Specific indication of hemoproteins in polyacrylamide gels using a double-staining process.
Hemoproteins were revealed in polyacrylamide gels in the presence of sodium dodecyl sulfate by staining with different benzidine derivatives. When the protein samples were treated with either beta-mercaptoethanol or dithiothreitol, a significant decrease in peroxidase activity of the proteins possessing noncovalently bound heme led to diminished staining. However, when Coomassie blue R-250 staining followed the hemespecific stain it was observed that the hemoprotein bands stained more intensely than duplicate sample bands that had been stained only with the Coomassie blue R-250. This staining property allows the indication of hemoproteins in gels even after the peroxidase yield has been significantly depleted by reducing agents.


Clinical values of creatine kinase and its isoenzymes in children and adolescents with vasovagal syncope.
Vasovagal syncope (VVS) in children and adolescents is a common disorder. There may be an internal relationship between creatine kinase (CK) and its isoenzymes (CKMB) and syncope. The aim of this study was to evaluate the changes of CK and CKMB in children and adolescents with VVS. The VVS group included 218 patients (93 male and 125 female). The control group included 129 healthy children (78 male and 51 female). Serum CK and CKMB levels were estimated. We founded ①Serum CK and CK-MB levels decreased in VVS group than that in control group (P < 0.05). ②The CK levels of female were significantly lower than those of male in VVS group (P < 0.05). ③Serum level of CK-MB were in negative correlation with age, height, weight, BMI whereas in positively correlation with HR. ④CK was effected by CK-MB (β = 0.147, P = 0.037) while CK-MB was independently influenced by age (β = -0.203, P = 0.002) and DBP (β = 0.171, P = 0.011). ⑤Both CK and CK-MB significantly influenced on VVS occurrence after adjusting for the effects of gender, age, height, weight, BMI and HR. The serum CK and CKMB levels decrease in children and adolescents with VVS. CK and CK-MB are the independent protective factors with VVS.


Systematic review and meta-analysis: importance of CagA status for successful eradication of Helicobacter pylori infection.
Some, but not all studies have provided evidence that the CagA status of Helicobacter pylori strains is a predictive factor for the outcome of eradication therapy. To clarify the association between CagA status and eradication outcome. We included studies reporting the numbers of successful and failed cases in H. pylori-eradication therapy according to the CagA status. Fourteen studies (1529 patients) were included of 325 articles identified in the search. The pooled risk ratio for H. pylori-eradication failure in CagA-negative relative to CagA-positive strains and the pooled risk difference in eradication success between the two groups were used as summary statistics. Meta-regression was used for examining the source of heterogeneity. The summary risk ratio for eradication failure in CagA-negative relative to CagA-positive was 2.0 (95% CI: 1.6-2.4, P < 0.001), corresponding with the summary risk difference for eradication success between the groups of 11% (95% CI: 3-19%, P = 0.011). Meta-regression analysis demonstrated that usage of polymerase chain reaction examination for CagA status and a high proportion of non-ulcer dyspepsia patients were factors for heterogeneity among studies. Our meta-analysis confirmed the importance of the presence of CagA as a predictor for successful eradication of H. pylori.


Multidisciplinary team approach in the oral rehabilitation of patients with cleidocranial dysplasia to achieve a functional aesthetic outcome.
Cleidocranial dysplasia is a hereditary congenital disorder that results in delayed ossification of midline structures, and is caused by mutations in the RunX2 (runt-related transcription factor 2) gene located on the short arm of chromosome 6. Successful treatment depends on multidisciplinary assessment and a comprehensive staged treatment plan. We present a case series of 12 patients who were managed with a specifically tailored combination of surgery, orthodontics, and prosthodontics to provide a functional dentition and restore their smile and facial contour. Successful dental rehabilitation can be challenging in this group because patients often have multiple dental anomalies and a reduced quantity and density of alveolar bone. Rehabilitation with early intervention and a carefully planned multidisciplinary approach has been successful in the long term.


Ventricle-directed migration in the developing cerebral cortex.
It is believed that postmitotic neurons migrate away from their sites of origin in the germinal zones to populate distant targets. Contrary to this notion, we found, using time-lapse imaging of brain slices, populations of neurons positioned at various levels of the developing neocortex that migrate towards the cortical ventricular zone. After a pause in this proliferative zone, they migrate radially in the direction of the pial surface to take up positions in the cortical plate. Immunohistochemical analysis together with tracer labeling in brain slices showed that cells showing ventricle-directed migration in the developing cortex are GABAergic interneurons originating in the ganglionic eminence in the ventral telencephalon. We speculate that combinations of chemoattractant and chemorepellent molecules are involved in this ventricle-directed migration and that interneurons may seek the cortical ventricular zone to receive layer information.


Fracture micromechanics of human dentin: A microscale numerical model.
In the present study, we investigate the effects of microstructural morphology and heterogeneity on the initiation and propagation of microcracks in dentin. We create 2D pre-cracked models of human dentin at the microscale level and use a brittle fracture framework of the phase-field method to analyze the crack growth. We discuss the influence of the microstructural features on crack deflection, microcracking, and uncracked ligament bridging through various regions in dentin. The results demonstrate that the difference between the critical energy release rates of peritubular (PTD) and intertubular dentin (ITD) has considerable impacts on microcracking. Our simulations reveal that tubules surrounded by PTDs play an important role in the crack deflection. Our results also indicate that the toughness of dentin increases from the inner to outer dentin. In conclusion, the findings in our study provide valuable insights into the fracture behavior in various regions of dentin.


Neogene speciation and Pleistocene expansion of the genus Pseudephebe (Parmeliaceae, lichenized fungi) involving multiple colonizations of Antarctica.
Widespread geographic distributions in lichens have been usually explained by the high dispersal capacity of their tiny diaspores. However, recent phylogenetic surveys have challenged this assumption and provided compelling evidence for cryptic speciation and more restricted distribution ranges in diverse lineages of lichen-forming fungi. To evaluate these scenarios, we focus on the fungal genus Pseudephebe (Parmeliaceae) which includes amphitropical species, a distribution pattern whose origin has been a matter of debate since first recognized in the nineteenth century. In our study, a six-locus dataset and a broad specimen sampling covering almost all Earth's continents is used to investigate species delimitation in Pseudephebe. Population structure, gene flow and dating analyses, as well as genealogical reconstruction methods, are employed to disentangle the most plausible transcontinental migration routes, and estimate the timing of the origin of the amphitropical distribution and the Antarctic populations. Our results demonstrate the existence of three partly admixed phylogenetic species that diverged between the Miocene and Pliocene, and whose Quaternary distribution has been strongly driven by glacial cycles. Pseudephebe minuscula is the only species showing an amphitropical distribution, with populations in Antarctica, whereas the restricted distribution of P. pubescens and an undescribed Alaskan species might reflect the survival of these species in European and North American refugia. Our microevolutionary analyses suggest a Northern Hemisphere origin for P. minuscula, which could have dispersed into the Southern Hemisphere directly and/or through "mountain-hopping" during the Pleistocene. The Antarctic populations of this species are sorted into two genetic clusters: populations of the Antarctic Peninsula were grouped together with South American ones, and the Antarctic Continental populations formed a second cluster with Bolivian and Svalbard populations. Therefore, our data strongly suggest that the current distribution of P. minuscula in Antarctica is the outcome of multiple, recent colonizations. In conclusion, our results stress the need for integrating species delimitation and population analyses to properly approach historical biogeography in lichen-forming fungi.


Opiate antagonists and rewarding brain stimulation.
This review examines the literature on the effects of opiate antagonists on brain stimulation (ICSS) reward. Antagonists should have predictable effects if endogenous opioids modulate ICSS. Naloxone is the antagonist most often used, and it has produced inconsistent results in some ICSS paradigms. When schedules of intermittent reinforcement are used, however, naloxone reliably reduces the rate of responding. It reverses the effects of opiate agonists on ICSS behavior, and it also attenuates the effects of psychomotor stimulants, such as amphetamine. The results produced by naloxone are consistent with a modulatory effect of endogenous opioid systems on reward, and suggest that the opiate and dopamine systems together exert significant control over ICSS. Further research is needed to characterize better the actions of the antagonists on ICSS behavior, and productive research directions are proposed. Data obtained in future studies might suggest how the endogenous opioid systems modulate both natural and brain stimulation reward.


Gastric metaplasia in duodenal bulb and Campylobacter-like organisms in development of duodenal ulcer.
Multiple biopsies were taken from 50 patients with endoscopic appearance of duodenitis in order to study the relationship between gastric metaplasia in duodenum and the presence of Campylobacter-like organisms (CLOs) and the development of duodenal ulcer disease. Metaplasia was found in at least one biopsy specimen from 46 of the 50 patients; CLOs were seen in 25 of the 50 patients, only in metaplastic areas. In four cases a "transitional epithelium" with the presence of cells containing few apical mucoid granules and absorbent-type brush border as well as goblet cells was seen. In two cases, this epithelium was colonized by a small number of CLOs. Gastric heterotopia appears to be a rare condition.


Predictors of mortality from pump failure and sudden cardiac death in patients with systolic heart failure and left ventricular dyssynchrony: results of the CARE-HF trial.
Determining a specific death cause may facilitate individualized therapy in patients with heart failure (HF). Cardiac resynchronization therapy (CRT) decreased mortality in the Cardiac Resynchronization in Heart Failure trial by reducing pump failure and sudden cardiac death (SCD). This study analyzes predictors of specific causes of death. Univariate and multivariate analyses used 8 baseline and 3-month post-randomization variables to predict pump failure and SCD (categorized as "definite," "probable," and "possible"). Of 255 deaths, 197 were cardiovascular. There were 71 SCDs with a risk reduction by CRT of 0.47 (95% confidence interval 0.29-0.76; P = .002) with similar reductions in SCD classified as definite, probable, and possible. Univariate SCD predictors were 3-month HF status (mitral regurgitation [MR] severity, plasma brain natriuretic peptide [BNP], end-diastolic volume, and systolic blood pressure), whereas randomization to CRT decreased risk. Multivariate SCD predictors were randomization to CRT 0.56 (0.53-0.96, P = .035) and 3-month MR severity 1.82 (1.77-2.60, P = .0012). Univariate pump failure death predictors related to baseline HF state (quality of life score, interventricular mechanical delay, end-diastolic volume, plasma BNP, MR severity, and systolic pressure), whereas randomization to CRT and nonischemic cardiomyopathy decreased risk; multivariate predictors of pump failure death were baseline plasma BNP and systolic pressure and randomization to CRT. CRT decreased SCD in patients with systolic HF and ventricular dyssynchrony. SCD risk was increased with increased severity of MR (including the 3-month value for MR as a time-dependent covariate) and reduced by randomization to CRT. HF death was increased related to the level of systolic blood pressure, log BNP, and randomization to CRT. These results emphasize the importance and interdependence of HF severity to mortality from pump failure and SCD.


