injector, as verified with injections at 250 and 350 °C. Two standard mixtures featuring over 150 components are used for accurate identification and a designed calibration standard accounts for compound discrimination at the injector and differing FID responses of various classes of compounds. This new method was used to identify and quantify over 250 species in OLPs generated from canola oil, soybean oil, and canola methyl ester (CME). In addition to hydrocarbons, the method was used to quantify polar (upon derivatization) and unidentified species, plus the unresolved complex mixture that has not typically been determined in previous studies. Repeatability of the analytical method was below 5% RSD for all individual components. Using this method, the mass balance was closed for samples derived from canola and soybean oil but only ca. 77 wt% of the OLP generated from CME could be characterized. The ability to close the mass balance depended on sample origin, demonstrating the need for an accurate quantification method for biofuels at various stages of production.


A Comparative Study on Aspiration Cytology and Histopathology in Diagnosis of Thyroid Nodule and Its Correlation.
Fine Needle Aspiration Cytology (FNAC) is an initial diagnostic test that has been found to be simple and cost effective but it is associated with pitfalls due to difference in correlations between FNAC and Histopathological Examination (HPE). A study was undertaken with a total of 112 patientspresenting with thyroid nodule who underwent thyroid surgeries. Preoperative FNAC and postoperative HPE were evaluated to calculate sensitivity, specificity, positive and negative predictive value and accuracy. In our study we found sensitivity to be 81.48%, specificity 95.29%, positive predictive value 84.61%, Negative predictive value 94.18% and accuracy to be 91.16%. Thus we can conclude that FNAC is a simple, safe and cost-effective diagnostic modality in the investigation of Thyroid nodule with high specificity and accuracy.


Comparison of GC-MS and TLC techniques for asarone isomers determination.
Two chromatographic methods (GC-MS and TLC) have been developed for separation and determination of alpha and beta asarone from essential oils and alcoholic extracts. The study has been performed on the Acorus calamus (I) and Asarum europaeum (II) essential oils of Romanian origin and the alcoholic extract of Acorus calcamus L (III) and it is a consequence of the International Boards exigency regarding the presence of beta asarone in food, beverages and pharmaceuticals. The isomers were determined using both internal and external standard methods. Both SIM and SCAN techniques were used and the results were compared regarding the chromatographic resolution and interference compounds. The method exhibits good repeatability and low detection limit but is expensive and time consuming. The two isomers concentrations are 5.2- 6.7 microg ml(-1) (I), 460-510 microg ml(-1) (II) and 2.7 5.7 microg ml (III) for alpha asarone and 91-98 microg ml(-1) (I), 24-29 microg ml(-1) (II) and 88 97 microg ml(-1) (III) for beta asarone. The TLC method was developed as an alternative for the GC method. The separation was performed on silica gel plates using toluene: ethyl acetate 8:2 as mobile phase. The evaluation of the chromatograms was made by densitometry using multiple wavelength. The sum of the two isomers are between 80-120 microg ml(-1) (I) and 127-145 microg ml(-1) (III) using spectrophotometric detection and between 73-93 microg ml(-1) (I) and 99-105 microg ml(-1) (III) using fluorimetric detection. The results of the two chromatographic methods were compared. Even the GC is more sensitive, mathematical computations for spots optimization and interference elimination could improves the TLC quality results.


Seasonal changes and inhibited development of the abomasal nematodes of sheep and goats in Saudi Arabia.
From August 1991 to July 1992, a total of 48 abomasa from sheep and 48 from goats were examined at the Jeddah abattoir for the presence of adult and larval nematodes. Twenty-three of the sheep (47.9%) and 21 of the goats (43.8%) were infected with nematodes, a total of eight species being found. Haemonchus contortus and Marshallagia marshalli appeared to be the most important parasites. Other species were less prevalent: Trichostrongylus axei, Camelostrongylus mentulatus, Parabronema skrjabini, Ostertagia circumcincta, Skrjabinagia lyrata, Ostertagia trifurcata. Overall worm counts and infection rates were lowest in the winter. Larval inhibition plays an important role in the Marshallagia life cycle, since this parasite survives the dry season as arrested larvae. By contrast, inhibition of Haemonchus was less pronounced and the parasite survived the same season as adults and larvae. Although the phenomenon of hypobiosis was observed, the number of inhibited larvae was too small to be expected to induce clinical symptoms after resumption of development.


