Spermidine application reduces fluoride uptake and ameliorates physiological injuries in a susceptible rice cultivar by activating diverse regulators of the defense machinery.
The manuscript illustrates the ameliorative effects of exogenously applied higher polyamine (PA), spermidine (Spd) in the susceptible indica rice cultivar IR-64 subjected to prolonged fluoride stress. The Spd treatment drastically reduced fluoride bioaccumulation by restricting entry of the anions through chloride channels and enabled better maintenance of the proton gradient via accumulation of P-H+/ATPase, thereby improving the root and shoot lengths, fresh and dry weights, RWC, chlorophyll content and activities of pyruvate dehydrogenase (PyrDH), α-amylase, and nitrate reductase (NR) in the Spd-treated, stressed plants. Expression of RuBisCo, PyrDH, α-amylase, and NR was stimulated. Spd supplementation reduced the molecular damage indices like malondialdehyde, lipoxygenase, protease activity, electrolyte leakage, protein carbonylation, H2O2, and methylglyoxal (detoxified by glyoxalase II). Mitigation of oxidative damage was facilitated by the accumulation and utilization of proline, glycine-betaine, total amino acids, higher PAs, anthocyanin, flavonoids, β-carotene, xanthophyll, and phenolics as verified from the expression of genes like P5CS, BADH1, SAMDC, SPDS, SPMS, DAO, PAO, and PAL. Spd treatment activated the ascorbate-glutathione cycle in the stressed seedlings. Expression and activities of enzymatic antioxidants showed that GPOX, APX, GPX, and GST were the chief ROS scavengers. Exogenous Spd promoted ABA accumulation by upregulating NCED3 and suppressing ABA8ox1 expression. ABA-dependent osmotic stress-responsive genes like Osem, WRKY71, and TRAB1 as well as ABA-independent transcription factor encoding gene DREB2A were induced by Spd. Thus, Spd treatment ameliorated fluoride-mediated injuries in IR-64 by restricting fluoride uptake, refining the defense machinery and activating the ABA-dependent as well as ABA-independent stress-responsive genes.


Submaximal fatiguing contractions reduce stability of voluntary postural control more than maximal fatiguing contractions.
It is well known that different exercise intensities for the ankle muscles can impair postural stability. However, it remains unclear whether it is low-intensity exercises (which primarily induce central fatigue) or high-intensity exercises (which primarily induce peripheral fatigue) that have a greater effect on voluntary postural control. The aim of this study was to compare the effects of fatiguing exercises that induce either central or peripheral fatigue on voluntary postural stability. Following isometric maximum voluntary contraction (IMVC) tests, 12 volunteers randomly performed submaximal (40% IMVC) and maximal (100% IMVC) fatiguing contractions of the plantar flexors on a dynamometer. Before and after the fatiguing protocols, postural stability was assessed by measuring the centre of pressure trajectory during voluntary sways on a force plate. The electromyography activity of the right gastrocnemius medialis was recorded. To assess central and peripheral fatigue, electrical stimulations were applied both during and after the IMVC. The effects of the fatiguing protocols and the differences between the protocols were tested with a two-way repeated measures ANOVA test (fatigue × time). Submaximal contractions induced a greater increase of sway area and medial-lateral sway cycle range than maximal contractions (P < 0.01). Voluntary activation decreased significantly more after submaximal contractions than maximal contractions (P < 0.01). Submaximal contractions induced a significantly greater reduction of IMVC than maximal contractions (P < 0.01). Resting twitch size was smaller after maximal contractions than submaximal contractions (P = 0.04). The outcome of this study suggests that training programmes for patients with balance issues should be based on training with maximal rather than submaximal load, to avoid deficits that might compromise postural stability.


Glycopeptide-Based Supramolecular Hydrogels Induce Differentiation of Adipose Stem Cells into Neural Lineages.
We applied a bottom-up approach to develop biofunctional supramolecular hydrogels from an aromatic glycodipeptide. The self-assembly of the glycopeptide was induced by either temperature manipulation (heating-cooling cycle) or solvent (DMSO to water) switch. The sol-gel transition was salt-triggered in cell culture media and resulted in gels with the same chemical compositions but different mechanical properties. Human adipose derived stem cells (hASCs) cultured on these gels under basal conditions (i.e., without differentiation factors) overexpressed neural markers, such as GFAP, Nestin, MAP2, and βIII-tubulin, confirming the differentiation into neural lineages. The mechanical properties of the gels influenced the number and distribution of the adhered cells. A comparison with gels obtained from the nonglycosylated peptide showed that glycosylation is crucial for the biofunctionality of the hydrogels by capturing and preserving essential growth factors, e.g., FGF-2.


Ovarian pregnancy torsion.
Ovarian-cyst torsion is often seen in early pregnancy due to progesterone stimulation. It is the fifth-most-common gynecological emergency, with a reported incidence of 3%. An ectopic pregnancy implanted in the ovary is rare, occurring in only 0.5-1% of such pregnancies. We presented a case of ovarian pregnancy with concomitant ipsilateral side tubal torsion managed by laparoscopy. Varying density distribution in the ovary can predispose to ovarian torsion. Emergency surgery may offer the possibility of avoiding the ablation of functional ovarian tissue. Early detection and prompt management can preserve fertility and ovarian function. The etiology, presenting syndrome and management are discussed.


Effect of aerobic exercise training on mtDNA deletion in soleus muscle of trained and untrained Wistar rats.
According to the theory of mitochondrial aging, oxidative stress plays a major role in aging and age related degenerative diseases. Since oxygen consumption and reactive oxygen species rate increase during aerobic exercise, we hypothesised that heavy aerobic training could lead to enhanced mitochondrial DNA (mtDNA) deletion in postmitotic tissues, leading in turn to premature aging and degenerative diseases. Sixty adult male 2 month old Wistar14848 rats were divided into six equal groups. Two groups were trained for 3 months by running on a treadmill (5 days/week, incline 6 degrees; group 1: 40 m/min, 20 min/day; group 2: 20 m/min, 40 min/day), while two sedentary groups participated in aerobic exercise only at the end of the study (incline 6 degrees; group 3: 40 m/min; group 4: 20 m/min). To control for physical and physiological parameters, two groups of untrained animals were killed at the beginning (group 6) and end (group 5) of the study. Expand long PCR was used to investigate mtDNA deletion in soleus muscle and a sequencing method was used to confirm the mtDNA deletion break point. Our results did not show any mtDNA deletion in untrained rats or in those that underwent moderate training (group 2) We only found mtDNA deletion (4.6 kb) in the soleus muscle of heavily trained rats (group 1). These results demonstrate that one session of aerobic exercise does not cause mtDNA deletion in skeletal muscle. The difference in results between heavy and moderate aerobic training may be due to low work rate or up-regulation of inducible antioxidant systems in moderate training.


Natural antimicrobials from plants: Recent advances and future prospects.
Plant-based antimicrobial substances have emerged as promising alternatives to conventional antibiotics and preservatives. Although many review studies have been done in this field, many of these reviews solely focus on specific compounds from particular perspectives. This paper aims to provide a comprehensive review on the various types of plant-based antimicrobial substances, the extraction and purification processes, as well as the application and safety issues. Combining different natural plant-derived substances shows promise in enhancing antimicrobial activities. Moreover, despite the existence of various methods (e.g., microwave-assisted extraction, supercritical fluid extraction) to extract and purify antimicrobial substances, isolating pure compounds remains a laborious process. Sustainability issues should also be considered when developing extraction methods. Additionally, the extraction process generates a significant amount of plant waste, necessitating proper utilization to ensure economic viability. Lastly, not all plant-derived substances are safe, and further research is needed to investigate their toxicity before widespread application.


Estimating the propagation of both reported and undocumented COVID-19 cases in Spain: a panel data frontier approximation of epidemiological models.
We use a stochastic frontier analysis (SFA) approach to model the propagation of the COVID-19 epidemic across geographical areas. The proposed models permit reported and undocumented cases to be estimated, which is important as case counts are overwhelmingly believed to be undercounted. The models can be estimated using only epidemic-type data but are flexible enough to permit these reporting rates to vary across geographical cross-section units of observation. We provide an empirical application of our models to Spanish data corresponding to the initial months of the original outbreak of the virus in early 2020. We find remarkable rates of under-reporting that might explain why the Spanish Government took its time to implement strict mitigation strategies. We also provide insights into the effectiveness of the national and regional lockdown measures and the influence of socio-economic factors in the propagation of the virus.


Accidental ingestion of foreign object: Systematic review, recommendations and report of a case.
One of the serious complications during a routine endodontic procedure is accidental ingestion/aspiration of the endodontic instruments, which can happen when proper isolation is not done. There are at present no clear guidelines whether foreign body ingestion in the gastrointestinal tract should be managed conservatively, endoscopically or surgically. A 5 year old boy reported to the Department of Pediatric and Preventive Dentistry, D.A. Pandu Memorial R.V. Dental College, Bangalore, India, with a complaint of pain and swelling in the lower right back teeth region. Endodontic therapy was planned for the affected tooth. During the course of treatment the child accidentally swallowed a 21 mm 15 size K file. Endoscopy was performed immediately but the instrument could not be retrieved. The instrument passed out uneventfully along with the stools 48 h after ingestion. Careful evaluation of the patient immediately after the accident helps in managing the patient effectively along with following the recommended guidelines.