A practical review of laser-assisted hair removal using the Q-switched Nd:YAG, long-pulsed ruby, and long-pulsed alexandrite lasers.
The recent development of numerous laser-assisted hair removal techniques has produced confusion in the field of cutaneous laser surgery. A systematic review of the hair removal laser systems currently available is attempted. Appropriate patient selection, treatment protocols, and discussion of each system's advantages, disadvantages, and complications is provided. Techniques involving the use of selective Q-switched Nd:YAG and long-pulsed alexandrite and ruby laser systems provide a significant delay in hair growth. With further technologic advances and understanding of proper treatment parameters and intervals, it is expected that permanent hair removal may eventually be possible.


Comparison of gut hormones and adipokines stimulated by glucagon test among patients with type II diabetes mellitus after metabolic surgery.
Laparoscopic Roux-en-Y gastric bypass (RYGB) achieves a higher remission of type 2 diabetes mellitus (T2DM) than laparoscopic sleeve gastrectomy (SG) in non-morbidly obese patients. However, the mechanisms of the higher remission are unknown. To compare glucagon-provoked acute insulin responses, as well as changes of gut and pancreatic hormones and adipokines between patients with T2DM after RYGB and SG at one year post-operatively, a total of 14 RYGB and 13 SG patients were followed-up and evaluated for glucose metabolism, gut and pancreatic hormones, and adipokines. One year after surgery, 1-mg intravenous glucagon tests were performed. The differences in each hormone at different time points and the area under the curve (AUC) were compared between the two groups. Glucagon-stimulated acute insulin responses were not different between the RYGB and SG groups, nor were they different between the remitters and non-remitters at one year after the metabolic surgery. Plasma des-acyl ghrelin and nesfatin-1 levels significantly decreased at 6 min after glucagon stimulation in the RYGB and SG groups, as well as in the remitters and non-remitters. The glucagon test did not affect intestinal hormones. Plasma resistin was suppressed after intravenous glucagon stimulation in both RYGB and SG groups. In conclusion, intravenous glucagon inhibited plasma levels of des-acyl ghrelin, nesfatin-1, and resistin in T2DM patients at one year after both RYGB and SG, whereas post-glucagon suppression of plasma obestatin and resistin was shown in the remitters but not in the non-remitters.


Inhibitory effects of furoquinoline alkaloids from Melicope confusa and Dictamnus albus against human phosphodiesterase 5 (hPDE5A) in vitro.
Eight furoquinoline alkaloids were purified from two plants belonging to the Rutaceae family. Kokusaginine, skimmianine, evolitrine, and confusameline were purified from Melicope confusa, and haplopine, robustine, dictamine, and gamma-fagarine from Dictamnus albus. In this study, the eight furoquinoline alkaloids were examined for inhibitory potency against human phosphodiesterase 5 (hPDE5A) in vitro. DNA encoding the catalytic domain of human PDE5A was amplified from the mRNA of T24 cells by RT-PCR and was fused to GST in an expression vector. GST-tagged PDE5A was then purified by glutathione affinity chromatography and used in inhibition assays. Of the eight alkaloids, gamma-fagarine was the most potent inhibitor of PDE5A, and its single methoxy group at the C-8 position was shown to be critical for inhibitory activity. These results clearly illustrate the relationship between PDE5A inhibition and the methoxy group position in furoquinoline alkaloids.


Impact of COVID-19 on the sleep-wake patterns of preschool children.
To explore the impact of the COVID-19 pandemic on the sleep-wake patterns of preschool children. A cohort of preschoolers established before the COVID-19 pandemic was invited to participate in this study. Data including children's demographics, their own and parental sleep-wake patterns, physical activities, and screen time were collected through an online questionnaire from August to September 2020. A comparison was made on the collected data from the same cohort of children before and during the pandemic. The cohort which was established before the pandemic consisted of 3720 preschoolers. For this current study, 642 (17%) participated, and 497 (13%) children who fulfilled the eligibility criteria were included in the final analysis. They showed a delay in their bedtime and wake time on both weekdays and weekends with a 15-30 min increase in nocturnal sleep duration. However, with a reduction in nap time, the average daily sleep duration was shortened by 16.3 ± 64.3 min (p < 0.001) and 27.5 ± 72.9 min (p < 0.001) during weekdays and weekends, respectively. Screen time was increased while outdoor activity duration was decreased. Parental sleep/wake times were also delayed with an increase in sleep duration. Children's sleep habits were associated with screen time and parental sleep/wake patterns. Despite school suspension during the COVID-19 pandemic, preschoolers were not sleeping longer. Screen time and parental sleep/wake patterns were the major factors driving the preschoolers' sleep habits. Health education is required to control screen time in children and to promote sleep hygiene among all family members.


Health Burden and Service Utilization in Texas Medicaid Deliveries from the Prenatal Period to 1 Year Postpartum.
To describe health burden and health service utilization from the prenatal period to 1 year postpartum among women with births covered by Texas Medicaid, focusing on the major contributors to maternal mortality after 60 days postpartum in Texas. We analyzed diagnoses and health service utilization during the prenatal, early postpartum (5-60 days postpartum), and late postpartum (> 60 days to 1 year postpartum) periods, using administrative medical claims data for women ages 18-44 years with a Medicaid-paid delivery in 2017 residing in selected regions in Texas (n = 49,302). Overall, 12.6% and 17.5% of women had diagnoses of cardiovascular/coronary conditions and substance use disorder, respectively. Mental health conditions affected 30% of women, with anxiety (47.1%) and depression (34.3%) accounting for the greatest proportion of diagnosed mental health conditions. The prevalence of these conditions was higher during the late (19.4%) versus early (9.9%) postpartum period. About 47.8% of women had other chronic health conditions, including obesity, diabetes mellitus, and hypertension. Among women with the selected health conditions, utilization of any health services was higher during the prenatal period compared to early and late postpartum periods (e.g., any mental health service utilization: prenatal period (57.4%) versus early postpartum (26.9%) and late postpartum (25.5%) periods). However, among women with the selected health conditions, there was a high utilization of emergency room services during the late postpartum period [e.g., emergency room service utilization among those with mental health conditions: prenatal period (35.6%); postpartum period: early (5.5%) and late (30.1%)]. Increasing access to the full range of recommended services during the prenatal period through 1 year postpartum has potential to help improve vulnerable women's birth outcomes.


Role of the harderian gland in immunoglobulin A production in chicken lacrimal fluid.
The role of the Harderian gland (HG) in the production of immunoglobulin, especially IgA, was investigated. Lacrimal immunoglobulin almost disappeared after surgical removal of the HG and immunoglobulin produced by HG cells was detected in saliva but not in the trachea. Immunoglobulin production occurred in HG cell culture in vitro and it consisted mostly of IgA; the production of IgM and IgG was very low. These findings indicate that lacrimal IgA is produced locally in the HG and lacrimal IgG and IgM are mostly transported from the blood.


Accuracy of suture techniques of surgeons with different surgical experience.
The pinpoint accuracy in the suturing techniques of 35 surgeons was measured by the distance deviated from an expected needle exit. The grip of the needle holder used by the surgeons was their habitual one. Accuracy was tested using two ways of suturing; namely with, and without wavering a needle to aim at an exit after suturing began. Where the surgeons did not waver the needle, accuracy improved both when the surgical experience was greater than one year (p less than 0.05) and again when the experience exceeded 10-15 years (not significant). Difference of the accuracy between individual surgeons disappeared in those surgeons with 15 to 20 years experience. Where the surgeons wavered the needle, improvement was more marked in the former period and less in the latter. Wavering improved the accuracy in each group (p less than 0.01), but with the possible danger of injuring tissue. In conclusion, accuracy was not as precise as most surgeons' confidence believed. A significant improvement was observed with 1-year experience, but the improvement thereafter was poor. Fifteen to 20 years experience was not enough to acquire a suturing technique not necessitating needle wavering in the tissue, still yielding a 1.7 mm deviation in a 2 cm suture-stride. It was however enough to eliminate surgeons' individual differences in accuracy.


Transcription factor Nrf2 protects the brain from damage produced by intracerebral hemorrhage.
Intracerebral hemorrhage (ICH) remains a major medical problem for which there is no effective treatment. Oxidative and cytotoxic damage plays an important role in ICH pathogenesis and may represent a target for treatment of ICH. Recent studies have suggested that nuclear factor-erythroid 2-related factor 2 (Nrf2), a pleiotropic transcription factor, may play a key role in protecting cells from cytotoxic/oxidative damage. This study evaluated the role of Nrf2 in protecting the brain from ICH-mediated damage. Sprague-Dawley rats and Nrf2-deficient or control mice received intracerebral injection of autologous blood to mimic ICH. Sulforaphane was used to activate Nrf2. Oxidative stress, the presence of myeloperoxidase-positive cells (neutrophils) in ICH-affected brains, and behavioral dysfunction were assessed to determine the extent of ICH-mediated damage. Sulforaphane activated Nrf2 in ICH-affected brain tissue and reduced neutrophil count, oxidative damage, and behavioral deficits caused by ICH. Nrf2-deficient mice demonstrated more severe neurologic deficits after ICH and did not benefit from the protective effect of sulforaphane. Nrf2 may represent a strategic target for ICH therapies.


Reduced inhibition underlies early life LPS exposure induced-cognitive impairment: Prevention by environmental enrichment.
Early life immune activation has negative effects on the development of central nervous system and cognitive function, yet the underlying mechanism remains unclear. Increasing evidence has demonstrated that inflammation induces changes in microglia morphology, which lead to excessive synaptic pruning and improper function of neural circuits. Therefore, we hypothesized that early immune activation induced microglia activation, contributing to synaptic and cognitive impairments in adolescent mice. To establish the animal model of early immune activation, pups received a single intraperitoneal injection of 100 μg/kg lipopolysaccharide (LPS) on postnatal 10 (P10). Environmental enrichment (EE) was conducted four hours per day during P10-P38. Behavioral tests were performed by open field (P39), elevated plus-maze (P40) and Y maze tests (P41). The protein levels of glutamic acid decarboxylas67 (GAD67), parvalbumin (PV), vesicular gaba amino acid transporter (vGAT) and vesicular glutamate transporters (vGLUT1) were determined in the hippocampi and medial prefrontal cortex (mPFC). The protein levels of nuclear factor κB (NF-κB)/p65, NF-κB/p50, interleukin-1β (IL-1β), tumor necrosis factor - ɑ (TNF-ɑ) were determined in the hippocampi. The dendritic spine density was evaluated in the CA1 of the hippocampus. In our study, we showed that early life LPS exposure induced microglia activation and excessive inhibitory synapse engulfment, decreased number of perisomatic puncta on both inhibitory PV interneurons and excitatory neurons, which might contribute to excitation/inhibition imbalance, dendritic spine loss, and cognitive impairment in adolescent mice. Notably, EE rescued most of these abnormalities and improved cognitive impairment. In conclusion, our study demonstrated that reduced inhibition might contribute to early life LPS exposure induced-cognitive impairment. We also provided the possibility of the protective role of EE in rescuing these long-term adverse effects.