[Application of molecular method in identification of bacterial endophthalmitis].
To search for a non-culture method to improve the efficiency of etiological diagnosis of infective endophthalmitis and other infectious diseases. Fifty clinical specimens of infective endophthalmitis were collected. DNA of bacterium was extracted directly from the aqueous fluid or vitreous fluid and then amplified with universal primers. The PCR products were sequenced subsequently. Twenty-two (44%) bacterial smears and 13 (26%) cultured specimens were positive. PCR positivity was seen in 28 of the 50 cases (56%) and 22 specimens (44%) were sequenced successfully. The results of 16s rRNA gene sequence analysis did not completely conform to the results obtained from culture-dependent approach. Molecular identification method is a rapid and efficient approach for the diagnosis of bacterial infective endophthalmitis.


Quinone transport in the closed light-harvesting 1 reaction center complex from the thermophilic purple bacterium Thermochromatium tepidum.
Redox-active quinones play essential roles in efficient light energy conversion in type-II reaction centers of purple phototrophic bacteria. In the light-harvesting 1 reaction center (LH1-RC) complex of purple bacteria, QB is converted to QBH2 upon light-induced reduction and QBH2 is transported to the quinone pool in the membrane through the LH1 ring. In the purple bacterium Rhodobacter sphaeroides, the C-shaped LH1 ring contains a gap for quinone transport. In contrast, the thermophilic purple bacterium Thermochromatium (Tch.) tepidum has a closed O-shaped LH1 ring that lacks a gap, and hence the mechanism of photosynthetic quinone transport is unclear. Here we detected light-induced Fourier transform infrared (FTIR) signals responsible for changes of QB and its binding site that accompany photosynthetic quinone reduction in Tch. tepidum and characterized QB and QBH2 marker bands based on their 15N- and 13C-isotopic shifts. Quinone exchanges were monitored using reconstituted photosynthetic membranes comprised of solubilized photosynthetic proteins, membrane lipids, and exogenous ubiquinone (UQ) molecules. In combination with 13C-labeling of the LH1-RC and replacement of native UQ8 by ubiquinones of different tail lengths, we demonstrated that quinone exchanges occur efficiently within the hydrophobic environment of the lipid membrane and depend on the side chain length of UQ. These results strongly indicate that unlike the process in Rba. sphaeroides, quinone transport in Tch. tepidum occurs through the size-restricted hydrophobic channels in the closed LH1 ring and are consistent with structural studies that have revealed narrow hydrophobic channels in the Tch. tepidum LH1 transmembrane region.


An unexpected new flightless dung beetle species (Coleoptera: Scarabaeidae: Scarabaeinae: Endroedyolini) from the Cederberg Mountains, South Africa.
Silvaphilus joselmae Daniel, Strmpher Deschodt, new species is diagnosed and described from the Cederberg Mountains, South Africa. The tribal affiliation of Silvaphilus Roets Oberlander, 2010 is briefly discussed. We revise some generic characters and update the key for the species and genera in the tribe Endroedyolini Davis, Deschodt Scholtz, 2019. We also provide a distribution map of the two Silvaphilus species.


Miniaturized sealed-tube allele-specific PCR.
DNA diagnostics has been progressively moving from expensive, low-throughput, multi-step methods towards inexpensive, robust, and high-throughput methods. Here we describe the further validation and refinement of a recently described novel genotyping method that has the latter characteristics. An evolved form of allele-specific PCR, the method generates a fluorescent signal through the use of universal labeled primers, which can be quantified directly from microplates using standard plate readers. We have applied the method successfully to a test set of 12 novel single nucleotide polymorphisms (SNPs) on a panel of 47 individuals using low reaction volumes. We demonstrate that the method is extremely accurate, robust, and can be optimized in a simple and predictable manner. By conducting the assay in closed-tube format, the potential for contamination is reduced to a minimum. By virtue of its simplicity, the method is versatile and cost-effective with potential for use in industrial-scale genetic studies or in the clinical diagnostic setting.


Lateralization reversal after intertectal commissurotomy in the pigeon.
After transection of the intertectal commissures, the lateralization of a visually controlled behaviour of pigeons was reversed to a degree proportional to the extent of the commissurotomy. Preoperatively, the pigeons produced higher pecking rates in a successive pattern discrimination task when seeing with the right eye. Postoperatively, this lateralization reversed to show a superiority with the left eye. Visual lateralization may depend on an asymmetrical intertectal interaction.


Muscle cellularity at cranial and caudal levels of the trunk musculature of commercial size sea bass, Dicentrarchus labrax (Linnaeus, 1758).
In eight specimens of Atlantic sea bass of commercial size (congruent with 350 g) muscle cellularity was studied at two selected sampling levels of the trunk axial musculature: caudal (anal opening) and cranial (fourth radius of the dorsal fin). The following parameters were quantified at both sampling levels: white muscle cross-sectional area, white muscle fibre diameter (900-1200 fibres), muscle fibre number and muscle fibre density. Results showed a higher total cross-sectional area at cranial than at caudal level (P < 0.05), what is related with their different gross morphology. However, the white muscle fibre size distribution, as well as the muscle fibre number and density did not show significant differences between them. This study contributes to typify muscle fibre sampling in sea bass of commercial size what is of great interest for morphometric studies where white muscle cellularity is commonly correlated with textural or organoleptic parameters.