Molecular profiling pleomorphic lobular carcinomas of the breast: evidence for a common molecular genetic pathway with classic lobular carcinomas.
Pleomorphic lobular carcinomas (PLC) of the breast display histological features associated with classic invasive lobular carcinoma (ILC), yet they also exhibit more conspicuous nuclear atypia and pleomorphism, and an aggressive clinical behaviour. From a breast cancer progression perspective, it is unclear whether PLC is a variant of ILC or is a high-grade invasive ductal carcinoma (IDC) that has lost E-cadherin. The molecular features of 26 PLC were studied using immunohistochemistry [oestrogen receptor (ER), progesterone receptor (PR), HER2, p53 and E-cadherin], 0.9 Mb resolution, microarray-based comparative genomic hybridization (aCGH), fluorescent (FISH) and chromogenic (CISH) in situ hybridization and loss of heterozygosity. Comparative analysis was performed with aCGH data from PLC with classic ILC (16 cases) and high grade IDC (35 cases). PLCs were frequently ER- and PR-positive, E-cadherin-negative and occasionally HER2- and p53-positive. Recurrent copy number changes identified by aCGH included gains on 1q, 8q, 11q, 12q, 16p and 17q and losses on 8p, 11q, 13q, 16q and Xq. Highly recurrent 1q+ (100% of cases), 16p+ (93%), 11q- (53%) and 16q- (93%) and evidence of the der(1;16)/der(16)t(1;16) rearrangement, as detected by FISH, suggested that PLC had a 'lobular genotype'. Focal amplifications were evident at 8p12-p11, 8q24, 11q13.1-q14.1, 12q14, 17q12 and 20q13. Loss of BRCA2 was detected in 40% of PLC by LOH. Comparative analysis of aCGH data suggested the molecular features of PLC (ER/PR-positive, E-cadherin-negative, 1q+, 11q(-), 16p+ and 16q(-)) were more closely related to those of ILC than IDC, implicating an overlapping developmental pathway for these lobular tumour types. Molecular alterations found in PLC that are more typical of high-grade IDC than ILC (p53 and HER2 positivity, 8q+, 17q24-q25+, 13q(-) and amplification of 8q24, 12q14, 17q12 and 20q13) are likely to drive the high-grade and more aggressive biology of PLC.


Discovery of hPRDX5-based peptide inhibitors blocking PD-1/PD-L1 interaction through in silico proteolysis and rational design.
The human peroxiredoxin-5 (hPRDX5) is a member of the family of antioxidant enzymes, which could resist immunosuppression by promoting immune organs development, lymphocyte proliferation and up-regulation of the levels of serum cytokines. However, being a recombinant protein, the hPRDX5 exhibits some problems including the high production cost and bad tissue penetration. Compared to macromolecular therapeutic agents, synthetic peptides have several advantages as drug candidates, such as lower manufacturing costs, reduced immunogenicity, and better organ or tumor penetration. The purpose of this research was to design the novel peptides come from hPRDX5 that can block the interaction of PD-1 and PD-L1. Herein in this work, we firstly confirmed the inhibitory activity of hPRDX5 on the binding of PD-L1 to PD-1 based on the previous observation, subsequently, in silico proteolysis and rational design (such as alanine scanning mutagenesis and truncation) were used to automate the design of new peptides derived from hPRDX5 with anti-tumour activity. We found that the most potent peptide could block the PD-1/PD-L1 interaction effectively with an IC50 of 0.646 μM, and could restore the function of Jurkat T cells which had been suppressed by stimulated HCT116 cells. Moreover, experiments with tumor-bearing mice models showed that the peptide IMB-P6-10 could effectively inhibit tumor growth and showed extraordinary low acute toxicity in vivo. The peptides described in this paper may provide novel low-molecular-weight drug candidates for cancer immunotherapy.


[Discriminant analysis applied to formants of vowels in Castellano dialect during the phonation with prosthesis and esophageal voice after total laryngectomy].
The objective of the present work consists, in analyzing if there is statistically significant differences between certain quantitative variables of acoustic parameters of the voice, in three samples of individuals, using a discriminative analysis. The study groups are formed by: 20 patients with total laringectomy and esophageal voice, 20 patients with total laringectomy and tracheoesophageal voice with phonatory prosthesis, and 20 normal individuals. We analyzed the energy, bandwidth and frequency of the four first formants of the five Castilian vowels in the context of an established phrase. Based on our results the method of discriminative analysis of the used variables is valid to establish differences between the three studied groups; that is they display their own characteristics from the acoustic point of view. This method allows us to affirm that the tracheoesophageal voice does not come closer to the normal voice that the esophageal voice does.


Assessing farmers' intention towards conservation agriculture by using the Extended Theory of Planned Behavior.
Conservation agriculture is considered as an agro-ecological approach to the sustainable and resource-saving crop production system. Previous studies on the adoption of agricultural innovation often consider socio-economic characteristics and overlook underlying psychological factors influencing farmers' behavioral intention. This study adopted three socio-psychological constructs, Attitude, Subjective Norm (SN), and Perceived Behavioral Control (PBC), which originated from the Theory of Planned Behavior (TPB), and proposed two new constructs, Knowledge (K) and Perceived Climatic Threats of Conventional Farming (PCTCV). Using the Extended Theory of Planned Behavior Model (ETPBM), this study analyzed factors affecting the intention of farmers to continue Conservation Agriculture (CA) for the following year (2020) in three districts located in the northern part of Bangladesh. This research found evidence from the hierarchical regression analysis that variables of ETPBM could explain 7% more variation (change in R2) in intention than variables of TPB, while the socio-economic characteristics of farmers can explain only 0.5% more than the variables of ETPBM. Structural Equation Modelling (SEM) was used to determine and examine the extended theory and indicated that farmers' knowledge (K) had the highest total impact, while attitude had the highest direct impact on intention. The results indicated that factors including Attitude, SN, PBC, Knowledge (K), and PCTCV are positively significant when attempting to understand farmers' intentions to adopt CA. This study presented evidence about the effectiveness of ETPBM when assessing the farmers' intentions towards conservation farming. Based on these findings, we concluded that the combination of extension services for upgrading the level of knowledge concerning the importance of CA could significantly affect farmers' attitudes on intention towards the application of CA.


Analysis of hiPSCs differentiation toward hepatocyte-like cells upon extended exposition to oncostatin.
The capability to produce and maintain functional human adult hepatocytes remains one of the major challenges for the use of in-vitro models toward liver cell therapy and industrial drug-screening applications. Among the suggested strategies to solve this issue, the use of human-induced pluripotent stem cells (hiPSCs), differentiated toward hepatocyte-like cells (HLCs) is promising. In this work, we propose a 31-day long protocol, that includes a final 14-day long phase of oncostatin treatment, as opposed to a 7-day treatment which led to the formation of a hepatic tissue functional for CYP1A2, CYP2B6, CYP2C8, CYP2D6, and CYP3A4. The production of albumin, as well as bile acid metabolism and transport, were also detected. Transcriptome profile comparisons and liver transcription factors (TFs) motif dynamics revealed increased expression of typical hepatic markers such as HNF1A and of important metabolic markers like PPARA. The performed analysis has allowed for the extraction of potential targets and pathways which would allow enhanced hepatic maturation in-vitro. From this investigation, NRF1 and SP3 appeared as transcription factors of importance. Complex epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET) patterns were also observed during the differentiation process. Moreover, whole transcriptome analysis highlighted a response typical of the one observed in liver regeneration and hepatocyte proliferation. While a complete maturation of hepatocytes was yet to be obtained, the results presented in this work provide new insights into the process of liver development and highlight potential targets aimed to improve in-vitro liver regeneration.


Update 2020: nomenclature and listing of celiac disease-relevant gluten epitopes recognized by CD4+ T cells.
Celiac disease is caused by an abnormal intestinal T cell response to cereal gluten proteins. The disease has a strong human leukocyte antigen (HLA) association, and CD4+ T cells recognizing gluten epitopes presented by disease-associated HLA-DQ allotypes are considered to be drivers of the disease. This paper provides an update of the currently known HLA-DQ restricted gluten T cell epitopes with their names and sequences.


What has high-throughput sequencing ever done for us?
This month's Genome Watch looks back over the past 10 years and highlights how the incredible advances in sequencing technologies have transformed research into microbial genomes.


Typing with mobile devices: A comparison of upper limb and shoulder muscle activities, typing performance and perceived workload under varied body postures, typing styles and device types.
This study aimed to examine the effects of body posture, typing style and device type on upper limb and shoulder muscle activities, typing performance and perceived workload while typing with mobile devices. Participants were asked to type with two mobile devices (i.e., a tablet and a smartphone) under three postures and in two typing styles. Muscle activity was recorded for four upper limb and shoulder muscles on both sides with surface electromyography. Results showed that body posture and typing style yielded significant effects on tying performance, perceived workload, and muscle activities in the forearm, upper arm and shoulder. Typing with a tablet was more accurate and had greater muscle activities in the upper arm and forearm on both sides than typing with a smartphone. The findings may be useful in developing evidence-based guidelines for the wise use of mobile devices and for the prevention of risks for musculoskeletal disorders.


Assessment of promoter methylation and expression of SIX2 as a diagnostic and prognostic biomarker in Wilms' tumor.
This study was designed to evaluate the utility of expression and DNA methylation patterns of the sine oculis homeobox homolog 2 (SIX2) gene in early diagnosis and prognosis of Wilms' tumor (WT). Methylation-specific polymerase chain reaction (MSP), real-time quantitative polymerase chain reaction (qRT-PCR), receiver operating characteristic (ROC), and survival curve analyses were utilized to measure the expression and DNA methylation patterns of SIX2 in a cohort of WT tissues, with a view to assessing their diagnostic and prognostic value. Relative expression of SIX2 mRNA was higher, while the promoter methylation level was lower in the WT than control group (P < 0.05) and closely associated with poor survival prognosis of WT children (P < 0.05). Increased expression and decreased methylation of SIX2 were correlated with increasing tumor size, clinical stage, vascular invasion, and unfavorable histological differentiation (P < 0.05). ROC curve analysis showed areas under the curve (AUCs) of 0.579 for methylation and 0.917 for expression in WT venous blood, indicating higher diagnostic yield of preoperative SIX2 expression. The preoperative venous blood SIX2 expression level serves as an underlying biomarker for early diagnosis of WT. SIX2 overexpression and concomitantly decreased promoter methylation are significantly associated with poor survival of WT children.