Cytometric and electron microscopic studies of the direct interaction of divalent nickel with intact and chemically modified HuT-78 lymphoblasts.
Cytometric and ultrastructural studies on 24 hr cultures of intact, 1.0 mM H5IO6, and 0.1 mM SeO2-oxidized HuT-78 lymphoblasts were performed after their direct, 30 min interaction with 1.0 mM NiCl2. Except for moderately depressed cell viability, divalent nickel did not alter the progression of intact and oxidized target cells through the phases of the cell cycle. Although the plasma membrane remained structurally intact, marked distortion of mitochondria structure and increased osmiophilia were an invariable attribute of all nickel-pulsed cells. Moreover, numerous electron-opaque, intracellular depositions were detected in SeO2-oxidized, nickel-pulsed cells. It is concluded that the initial state of plasma membrane, and the interaction of nickel with other trace elements, have jointly determined the response of HuT-78 cells to brief and direct, divalent nickel pulses.


Antileukemic activity against L1210 leukemia, pharmacokinetics and hematological side effects of 5-hydroxymethyl-2'-deoxyuridine.
The chemotherapeutic potential of 5-hydroxymethyl-2'-deoxyuridine (5HmdUrd) was examined in vitro and in vivo. The compound was toxic in 2-day cultures; 7, 66 and 88% inhibition in the growth of L1210 cells was achieved with 1, 10 and 100 microM 5HmdUrd, respectively. The maximal plasma concentration of 5HmdUrd at 15 min after a single i.p. injection (100 mg/kg) in DBA/2 mice was 193-244 mumol./l and the compound had a logarithmic disappearance curve with a half-life of 20 min. Chemotherapy given as two daily i.p. injections of 5HmdUrd (100 mg/kg) for five successive days resulted in a 239% increase in median lifespan and 2/6 long-term survivals among DBA/2 mice bearing leukemia L1210. This treatment resulted in temporary neutropenia and thrombocytopenia, which were followed by rebound thrombocytosis and neutrophilia of short duration. Our data indicate that 5HmdUrd can successfully be used in experimental cancer chemotherapy in vivo.


Temporally resolved SMLM (with large PAR shift) enabled visualization of dynamic HA cluster formation and migration in a live cell.
The blinking properties of a single molecule are critical for single-molecule localization microscopy (SMLM). Typically, SMLM techniques involve recording several frames of diffraction-limited bright spots of single-molecules with a detector exposure time close to the blinking period. This sets a limit on the temporal resolution of SMLM to a few tens of milliseconds. Realizing that a substantial fraction of single molecules emit photons for time scales much shorter than the average blinking period, we propose accelerating data collection to capture these fast emitters. Here, we put forward a short exposure-based SMLM (shortSMLM) method powered by sCMOS detector for understanding dynamical events (both at single molecule and ensemble level). The technique is demonstrated on an Influenza-A disease model, where NIH3T3 cells (both fixed and live cells) were transfected by Dendra2-HA plasmid DNA. Analysis shows a 2.76-fold improvement in the temporal resolution that comes with a sacrifice in spatial resolution, and a particle resolution shift PAR-shift (in terms of localization precision) of [Formula: see text] 11.82 nm compared to standard SMLM. We visualized dynamic HA cluster formation in transfected cells post 24 h of DNA transfection. It is noted that a reduction in spatial resolution does not substantially alter cluster characteristics (cluster density, [Formula: see text] molecules/cluster, cluster spread, etc.) and, indeed, preserves critical features. Moreover, the time-lapse imaging reveals the dynamic formation and migration of Hemagglutinin (HA) clusters in a live cell. This suggests that [Formula: see text] using a synchronized high QE sCMOS detector (operated at short exposure times) is excellent for studying temporal dynamics in cellular system.


Prehypertension and chronic kidney disease: the ox or the plow?
Nearly ten years ago, practice recommendations supported use of the clinical classification of 'prehypertension' for people with systolic blood pressure of 120-139 mm Hg or diastolic pressure of 80-89 mm Hg. This recommendation was based on observations that these ranges of blood pressure were associated with enhanced cardiovascular and cerebrovascular risks compared with blood pressure less than 120/80 mm Hg. Recent observations, including the report by Yano and colleagues, also suggest that prehypertension is an important risk factor for the development of chronic kidney disease.


Acquired total immunoglobulin G deficiency in a patient with primary biliary cirrhosis.
A 38-year-old Japanese woman was hospitalized for susceptibility to respiratory tract infections. Clinical examinations revealed asymptomatic primary cholestasis, abnormally elevated immunoglobulin M (IgM) and antimitochondrial antibody, being consistent with asymptomatic primary biliary cirrhosis. Three years later her serum immunoglobulin G (IgG) decreased remarkably, whereas other immunoglobulins were unchanged. Immunological examinations on the peripheral blood lymphocytes demonstrated spontaneous over-synthesis of serum IgM and decreased synthesis of IgG due to abnormal function of both T and B cells. Our case suggests a new possible association between primary biliary cirrhosis and IgG deficiency.


Association of Umbilical Cord Milking vs Delayed Umbilical Cord Clamping With Death or Severe Intraventricular Hemorrhage Among Preterm Infants.
Umbilical cord milking as an alternative to delayed umbilical cord clamping may provide equivalent benefits to preterm infants, but without delaying resuscitation. To determine whether the rates of death or severe intraventricular hemorrhage differ among preterm infants receiving placental transfusion with umbilical cord milking vs delayed umbilical cord clamping. Noninferiority randomized clinical trial of preterm infants (born at 23-31 weeks' gestation) from 9 university and private medical centers in 4 countries were recruited and enrolled between June 2017 and September 2018. Planned enrollment was 750 per group. However, a safety signal comprising an imbalance in the number of severe intraventricular hemorrhage events by study group was observed at the first interim analysis; enrollment was stopped based on recommendations from the data and safety monitoring board. The planned noninferiority analysis could not be conducted and a post hoc comparison was performed instead. Final date of follow-up was December 2018. Participants were randomized to umbilical cord milking (n = 236) or delayed umbilical cord clamping (n = 238). The primary outcome was a composite of death or severe intraventricular hemorrhage to determine noninferiority of umbilical cord milking with a 1% noninferiority margin. Among 540 infants randomized, 474 (88%) were enrolled and completed the trial (mean gestational age of 28 weeks; 46% female). Twelve percent (29/236) of the umbilical cord milking group died or developed severe intraventricular hemorrhage compared with 8% (20/238) of the delayed umbilical cord clamping group (risk difference, 4% [95% CI, -2% to 9%]; P = .16). Although there was no statistically significant difference in death, severe intraventricular hemorrhage was statistically significantly higher in the umbilical cord milking group than in the delayed umbilical cord clamping group (8% [20/236] vs 3% [8/238], respectively; risk difference, 5% [95% CI, 1% to 9%]; P = .02). The test for interaction between gestational age strata and treatment group was significant for severe intraventricular hemorrhage only (P = .003); among infants born at 23 to 27 weeks' gestation, severe intraventricular hemorrhage was statistically significantly higher with umbilical cord milking than with delayed umbilical cord clamping (22% [20/93] vs 6% [5/89], respectively; risk difference, 16% [95% CI, 6% to 26%]; P = .002). In this post hoc analysis of a prematurely terminated randomized clinical trial of umbilical cord milking vs delayed umbilical cord clamping among preterm infants born at less than 32 weeks' gestation, there was no statistically significant difference in the rate of a composite outcome of death or severe intraventricular hemorrhage, but there was a statistically significantly higher rate of severe intraventricular hemorrhage in the umbilical cord milking group. The early study termination and resulting post hoc nature of the analyses preclude definitive conclusions. ClinicalTrials.gov Identifier: NCT03019367.


Public values and guiding principles for implementing epitope compatibility in kidney transplantation allocation criteria: results from a Canadian online public deliberation.
Epitope compatibility in deceased donor kidney allocation is an emerging area of precision medicine (PM), seeking to improve compatibility between donor kidneys to transplant candidates in the hope of avoiding kidney rejection. Though the potential benefits of using epitope compatibility are promising, the implied modification of deceased organ allocation criteria requires consideration of significant clinical and ethical trade-offs. As a matter of public policy, these trade-offs should consider public values and preferences. We invited members of the Canadian public to participate in a deliberation about epitope compatibility in deceased donor kidney transplantation; to identify what is important to them and to provide recommendations to policymakers. An online public deliberation was conducted with members of the Canadian public, in which participants were asked to construct recommendations for policymakers regarding the introduction of epitope compatibility to kidney allocation criteria. In the present paper, a qualitative analysis was conducted to identify the values reflected in participants' recommendations. All virtual sessions were recorded, transcribed, and analyzed using NVivo 12 software. Thirty-two participants constructed nine recommendations regarding the adoption of epitope compatibility into deceased donor kidney allocation. Five values were identified that drove participants' recommendations: Health Maximization, Protection/Mitigation of Negative Impacts, Fairness, Science/Evidence-based Healthcare, and Responsibility to Maintain Trust. Conflicts between these values were discussed in terms of operational principles that were required for epitope compatibility to be implemented in an acceptable manner: the needs for Flexibility, Accountability, Transparent Communication and a Transition Plan. All nine recommendations were informed by these four principles. Participant deliberations were often dominated by the conflict between Health Maximization and Fairness or Protection/Mitigation of Negative Impacts, which was discussed as the need for Flexibility. Two additional values (Efficient Use of Resources and Logic/Rationality) were also discussed and were reasons for some participants voting against some recommendations. Public recommendations indicate support for using epitope compatibility in deceased donor kidney allocation. A flexible approach to organ allocation decision-making may allow for the balancing of Health Maximization against maintaining Fairness and Mitigating Negative Impacts. Flexibility is particularly important in the context of epitope compatibility and other PM initiatives where evidence is still emerging.