Pregnancy-induced hypertension: development of a model in the pregnant primate (Papio anubis).
Experiments were performed on two groups of pregnant baboons. In the experimental group, the subrenal aortic blood flow was reduced by 58% of its original value at 100 days of gestational age. In the control group, the blood flow was measured but not restricted. In the experimental group fetal death occurred in three of 12 animals following the use of a single left-flank incision to approach both the renal artery and the abdominal aorta. In the control group, pregnancies in eight of nine animals went successfully to 165 days. In the experimental group the development of hypertension, a decrease in plasma renin activity, an increase in renal resistance, an increase in serum uric acid, and the development of glomerular changes consistent with those seen in human pregnancy-induced hypertension were noted. These studies demonstrate that pregnancy-induced hypertension can be produced experimentally in a pregnant baboon, and this model should prove useful in expediting studies on the pathophysiologic features and treatment of this condition.


Angiogenic factors and angiogenesis inhibitors in exudative pleural effusions.
The angiogenesis system has been implicated in inflammatory and neoplastic processes; nevertheless, it has been little studied in relation to the pleural space. Our aim is to analyze pleural and plasma levels of the activators--vascular endothelial growth factor, basic fibroblastic growth factor, and inhibitors--endostatin and thrombospondin-1 and to estimate the association between these factors and related biochemical markers. We analyzed pleural fluid from 105 patients with one of the following types of pleural effusion: empyema or complicated parapneumonic, non-complicated parapneumonic, tuberculous, neoplastic and transudative. Angiogenesis activators were higher in exudates than in transudates (p < 0.001) and in empyema than in non-complicated parapneumonic patients (p < 0.001). Endostatin showed no significant differences. Trombospondin-1 showed higher levels in exudates than in transudates and in empyema than in non-complicated parapneumonic effusions (p < 0.001). In pleural exudates there was a positive correlation of angiogenesis activators and trombospondin-1 with low glucose and pH and high LDH. There was no correlation between pleural and plasma levels of the angiogenesis factors. We conclude that exudative pleural effusions showed higher vascular endothelial growth factor, basic-fibroblastic growth factor and trombospondin-1 values than transudative effusions that associated to low glucose and pH, and high LDH. There was no correlation between pleural and plasma concentrations, suggesting a compartmentalized response.


Extracting electrophysiological correlates of functional magnetic resonance imaging data using the canonical polyadic decomposition.
The relation between electrophysiology and BOLD-fMRI requires further elucidation. One approach for studying this relation is to find time-frequency features from electrophysiology that explain the variance of BOLD time-series. Convolution of these features with a canonical hemodynamic response function (HRF) is often required to model neurovascular coupling mechanisms and thus account for time shifts between electrophysiological and BOLD-fMRI data. We propose a framework for extracting the spatial distribution of these time-frequency features while also estimating more flexible, region-specific HRFs. The core component of this method is the decomposition of a tensor containing impulse response functions using the Canonical Polyadic Decomposition. The outputs of this decomposition provide insight into the relation between electrophysiology and BOLD-fMRI and can be used to construct estimates of BOLD time-series. We demonstrated the performance of this method on simulated data while also examining the effects of simulated measurement noise and physiological confounds. Afterwards, we validated our method on publicly available task-based and resting-state EEG-fMRI data. We adjusted our method to accommodate the multisubject nature of these datasets, enabling the investigation of inter-subject variability with regards to EEG-to-BOLD neurovascular coupling mechanisms. We thus also demonstrate how EEG features for modelling the BOLD signal differ across subjects.


Three-dimensional reconstruction of the musculature of various life cycle stages of the cycliophoran Symbion americanus.
Cycliophora is a very recently described phylum of acoelomate metazoans with a complex life cycle and a phylogenetic position that has been under debate ever since its discovery in 1995. Symbion americanus, which lives attached to the mouthparts of the American lobster, Homarus americanus, represents the second species described for the phylum. Aiming to increase the morphological knowledge about this cryptic clade, the present study describes the muscle arrangement of the feeding stage, the attached Prometheus larva with the dwarf male inside, the free living male, the Pandora larva, and the chordoid larva of S. americanus using actin staining and confocal laser scanning microscopy. 3D reconstructions of the muscular systems are presented. In the feeding stage, circular muscles compose the buccal funnel aperture. In addition, a pair of muscles runs longitudinally in the buccal funnel. A complex sphincter was found just proximally to the anus, and six longitudinal muscles run from the trunk constriction ("neck") in basal direction. The musculature of the larval stages and the dwarf male is very complex and includes longitudinal muscles that run dorsally and ventrally. In addition, we found dorso-ventral muscles. The male has a complex posterior muscle apparatus in the vicinity of the penis. In this stage, X- and V-shaped structures were identified on the dorsal and the ventral side, respectively. Pandora and chordoid larvae possess additional circular muscles. We discuss our findings with respect to muscle elements of other metazoan groups and the chordoid larva of Symbion pandora.