Evaluation of Symptoms of Depression and Anxiety in Adolescents with Epilepsy in a Tertiary-Level Medical Center in Serbia.
Even though epilepsy is the most common neurological disorder in adolescence; studies of adolescents with epilepsy are scarce. To evaluate whether adolescents with epilepsy are more likely to develop anxiety and depressive symptoms than their healthy peers and to determine the relationship of depression and anxiety scores to epilepsy-related variables. Cross-sectional observational study. Ninety adolescents with epilepsy aged 13-19 years were compared with healthy controls using the Beck Depression Inventory II (BDI II) and the Stait Trait Anxiety Inventory (STAI) for assessment of depressive and anxiety symptoms. Within the study group, epilepsy-related variables were also determined using the stated measures. The independent sample t-test, One-Way ANOVA, Pearson's correlation coefficient. 23.3% of the study group and 8.3% of the control group had mild to moderate depression. The mean BDI II score was significantly higher in the group of adolescents with epilepsy. Comparing the STAI scores, results on both STAI scales were higher in the study group. Regarding the epilepsy-related variables, a high frequency of seizures, symptomatic etiology and polytherapy showed strong associations with high depression and anxiety scores. Symptoms of depression and anxiety are not uncommon among adolescents with epilepsy. BDI II and STAI may be used as screening tools to provide useful information to clinicians when assessing adolescents' mental status.


Genetics in rheumatoid arthritis.
This chapter reviews the latest original research on the genetics of rheumatoid arthritis (RA), with a focus on its relevance for the clinical rheumatologist. The following questions will be dealt with in order to appreciate the recent progress in this field. * Why is a knowledge of genetics useful for an understanding of the pathogenesis of RA? * Is a knowledge of genetic risk factors relevant for day-to-day clinical practice? * What methods are used for identifying genetic risk factors? * Which genetic regions have been identified in susceptibility to RA? * What risk factors have been identified? * What are the future prospects and research agenda?


Rectus abdominis as a source of myopotentials inhibiting demand pacemakers.
We examined four patients with a demand pacemaker who exhibited transient symptoms of vertigo while trying to rise from the supine position. In two of these there was an epicardial pacing system with the pacemaker in the abdominal wall superficial to the rectus abdominis muscle; in the other two cases the pacing system was transvenous with the pacemaker superficial to the right pectoralis major muscle. It was found that the "pseudovertigo" was due to pacemaker inhibition caused by rectus abdominis myopotentials in all four patients. Changing the pacing mode from the demand to fixed-rate resulted in the disappearance of all symptoms at rest and during exercise.


Accumulation and distribution of mercury in fruiting bodies by fungus Suillus luteus foraged in Poland, Belarus and Sweden.
Presented in this paper is result of the study of the bioconcentration potential of mercury (Hg) by Suillus luteus mushroom collected from regions within Central, Eastern, and Northern regions of Europe. As determined by cold-vapor atomic absorption spectroscopy, the Hg content varied from 0.13 ± 0.05 to 0.33 ± 0.13 mg kg(-1) dry matter for caps and from 0.038 ± 0.014 to 0.095 ± 0.038 mg kg(-1) dry matter in stems. The Hg content of the soil substratum (0-10 cm layer) underneath the fruiting bodies showed generally low Hg concentrations that varied widely ranging from 0.0030 to 0.15 mg kg(-1) dry matter with mean values varying from 0.0078 ± 0.0035 to 0.053 ± 0.025 mg kg(-1) dry matter, which is below typical content in the Earth crust. The caps were observed to be on the richer in Hg than the stems at ratio between 1.8 ± 0.4 and 5.3 ± 2.6. The S. luteus mushroom showed moderate ability to accumulate Hg with bioconcentration factor (BCF) values ranging from 3.6 ± 1.3 to 42 ± 18. The consumption of fresh S. luteus mushroom in quantities up to 300 g week(-1) (assuming no Hg ingestion from other foods) from background areas in the Central, Eastern, and Northern part of Europe will not result in the intake of Hg exceeds the provisional weekly tolerance limit (PTWI) of 0.004 mg kg(-1) body mass.


Reporting and utilization of Patient-Reported Outcomes Measurement Information System® (PROMIS®) measures in orthopedic research and practice: a systematic review.
The Patient-Reported Outcomes Measurement Information SystemⓇ (PROMISⓇ) is a dynamic system of psychometrically sound patient-reported outcome (PRO) measures. There has been a recent increase in the use of PROMIS measures, yet little has been written about the reporting of these measures in the field of orthopedics. The purpose of this study was to conduct a systematic review to determine the uptake of PROMIS measures across orthopedics and to identify the type of PROMIS measures and domains that are most commonly used in orthopedic research and practice. We searched PubMed, Embase, and Scopus using keywords and database-specific subject headings to capture orthopedic studies reporting PROMIS measures through November 2018. Our inclusion criteria were use of PROMIS measures as an outcome or used to describe a population of patients in an orthopedic setting in patients ≥ 18 years of age. We excluded non-quantitative studies, reviews, and case reports. Our final search yielded 88 studies published from 2013 through 2018, with 57% (50 studies) published in 2018 alone. By body region, 28% (25 studies) reported PROMIS measures in the upper extremity (shoulder, elbow, hand), 36% (32 studies) reported PROMIS measures in the lower extremity (hip, knee, ankle, foot), 19% (17 studies) reported PROMIS measures in the spine, 10% (9 studies) reported PROMIS measures in trauma patients, and 6% (5 studies) reported PROMIS measures in general orthopedic patients. The majority of studies reported between one and three PROMIS domains (82%, 73 studies). The PROMIS Computerized Adaptive Test (CAT) approach was most commonly used (81%, 72 studies). The most frequently reported PROMIS domains were physical function (81%, 71 studies) and pain interference (61%, 54 studies). Our review found an increase in the reporting of PROMIS measures over the recent years. Utilization of PROMIS measures in orthopedic populations is clinically appropriate and can facilitate communication of outcomes across different provider types and with reduced respondent burden. The protocol for this systematic review was designed in accordance with the PRISMA guidelines and is registered with the PROSPERO database (CRD42018088260).


Novel spirohydantoin derivative as a potent multireceptor-active antipsychotic and antidepressant agent.
A series of novel spirohydantoin derivatives with arylpiperazinylbutyl moiety were synthesized and evaluated for serotonin 5-HT1A, 5-HT2A, 5-HT7 and dopamine D2 receptors. Based on these data, four compounds were selected for further binding affinity assays on dopamine D1, D3, D4, and 5-HT2C, 5-HT6 as well as adrenergic α1 and α2C receptors, which are involved in various CNS diseases such as schizophrenia, anxiety and/or depression. The compound 14, 1-{4-[4-(2-metoxyphe-nyl)piperazin-1-yl]butyl}-3',4'-dihydro-2H,2'H,5H-spiro[imidazolidine-4,1'-naphthalene]-2,5-dione, with the most promising functional profile, mixed 5-HT2A/D2 antagonist and 5-HT1A partial agonist, was selected. In the mouse d-amphetamine-induced locomotor hyperactivity model, compound 14 produced antipsychotic-like activity, which is devoid of cataleptogenic effects and in the forced swim test in mice, it showed a significant antidepressant-like effect unlike the reference drug aripiprazole.


Both H4K20 mono-methylation and H3K56 acetylation mark transcription-dependent histone turnover in fission yeast.
Nucleosome dynamics facilitated by histone turnover is required for transcription as well as DNA replication and repair. Histone turnover is often associated with various histone modifications such as H3K56 acetylation (H3K56Ac), H3K36 methylation (H3K36me), and H4K20 methylation (H4K20me). In order to correlate histone modifications and transcription-dependent histone turnover, we performed genome wide analyses for euchromatic regions in G2/M-arrested fission yeast. The results show that transcription-dependent histone turnover at 5' promoter and 3' termination regions is directly correlated with the occurrence of H3K56Ac and H4K20 mono-methylation (H4K20me1) in actively transcribed genes. Furthermore, the increase of H3K56Ac and H4K20me1 and antisense RNA production was observed in the absence of the histone H3K36 methyltransferase Set2 and histone deacetylase complex (HDAC) that are involved in the suppression of histone turnover within the coding regions. These results together indicate that H4K20me1 as well as H3K56Ac are bona fide marks for transcription-dependent histone turnover in fission yeast.


Repetitive Knee Bending and Synovitis in Individuals at Risk of and With Knee Osteoarthritis: Data From the Foundation for the National Institutes of Health Osteoarthritis Biomarkers Consortium.
To investigate associations between engagement in knee bending (stair climbing, kneeling, squatting, heavy lifting, getting in/out of a squatting position) and synovitis prevalence on noncontrast magnetic resonance imaging (MRI) in individuals at risk of and with knee osteoarthritis. We included baseline data from 594 participants (mean ± SD age 61.5 ± 8.9 years, 61% had Kellgren/Lawrence grade ≥2; 59% were female; mean ± SD body mass index was 30.7 ± 4.8 kg/m2 ) of the Osteoarthritis Biomarker Consortium Foundation for the National Institutes of Health project. Knee bending activities were queried by a standard questionnaire, and the severity of Hoffa synovitis and effusion synovitis (surrogate outcomes of synovitis) were graded using the MRI OsteoArthritis Knee Scoring system. Logistic regression was used, unadjusted and adjusted, for metabolic syndrome, physical activity level, and sex. A grade ≥1 defined synovitis prevalence, with a grade ≥2 cutoff implemented in sensitivity analyses. The prevalence of grade ≥1 Hoffa synovitis and effusion synovitis equaled 59% (n = 353) and 62% (n = 366), respectively. Adjusted for confounders, kneeling for ≥30 minutes during a single day was associated with grade ≥1 Hoffa synovitis prevalence (odds ratio [OR] 1.65 [95% confidence interval (95% CI) 1.11-2.47]). Participants engaging in this activity ≤1 day per week had greater odds for prevalent Hoffa synovitis than those who did not perform the activity (OR 1.88 [95% CI 1.11-3.18]). No other significant associations were found. Sensitivity analyses yielded similar findings. In this selected sample with a preponderance of grade ≥1 Hoffa and/or effusion synovitis on noncontrast MRI, only prolonged kneeling was associated with Hoffa synovitis prevalence. Replication in other samples is warranted.