Palliative care in advanced Huntington's disease: a scoping review.
As Huntington's disease (HD) is a progressive disease for which there is no cure yet, patients in the advanced stage of HD may benefit from palliative care. To review the literature focusing on palliative care in advanced stage HD, and the level of evidence. Publications between 1993 and October 29th, 2021 from 8 databases (Embase, Web of Science, Cochrane, Emcare, PsycINFO, Academic Search Premier, PMC PubMed Central and Pubmed) were included. The literature was deductively classified based on topics that are part of the definition of palliative care, or as care-related topics that emerged from the literature. Levels of evidence I (high) - V (low) were determined as defined by the Joanna Briggs Institute. Our search resulted in 333 articles, 38 of which were included. The literature covered four domains of palliative care: physical care, psychological care, spiritual care, and social care. Four other topics in the literature were: advance care planning, end-of-life needs assessments, pediatric HD care, and need for health care services. Most literature was underpinned by a low level of evidence, except for the topics on social care (Level III-V), advance care planning (Level II-V) and end-of-life needs assessments (Level II-III). To deliver adequate palliative care in advanced HD, both general and HD-specific symptoms and problems need to be addressed. As the level of evidence in existing literature is low, further research is essential to improve palliative care and to meet patient's wishes and needs.


Dietary regulation of dipalmitoyl phosphatidylcholine in the lung. Effects of essential fatty acid deficiency.
An essential fatty acid deficiency resulted in a significant decrease of saturated phosphatidylcholine in rat lung tissue, with the reduction being mainly due to that of dipalmitoyl species. This decrease was almost completely reversed by administration of a diet containing linoleate from seven days. The features of saturated phosphatidylcholine synthesis in the deficient rat lung slices were studied with labeled precursors in the presence of different concentrations of linoleate and oleate in the medium. Labeling patterns of saturated phosphatidylcholine using [2-3H]glycerol and [Me-14C]choline were unaffected in all ratios of linoleate to oleate examined. The incorporation rates of both [3H]acetate and [1-14C]palmitate into saturated phosphatidylcholine decreased in the deficient state, but that of the palmitate label increased markedly with increase in the concentration of linoleate in the medium. This stimulatory effect due to the increased linoleate concentration, however, was not found with the acetate label. While the utilization of 1-[1-14C]palmitoyl glycerophosphocholine by lung tissue was enhanced in the deficient state, in the presence of linoleate it showed almost the same level as that of the controls. The essential fatty acid deficiency resulted in an increase in the activity of liver choline kinase by 46%. However, other enzyme activities involved in phosphatidylcholine synthesis in the lung and liver were unaffected by the deficient state.


The vasoconstrictor activities of prostaglandin D2 via the thromboxane prostanoid receptor and E prostanoid receptor-3 outweigh its concurrent vasodepressor effect mainly through D prostanoid receptor-1 ex vivo and in vivo.
Prostaglandin (PG) D2, a commonly considered vasodilator through D prostanoid receptor-1 (DP1), might also evoke vasoconstriction via acting on the thromboxane (Tx)-prostanoid receptor (the original receptor of TxA2; TP) and/or E prostanoid receptor-3 (one of the vasoconstrictor receptors of PGE2; EP3). This study aimed to test the above hypothesis in the mouse renal vascular bed (main renal arteries and perfused kidneys) and/or mesenteric resistance arteries and determine how the vasoconstrictor mechanism influences the overall PGD2 effect on systemic blood pressure under in vivo conditions. Experiments were performed on control wild-type (WT) mice and mice with deficiencies in TP (TP-/-) and/or EP3 (EP3-/-). Here we show that PGD2 indeed evoked vasoconstrictor responses in the above-mentioned tissues of WT mice, which were however not only reduced by TP-/- or EP3-/-, but also reversed by TP-/-/EP3-/- in some of the above tissues (mesenteric resistance arteries or perfused kidneys) to dilator reactions that were reduced by non-selective DP antagonism. A slight or mild pressor response was also observed with PGD2 under in vivo conditions, and this was again reversed to a depressor response in TP-/- or TP-/-/EP3-/- mice. Non-selective DP antagonism reduced the PGD2-evoked depressor response in TP-/-/EP3-/- mice as well. These results thus demonstrate that like other PGs, PGD2 activates TP and/or EP3 to evoke vasoconstrictor activities, which can outweigh its concurrent vasodepressor activity mediated mainly through DP1, and hence result in a pressor response, although the response might only be of a slight or mild extent.


Multiple acyl-CoA dehydrogenase deficiency occurring in pregnancy and caused by a defect in riboflavin metabolism in the mother. Study of a kindred with seven deaths in infancy: Value of riboflavin therapy in preventing this syndrome.
Seven infants in one kindred died: one was stillborn; the others, who were floppy at birth and were breast-fed, developed a disorder with the odor of sweaty feet and died in early infancy. In two further pregnancies, 3-hydroxvisovaleric, glutaric, and C6-C10-dicarboxylic acids were demonstrated in the mother's urine during the seventh month. Riboflavin therapy in the last trimester of pregnancy and a riboflavin-rich diet given the infants prevented this syndrome. The presence of abnormal erythrocyte glutathione-reductase activity in the mother while she excreted normal amounts of riboflavin in her urine indicates a probable disorder of riboflavin metabolism resulting in multiple acyl-CoA dehydrogenase deficiency.


A Generalization of the Concavity of Rényi Entropy Power.
Recently, Savaré-Toscani proved that the Rényi entropy power of general probability densities solving the p-nonlinear heat equation in Rn is a concave function of time under certain conditions of three parameters n,p,μ, which extends Costa's concavity inequality for Shannon's entropy power to the Rényi entropy power. In this paper, we give a condition Φ(n,p,μ) of n,p,μ under which the concavity of the Rényi entropy power is valid. The condition Φ(n,p,μ) contains Savaré-Toscani's condition as a special case and much more cases. Precisely, the points (n,p,μ) satisfying Savaré-Toscani's condition consist of a two-dimensional subset of R3, and the points satisfying the condition Φ(n,p,μ) consist a three-dimensional subset of R3. Furthermore, Φ(n,p,μ) gives the necessary and sufficient condition in a certain sense. Finally, the conditions are obtained with a systematic approach.


MRI of peroneal nerve entrapment due to a ganglion cyst.
Ganglion cysts are relatively common entities, but impingement upon the peripheral nerves is rare. We describe a case in which peroneal nerve palsy was caused by a ganglion cyst demonstrated by MRI.


Loss of Paip1 causes translation reduction and induces apoptotic cell death through ISR activation and Xrp1.
Regulation of protein translation initiation is tightly associated with cell growth and survival. Here, we identify Paip1, the Drosophila homolog of the translation initiation factor PAIP1, and analyze its role during development. Through genetic analysis, we find that loss of Paip1 causes reduced protein translation and pupal lethality. Furthermore, tissue specific knockdown of Paip1 results in apoptotic cell death in the wing imaginal disc. Paip1 depletion leads to increased proteotoxic stress and activation of the integrated stress response (ISR) pathway. Mechanistically, we show that loss of Paip1 promotes phosphorylation of eIF2α via the kinase PERK, leading to apoptotic cell death. Moreover, Paip1 depletion upregulates the transcription factor gene Xrp1, which contributes to apoptotic cell death and eIF2α phosphorylation. We further show that loss of Paip1 leads to an increase in Xrp1 translation mediated by its 5'UTR. These findings uncover a novel mechanism that links translation impairment to tissue homeostasis and establish a role of ISR activation and Xrp1 in promoting cell death.


First molecular identification of Buxtonella ciliates from captive-bred mangabeys (Cercocebus torquatus) from China.
Buxtonella species are large cyst-forming ciliates that infect ruminants and monkeys, and are morphologically similar to Balantidium coli ciliates that infect pigs, humans, monkeys, and other animals. In this study, we isolated spherical cysts of ciliates that were similar to those of Balantidium and Buxtonella species within collared mangabeys (Cercocebus torquatus) from the Wangcheng Zoo of Luoyang in the Henan Province of central China. The cysts were further identified and designated as belonging to the Buxtonella monkey genotype based on molecular analyses of 18S rRNA, 5.8S rRNA, and ITS1-5.8S rRNA-ITS2 genetic markers. To our knowledge, this is the first report of Buxtonella monkey genotype within monkeys in China. These results will help clarify the classification of species of cyst-forming ciliate infections in monkeys.


Positron emission tomography study of regional cerebral blood flow and flow-metabolism coupling during general anaesthesia with xenon in humans.
The effects of xenon on regional cerebral blood flow (rCBF) are controversial. Moreover, the precise sites of action at which xenon exerts its effects in the human brain remain to be established. rCBF was sequentially assessed by H(2)(15)O positron emission tomography in six volunteers. rCBF was determined at baseline and during general anaesthesia induced with propofol and maintained with one minimum alveolar concentration xenon. rCBF measurements were started after the calculated plasma concentration of propofol had decreased to subanaesthetic levels (<1.0 microg ml(-1)). Changes in rCBF were calculated for 13 cerebral volumes of interest by measurement of a semi-quantitative perfusion index (PI). In addition, voxel-wise changes in rCBF were analysed using statistical parametric mapping. Xenon had only minor effects on PI in grey matter volumes of interest. In contrast, PI was increased in white matter [from 1.01 (0.11) to 1.24 (0.15) kcnt ml(-1) MBq(-1), P=0.05, mean (SD)]. Voxel-based analysis showed an increase of rCBF in white matter and a relative decrease of rCBF during xenon anaesthesia in distinct grey matter regions, particularly the orbito- and mesiofrontal cortex, cingulate gyrus, thalamus, hippocampus and bilateral cerebellum (P<0.05 corrected). When correlating PI with cerebral metabolic rate of glucose (previously obtained in another group of six volunteers using (18)F-fluorodeoxyglucose as tracer), the flow-metabolism coupling was preserved during xenon anaesthesia. Xenon exerted distinct regional effects on CBF: relative decreases in several cortical, subcortical, and cerebellar areas were accompanied by an increase in white matter. Flow-metabolism coupling was not impaired during xenon anaesthesia.


Weight Gain, Overweight, and Obesity: Determinants and Health Outcomes from the Australian Longitudinal Study on Women's Health.
Recent estimates suggest that 35.3 % of adult Australians are overweight and a further 27.5 % are obese. The Australian Longitudinal Study on Women's Health (ALSWH) is a prospective study of women's health that commenced in Australia in 1996. The study recruited approximately 40,000 women in three birth cohorts, 1973-1978, 1946-1951 and 1921-1926, who have since been followed up approximately every three years using self-report surveys. Six surveys have been completed to date. This review aims to describe the changes in weight and weight status over time in the three ALSWH cohorts, and to review and summarise the published findings to date relating to the determinants and health consequences of weight gain, overweight and obesity. Future plans for the ALSWH include on-going surveys for all cohorts, with a seventh survey in 2013-2015, and establishment of a new cohort of women born in 1990-1995, which is currently being recruited.