Biological interaction of Pt complex with imidazole derivative as an anticancer compound with DNA: Experimental and theoretical studies.
This investigation is applied to find out interesting information on DNA binding mode with Pt(II) derivative of two N, N bidentate ligands in treating cancer. Thus, one new water-soluble platinum complex with FIP and phen with a new formula of [Pt(phen)(FIP)](NO3)2 was prepared and specified. DFT data can be used to evaluate geometry parameters. Based on the ADMET prediction, this complex can be considered a drug-like agent. Cytotoxicity property was evaluated against some human cancerous MCF7, A549, and HCT116 cell lines. Accumulation of Pt complex, cisplatin, and oxaliplatin in each cancerous cell was determined, which is probably related to their lipophilicity and solubility properties. The binding mode of the complex to ct-DNA was investigated by fluorescence spectroscopy, circular dichroism, and molecular docking simulation. The viscosity of DNA by different concentrations of EB and Pt complex titration shows Pt complex interacts with DNA via groove binding like the spectroscopic binding result. In the MD study, DNA helix, RMSD, and RMSF analysis showed that DNA stability decreased and that the majority of residues left the initial state. DNA increased residual deviations and flexibility are linked to an increase in its gyratory radius, which is consistent with the findings of the experiments.


Optimizing the solid-phase immunofiltration assay. A rapid alternative to immunoassays.
The technical variables of the solid-phase immunofiltration assay (SPIA) for the detection of antibodies bound to antigens on a solid-phase filter have been investigated. The binding to solid-phase filters of 125I-labelled axial filament proteins derived from Treponema phagedenis and the optimal conditions for blocking non-specific protein binding were analysed. Axial filament was applied to nitrocellulose, Hybond Nylon and Zeta Probe. After extensive rinsing, the highest amount (68%) of axial filament was observed bound to Zeta Probe. However, blocking non-specific protein binding by pre-wetting the filter with rinsing buffer containing 0.5% Tween 20, prevented the binding of protein to the filter only when nitrocellulose was used as solid phase. Tween 20 (0.5%) in the rinsing and incubation solutions was found to be necessary for the reduction of non-specific binding of contaminants in turbid sera. However, the use of such solutions resulted in a substantial leakage of antigen (47%) during rinsing procedures. Binding of antigen-specific antibody was analysed using 125I-labelled protein A. The maximal possible binding of the antibody occurred within 5 min when the antibody solution was filtered. For optimal binding of 125I-labelled protein A an incubation time of 1 h was needed. It is suggested that solid-phase immunofiltration may provide a rapid alternative for radioimmunoassays or enzyme immunoassays for the detection of specific antibodies.


Fractures of the phalanges of the hand in children.
Fractures of the phalanges of the hand were studied in 203 children. Fingertip fractures caused disability for three weeks but usually had good results. Comminuted fractures often took the form of longitudinal splitting. Epiphyseal fractures were mostly of the Salter type 2 and metaphyseal types and usually had good results: remodelling occurred in both planes at the base of the proximal phalanx but only in the sagittal plane at the base of the middle phalanx. Epiphyseal fractures at the base of the distal phalanx often had unsatisfactory results. Fractures of the neck of phalanx showed no remodelling at all and if deformity is not corrected it is probably permanent.


Measuring IgG anti-A/B titres using dithiothreitol (DTT).
In comparing the use of the sulphydryl compounds 2-mercaptoethanol (2ME) and dithiothreitol (DTT) a rapid one-stage technique for titrating IgG anti-A and anti-B using DTT in an antiglobulin test was tried. The results correlated well with those obtained by either of the two-stage methods using 2ME or DTT.