Unusual presentations of cutaneous tuberculosis.
Cutaneous tuberculosis occurs in 1-2% of world cases of tuberculosis and more common in tropical countries. It presents with different clinical forms. Unusual clinical presentations are not uncommon and awareness of these will help in suspecting and managing these patients successfully. Lupus pernio like lupus vulgaris, tuberculosis of glans penis and lichen scrofulosorum on the distal parts of limbs are presented here because of their unusual clinical presentation.


Planar cell polarity protein localization in the secretory ameloblasts of rat incisors.
The localization of the planar cell polarity proteins Vang12, frizzled-3, Vang11, and Celsr1 in the rat incisors was examined using immunocytochemistry. The results showed that Vang12 was localized at two regions of the Tomes' processes of inner enamel-secretory ameloblasts in rat incisors: a proximal and a distal region. In contrast, frizzled-3 was localized at adherens junctions of the proximal and distal areas of inner enamel- and outer enamel-secretory ameloblasts, where N-cadherin and β-catenin were localized. frizzled-3 was also localized in differentiating inner enamel epithelial cells. Vang11 was localized sparsely in differentiating preameloblasts and extensively at the cell boundary of stratum intermedium. Celsr1 was not localized in ameloblasts but localized in odontoblasts extensively. These results suggest the involvement of planar cell polarity proteins in odontogenesis.


The electrical coupling of two simple oscillators: load and acceleration effects.
We consider two electrically coupled oscillators described by modified Fitzhugh-Nagumo equations. We study the relative influence of the individual cellular characteristics and the electrical coupling on the behavior of the coupled system. We show that, for similar oscillators, the load effect of the slow oscillator increases with the coupling strength. We prove that an asymmetry between the uncoupled bursters can accelerate the system with respect to the free cells, this effect depending on the characteristics of the coupling.


Outcomes After Arthroscopic Revision Bankart Repair: An Updated Systematic Review of Recent Literature.
To provide an update of recent literature with a specialized focus on clinical outcomes following arthroscopic revision Bankart repair (ARBR) by performing a systematic review of all available literature published between 2013 and 2020. A literature search reporting clinical outcomes after ARBR was performed. Criteria for inclusion consisted of original studies; Level of Evidence of I-IV; studies focusing on clinical outcomes after ARBR published between January 1, 2013, and January 4, 2021; studies reporting recurrent dislocation or instability rate after ARBR; reoperation/revision following ARBR, return to sport rates following ARBR; and patient-reported outcomes. The primary outcomes of interest were failure defined as recurrent instability or dislocation, return to sport rates, and patient-reported outcomes at follow-up. A large proportion of patients undergoing arthroscopic revision Bankart repair were male, ranging between 67.7% and 93.8%. Failure rate and return to sports rate ranged between 6.1% and 46.8% and 25.9% and 88.3%, respectively, when patients with significant or greater than 20% glenoid bone loss was excluded. Patient-reported outcome scores, which included American Shoulder and Elbow Surgeons, Simple Shoulder Test, and visual analog scale, saw significant improvement over mean follow-up of ranging 21.64 to 60 months. Both the failure rate and RTS rates after ARBR had a wide range, given the heterogeneity of the studies included, which varied in patient selection criteria pertaining to patients with greater than 20% glenoid bone. Although there have been advancements in arthroscopic techniques and a trend favoring arthroscopic stabilization procedures, there is a lack of consensus in recent literature for careful patient selection criteria that would minimize failure rates and maximize RTS rates after ARBR. Level IV, a systematic review of Level III-IV studies.


Evaluating the design and implementation fidelity of an adapted Plan-Do-Study-Act approach to improve health system performance in a Nigerian state.
The Plan-Do-Study-Act (PDSA) cycle is fundamental to many quality improvement (QI) models. For the approach to be effective in the real-world, variants must align with standard elements of the PDSA. This study evaluates the alignment between theory, design and implementation fidelity of a PDSA variant adapted for Nigeria's health system performance improvement. An iterative consensus building approach was used to develop a scorecard evaluating new conceptual indices of design and implementation fidelity of QI interventions (design and implementation index, defects and gaps) based on Taylor's theoretical framework. Design (adaptation) scores were optimal across all standard features indicating that design was well adapted to the typical PDSA. Conversely, implementation fidelity scores were only optimal with two standard features: prediction-based test of change and the use of data over time. The other features, use of multiple iterative cycles and documentation had implementation gaps of 17 % and 50 % respectively. This study demonstrates how both adaptation and implementation fidelity are important for success of QI interventions. It also presents an approach for evaluating other QI models using Taylor's PDSA assessment framework as a guide, which might serve to strengthen the theory behind future QI models and provide guidance on their appropriate use.


Improvement in Lesion Formation with Radiofrequency Energy and Utilization of Alternate Energy Sources (Cryoablation and Pulsed Field Ablation) for Ventricular Arrhythmia Ablation.
Current ablation systems rely on thermal energy to produce ablation lesions (heating: RF, laser and ultrasound, and cooling: cryo-thermia). While thermal ablation has been proven to be effective, there are several limitations: 1) relatively long procedural times; 2) high recurrence rate of ventricular arrhythmias; and 3) excessive heating potentially leading to serious complications, including steam pop (perforation), coronary arterial injury and thrombo-embolism. Pulsed field ablation (PFA)/irreversible electroporation (IRE) offers a unique non-thermal ablation strategy which has the potential to overcome these limitations. Recent pre-clinical studies suggest that PFA/IRE might be effective and safe for the treatment of cardiac arrhythmias.


Placenta percreta causing spontaneous uterine rupture and intrauterine fetal death in an unscared uterus: A case report.
Placenta percreta is a rare; a life-threatening disorder of placentation and one of the components of the placenta accreta spectrum. It can lead to uterine rupture, an obstetric catastrophe that can be associated with increased maternal and fetal morbidity and mortality. We present an unusual case of spontaneous uterine rupture due to placenta percreta in an unscarred uterus of a multiparous woman leading to spontaneous intrauterine fetal death. She presented with hypovolaemic shock following spontaneous rupture of the uterus and subsequent intra-peritoneal bleeding. Uterine rupture occurs commonly in a scarred uterus from some form of trauma or injudicious use of oxytocics. However, uterine rupture occurring in the absence of prior scar or use of oxytocics is a rarity. Placenta percreta is an unusual cause of uterine rupture and subsequent intra-uterine fetal death. Placenta percreta occurs when the uterine wall is invaded by the placenta up to the level of the serosa. A high index of suspicion and thorough review of the patient is required for making this diagnosis. Misdiagnosis is associated with dare consequences of increased maternal morbidity and mortality. Placenta percreta is a rare disorder of placentation that can cause uterine rupture which can easily be misdiagnosed. Prompt diagnosis and institution of the appropriate care can help prevent catastrophic outcomes as demonstrated in the case reported.


HDAC6 promotes cell proliferation and confers resistance to temozolomide in glioblastoma.
Histone deacetylases are considered to be among the most promising targets in drug development for cancer therapy. Histone deacetylase 6 (HDAC6) is a unique cytoplasmic enzyme that regulates many biological processes involved in tumorigenesis through its deacetylase and ubiquitin-binding activities. Here, we report that HDAC6 is overexpressed in glioblastoma tissues and cell lines. Overexpression of HDAC6 promotes the proliferation and spheroid formation of glioblastoma cells. HDAC6 overexpression confers resistance to temozolomide (TMZ) mediated cell proliferation inhibition and apoptosis induction. Conversely, knockdown of HDAC6 inhibits cell proliferation, impairs spheroid formation and sensitizes glioblastoma cells to TMZ. The inhibition of HDAC6 deacetylase activity by selective inhibitors inhibits the proliferation of glioblastoma cells and induces apoptosis. HDAC6 selective inhibitors can sensitize glioblastoma cells to TMZ. Moreover, we showed that HDAC6 mediated EGFR stabilization might partly account for its oncogenic role in glioblastoma. TMZ resistant glioblastoma cells showed higher expression of HDAC6 and more activation of EGFR. HDAC6 inhibitors decrease EGFR protein levels and impair the activation of the EGFR pathway. Taken together, our results suggest that the inhibition of HDAC6 may be a promising strategy for the treatment of glioblastoma.