Electrophoresis techniques to investigate defects in oxidative phosphorylation.
Defects in mitochondrial oxidative phosphorylation (OXPHOS) are a frequent cause of severe inherited metabolic disorders and also contribute to aging. The OXPHOS system constitutes five multi-subunit complexes embedded in the mitochondrial inner membrane. Correct function of this system requires proper assembly of the approximately 80 proteins in the complexes, as well as numerous assembly factors. Blue native electrophoresis has become a crucial tool to investigate OXPHOS-related defects in mitochondrial disease patients. In addition, OXPHOS-assembly profiles can be obtained by two dimensional blue native/SDS gel electrophoresis, which provides additional information for identifying disease-causing mutations and insight in the role of specific proteins in the biogenesis of the OXPHOS system. Here we provide a practical guide on how to set-up the basic technique to study OXPHOS defects in patient-derived cells and tissues.


Protein transduction technology.
Intracellular delivery of macromolecules remains problematic because of the bioavailability restriction imposed by the cell membrane. Recent studies on protein transduction domains have circumvented this barrier, however, and have resulted in the delivery of peptides, full-length proteins, iron beads, liposomes, and radioactive isotopes into cells in culture and animal models in vivo.


Damage sensitivity of dental zirconias to simulated occlusal contact.
Mechanical damages can occur from dental restoration processing and fitting, or while it is in-service. This study evaluates the damage sensitivity of translucent zirconia (5Y-PSZ) relative to conventional 3Y-PSZ following mouth-motion simulations at various loads. 5Y-PSZ and 3Y-PSZ discs were adhesively bonded to a dentin-like substrate and divided into groups according to the load (50 N or 200 N) and number of cycles (up to 106) used in the chewing simulation. Specimens were mounted with 30° inclination in an electrodynamic mouth-motion simulator, and subjected to contact-slide-liftoff cyclic loading in water. Surface and sub-surface damages were analyzed using a sectioning technique. After the simulation, specimens were removed from the substrate and loaded with the damaged surface in tension for biaxial strength testing to assess their damage tolerance. The strength of both ceramics underwent significant degradation after mouth-motion simulations. For 5Y-PSZ, the strength degradation was greater (∼60%) and occurred at a lower number of cycles than 3Y-PSZ. Herringbone cracks emerged on 3Y-PSZ and 5Y-PSZ surfaces under a 200-N load after 50 and 10 cycles, respectively. Meanwhile at a 50-N load, cracks formed at ∼1000 cycles in both ceramics. Further increasing the number of cycles only had moderate effects on the strength of both ceramics, despite an increase in surface and sub-surface damage. More significantly, a 50-N occlusal load can debase the zirconia strengths as much as a 200-N load. Surface flaws produced during the chewing simulation are capable of significant strength degradation in zirconia, even after a small number of low-load cycles.


Nonstimulant Medications for Attention-Deficit/Hyperactivity Disorder (ADHD) in Adults: Systematic Review and Meta-analysis.
For some adults with Attention-Deficit/Hyperactivity Disorder (ADHD), nonstimulants need to be considered either as a monotherapy or as an adjunct to stimulants. The objectives of this systematic review and meta-analysis were to assess the efficacy, acceptability, and tolerability of nonstimulants in adults with ADHD. Data sources, searches, and study selection were based on a previously published network meta-analysis of randomized clinical trials (RCTs) by Cortese at al. (Lancet Psychiatry 5(9):727-738, 2018), which we updated in March 2022. Specifically, we searched PubMed, BIOSIS Previews, CINAHL, the Cochrane Central Register of Controlled Trials, EMBASE, ERIC, MEDLINE, PsycINFO, OpenGrey, Web of Science Core Collection, ProQuest Dissertations and Theses (UK and Ireland), ProQuest Dissertations and Theses (abstracts and international), and the WHO International Trials Registry Platform, including ClinicalTrials.gov for double-blind RCTs with a placebo arm, lasting at least one week, including adults with a diagnosis of ADHD based on DSM-III, DSM-III-R, DSM-IV(TR), DSM-5 or ICD-9- or 10, and reporting data on efficacy, tolerability (drop-out due to side effects) and acceptability (drop-out due to any cause) of guanfacine, clonidine, or atomoxetine. Additionally, we searched for RCTs of viloxazine extended release (ER), approved for ADHD in 2021. Random-effects meta-analyses were conducted, and the risk of bias for individual RCTs was assessed using the Cochrane Risk of Bias tool. We included 18 studies in the meta-analyses (4308 participants) plus one additional study in the narrative synthesis (374 participants). The meta-analysis showed that atomoxetine (15 RCTs) (Hedge's g = - 0.48, 95% CI [- 0.64; - 0.33]), guanfacine (two RCTs) (Hedge's g = - 0.66, 95% CI [- 0.94; - 0.38]) and viloxazine ER (one RCT) were significantly more efficacious than placebo. Atomoxetine was less well tolerated than placebo, while tolerability of guanfacine and viloxazine ER could not be meta-analysed, since only one study, for each medication, reported on it. All investigated nonstimulants were more efficacious in the treatment of ADHD in adults, than placebo, while the placebo had better acceptability and tolerability. https://osf.io/5vnmt/?view_only=2bf87ed12ba94645babedceeee4c0120 .


How much physical activity do people with schizophrenia engage in? A systematic review, comparative meta-analysis and meta-regression.
Physical activity (PA) improves health outcomes in people with schizophrenia. It is unclear how much PA people with schizophrenia undertake and what influences PA participation. We conducted a meta-analysis to investigate PA levels and predictors in people with schizophrenia. Major databases were searched from inception till 02/2016 for articles measuring PA (self-report questionnaire (SRQ) or objective measure (e.g. accelerometer)) in people with schizophrenia, including first episode psychosis (FEP). A random effects meta-analysis and meta-regression analysis were conducted. 35 studies representing 3453 individuals with schizophrenia (40.0years; 64.0% male) were included. Engagement in light PA was 80.44min (95% CI 68.32-92.52, n=2658), 47.1min moderate-vigorous PA (95% CI 31.5-62.8, n=559) and 1.05min (95% CI 0.48-1.62, n=2533) vigorous PA per day. People with schizophrenia engaged in significantly less moderate (hedges g=-0.45, 95% CI -0.79 to -0.1, p=0.01) and vigorous PA (g=-0.4, 95% CI -0.60 to -0.18) versus controls. Higher light to moderate, but lower vigorous PA levels were observed in outpatients and in studies utilizing objective measures versus SRQ. 56.6% (95% CI 45.8-66.8, studies=12) met the recommended 150min of moderate physical activity per week. Depressive symptoms and older age were associated with less vigorous PA in meta-regression analyses. Our data confirm that people with schizophrenia engage in significantly less moderate and vigorous PA versus controls. Interventions aiming to increase PA, regardless of intensity are indicated for people with schizophrenia, while specifically increasing moderate-vigorous PA should be a priority given the established health benefits.


The Role of Immersion for Improving Extended Reality Analysis of Personalized Flow Simulations.
Computational models of flow in patient-derived arterial geometries have become a key paradigm of biomedical research. These fluid models are often challenging to visualize due to high spatial heterogeneity and visual complexity. Virtual immersive environments can offer advantageous visualization of spatially heterogeneous and complex systems. However, as different VR devices offer varying levels of immersion, there remains a crucial lack of understanding regarding what level of immersion is best suited for interactions with patient-specific flow models. We conducted a quantitative user evaluation with multiple VR devices testing an important use of hemodynamic simulations-analysis of surface parameters within complex patient-specific geometries. This task was compared for the semi-immersive zSpace 3D monitor and the fully immersive HTC Vive system. The semi-immersive device was more accurate than the fully immersive device. The two devices showed similar results for task duration and performance (accuracy/duration). The accuracy of the semi-immersive device was also higher for arterial geometries of greater complexity and branching. This assessment demonstrates that the level of immersion plays a significant role in the accuracy of assessing arterial flow models. We found that the semi-immersive VR device was a generally optimal choice for arterial visualization.


C3 polymorphism in a Danish cystic fibrosis population and its possible association with antibody response.
The C3 types of human serum are reported for a material of 113 Danish cystic fibrosis patients, age 0-30 years. The frequency of the C3F gene was 0.2832 which was significantly higher (p less than 0.0005) than the frequency found in a control group of 224 healthy babies (C3F = 0.1585). It also differed significantly (p less than 0.01) from the C3F gene frequency of 0.1780 found in 177 blood donors, age 20-24 years. A significant association between any of the C3 phenotypes and the most serious infection in cystic fibrosis, chronic mucoid P. aeruginosa infection, or the antibody response against these bacteria was not found.


Accuracy and outcomes of virtual surgical planning and 3D-printed guides for osseous free flap reconstruction of mandibular osteoradionecrosis.
Virtual surgical planning, 3D-printed osteotomy guides and preoperatively-bent or custom-milled mandibular reconstruction plate (VSP/3Dprinted-guide/plate) have been shown to ease intraoperative decision making and reduce operative time. Few studies have examined outcomes of VSP/3Dprinted-guide/plate specifically for mandibular osteoradionecrosis (mORN) cases, which pose unique challenges. We aimed to examine reconstruction accuracy, functional outcomes, and postoperative complications following osseous-free-flap reconstruction with VSP/3Dprinted-guide/plate for mORN. Single academic medical center retrospective case series of ORN-related osseous-free-flap mandibular reconstructions with VSP/3Dprinted-guide/plate between January 2015 and March 2021. Most cases were performed by the same two-surgeon team. Outcomes include reconstruction accuracy (assessed by 3D-overlay computer models with cephalometric and donor-bone segment length measurements), complications and function. Twenty-six cases were identified with a mean follow-up of 85 weeks. Most patients were male (69 %); mean age was 64 years. 3D-model-overlay demonstrated minimal deviation between planned and actual reconstruction among 18 evaluable cases: intercondylar distance = 1.46 mm (SD 2.4); intergonial distance = 1.82 mm (SD 2.0); anterior-posterior distance = 2.14 mm (SD 1.9); gonial angle = 3.33 degrees (SD 2.4). Mean change donor-bone segment length inferiorly 4.39 mm (SD 4.3) and superiorly 3.43 mm (SD 4.0). returned to operating room (N = 2), minor primary/neck site infection/dehiscence (N = 11). Function improved postoperatively: 20/21 (95 %) cases with preoperative pain, resolved; 13/20 (65 %) with preoperative trismus, improved; 21/24 (87 %) with preoperative malocclusion/jaw malignment, improved. This is the largest series of VSP/3Dprinted-guide/plate surgery for mORN to date. Mandibular reconstruction for ORN is aided by VSP/3Dprinted-guide/plate with accurate results, acceptable complications, and improved function.