Visual cancellation of the torsional vestibulo-ocular reflex in humans.
Using the eye-coil/magnetic field method, we measured the torsional vestibulo-ocular reflex (VOR) in ten subjects during active head rotations in roll at about 0.5 Hz. In the dark, regardless of instructions or mental effort, the gains (eye velocity/head velocity) had a mean value of around 0.61. When they viewed a visual display that was stationary, gains rose to 0.72. When viewing a visual display that moved in roll with their heads, subjects could decrease their gains to a mean of 0.46. Separate experiments showed that, as expected at this frequency, the optokinetic system made only a weak contribution. It has been proposed that the horizontal VOR is cancelled by the smooth pursuit system. Since there is no torsional pursuit system, some other mechanism must be used to augment or partially cancel the torsional VOR. Attempts to show that imagination could change this gain showed only weak effects. When asked to imagine an earth-fixed scene, gains were around 0.63; when asked to imagine a subject-fixed scene, gains decreased to only 0.60. When allowed to use a tactile contribution to aid the imagination in cancelling the VOR, the gain dropped further but only to 0.57. We conclude that mental effort in the dark has little influence on the torsional VOR but vision does by a mechanism that is not optokinetic or pursuit.


Prophylactic anticonvulsants for prevention of immediate and early postcraniotomy seizures.
Phenytoin (15 mg/kg) was administered intravenously to 189 patients shortly before their intracranial, supratentorial surgery was completed. Intravenous phenytoin of 5-6 mg/kg/day in three divided doses was administered daily for the first 3 postoperative days. Therapeutic serum levels (10-20 micrograms/mL) were achieved in 113 (59.8%) patients. An equally constituted, randomized control group of 185 patients received a placebo under identical conditions. The group receiving phenytoin had only one immediate and two early postoperative seizures. The 185 controls had four immediate and nine early postoperative seizures. None of the follow-up computed tomography scans of the patients with seizures showed postoperative hematoma. One patient had a significant tension pneumocranium, a possible cause of postoperative seizures. To avoid a decrease in the serum anticonvulsant level due to intraoperative blood loss, it is suggested that for patients who need an urgent or emergent craniotomy, prophylatic anticonvulsant medication should be given at least 20 minutes before completion of wound closure.


A Potential Role of Interleukin 10 in COVID-19 Pathogenesis.
A unique feature of the cytokine storm in coronavirus disease 2019 (COVID-19) is the dramatic elevation of interleukin 10 (IL-10). This was thought to be a negative feedback mechanism to suppress inflammation. However, several lines of clinical evidence suggest that dramatic early proinflammatory IL-10 elevation may play a pathological role in COVID-19 severity.


Lipid A with 2,3-diamino-2,3-dideoxy-glucose in lipopolysaccharides from slow-growing members of Rhizobiaceae and from "Pseudomonas carboxydovorans".
Lipid A's from two Bradyrhizobium species and from the phylogenetically closely related species "Pseudomonas carboxydovorans" were found to contain 2,3-diamino-2,3-dideoxy-glucose as lipid A backbone sugar. In contrast, three representatives of the genus Rhizobium, as well as the phylogenetically related species Agrobacterium tumefaciens, contain solely glucosamine as lipid A backbone sugar. These findings support independent studies on the phylogenetical relatedness based on 16S rRNA-data of the genus Bradyrhizobium with "Pseudomonas carboxydovorans" and Rhodopseudomonas palustris, which form a tight phylogenetical cluster and which all contain the 2,3-diamino-2,3-dideoxy-glucose-containing lipid A. The relatedness of these species to the glucosamine-containing species of the genus Rhizobium and to Agrobacterium tumefaciens is rather distant as documented by 16S rRNA studies.


Mesocoelium malayanum sp. n. (Digenea: Brachycoeliidae) in a frog, Rana macrodon Dumeril and Bibron 1941, from Malaysia.
Mesocoelium malayanum sp. n. is described from the frog Rana macrodon, in Malaysia. Elongate body, broader anteriorly, measuring 1.900 (1.679-2.070) mm long by 0.404 (0.380-0.437) wide, tegument aspinose oral sucker 0.212 (0.200-0.228) by 0.202 (9.191-0.205), acetabulum 0.141 (0.132-0.150) by 0.139 (0.123-0.146), prepharynx present, oesophagus 0.115 (0.096-0.137), caeca reaching posterior 1/3 of body, anterior testis 0.097 (0.087-0.110) by 0.091 (0.087-0.100) dorsal to acetabulum, posterior testis 0.094 (0.087-0.101) by 0.092 (0.091-0.100), cirrus pouch 0.121 (0.111-0.130) by 0.047 (0.041-0.055), genital pore at left of midline of oesophagus just anterior to intestinal bifurcation, ovary 0.110 (0.091-0.127) by 0.089 (0.085-0.096) on left of body and posterior to acetabulum, vitelline glands with single follicles extending from intestinal bifurcation to ends of caeca, excretory vesicle I-shaped and eggs 0.040 (0.037-0.046) by 0.023 (0.022-0.024). Although morphologically related to M. maroccanum and M. meggitti, M. malayanum is considered to be a new species.