Isolation and characterization of neural stem/progenitor cells in the subventricular zone of the naked mole-rat brain.
The naked mole-rat (NMR) is the longest-lived rodent with a maximum lifespan of more than 37 years and shows a negligible senescence phenotype, suggesting that tissue stem cells of NMRs are highly capable of maintaining homeostasis. However, the properties of NMR tissue stem cells, including neural stem cells (NSCs), are largely unclear. Neural stem/progenitor cells (NS/PCs) were isolated from the subventricular zone of the neonate NMR brain (NMR-NS/PCs) and cultured in neurosphere and adherent culture conditions. Expression of NSC markers and markers of neurons, astrocytes, and oligodendrocytes was analyzed by immunocytochemistry. In adherent culture conditions, the proliferation rate and cell cycle of NMR-NS/PCs were assessed and compared with those of NS/PCs from mice (mouse-NS/PCs). The DNA damage response to γ-irradiation was analyzed by immunocytochemistry and reverse transcription-quantitative PCR. NMR-NS/PCs expressed several NSC markers and differentiated into neurons, astrocytes, and oligodendrocytes. NMR-NS/PCs proliferated markedly slower than mouse-NS/PCs, and a higher percentage of NMR-NS/PCs than mouse-NS/PCs was in G0/G1 phase. Notably, upon γ-irradiation, NMR-NS/PCs exhibited a faster initiation of the DNA damage response and were less prone to dying than mouse-NS/PCs. NMR-NS/PCs were successfully isolated and cultured. The slow proliferation of NMR-NS/PCs and their resistance to DNA damage may help to prevent stem cell exhaustion in the brain during the long lifespan of NMRs. Our findings provide novel insights into the mechanism underlying delayed aging of NMRs. Further analysis of NMR tissue stem cells may lead to the development of new strategies that can prevent aging in humans.


Uptake, localization and clearance of quantum dots in ciliated protozoa Tetrahymena thermophila.
Protozoa as phagocytizing cells have been shown to integrate engineered nanoparticles (NPs), while the mechanism, dynamics and extent of such uptake are unclear. Here our fluorescence microscopy data showed that CdSe/ZnS quantum dots (QDs) with primary size of 12 nm were readily phagocytized into the food vacuoles of Tetrahymena thermophila in a time- and dose-dependent manner. Twenty hours after the exposure to QDs in sublethal concentration the clearance of the QDs from the cells was incomplete suggesting that phagocytosis of QDs into food vacuoles was not the only pathway of uptake by T. thermophila. This was further proven by the results that the inhibition of phagocytosis did not block the internalization of QDs into protozoans. This study provides a new insight into uptake and cellular trafficking of subtoxic concentrations of nanoparticles that may, due to prolonged retention times in the cells, pose risks by potentially becoming available to higher trophic levels.


Elicitation of jasmonate-mediated defense responses by mechanical wounding and insect herbivory.
Many plant immune responses to biotic stress are mediated by the wound hormone jasmonate (JA). Functional and mechanistic studies of the JA signaling pathway often involve plant manipulations that elicit JA production and subsequent changes in gene expression in local and systemic tissues. Here, we describe a simple mechanical wounding procedure to effectively trigger JA responses in the Arabidopsis thaliana rosette. For comparison, we also present a plant-insect bioassay to elicit defense responses with the chewing insect Trichoplusia ni. This latter procedure can be used to determine the effect of JA-regulated defenses on growth and development of insect herbivores.


Destruction of OPA from munitions demilitarization in supercritical water oxidation: kinetics of total organic carbon disappearance.
The destruction of OPA from munitions demilitarization has been accomplished in supercritical water oxidation (SCWO) with oxygen as oxidant in an isothermal continuous-flow reactor. The experiments were conducted at a temperature of 689-887 K and a fixed pressure of 25 MPa, with a residence time that ranged from 7 s to 14 s. The destruction efficiency was measured by total organic carbon (TOC) conversion. At the reaction condition, the initial TOC concentrations of OPA were varied from 1.41 mmol/L to 19.57 mmol/L and the oxygen concentrations were varied from 15.03 mmol/L to 81.85 mmol/L. Experimental data showed that all the TOC conversions were >80% under the above experimental conditions. The kinetics of TOC disappearance, which is essential for the design, optimization, and control of reliable commercial SCWO reactor was developed by taking into account the dependence of the oxidant and TOC concentration on the reaction rate. A global TOC disappearance rates expression was regressed from the data of 38 experiments, to a 95% confidence level. The resulting activation energy was determined to be 44.01 +/- 1.52 kJ/mol, and the pre-exponential factor was (1.67 +/- 0.45) x 10(2) L(1.14) mmol(-0.14) s(-1). The reaction orders for the TOC and the oxidant were 0.98 +/- 0.01 and 0.16 +/- 0.02, respectively.


Echocardiographic outcomes of self-expandable CoreValve versus balloon-expandable Edwards SAPIEN XT valves: the comparison of two bioprosthesis implanted in a single centre.
Transcatheter aortic valve implantation (TAVI) with self-expandable Medtronic CoreValve (MCV; Medtronic, Minneapolis, MN) or balloon-expandable Edwards SAPIEN XT valve (ESV; Edwards Lifesciences, Irvine, CA) has been widely used for treatment of high-risk patients with severe aortic stenosis (AS). There is limited data comparing the long-term hemodynamic performance of these two valves. Therefore, this study aimed to compare the short and long-term hemodynamic performance of TAVI with either MCV or ESV. A total of 78 patients who underwent TAVI in our center between June 01, 2012 and January 01, 2014 were enrolled in this retrospective study. For each of the patients we recorded the preprocedural echocardiographic data as well as the post-TAVI echocardiographic outcomes at day one, 6 months and 1 year. The MCV group had lower transaortic gradients than the ESV group, with respect to both maximum (13.4 ± 5.8 vs 18.7 ± 8.1 mmHg, p = 0.001) and mean values (6.5 ± 3.2 vs 9.4 ± 4.3 mmHg, p < 0.001) at post-TAVI day one. These values continued to be significantly lower in the MCV group during post-TAVI 6 months (p < 0.001) and post-TAVI 1 year follow-up (p < 0.05). A paravalvular leak (PVL, grade ≥2) was observed in 6.4 % of patients after TAVI; however, this value decreased over time, and there was no significant difference between the MVC and ESV groups (8.2 vs 3.4 %, p = 0.646). The MCV bioprosthesis was associated with lower transaortic gradients than those of the ESV throughout 1 year of follow-up. The incidence of PVL grade ≥2 in MCV and ESV was comparable.


The effect of distal arterial anatomy on the success of popliteal aneurysm repair.
The success of bypass grafting for popliteal aneurysms may be compromised by the natural history of these lesions. During a 9-year period 35 patients had repair of 48 popliteal aneurysms. Elective repair was performed in 26 limbs and the remaining 22 limbs required urgent or emergent treatment after aneurysm thrombosis (16) or embolic digital ischemia (six). Retrospective review of preoperative and intraoperative arteriograms and CT or ultrasound scans was done to define the relationship of distal runoff anatomy to initial symptoms and long-term grafting success during a mean follow-up of 48 months. Only five of 48 limbs (10%) had three continuous tibial vessels and a patent pedal arch. Twenty-six limbs (55%) had one or no named calf vessels and 11 limbs had incomplete pedal arches. When thrombosis or symptomatic embolism occurred, the anatomy was significantly worse; 15 of 22 such limbs (68%) had one or no patent tibial vessels and seven had incomplete pedal arches. Nineteen of 28 aneurysms examined by CT or ultrasound scanning showed intraluminal thrombus. Eight of 13 patent aneurysms with intraluminal thrombus had one or no patent tibial vessels. The 5-year graft patency rate was 74% for the entire study group. Bypass for repair of asymptomatic aneurysms had a patency rate of 91% at 5 years vs 54% for symptomatic patients (p less than 0.05). Patients with popliteal aneurysms often have advanced tibial disease that appears worse in those with symptoms. The occurrence of a patent popliteal aneurysm with intraluminal thrombus and advanced runoff disease suggests that chronic microembolism may be an etiologic factor in the tibial disease observed.(ABSTRACT TRUNCATED AT 250 WORDS)


Time for change? A national audit on bereavement care in intensive care units.
Bereaved ICU family members frequently experience anxiety, depression and post-traumatic stress disorder, which have been associated with significantly impaired quality of life. Recognising that their needs extend beyond the support provided by their friends and family, the Intensive Care Society had published in 1998 recommendations around bereavement care. The aim of the present national audit was to compare bereavement services in England against the nine recommendations set out by the Intensive Care Society guidelines. A telephone audit was carried out in all adult ICUs in England. A total of 144 NHS Trusts (179 ICUs) met the inclusion criteria and 113 responses were collected (78% of Trusts, 63% of individual ICUs). Although most ICUs provided administrative information (96% had an information booklet), training (53%), auditing (19%) and adequate facilities (27%) did not meet the recommended standards. Bereavement care is underdeveloped in English ICUs. This important but underreported topic should be prioritised in the critical care research agenda.


A diagnostic time-out to improve differential diagnosis in pediatric abdominal pain.
Pediatric abdominal pain is challenging to diagnose and often results in unscheduled return visits to the emergency department. External pressures and diagnostic momentum can impair physicians from thoughtful reflection on the differential diagnosis (DDx). We implemented a diagnostic time-out intervention and created a scoring tool to improve the quality and documentation rates of DDx. The specific aim of this quality improvement (QI) project was to increase the frequency of resident and attending physician documentation of DDx in pediatric patients admitted with abdominal pain by 25% over 6 months. We reviewed a total of 165 patients admitted to the general pediatrics service at one institution. Sixty-four history and physical (H&P) notes were reviewed during the baseline period, July-December 2017; 101 charts were reviewed post-intervention, January-June 2018. Medical teams were tasked to perform a diagnostic time-out on all patients during the study period. Metrics tracked monthly included percentage of H&Ps with a 'complete' DDx and quality scores (Qs) using our Differential Diagnosis Scoring Rubric. At baseline, 43 (67%) resident notes and 49 (77%) attending notes documented a 'complete' DDx. Post-intervention, 59 (58%) resident notes and 69 (68%) attending notes met this criteria. Mean Qs, pre- to post-intervention, for resident-documented differential diagnoses increased slightly (2.41-2.47, p = 0.73), but attending-documented DDx did not improve (2.85-2.82, p = 0.88). We demonstrated a marginal improvement in the quality of resident-documented DDx. Expansion of diagnoses considered within a DDx may contribute to higher diagnostic accuracy.