Modulation of angiogenesis by human glioma xenograft models that differentially express vascular endothelial growth factor.
To evaluate the potential actions of vascular endothelial growth factor (VEGF) on capillary permeability and drug transport, tumorigenic human glioma cell lines were developed that expressed different levels of VEGF. Three human glioma cell lines (i.e. U373, SF126, SF188) were screened for VEGF under normoxic and hypoxic (i.e. induced by CoCl2) conditions by Western blot analysis. Subsequent to these results, sense and antisense VEGF164 cDNA transfections were conducted. It was found that parental SF188 (SF188/V-) and SF188/V+ (sense transfected) cells could serve as an appropriate in vivo model based on their divergent levels of VEGF expression. Media derived from SF188/V+ cells stimulated endothelial cell growth by 30-60%, and enhanced endothelial cell clonogenicity by 5-10-fold compared to SF188/V- or empty vector transfected cells. Nude rats implanted with either SF188/V- or SF188/V+ cells subcutaneously and intracerebrally formed tumors, with those derived from SF188/V+ cells growing at a faster rate. Immunohistochemistry analysis indicated that the expression of VEGF and number of capillaries (factor VIII assay) were approximately 3-fold greater in SF188/V+ tumors compared to SF188/V- tumors. Pharmacological assays, such as measurements of cytotoxicity and DNA adducts, in SF188/V- and SF188/V+ cells treated with carmustine or temozolomide were similar. Therefore, other than differences in VEGF expression and growth in vivo, SF188/V- and SF188/V+ cells possess a similar phenotype, and can serve as models to evaluate the influence of VEGF on drug transport.


Corrigendum: Correction of Acknowledgments: Epidemiology of Childhood Obesity in Korea.
This corrects the article on p. 510 in vol. 31, PMID: 27834078.


Canola oil/candelilla wax oleogel improves texture, retards staling and reduces in vitro starch digestibility of maize tortillas.
Maize tortilla staling is a major drawback that affects its commercialization and consumption, and so novel methods for retarding staling are continuously being explored. The present study evaluated the effect of adding a canola oil/candelilla wax oleogel (CWO; 0. 2, 4 and 6 g 100 g-1 ) to a basic masa formulation (water, 60 g 100 g-1 ; nixtamalized maize flour, 40 g 100 g-1 ) on the texture, staling and in vitro starch digestibility of maize tortillas made using a hot plate (200 °C). Textural analysis showed that CWO reduced hardness and increased the tensile strength of tortillas. Fourier transform infrared analysis indicated that the retrogradation of starch chains, quantified in terms of the intensity ratio 1047/1022, was reduced by oleogel incorporation. In vitro starch digestibility tests showed that tortillas containing CWO had lower readily digestible and slowly digestible starch fractions compared to the control tortilla without oleogel. The formation of amylose-lipid inclusion complexes and the formation of an oily physical barrier around starch granules were postulated as mechanisms underlying the reduced starch digestibility. © 2019 Society of Chemical Industry.


Scale-dependent and driving relationships between spatial features and carbon storage and sequestration in an urban park of Zhengzhou, China.
Research indicates that urban ecosystems can store large amounts of carbon. However, few studies have examined how the spatial features of park greenspace affect its carbon-carrying capacity, and how those effects vary with the spatial scale. Lidar point clouds and remote sensing images were extracted for the 196 ha green space in the China Green Expo to study carbon storage and sequestration in parks. Full subset regression, stepwise regression, HP analysis, and structural equation modeling were used to examine the scale dependency and the driving relationship between carbon storage and carbon sequestration in parks. The results show that the optimal statistical sample diameters for carbon density and carbon sequestration density in parks are 100 m. Under the influence of impermeable surfaces and water bodies, the statistical values of carbon density were minimized when the sample plot diameter was 700 m. Biodiversity and forest structure are the main drivers of carbon density, with the influence of water bodies being more prominent on a larger scale. Texture characteristics explain more carbon density than the vegetation index, and RVI could better explain the variation of carbon sequestration than NDVI. This study explores scaled changes in carbon density, carbon sequestration density in parks, and their driving relationships, which can aid in developing carbon sequestration strategies based on parks.


Synthesis, analysis, and multi-faceted applications of solid wastes-derived silica nanoparticles: a comprehensive review (2010-2022).
The synthesis of silica nanoparticles (SiNPs) has emerged as an extensive area of research in the last century. Owing to their instinctive properties like modifiable mesoporous structure, high surface area, adjustable pore size, and pore volume, SiNPs could be utilized in numerous fields like chemical, biochemical, catalysis, adsorption, and pollution control. Conventionally, SiNPs are produced by tetraethylorthosilicate (TEOS), tetramethylorthosilicate (TMOS), and sodium silicate, which are toxic and expensive. Therefore, the development of green, cost-effective approaches for the synthesis of SiNPs is highly desirable. In this course, during the last decade, silica-rich solid wastes (rice husk, corn cob, sugarcane bagasse, palm ash, fly ash, waste glass, waste packaging materials, photonic industrial wastes, etc.) were acknowledged as economical precursors to produce green SiNPs. In this respect, the present review focuses on reviewing several solid waste materials used for the synthesis of SiNPs, their properties, and different characterization techniques used for the analysis of SiNPs. The present review also accounts for the potential applications of such green SiNPs in several fields like catalysis, adsorption, biomedical applications, and energy storage. Moreover, despite the potential applications of SiNPs, still there is a lot to explore about their synthesis and utilization. Hence, in the last section of this review, future scope, challenges, and risk assessment of SiNPs have been discussed.


Modular flexible ureteroscopy and holmium laser lithotripsy for the treatment of renal and proximal ureteral calculi: A single-surgeon experience of 382 cases.
To determine the safety and efficacy of modular flexible ureteroscopy and holmium laser lithotripsy for the treatment of renal and proximal ureteral calculi, a retrospective chart review of a single surgeon's 3-year modular flexible ureteroscopy experience was performed. All of the patients were treated with modular flexible ureteroscopy and holmium laser lithotripsy by a single surgeon. Stone-free status was defined as no fragments or a single fragment ≤4 mm in diameter at the 3-month follow-up. The procedure number, operative time, stone-free rates, repeat usage of the multilumen catheter, and perioperative complications were documented. The present study included 215 male patients and 167 female patients, with an average age of 48.5±13.7 years (range, 17-84 years). The mean stone size was 11.5±4.1 mm (range, 4-28 mm), and the mean total stone burden was 17.5±5.7 mm (range 15-46 mm). A total of 305 patients (79.8%) had a stone burden ≤20 mm, and 77 patients (20.2%) had a stone burden >20 mm. The mean number of primary procedures was 1.3±0.2 (range, 1-3). The stone-free rate following the first and the second procedure was 73.4 and 86.9%, respectively. The mean postoperative hospital stay was 3.1±1.2 days (range, 2-6 days). The highest clearance rates were observed for proximal ureteral stones (100%) and renal pelvic stones (88.7%), whereas the lowest clearance rates were observed for lower calyx stones (76.7%) and multiple calyx stones (77.8%). The higher the initial stone burden, the lower the postoperative stone-free rate (≤20 vs. >20 mm; 89.8 vs. 75.3%). The overall complication rate was 8.1%. The results of the present study suggest that modular flexible ureteroscopy with holmium laser lithotripsy may be considered the primary method for the treatment of renal and proximal ureteral calculi in select patients, due to its acceptable efficacy, low morbidity, and relatively low maintenance costs.


Prevalence of painful diabetic peripheral neuropathy and its impact on quality of life among diabetic patients in Western region, Saudi Arabia.
Diabetic neuropathy is the most common long-term complications of diabetes, frequently presenting as painful diabetic peripheral neuropathy (PDPN), which can significantly impair patients' quality of life (QOL). This study set to estimate the prevalence of PNPD and health-related quality of life (HRQoL) in the setting of primary health care in Saudi Arabia. This study was conducted in primary health-care centers affiliated with the National Guard Health Affairs in Western Saudi Arabia. Arabic version of the Douleur Neuropathique 4 questionnaire was administered on diabetic patients to screen for neuropathic pain and short-form 12 questionnaire to assess HRQoL. The study screened (n = 349) Type 2 diabetic patients. The prevalence of PDPN was 33.2%. PDRN was more likely to affect females (adjusted odds ratio ["AOR"] =1.96, P = 0.024), and those living with diabetes for over 15 years (AOR = 2.26, P = 0.039), and those on insulin treatment (AOR: 2.33, P = 0.010) alone or in combination (AOR = 1.78, P = 0.034). Both physical and mental components (MCs) of QOL scores were significantly higher in diabetic patients without PDPN compared to those with it; 49.57 ± 9.31 versus 40.77 ± 8.14 for physical component QOL and 51.72 ± 9.36 versus 44.35 ± 8.12 for MC QOL, P < 0.001. Painful peripheral neuropathy is relatively common among type 2 diabetic patients in Western Saudi Arabia and impacts both physical and MCs of the QOL of affected patients.


Utility of leaf senescence-associated gene homologs as developmental markers in common wheat.
Senescence is the final stage of development in plant tissues. In the senescence process, many senescence-associated genes (SAGs) are reportedly transcriptionally up-regulated. Here, we reported the isolation of nine wheat SAG cDNA clones named TaSAG1 to TaSAG9, and evaluated the usefulness of the SAG homologs for wheat developmental molecular markers based on their expression patterns. The nine wheat SAGs were identified in wheat EST libraries based on their homology to rice SAGs. All wheat SAG transcripts were up-regulated during natural senescence as well as during dark-induced senescence in seedling leaves. However, the expression patterns of wheat SAGs in the flag leaf did not necessarily correspond to those in seedling leaves during senescence. The nine wheat SAGs also showed variable expression patterns in developing and ripening seeds. The transcript accumulation patterns of TaSAG5 and TaSAG6 increased linearly during the period examined in the flag leaf and seed, and are therefore available as molecular markers to respectively evaluate the degree of wheat flag leaf senescence and seed maturation. Transcript accumulation levels of the six SAGs were increased before apparent necrotic cell death of seeding leaves exhibiting wheat hybrid necrosis. These results suggested that necrotic cell death in wheat hybrid necrosis could be closely related not only to senescence but also to defense responses.


[Expression of aldolase of Toxoplasma gondii and preparation of the polyclonal antibodies against this protein].
To produce the aldolase protein and its polyclonal antibody. Aldolase gene was obtained from cDNA library by PCR amplification and subcloned to vector pET30a. The recombinant protein aldolase-His(6); was expressed in E.coli upon IPTG induction and then purified with affinity chromatography. The purified protein mixed with adjuvant was used to immunize SD rats to produce the polyclonal antibodies. The recombinant plasmid aldolase/pET30a was constructed successfully and expressed as a fusion protein aldolase-His(6);; Polyclonal antibody against aldolase-His(6); was obtained from rat, and antibody titer was 1:4 000. The purified protein aldolase-His(6); and its polyclonal antibodies were obtained, which may provide the foundation for the further studies on the function of aldolase.