General and Abdominal Obesity as Risk Factors for Late-Life Mobility Limitation After Total Knee or Hip Replacement for Osteoarthritis Among Women.
To investigate associations of body mass index (BMI), waist circumference (WC), and waist/hip ratio (WHR) with survival to age 85 years with mobility limitation or death before age 85 years among older women with total knee replacement (TKR) or total hip replacement (THR) for osteoarthritis (OA). This was a prospective study of women (ages 65-79 years at baseline) from the Women's Health Initiative, recruited during 1993-1998 and followed through 2012. Women's Health Initiative data were linked to Medicare claims data to determine TKR (n = 1,867) and THR (n = 944) for OA. Women were followed for up to 18 years after undergoing THR or TKR to determine mobility status at age 85 years. Compared with normal-weight women, overweight, obese I, and obese II women with THR had significantly increased risk of survival to age 85 years with mobility limitation (P < 0.001 for linear trend), with the strongest risk among obese II women (odds ratio [OR] 4.37 [95% confidence interval (95% CI) 1.96-9.74]). Obese II women with THR also had increased risk of death before age 85 years. Women with THR and WC >88 cm relative to ≤88 cm had increased risk of survival to age 85 years with mobility limitation (OR 1.65 [95% CI 1.17-2.33]) but not death before age 85 years. High BMI, WC, and WHR were associated with significantly increased risk of late-life mobility limitation and death among women with TKR for OA. Among older women who underwent THR or TKR for OA, baseline general and abdominal obesity were associated with increased risk of late-life mobility limitation.


Identification of SAA and ACTB as potential biomarker of patients with severe HFMD using iTRAQ quantitative proteomics.
Hand, foot and mouth disease (HFMD) is an infectious disease caused by a variety of enterovirus infections, and the most common types of virus infections are the newenterovirus71 (EV71) and coxsackievirus A group 16 (CoxA16). A small fraction of HFMD will cause further severe HFMD. A rapid and accurate diagnosis biomarker of severe HFMD is important for the timely treatment. In the study, we conducted a clinical biomarker discovery study using iTRAQ combined with MS. Serum proteome alterations in severe HFMD group (n = 32) and health control group (n = 32) were analyzed. 47 proteins were upregulated (fold change > 1.5) between the severe HFMD group and HC group. The identified proteins were classified into different groups according to the molecular function, biology processes, cellular component. During the up-regulated proteins, serum amyloid A (SAA) and human β-actin (ACTB), were confirmed in the serum of the severe HFMD and HC by ELISA assay. SAA and ACTB levels were significantly higher in the sever HFMD patients (P < .01), consistent with iTRAQ-LC-MS/MS analysis. In summary, Our results showed that SAA and human β-actin (ACTB) may be served as a potential biomarker of the clinical diagnosis of severe HFMD.


Fermentation of wort containing deoxynivalenol and zearalenone.
Wort containing deoxynivalenol and zearalenone, each added at a level of 1.9 μg/mL, was fermented by 3 strains ofSaccharomyces cerevisiae for 7 or 9 days to make beer. Analysis showed that deoxynivalenol was stable during this process. The major metabolite of zearalenone was β - zearalenol, which formed in up to 69% of the initial zearalenone concentration, while up to 8.1% of the initial zearalenone was converted to α - zearalenol. The major part of the metabolism of zearalenone occurred by 1 - 2 days. Control experiments, where the yeasts were omitted and deoxynivalenol, zearalenone and α - and β - zearalenol were added, showed good recovery and stability of the mycotoxins over the 7-9 day time period. No deoxynivalenol, zearalenone, α-zearalenol or β-zearalenol was detected in control yeast fermentations where they were not added to the wort.


Longitudinal studies of drug abuse in a fifteen-year-old population. 4. Chronic drug abusers.
In the Gothenburg year cohort of 1953 1% of the men and 0.2% of the women were registered for chronic drug abuse through 1979. An early début and a polydrug abuse with emphasis on intravenous abuse of central stimulants but not opiates were found. Compared with matched individuals who were unregistered for drug abuse, chronic drug abusers had a large over-consumption of psychiatric and social care. Their registered criminality was higher, they were more often sick-listed and had lower incomes than the controls. The prevalence and overall impairment of chronic abusers are similar to those of schizophrenics.