Preparation in the business and practice of medicine: perspectives from recent gynecologic oncology graduates and program directors.
Preparation in the business of medicine is reported to be poor across a number of specialties. No data exist about such preparation in gynecologic oncology training programs. Our objectives were to evaluate current time dedicated to these initiatives, report recent graduate perceptions about personal preparedness, and assess areas where improvements in training can occur. Two separate surveys were created and distributed, one to 183 Society of Gynecologic Oncology candidate members and the other to 48 gynecologic oncology fellowship program directors. Candidate member surveys included questions about perceived preparedness for independent research, teaching, job-hunting, insurance, and billing. Program director surveys assessed current and desired time dedicated to the topics asked concurrently on the candidate survey. Statistical analysis was performed using Chi-squared (or Fisher's exact test if appropriate) and logistic regression. Survey response rates of candidate members and program directors were 28% and 40%, respectively. Candidate members wanted increased training in all measures except retrospective protocol writing. Female candidates wanted more training on writing letters of intent (LOI) (p = 0.01) and billing (p < 0.01). Compared to their current schedules, program directors desired more time to teach how to write an investigator initiated trial (p = 0.01). 94% of program directors reported having career goal discussions with their fellows, while only 72% of candidate members reported that this occurred (p = 0.05). Recent graduates want more preparation in the non-clinical aspects of their careers. Reconciling program director and fellow desires and increasing communication between the two may serve to achieve the educational goals of each.


Subjective visibility depends on level of processing.
Is visual awareness graded or binary? Experimental work has provided support for both possibilities, leading to two coexisting but contradictory theoretical accounts. Here we propose a promising candidate factor through which to integrate both accounts: the depth of stimulus processing required by the task. We compared color identification (a low-level task) with numerical judgements (a high-level task) performed on the very same colored number stimuli. Psychophysical curves were analyzed for both objective discrimination performance and subjective visibility ratings on a trial-by trial basis. We observed a graded relationship between stimulus duration and visibility in the low-level task, but a more non-linear relationship in the high-level task. Both patterns of results have previously been consistently associated with the graded and the dichotomous account, respectively. Follow-up experiments that manipulate the level of processing can further unify previously inconsistent results, thus integrating two major theories of visual awareness.


[Professor V. A. Arnol'dov--a hygienist, scientist, university teacher, and initiator of sanitation in Russia (on his 150th birthday)].
The paper gives Professor V. A. Arnoldov's brief bibliographic data pertaining to the development of sanitation and zemstvo medicine in Bashkiria, Tatarstan, and the Saratov Province. It shows V. A. Arnoldov's scientific and pedagogical potential in the development of preventive medicine in Russia.


Gas volume corrections in intensive care unit: needed or pointless?
The conditions of temperature, pressure, and saturation in which respiratory gas volumes are expressed [standard temperature and pressure, dry (STPD), ambient temperature and pressure, saturated (ATPS), or body temperature and pressure, saturated (BTPS)] are physiologically relevant, but often ignored or unknown in clinical practice. In this study, we aimed to investigate whether and at which extent the gas volume corrections, either in natural or artificial lung, may alter key respiratory and metabolic variables and the possible clinical consequences. We primarily referred to the effects of gas volume corrections on three physiological variables: physiological dead space, venous admixture, and total CO2 production (V̇co2) during extracorporeal support. We used three physiological models in which calculations of these variables have been performed with and without correction of gas volumes, both in a theoretical model and in 448 patients. The lack of gas volume correction leads to an error in the computation of physiological dead space fraction between 0.05 and 0.15, both in the theoretical model and in the patient population. The venous admixture was minimally affected by the absence of correction (0.01-0.04 error). During extracorporeal support, if the V̇co2 of natural and membrane lung is expressed in different conditions, potentially large errors (0%-18.4%) may occur in the computation of total V̇co2 (V̇co2tot = V̇co2ML + V̇co2NL). This may lead to inappropriate settings of mechanical ventilation with higher plateau pressure. As the dead space and the CO2 sharing between natural and artificial lung are relevant both as prognostic index and as a guide for appropriate mechanical ventilation, their inappropriate computation may lead to erroneous categorization of the patients and inappropriate mechanical treatment.NEW & NOTEWORTHY Gas volume conditions are often ignored or unknown in the clinical practice. However, they could have relevance for the calculation of some key variables in ICU setting. This study shows that gas volume corrections are mostly relevant when assessing CO2 clearance, both in mechanical ventilation and during extracorporeal support, whereas irrelevant for oxygenation assessment of patients. Knowing when the appropriate corrections are needed allows to better understand patients' clinical conditions and to tailor the treatment.


Embedding clinical interventions into observational studies.
Novel approaches to observational studies and clinical trials could improve the cost-effectiveness and speed of translation of research. Hybrid designs that combine elements of clinical trials with observational registries or cohort studies should be considered as part of a long-term strategy to transform clinical trials and epidemiology, adapting to the opportunities of big data and the challenges of constrained budgets. Important considerations include study aims, timing, breadth and depth of the existing infrastructure that can be leveraged, participant burden, likely participation rate and available sample size in the cohort, required sample size for the trial, and investigator expertise. Community engagement and stakeholder (including study participants) support are essential for these efforts to succeed.


HIV-Associated Cryptococcal Meningitis: Bridging the Gap Between Developed and Resource-Limited Settings.
Cryptococcal meningitis is a major cause of HIV-associated morbidity and mortality worldwide. Most cases occur in low-income countries, where over half of patients die within 10 weeks of diagnosis compared to as few as 10 % of patients from developed countries. A host of factors, spanning the HIV care continuum, are responsible for this gap in treatment outcomes between developed and resource-limited settings. We explore factors responsible for this outcomes gap and describe low-cost, highly effective measures that can be implemented immediately to improve outcomes in resource-limited settings. We also explore health-system challenges that must be addressed to reduce mortality further, recent research in disease prevention, and novel short-course treatment regimens that, if efficacious, could be implemented in resource-limited settings where the cost of standard treatment regimens is currently prohibitive.


Temporal variation of airborne fungi in university library rooms and its relation to environmental parameters and potential confounders.
Indoor airborne fungi have been associated with adverse human health effects. Therefore, it is important to understand the causes of underlying variation in airborne fungi in indoor environments. This study consequently aimed to investigate the association between indoor fungi with temporal variation, environmental parameters, and potential confounders over 10 months in four library rooms using Andersen samplers. Indoor fungal concentrations peaked in October and were lowest in March in both stack rooms, whereas the highest concentrations in both reading rooms were observed in September with lowest concentrations in July. Nonparametric analyses revealed higher fungal concentrations in the rooms that were significantly associated with relative humidity ≥ 60%, PM2.5 ≥ 35 μg/m3, number of people ≥ 16, open windows, working air conditioners, and room area < 400 m2. Multiple linear regression modeling for the library building considering only continuous variables revealed that relative humidity, PM2.5, and the number of people were significant predictors of fungal concentrations. Additionally, the model with continuous and categorical variables suggested that relative humidity, PM2.5, the number of people, ceiling fan condition, window state, and air conditioner operating status were significant predictor variables of concentrations. Outdoor fungal concentrations were a significant predictor for the two models of indoor fungal concentrations for each room. Ceiling fan or air conditioner operation was associated with altered fungal particle concentrations. These results provide a deeper understanding of indoor air fungal quality.


Surgery of obstructive urolithiasis in ruminants.
Most cases of obstructive urolithiasis will require surgical intervention at some point during the treatment process. Fluid, anti-inflammatory, antibiotic, and acidifying therapies should be used in support of surgical intervention. Surgical technique may be chosen based on the characteristics of the individual case, including site of obstruction, location of the rupture, and value of the animal. Prevention remains the mainstay of urolithiasis management. Identification of a case of obstructive urolithiasis should trigger action for the affected individual and the entire herd or flock of origin.


Oligoclonal IgG in rabbits with experimental allergic encephalomyelitis: non-reactivity of the bands with sensitizing neural antigens.
Oligoclonal IgG bands have recently been reported to occur in cerebrospinal fluid (CSF) and serum of rabbits with experimental allergic encephalomyelitis (EAE). To examine the specificity of these bands, a) individual bands eluted from rabbit CSF and sera were tested by radioimmunoassay (RIA) for anti-MBP activity and b) rabbit sera were absorbed with the neuroantigens used for sensitization. RIA of eluates from sequential agar gel slices of the entire IgG region of serum or CSF from MBP sensitized rabbits showed that anti-MBP activity occurred throughout the IgG region and did not localize to specific band-containing fractions. Furthermore, there was no change in banding patterns following absorption of EAE rabbit sera with washed brain homogenates, soluble MBP or MBP conjugated to Sepharose beads. Therefore, our results indicate that the oligoclonal IgG response in EAE is not preferentially directed against the sensitizing neuroantigen, and we raise the possibility of nonspecific B cell activation.


Transcriptome data on maternal RNA of 24 individual zebrafish eggs from five sibling mothers.
Maternal mRNA that is present in the mature oocyte plays an important role in the proper development of the early embryo. To elucidate the role of the maternal transcriptome we recently reported a microarray study on individual zebrafish eggs from five different clutches from sibling mothers and showed differences in maternal RNA abundance between and within clutches, "Mother-specific signature in the maternal transcriptome composition of mature, unfertilized Eggs" [1]. Here we provide in detail the applied preprocessing method as well as the R-code to identify expressed and non-expressed genes in the associated transcriptome dataset. Additionally, we provide a website that allows a researcher to search for the expression of their gene of interest in this experiment.