Cell responses regulated by early reorganization of actin cytoskeleton.
Microfilaments exist in a dynamic equilibrium between monomeric and polymerized actin and the ratio of monomers to polymeric forms is influenced by a variety of extracellular stimuli. The polymerization, depolymerization and redistribution of actin filaments are modulated by several actin-binding proteins, which are regulated by upstream signalling molecules. Actin cytoskeleton is involved in diverse cellular functions including migration, ion channels activity, secretion, apoptosis and cell survival. In this review we have outlined the role of actin dynamics in representative cell functions induced by the early response to extracellular stimuli.


A clinical evaluation of fixed partial denture impressions.
Providing the dental laboratory with an accurate replication of the hard and soft tissue of a patient is important. Therefore, it is essential to examine whether clinicians critically evaluate impressions routinely before sending them to the laboratory. This study evaluated the quality of impressions sent to commercial laboratories for the fabrication of fixed partial dentures (FPD) by describing the frequency of clinically detectable errors and by analyzing correlations between the various factors involved. A total of 193 FPD impressions were evaluated, immediately after arrival at 11 dental laboratories, by 3 calibrated examiners. The impression technique and material used, tray type, and number of prepared units were recorded for each impression. Data relating to errors and faults, including defects in material polymerization, retention to tray, tissue contact by tray, crucial areas beyond tray borders, heavy-bodied material exposure through the wash material (for double-step impressions), inadequate union of materials, retraction cords embedded in impressions, and air bubbles, voids, or tears along the margin were also documented. The data were analyzed with the Pearson chi-square test (alpha = .05). Of the impressions, 89.1% had 1 or more observable errors. Significant correlations were found between material type and voids or tears at the finish line (Rv = 0.17, P < .025) and between material type and polymerization problems (Rv = 0.223, P < .004). Within the limitations of this study, impressions made with polyethers had the most detectable errors, followed by condensation-type silicones. The high frequency of detectable errors found in impressions sent for FPD fabrication is of concern.


Intron sequences involved in lariat formation during pre-mRNA splicing.
We have shown that lariat formation during in vitro splicing of several RNA precursors, from Drosophila to man, occurs at a unique and identifiable but weakly conserved site, 18 to 37 nucleotides proximal to the 3' splice site. Lariat formation within an artificial intron lacking a normal branch-point sequence occurs at a cryptic site a conserved distance (approximately 23 nucleotides) from the 3' splice site. Analysis of beta-thalassemia splicing mutations revealed that lariat formation in the first intron of the human beta-globin gene occurs at the same site in normal and mutant precursors, even though alternate 5' and 3' splice sites are utilized in the mutants. Remarkably, cleavage at the 5' splice site and lariat formation do not occur when the precursor contains a beta-thalassemia deletion removing the polypyrimidine stretch and AG dinucleotide at the 3' splice site. In contrast, a single base substitution in the AG dinucleotide blocks cleavage at the 3' splice site but not at the 5' site.


Innate Immunity as an Executor of the Programmed Death of Individual Organisms for the Benefit of the Entire Population.
In humans, over-activation of innate immunity in response to viral or bacterial infections often causes severe illness and death. Furthermore, similar mechanisms related to innate immunity can cause pathogenesis and death in sepsis, massive trauma (including surgery and burns), ischemia/reperfusion, some toxic lesions, and viral infections including COVID-19. Based on the reviewed observations, we suggest that such severe outcomes may be manifestations of a controlled suicidal strategy protecting the entire population from the spread of pathogens and from dangerous pathologies rather than an aberrant hyperstimulation of defense responses. We argue that innate immunity may be involved in the implementation of an altruistic programmed death of an organism aimed at increasing the well-being of the whole community. We discuss possible ways to suppress this atavistic program by interfering with innate immunity and suggest that combating this program should be a major goal of future medicine.


Stimulation of glucose uptake by glycosides from Alectra parasitica subsp. chitrakutensis (M.A. Rau) K.K. Khanna & An. Kumar: An in-vitro study of TNF-α induced insulin resistance in L6 myoblasts.
Alectra parasitica subsp. chitrakutensis (M.A. Rau) K.K. Khanna & An. Kumar (Orobanchaceae) is a parasitic plant indigenous to India. Locally, the plant is known as 'Midaki and Nirgundikand'. It is used to treat fever, piles, cardiovascular disorders, and blood-borne non-infectious diseases by ethnic communities. The phytochemical investigation of A. parasitica subsp. chitrakutensis rhizome led to the isolation of azafrin (1), rehmaionoside-C (2), and mussaenoside (3). Compounds (2) and (3) are being reported for the first time from this plant. Compounds were evaluated for their intercellular glucose uptake activity in basal and insulin-TNF-α-stimulated L6 muscle cells. In particular, rehmaionoside C exhibited activity comparative to metformin, increasing uptake by basal- and insulin-TNF-α-stimulated cells by 4.88- and 3.90-fold and 5.04- and 4.04-fold. While azafrin and mussaenoside have produced 3.03- and 2.36-fold; 4.03- and 3.22-fold increase in intercellular glucose uptake. Compounds did not show toxicities in rat L6 myoblast cells. The study suggests that rehmaionoside-C from A. parasitica subsp. chitrakutensis might activate glucose uptake by insulin mimics and could be a nontoxic anti-diabetes lead for drug discovery.


A current view of the human Ia system based on serological and immunochemical analysis.
We have approached the analysis of the human Ia system by localizing antigenic determinants on subsets of Class II molecules. For this purpose two classes of reagents have been used: alloantisera and mouse anti human monoclonal antibodies (moabs). We summarize here the most recent data obtained by these two approaches.


Feasibility of biodiesel production by microalgae Chlorella sp. (FACHB-1748) under outdoor conditions.
Chlorella sp. (FACHB-1748) was cultivated outdoors under natural sunlight to evaluate its potential for biofuel production. Urea was selected as nitrogen source, and the concentration was optimized. When the culture reached the late exponential stage, a triggering lipid accumulation test was conducted using different concentrations of sodium chloride and acetate. A scaling-up experiment was also conducted in a 70L photobioreactor. The highest biomass productivity (222.42, 154.48 mg/L/d) and lipid productivity (64.30, 33.69mg/L/d) were obtained with 0.1g/L urea in 5 and 70 L bioreactors, respectively. The highest lipid content (43.25%) and lipid yield (1243.98 mg/L) were acquired with the combination of 10 g/L sodium chloride and acetate. Moreover, the qualities of biodiesel, cetane number, saponification value, iodine value, and cold filter plugging point complied with the standards set by the National Petroleum Agency (ANP255), Standard ASTMD6751, and European Standard (EN 14214).


Telomerase as a therapeutic target for malignant gliomas.
Telomerase, a ribonucleoprotein enzyme, is considered as a potential target of cancer therapy because of its preferential expression in tumors. In particular, malignant gliomas are one of the best candidates for telomerase-targeted therapy. It is because malignant gliomas are predominantly telomerase-positive, while normal brain tissues do not express telomerase. In theory, there are two telomerase-associated therapeutic approaches for telomerase-positive tumors. One approach is the anti-telomerase cancer therapy to directly inhibit telomerase activity, resulting in apoptotic cell death or growth arrest. Two major components of the telomerase holoenzyme complex, the RNA template (hTER) and catalytic subunit (reverse transcriptase, hTERT) are well considered as therapeutic targets. The other approach is the telomerase-specific cancer therapy by targeting telomerase-expressing tumor cells as a means to directly kill the cells. Strategies using the transfer of therapeutic gene under the hTERT promoter system as well as immunotherapy directed against telomerase-positive cells are generally included. These telomerase-associated therapies are very promising for the treatment of malignant gliomas.


Radical versus conservative surgical treatment of liver hydatid cysts: A paired comparison analysis.
The management of liver hydatid cysts (LHC) is complex and includes surgery, percutaneous drainage, chemotherapy and observation. Broadly, there are two types of surgical treatment for LHC - conservative surgery (CS) and radical surgery (RS). The purpose of this study was to compare the outcome of RS and CS. Data from all patients with LHC treated in Sahloul Hospital, between January 2000 and December 2019, were retrieved. To minimize selection bias, paired comparison analysis (PCA) was performed. A total of 914 patients were included in this study. RS and CS were performed in 284 and 630 patients, respectively. After PCA, 206 patients were included in each group. The incidence of intraoperative bleeding was significantly higher in the RS group. The overall morbidity was significantly lower in the RS group. Thity-four patients developed recurrence with significantly higher recurrence in CS group. RS is associated with fewer postoperative complications and lower recurrence rate compared to CS. RS may be the preferred procedure for LHC if the expertise is available.


Solution synthesis of [ASN76]-human parathyroid hormone (1-84).
Human parathyroid hormone, hPTH(1-84), was synthesized by the conventional solution procedure applying the maximal-protection approach. All protecting groups were removed simultaneously by the HF method. The product was purified by CM-cellulose column chromatography, gel-filtration on Sephadex G-50 and in the final stage, by reversed phase HPLC. The structure of the final product was confirmed not only by HPLC analysis but also by peptide mapping of tryptic digests on HPLC. The present product showed 350(249-480) IU/mg on in vitro rat renal adenylate cyclase assay.


Neonatal Gut Microbiota and Human Milk Glycans Cooperate to Attenuate Infection and Inflammation.
Glycans of the intestinal mucosa and oligosaccharides of human milk influence the early colonization of the infant gut and establishment of mucosal homeostasis, and differences in colonization of the gut influence the ontogeny of glycans on the surface of the intestinal mucosa, proinflammatory signaling, homeostasis, and resilience to insult. This interkingdom reciprocal interaction is typical of a mutualistic symbiotic relationship. The period in which the infant gut most needs protection from hypersensitive inflammation overlaps with the recommended period of exclusive nursing; electively substituting artificial formula that lacks human milk protective glycans seems ill advised, especially for premature infants.


Unilateral amblyopia: Optical coherence tomography findings.
This study was performed to measure the macular and the retinal nerve fiber layer (RNFL) thicknesses using optical coherence tomography (OCT) in patients with unilateral amblyopia. Measurement of the Retinal nerve fiber layer and Macular Retinal Layer thickness for both amblyopic and normal fellow eyes by (OCT) was carried out at king Abdulaziz University Hospital, Riyadh, Saudi Arabia. Ninety-three patients with unilateral amblyopia between the ages of 5 years and 12 years were included. The macular retinal thickness and the RNFL thickness were measured using OCT. The mean macular retinal thickness was 259.3 μm and 255.6 μm, and the mean RNFL thickness was 112.16 μm and 106 μm, in the amblyopic eye and the normal eye, respectively. OCT assessment of RNFL thickness revealed a significantly thicker RNFL in amblyopic eye (P < 0.0001), but no statistically significant difference was found in macular retinal thickness (P = 0.195). The amblyopic process may involve the RNFL, but not the macula. However, further evaluation is needed.