Nuclear Exosome Targeting Complex Core Factor Zcchc8 Regulates the Degradation of LINE1 RNA in Early Embryos and Embryonic Stem Cells.
The nuclear exosome targeting (NEXT) complex is responsible for specific nuclear RNA degradation in mammalian cells. However, its function in development remains unknown. Here, we find that the depletion of a central factor of the NEXT complex, Zcchc8, in mouse results in developmental defects, a shortened lifespan, and infertility. We find that Zcchc8-deficient embryonic stem cells (ESCs) exhibit proliferation abnormalities and reduced developmental potencies. Importantly, the transcripts of retrotransposon element LINE1 are found to be targeted by Zcchc8 and degraded by a Zcchc8-mediated mechanism. We further find that sustained expression of higher levels of LINE1 RNA is detected in maternal Zcchc8-depleted oocytes and embryos. Zcchc8-depleted oocytes show higher chromatin accessibility and developmental defects in both meiotic maturation and embryogenesis after fertilization. Collectively, our study defines Zcchc8-mediated RNA degradation as an important post-transcription regulation of LINE1 transcripts in early embryos and ESCs, which play vital roles in the pluripotency and early development.


Determination of relative UV response factors for HPLC by use of a chemiluminescent nitrogen-specific detector.
Ultraviolet (UV) absorbance is the most widely used detection method for high-performance liquid chromatography (HPLC) separations. In pharmaceutical analysis, purity determinations often include quantitation of related impurities based on relative HPLC peak areas obtained at a specific wavelength. In order for this quantitation to accurately reflect weight percentages of impurities, the relative UV response factors (absorptivities) at the given wavelength must be known. In this work, we present a convenient method for determining relative UV response factors on-line, without isolation or purification of impurities, without standards, and without requiring known analyte concentrations. The procedure described makes use of a chemiluminescent nitrogen-specific HPLC detector (CLND) in conjunction with a UV detector. The CLND response is directly proportional to the number of moles of nitrogen in each eluting peak, and can, therefore, be used to determine relative amounts of each nitrogen-containing impurity present in the sample, provided the molecular formulas are known (e.g. from exact mass LC-MS). It is a simple matter, then, to determine the relative UV response factors from the UV area ratios obtained for the same sample. The feasibility and accuracy of this method is demonstrated for gradient HPLC separations of commercially available compounds of widely varying structures. Finally, the method's utility in obtaining accurate mass balance is demonstrated by application to photodegradation of nifedipine.


Identification of an essential cysteine residue in pyridoxal phosphatase from human erythrocytes.
Pyridoxal-specific phosphatase purified from human erythrocytes was inactivated by a variety of thiol-specific reagents in a time- and concentration-dependent manner. The presence of pyridoxal phosphate, a substrate, or inorganic phosphate, a competitive inhibitor, protected the enzyme from inactivation. Phosphatase inactivated by disulfide reagents was reactivated by the addition of excess dithiothreitol, indicating that the inactivation was due to formation of a mixed disulfide between the reagent and a free cysteinyl residue at or near the active site of the enzyme. Incorporation of either 1 mol of 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), 0.6 mol of iodo[3H]acetate, or 0.6 mol of N-[3H]ethylmaleimide per mol of subunit led to complete inactivation of the enzyme. High concentration of phosphate prevented the incorporation of DTNB and iodo[3H]acetate. Amino acid analysis of carboxymethylated enzyme and DTNB titration of the denatured phosphatase indicated that there may be only 1 cysteinyl residue per subunit. Modification by iodoacetate did not affect the quaternary structure of the enzyme. The phosphatase modified by iodo[3H]acetate was subjected to trypsin digestion, and the resulting peptides were separated on a reverse phase C18 column. Two radioactive peaks were obtained and contained a peptide with the N-terminal sequence of Ala-Gln-Gly-Val-Leu-Phe-Asp-Cys(Cm)-Asp-Gly-Val-Leu-X-Asn-Gly. Most of the radioactivity was released with Cys(Cm). These results indicate that the cysteinyl residue in this sequence is at or near the active site and is essential for activity. Residues 5-12 and 15 of this peptide are identical with a sequence of a yeast alkaline p-nitrophenylphosphatase, and the peptide has little homology with other mammalian phosphatases.


Cumulative Evidence for Association between IL-10 Polymorphisms and Kawasaki Disease Susceptibility: A Systematic Review and Meta-Analysis.
This meta-analysis was carried out to evaluate the associations between IL-10 polymorphisms and Kawasaki disease (KD) risk. A comprehensive literature search was performed using PubMed, EMBASE, China National Knowledge Infrastructure and SciELO for all relevant studies evaluating IL-10 polymorphism and susceptibility to KD. The associations were measured by odds ratios (ORs) and its corresponding 95% confidence intervals (CIs). A total of 13 studies including four studies on -1082 A > G, four studies on -819 T > C and five studies on -592 A > C polymorphism were selected. Pooled data revealed that IL-10 -592 A > C polymorphism was significantly associated with an increased risk of KD (C vs. A: OR = 0.402, 95% CI 0.194-0.832, p = 0.014). However, IL-10 -1082 A > G and -819 T > C polymorphisms were not significantly associated with risk of KD under all five genetic models. Our results revealed that IL-10 -592 A > C polymorphism was associated with risk of KD, while IL-10 -1082 A > G and -819 T > C polymorphisms were not involved in the development of KD.