[11C]olanzapine, radiosynthesis and lipophilicity of a new potential PET 5-HT2 and D2 receptor radioligand.
Olanzapine and its precursor desmethyl-Olanzapine were synthesized from malononitrile, propionaldehyde, 1-fluoro-2-nitrobenzene, and substituted piperazine in 4, 4, 5, and 5 steps with 35%, 32%, 26%, and 32% overall chemical yield, respectively. [(11)C]Olanzapine was prepared from desmethyl-Olanzapine with [(11)C]CH3OTf through N-[(11)C]methylation and isolated by HPLC combined with solid-phase extraction (SPE) in 40-50% radiochemical yield based on [(11)C]CO2 and decay corrected to end of bombardment (EOB), with 370-740 GBq/μmol specific activity at EOB. The calculated LogP (CLogP) value of [(11)C]Olanzapine is 3.39.


Antibacterial activity of the bladder mucosa.
Antibacterial activity of bladder mucosa is believed to be a host defense against infection. In this study we examined the antibacterial activity of the bladder mucosa without the effect of voiding. In addition we examined whether the property of adherence was advantageous for the organism in contact with the bladder mucosa. For this, three adhering and three nonadhering strains of E. coli were placed in contact with the bladder mucosa for 4 h in an in vivo rabbit model. E. coli grown in broth and applied to 32 bladders increased in titer by 1 log. E. coli grown in rabbit urine and applied to seven bladders increased in titer by 1.2 log. In contrast E. coli inoculated into control vials containing broth increased in titer by 2.3 log (P = 0.01). There was no significant difference in the titer between adhering and nonadhering strains of E. coli after 4 h of contact with the bladder mucosa. Bladder mucosa may have an inhibitory effect on bacterial growth (regardless of adherence characteristics) and with urine flow serves to prevent infection of the bladder urine.


Pulmonary embolization of needle fragments resulting from intravenous drug abuse.
We present two case reports of central embolization of needle fragments as a complication of intravenous drug abuse. In both cases the needle embolization occurred from a peripheral venous site to the right distal pulmonary vasculature. No attempts at removal were made. No complications developed as a result of these needle emboli.


Success Rates of Sentinel Lymph Node Mapping for Endometrial Cancer in Patients with Body Mass Index < 45 Compared with Body Mass Index ≥ 45.
The objective is to evaluate the rate of sentinel lymph node (SLN) mapping in patients with body mass index (BMI [kg/m2]) BMI ≥ 45 compared with < 45. A retrospective chart review. Three urban referral-based settings-1 academic and 2 community based. Patients age ≥ 18 years, with endometrial intraepithelial neoplasia or clinical stage 1 endometrial cancer who underwent robot-assisted total laparoscopic hysterectomy with attempted SLN mapping between January 2015 and December 2021. Robot-assisted total laparoscopic hysterectomy with attempted SLN mapping. A total of 933 subjects were included: 795 (85.2%) with BMI < 45 and 138 (14.8%) with BMI ≥ 45. Comparing the BMI < 45 with BMI ≥ 45 group, bilateral mapping was successful in 541 (68.1%) vs 63 (45.7%), respectively. Unilateral mapping was successful in 162 (20.4%) vs 33 (23.9%), respectively. Failure to map occurred in 92 (11.6%) vs 42 (30.4%) (p <.001), respectively. Exploratory analysis also suggested an inverse relationship between success rate of bilateral SLN mapping and BMI, with patients with BMI < 20 having bilateral SLN mapping rates of 86.5% and patients with BMI ≥ 61 having rates of 20.0%. The steepest decline in bilateral SLN mapping rates was from BMI group 46 to 50 compared to 51 to 55, at 55.4% to 37.5%, respectively. Adjusted odds ratio (compared with those with BMI < 30) for those in the BMI 30 to 44 group was 0.36 (95% confidence interval 0.21-0.60) and for those in the BMI ≥ 45 group was 0.10 (95% confidence interval 0.06-0.19). There is a statistically significant lower rate of SLN mapping in patients with a BMI ≥ 45 than BMI < 45. Understanding the success of SLN mapping in patients with morbid obesity is essential for preoperative counseling, surgical planning, and developing a risk-appropriate postoperative treatment plan.


Short-term plasticity of central benzodiazepine receptors in status epilepticus: case report.
(123)I-iomazenil SPECT is of value in determining an epileptogenic focus, however, transient uptake change has been rarely reported in epileptic disorders. A 78-year-old woman diagnosed as status epilepticus (SE) showed transient reduction in (123)I-iomazenil uptake within the epileptic foci on SPECT images during a couple of weeks. It suggests a seizure-related 'short-term' plasticity in the central benzodiazepine receptors and dynamic change in the regulatory mechanisms of inhibitory neurotransmitter system within the epileptic foci in patients with SE.


Fundoplication for gastroesophageal reflux and factors associated with the outcome 6 to 10 years after the operation: multivariate analysis of prognostic factors using the propensity score.
The impact from the mode of operation (partial vs total fundoplication) on long-term outcome after fundoplication still is unknown, although short-term randomized studies have not shown significant differences in the efficacy of reflux control. To obtain some insight concerning the long-term results, the data of a nonrandomized cohort were analyzed using propensity score statistics. For 134 patients who underwent laparoscopic fundoplication for gastroesophageal reflux disease (GERD), the time until recurrence of reflux symptoms was assessed. The impact of putative prognostic factors and the mode of operation (partial vs total fundoplication) on outcome were tested for significance using univariate and multivariate statistics, including the propensity score, correcting for nonrandomized treatment groups. The follow-up period was 60 to 123 months (median, 93 months). In this study, 45 patients had a partial (Toupet) fundoplication, and 89 patients underwent a total (Nissen) fundoplication. The rate of recurrence after 93 months (the median follow-up interval) was 14% after Nissen and 9% after Toupet fundoplication (nonsignificant difference) as estimated according to Kaplan and Meier. Massive acid exposure to the esophagus was associated with an increased risk of recurrence for 23% of the patients with a DeMeester score of 50 or higher, but only for 9% of the patients with less severe reflux (DeMeester score <50; p < 0.05). Multiple proportional hazard regression using the propensity score did not show additional significance for the variables of age, gender, presence of a Barrett esophagus, and mode of operation. The operation method did not have a significant impact on the efficacy of laparoscopic fundoplication in a cohort during a follow-up period of 60 to 123 months (median, 93 months).


Temporal Unfolding of Racial Ingroup Bias in Neural Responses to Perceived Dynamic Pain in Others.
In this study, we investigated how empathic neural responses unfold over time in different empathy networks when viewing same-race and other-race individuals in dynamic painful conditions. We recorded magnetoencephalography signals from Chinese adults when viewing video clips showing a dynamic painful (or non-painful) stimulation to Asian and White models' faces to trigger painful (or neutral) expressions. We found that perceived dynamic pain in Asian models modulated neural activities in the visual cortex at 100 ms-200 ms, in the orbitofrontal and subgenual anterior cingulate cortices at 150 ms-200 ms, in the anterior cingulate cortex around 250 ms-350 ms, and in the temporoparietal junction and middle temporal gyrus around 600 ms after video onset. Perceived dynamic pain in White models modulated activities in the visual, anterior cingulate, and primary sensory cortices after 500 ms. Our findings unraveled earlier dynamic activities in multiple neural circuits in response to same-race (vs other-race) individuals in dynamic painful situations.


Helping social workers address the educational needs of foster children.
The main aim was to evaluate the effectiveness of the Education Initiative, an intervention program in one of the largest urban counties in the US seeking to increase the responsiveness of social workers to the educational needs of foster children. A pre-post test control group design was used. Data from case files and social workers were examined at the start of the project and 18 months later to determine changes in social workers' (a) knowledge and practices regarding school programs and services and (b) maintenance of educational records for children on their caseloads. Data were analyzed from approximately 300 case files and over 200 questionnaires completed by social workers. Findings indicated that social workers who received training and had access to an education liaison (1) increased their knowledge about the school system and (2) were more likely to gather current educational data and comment on schooling needs in the case files. On questionnaires measuring knowledge and practices, supervisory social workers showed no gains while case workers increased their levels of knowledge and involvement. Evaluative data from both sources support the effectiveness of this collaborative model between the school and child welfare agency for addressing the educational needs of foster youth. Social workers in the pilot offices knew more and focused more on the school experience of youth on their caseloads than workers in the control offices. They reported more educational information in the case files and solicited current progress reports from the schools. Discrepancies between worker and school reported performance data raise questions as to whether social workers by themselves are the most effective advocates for foster children.


Identification of sucrose-regulated genes in cultured rice cells using mRNA differential display.
In order to get more information about carbon metabolite regulation pathways, cloning and sequence analysis of sucrose-regulated genes from rice-suspension-cultured cells were performed. We used a new method, mRNA differential display, to screen differentially expressed genes under conditions of 3% and no sucrose in the cultured medium. Six candidate clones were identified and sequenced. Clones SI1 and SI2 were repressed by sucrose starvation, while clones SR1, SR2, SR3 and SR4 were induced by sucrose starvation. Nucleotide sequence analysis showed that clone SR2 has 94.8% homology to the salT gene, and clones SI1 and SR3 show 88.3 and 96.9% identity, respectively, to partial cDNA sequences in the GenBank database. The results suggest that mRNA differential display provides an easy and quick way to clone genes involved in the carbon metabolite regulation pathway.