A large-scale gene discovery for the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae).
The red palm weevil (RPW; Rhynchophorus ferrugineus) is a devastating pest of palms, prevalent in the Middle East as well as many other regions of the world. Here, we report a large-scale de novo complementary DNA (cDNA) sequencing effort that acquired ∼5 million reads and assembled them into 26 765 contigs from 12 libraries made from samples of different RPW developmental stages based on the Roche/454 GS FLX platform. We annotated these contigs based on the publically available known insect genes and the Tribolium castaneum genome assembly. We find that over 80% of coding sequences (CDS) from the RPW contigs have high-identity homologs to known proteins with complete CDS. Gene expression analysis shows that the pupa and larval stages have the highest and lowest expression levels, respectively. In addition, we also identified more than 60 000 single nucleotide polymorphisms and 1 200 simple sequence repeat markers. This study provides the first large-scale cDNA dataset for RPW, a much-needed resource for future molecular studies.


Puma and p53 play required roles in death evoked in a cellular model of Parkinson disease.
6-Hydroxydopamine (6-OHDA) is widely used in vivo and in vitro to mimic the selective neuronal degeneration that characterizes Parkinson disease (PD). To uncover candidate genes that may mediate neuron death in PD, we previously used SAGE to identify transcripts that are rapidly induced by 6-OHDA in neuronally differentiated PC12 cells. Among induced pro-apoptotic genes was that encoding the BH3-only protein PUMA. Here, we confirm that 6-OHDA induces both PUMA mRNA and protein. 6-OHDA additionally induced Bim, another pro-apoptotic BH3-only protein. Using specific siRNAs, we demonstrate that PUMA, but not Bim, is required for death evoked by 6-OHDA. PUMA is a target of p53, a transcription factor activated by 6-OHDA. Involvement of p53 in 6-OHDA evoked death was confirmed by the protective actions of a DN p53 and pifithrin alpha, inhibitors of p53 signaling. Our findings thus indicate that p53 and PUMA play required roles in a cellular model of PD.


Event-based modeling in temporal lobe epilepsy demonstrates progressive atrophy from cross-sectional data.
Recent work has shown that people with common epilepsies have characteristic patterns of cortical thinning, and that these changes may be progressive over time. Leveraging a large multicenter cross-sectional cohort, we investigated whether regional morphometric changes occur in a sequential manner, and whether these changes in people with mesial temporal lobe epilepsy and hippocampal sclerosis (MTLE-HS) correlate with clinical features. We extracted regional measures of cortical thickness, surface area, and subcortical brain volumes from T1-weighted (T1W) magnetic resonance imaging (MRI) scans collected by the ENIGMA-Epilepsy consortium, comprising 804 people with MTLE-HS and 1625 healthy controls from 25 centers. Features with a moderate case-control effect size (Cohen d ≥ .5) were used to train an event-based model (EBM), which estimates a sequence of disease-specific biomarker changes from cross-sectional data and assigns a biomarker-based fine-grained disease stage to individual patients. We tested for associations between EBM disease stage and duration of epilepsy, age at onset, and antiseizure medicine (ASM) resistance. In MTLE-HS, decrease in ipsilateral hippocampal volume along with increased asymmetry in hippocampal volume was followed by reduced thickness in neocortical regions, reduction in ipsilateral thalamus volume, and finally, increase in ipsilateral lateral ventricle volume. EBM stage was correlated with duration of illness (Spearman ρ = .293, p = 7.03 × 10-16 ), age at onset (ρ = -.18, p = 9.82 × 10-7 ), and ASM resistance (area under the curve = .59, p = .043, Mann-Whitney U test). However, associations were driven by cases assigned to EBM Stage 0, which represents MTLE-HS with mild or nondetectable abnormality on T1W MRI. From cross-sectional MRI, we reconstructed a disease progression model that highlights a sequence of MRI changes that aligns with previous longitudinal studies. This model could be used to stage MTLE-HS subjects in other cohorts and help establish connections between imaging-based progression staging and clinical features.


Relationship between PTEN, DNA mismatch repair, and tumor histotype in endometrial carcinoma: retained positive expression of PTEN preferentially identifies sporadic non-endometrioid carcinomas.
Loss of PTEN (phosphatase and tensin homolog) expression and microsatellite instability are two of the more common molecular alterations in endometrial carcinoma. From the published literature, it is controversial as to whether there is a relationship between these different molecular mechanisms. Therefore, a cohort of 187 pure endometrioid and non-endometrioid endometrial carcinomas, carefully characterized as to clinical and pathological features, was examined for PTEN sequence abnormalities and the immunohistochemical expression of PTEN and the DNA mismatch repair proteins MLH1, MSH2, MSH6, and PMS2. MLH1 methylation analysis was performed when tumors had loss of MLH1 protein. Mismatch repair protein loss was more frequent in endometrioid carcinomas compared with non-endometrioid carcinomas, a difference primarily attributable to the presence of MLH1 methylation in a greater proportion of endometrioid tumors. Among the non-endometrioid group, mixed endometrioid/non-endometrioid carcinomas were the histotype that most commonly had loss of a mismatch repair protein. In endometrioid tumors, the frequency of PTEN loss measured by immunohistochemistry and mutation did not differ significantly between the mismatch repair protein intact or mismatch repair protein loss groups, suggesting that PTEN loss is independent of mismatch protein repair status in this group. However, in non-endometrioid carcinomas, both intact positive PTEN immunohistochemical expression and PTEN wild type were highly associated with retained positive expression of mismatch repair proteins in the tumor. Relevant to screening endometrial cancers for Lynch Syndrome, an initial PTEN immunohistochemistry determination may be able to replace the use of four mismatch repair immunohistochemical markers in 63% of patients with non-endometrioid endometrial carcinoma. Therefore, PTEN immunohistochemistry, in combination with tumor histotype, is a useful adjunct in the clinical evaluation of endometrial carcinomas for Lynch Syndrome.


Five- and 10-Year Cause-Specific Survival Rates in Carcinoma of the Minor Salivary Gland.
Previous studies of prognostic factors of carcinoma of the minor salivary gland (MSG) have been limited to single-institution studies and small case series. Thus, limited data are available to guide the head and neck oncologist in counseling patients on the prognosis and management of these malignant neoplasms. To examine 5- and 10-year cause-specific survival (CSS) rates of MSG carcinomas across all histologic subtypes and head and neck tumor subsites. Retrospective, population-based study using National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) data from January 1, 1988, through December 31, 2009. The study included 5334 patients diagnosed as having MSG carcinoma and registered in the SEER database. Patients without follow-up, diagnostic confirmation, and/or race designation were excluded from the analysis (131 [2.4%]). Final follow-up was completed on December 31, 2009, and data were analyzed from August 5, 2013, to July 1, 2014. Five- and 10-year CSS rates for US patients with MSG carcinoma. Cox proportional hazard models were used to estimate adjusted hazard ratios (HRs) and 95% CIs. Among the 5334 patients with MSG carcinoma included, the most common histologic subtypes included mucoepidermoid carcinoma (1568 [29.4%]), adenoid cystic carcinoma (1228 [23.0%]), and adenocarcinoma (1313 [24.6%]). The most frequent sites of primary tumor were the oral cavity (3132 [58.7%]) and pharynx (1130 [21.2%]). Five-year CSS rate was significantly worse for MSG malignant neoplasms located in the larynx (HR, 2.42; 95% CI, 1.67-3.50) and nasal cavity and/or paranasal sinus (HR, 1.73; 95% CI, 1.29-2.32). Being older than 75 years was associated with a significantly worse 5-year CSS rate (HR, 2.88; 95% CI, 2.05-4.06). Compared with no surgery, local tumor destruction (HR, 0.44; 95% CI, 0.30-0.64), partial surgery (HR, 0.33; 95% CI, 0.23-0.47), and total surgery (HR, 0.55; 95% CI, 0.41-0.74) were each found to be a significant positive prognostic factor. No differences were observed in the 5-year hazard of death for race/ethnicity, sex, diagnosis year, or socioeconomic status, and 10-year adjusted HRs were similar to the 5-year patterns. This study, to date, represents the largest US survival analysis of carcinoma of the MSG. Prognosis is associated with histologic subtype, tumor subsite, age at diagnosis, grade, and surgical therapy.


REDD1 is essential for stress-induced synaptic loss and depressive behavior.
Major depressive disorder (MDD) affects up to 17% of the population, causing profound personal suffering and economic loss. Clinical and preclinical studies have revealed that prolonged stress and MDD are associated with neuronal atrophy of cortical and limbic brain regions, but the molecular mechanisms underlying these morphological alterations have not yet been identified. Here, we show that stress increases levels of REDD1 (regulated in development and DNA damage responses-1), an inhibitor of mTORC1 (mammalian target of rapamycin complex-1; ref. 10), in rat prefrontal cortex (PFC). This is concurrent with a decrease in phosphorylation of signaling targets of mTORC1, which is implicated in protein synthesis-dependent synaptic plasticity. We also found that REDD1 levels are increased in the postmortem PFC of human subjects with MDD relative to matched controls. Mutant mice with a deletion of the gene encoding REDD1 are resilient to the behavioral, synaptic and mTORC1 signaling deficits caused by chronic unpredictable stress, whereas viral-mediated overexpression of REDD1 in rat PFC is sufficient to cause anxiety- and depressive-like behaviors and neuronal atrophy. Taken together, these postmortem and preclinical findings identify REDD1 as a critical mediator of the atrophy of neurons and depressive behavior caused by chronic stress exposure.


The effects of surgical margin and edge cryotherapy after liver resection for colorectal cancer metastases.
A 1 cm margin seen at operation is typically the minimally acceptable margin for liver resections. Patients who fail to achieve this margin are routinely treated with edge cryotherapy at our unit. This paper aims to assess the benefit of edge cryotherapy on survival in patients with such suboptimal margins. Between January 1990 and February 2006, 608 patients underwent liver resection and/or cryotherapy for colorectal cancer metastases. All liver resections were performed using the CUSA transection method. Data on marginal status were available for 398 patients. Patient demographics, number and size of liver lesions, preoperative and postoperative carcinoembryonic antigen (CEA), extent of liver resection, margin status, site and date of recurrence, date of last follow-up and death were examined. There were 175 patients in the R0 group (>1 cm macroscopic and > or = 1 mm microscopic margin), 103 patients in the R1 group (>1 cm macroscopic and <1 mm microscopic margin) and 120 patients in the R2 group (< or = 1 cm macroscopic margin and received edge cryotherapy). After a median follow-up of 63 months, there were no significant difference between the 5-year survival rates for R0, R1 and R2 (40%, 30% and 28%, respectively). As long as the surgical margin is clear macroscopically, the microscopic margin width does not affect survival. In patients with suboptimal margins, the addition of edge cryotherapy improves the prospect for long-term survival and may lower recurrence risk.