Is copper effect on glucose incorporation mediated by the insulin receptor in rat adipose tissue?
The insulin receptor is not the site for the stimulatory effect of copper on glucose incorporation into total lipids by adipose tissue; prewashing of adipose tissue of rats fed a stock diet in an insulin-free medium increases the glucose incorporation into total lipids in the presence of 0.1 unit insulin from 220 above control to 430% in the nondiabetic and from 154 to 230% in the streptozotocin-diabetic rat. In contrast, glucose incorporation in the presence of CuCl2 X 2H2O is unaffected by prewashing, being the same in the prewashed as in the unwashed adipose tissue. On the other hand, mild trypsin digestion of adipocytes decreases the glucose incorporation in the presence of 28.5 mU insulin, from 201 to 126% - whereas the effect of copper on glucose incorporation is the same in the trypsin-treated or untreated adipocyte. In order to obtain maximal copper effect the adipocyte has to be preconditioned by insulin; preincubation of diabetic adipose tissue first for one hour with 0.1 unit insulin, and another hour with 100 micrograms CuCl2 X 2H2O added to the medium, results in greater glucose incorporation (230% above control) than when incubated alone with either CuCl2 X 2H2O (125%) or insulin (154%) for two hours. In addition to its previously noted effect on the in vivo insulin release, copper increased the number of the insulin receptor sites in adipocytes.


Fine needle aspiration biopsy in ophthalmology.
When noninvasive techniques fail to confirm or rule out the suspicion of a malignant lesion, fine needle aspiration biopsy may provide an efficient, economical and relatively safe method of obtaining material for cytological study. The technique may also be valuable for intraoperative morphological evaluation. Traumatic complications produced by fine (21-25 gauge) needles are infrequent and almost never serious, and concerns about tumor seeding through the procedure have been largely dispelled by recent studies. Reliable results require a high level of skill in performing the aspiration procedure and in cytologically examining the small amount of material obtained. The authors review the history, applications, techniques and complications of fine needle aspiration biopsy, presenting guidelines for and illustrations of its use in specific ophthalmic situations.


Neuroprotective effects of indole-3-carbinol on the rotenone rat model of Parkinson's disease: Impact of the SIRT1-AMPK signaling pathway.
Parkinson's disease (PD) is the second most common progressive neurodegenerative disorder. Although mounting studies have been conducted, no effective therapy is available to halt its progression. Indole-3-carbinol (I3C) is a naturally occurring compound obtained by β-thioglucosidase-mediated autolysis of glucobrassicin in cruciferous vegetables. Besides its powerful antioxidant activity, I3C has shown neuroprotection against depression and chemically induced neurotoxicity via its anti-inflammatory and antiapoptotic effects. This study aimed to investigate the neuroprotective effects of I3C against rotenone (ROT)-induced PD in male albino rats. The possible protective mechanisms were also explored. PD was induced by subcutaneous administration of ROT (2 mg/kg) for 28 days. The effects of I3C (25, 50, and 100 mg/kg/day) were assessed by catalepsy test (bar test), spontaneous locomotor activity, rotarod test, weight change, tyrosine hydroxylase (TH) expression, α-synuclein (α-Syn) expression, striatal dopamine (DA) content, and histological examination. The highest dose of I3C (100 mg/kg) was the most effective to prevent ROT-mediated motor dysfunctions and amend striatal DA decrease, weight loss, neurodegeneration, TH expression reduction, and α-Syn expression increase in both the midbrain and striatum. Further mechanistic investigations revealed that the neuroprotective effects of I3C are partially attributed to its anti-inflammatory and antiapoptotic effects and the activation of the sirtuin 1/AMP-activated protein kinase pathway. Altogether, these results suggested that I3C could attenuate biochemical, molecular, and functional changes in a rat PD model with following repeated rotenone exposures.


Flow-induced molecular orientation of amphiphile monolayers: incorporation of hexatic elasticity into Ericksen-Leslie theory.
By introducing the local lattice elasticity into the pure Ericksen-Leslie (EL) theory which only considers the deformation of the director, we developed a more general theory for understanding the behaviors of hexatic liquid crystal under flow. In the cases of amphiphilic monolayers in tilted phases (L2 and L'2), the exact solutions of the new EL equation in two types of flow, pure extension and simple shear, explain well most of the features of flow-induced tilt azimuth orientation observed by Fuller's group [Science, 274, 233 (1996)] and Schwartz's group [Nature (London) 410, 348 (2001)]. In particular, the "shear band" domain generated by flow discovered by the former is proved theoretically as the result of two-dimensional Wulf construction in L2 and L'2 phases.