Paraneoplastic syndrome and intrathoracic Castleman disease.
We report two cases of intrathoracic Castleman disease presenting with paraneoplastic syndrome. Patient 1 was a 10-year-old girl with short stature. She was found to have delayed bone age, slow growth velocity, and iron-deficiency anemia, which was refractory to treatment. Thrombocytosis and hypergammaglobulinemia were later detected. Chest X-ray revealed a hilar mass. Patient 2 was a 14-year-old boy who had severe cough, progressive mucocutaneous erosion, and dermatitis. Chest X-ray showed a mediastinal mass. Sections of skin biopsy showed findings consistent with pemphigus disease. In each case, the histological diagnosis of Castleman disease was made.


Phosphorus release from intact soil monoliths of manure-amended fields under simulated snowmelt flooding.
Anaerobic conditions developed in soils with flooding can enhance the release of soil P to overlying water, but little information is available for soils with a long history of manure application. We examined the P release from manure-amended soils under simulated snowmelt flooding. Intact monoliths from manured (solid swine manure [SSM] or liquid swine manure [LSM]) and unamended (control) field plots were collected from Carman, Manitoba. Monoliths were frozen for 7 d, thawed, flooded, and incubated at 4 ± 1 °C. Redox potential, pH, and concentrations of dissolved reactive P (DRP), Ca, Mg, Fe, and Mn in pore water and floodwater were determined weekly up to 56 d after flooding (DAF) and at 84 DAF. Redox potential decreased with DAF with a greater and more rapid decrease in SSM (from ∼300 to <0 mV by 84 DAF) compared with LSM and control (∼100 mV by 84 DAF). Pore water and floodwater DRP concentrations were significantly greater in manured treatments than in the control at all DAFs and in SSM than in LSM for most DAF. Whereas floodwater DRP concentrations remained relatively stable in the control treatment, concentrations in manured treatments increased substantially from the onset of flooding to 35-42 DAF (threefold to fourfold increase) and remained relatively stable thereafter. Significantly greater P release from SSM- than from LSM-treated monoliths was due to greater input of P and the higher organic matter content in SSM-treated soils. These favored the rapid development of anaerobic conditions that further induced P release.


Response to hepatitis A vaccine in HIV patients in the HAART era.
We evaluated the development of hepatitis A antibody after vaccination in a large cohort of patients studied in a clinical setting after the introduction of HAART. Overall, 130 of 214 vaccinated individuals developed hepatitis A antibody. In a multivariate analysis, only the CD4 cell count at the time of vaccination was associated with an absence of response. The lack of association with the nadir CD4 cell count suggests that patients will respond to vaccine after immunological reconstitution in response to HAART.


Melatonin modulates endometrial decidualization via NOTCH1-NRF2-FOXO1-GSH pathway†.
Melatonin is important for oocyte maturation, fertilization, early embryonic development, and embryo implantation, but less knowledge is available regarding its role in decidualization. The present study found that melatonin did not alter the proliferation of human endometrial stromal cells (ESCs), as well as cell cycle progress, but suppressed stromal differentiation after binding to the melatonin receptor 1B (MTNR1B), which was visualized in decidualizing ESCs. Further analysis evidenced that application of melatonin resulted in the diminishment for NOTCH1 and RBPJ expression. Supplementation of recombinant NOTCH1 protein (rNOTCH1) counteracted the impairment of stromal differentiation conferred by melatonin, while the addition of the NOTCH signaling pathway inhibitor DAPT aggravated the differentiation progress. Meanwhile, melatonin might restrain the expression and transcriptional activity of nuclear factor erythroid 2-related factor 2 (NRF2), whose blockage accelerated the fault of stromal differentiation under the context of melatonin, but this restraint was subsequently ameliorated by rNOTCH1. Forkhead box O 1 (FOXO1) was identified as a downstream target of melatonin in decidualization. Repression of NRF2 antagonized the retrieval of rNOTCH1 due to aberrant FOXO1 expression elicited by melatonin. Moreover, melatonin brought about the occurrence of oxidative stress accompanied by an obvious accumulation of intracellular reactive oxygen species and a significant reduction in glutathione (GSH) content, as well as enzymatic activities of glutathione peroxidase and glutathione reductase, whereas supplementation of rNOTCH1 improved the above-mentioned effects. Nevertheless, this improvement was disrupted by the blockage of NRF2 and FOXO1. Furthermore, addition of GSH rescued the defect of stromal differentiation by melatonin. Collectively, melatonin might impair endometrial decidualization by restraining the differentiation of ESCs dependent on NOTCH1-NRF2-FOXO1-GSH pathway after binding to the MTNR1B receptor.


Acute leukemias of childhood: pathologic features.
Major advances have been made in the diagnosis and therapy of childhood leukemia over the last two decades (Zuelzer and Flatz, 1960; Miller, 1980; Steinhorn and Myers, 1981; Kobrinsky et al., 1980). Sophisticated phenotypic analysis of these leukemias has partially revealed their complexity; there are at least three to five distinct clinical pathological entities in the childhood leukemia groups. Improved therapy has resulted in marked increases in survival and cures. Less attention has been given to the pathology of childhood leukemia, although the frequency and complexity of tissue examinations have greatly increased. In particular, the pathologic manifestation of relapses, detection of focal leukemia after treatment and the pathology of complications have not been systematically reviewed. Our goal was to look at all of the pathologic manifestations of leukemia and to relate these manifestations to homogeneous patient populations grouped by their type of leukemia. Therefore all of the cytologic, hematologic, histopathologic, ultrastructural and immunologic materials on a consecutive group of leukemic children seen at Vanderbilt from 1970 to 1981 were reviewed; pathologic manifestations of leukemia were then analysed in relationship to diagnostic categories.


Assessment of the impact of isolated coronary artery ectasia on left ventricular functions with 3D speckle-tracking echocardiography.
Coronary artery ectasia (CAE) is an angiographic definition of coronary artery pathology in which the diameter of the ectatic segment measures more than 1.5 times the diameter of an adjacent healthy reference segment. No previous study has reported on the use of 3D-STE for assessing the left ventricular (LV) functions in patients with isolated CAE. As a result of this, we aimed to evaluate the effects of isolated CAE on LV functions using 3D-STE in the present study. Ninety-one patients with isolated CAE and 90 controls who proved to have normal coronary angiograms were enrolled to the study. 3D-STE was performed and GLS, GCS, GAS, and GRS were obtained for every subject after coronary angiography. The mean age of the patients was 61.75 ± 10.02 years, and 71.8% were male. GLS, GCS, GAS, and GRS were significantly depressed in the isolated CAE group than in the control group (P < .001; P < .001; P = .001; and P = .001, respectively). ROC analyses were performed to find out the ideal strain cut off values to predict the presence of isolated CAE. A GLS value of >-16 has 92.1 % sensitivity, 88.5 % specificity; and a GCS value of >-20 has 86.7 % sensitivity, 89.2 % specificity to detect the presence of isolated CAE. Isolated CAE has a considerable negative effect on LV functions as evaluated by 3D-strain parameters, and 3D-STE could be an effective method to detect early stage myocardial impairment in patients with isolated CAE.


Complete endocardial cushion defect: the late result of repair using the single-patch technique.
Twenty-two patients have been followed for between 1 and 105 months after repair of a complete endocardial cushion defect. The mean period of follow-up is 3.5 years. The single-patch technique was used in every patient. The mitral valve was repaired with buttressed sutures in those seen more recently. The mean age at the time of operation was 15 months. Early in this experience, 4 patients had severe mitral valve incompetence after an initially satisfactory repair. In none of those patients had the mitral valve been repaired with pledgeted sutures. Two of those patients survived reoperation, and 2 died before a second operation could be performed. Every mitral valve is now repaired with pledgeted sutures, and there have been no failures of the mitral valve reconstruction. Each patient has been followed by the same pediatric cardiologist every 6 to 12 months after operation. The vast majority (17 of 20) are asymptomatic. Twelve have no mitral valve incompetence, and the remainder have only trivial or mild incompetence. Clinically, the pulmonary artery hypertension has resolved in 19 of 20 patients. Each patient remains in normal sinus rhythm. The long-term results following repair of complete endocardial cushion defect with the single-patch technique are excellent, but pledgeted sutures should be used in the mitral valve repair to insure its integrity.


Thermal fluctuations between conformational substates of the Fe(2+)-HisF8 linkage in deoxymyoglobin probed by the Raman active Fe-N epsilon (HisF8) stretching vibration.
We have measured the VFe-His Raman band of horse heart deoxymyoglobin dissolved in an aqueous solution as a function of temperature between 10 and 300 K. The minimal model to which these data can be fitted in a statistically significant and physically meaningful way comprises four different Lorentzian bands with frequencies at 197, 209, 218, and 226 cm-1, and a Gaussian band at 240 cm-1, which exhibit halfwidths between 10 and 12.5 cm-1. All these parameters were assumed to be independent of temperature. The temperature dependence of the apparent total band shape's frequency is attributed to an intensity redistribution of the subbands at omega 1 = 209 cm-1, omega 2 = 218 cm-1, and omega 3 = 226 cm-1, which are assigned to Fe-N epsilon (HisF8) stretching modes in different conformational substrates of the Fe-HisF8 linkage. They comprise different out-of-plane displacements of the heme iron. The two remaining bands at 197 and 240 cm-1 result from porphyrin modes. Their intensity ratio is nearly temperature independent. The intensity ratio I3/I2 of the vFe-His subbands exhibits a van't Hoff behavior between 150 and 300 K, bending over in a region between 150 and 80 K, and remains constant between 80 and 10 K, whereas I2/I1 shows a maximum at 170 K and approaches a constant value at 80 K. These data can be fitted by a modified van't Hoff expression, which accounts for the freezing into a non-equilibrium distribution of substates below a distinct temperature Tf and also for the linear temperature dependence of the specific heat of proteins. The latter leads to a temperature dependence of the entropic and enthalpic differences between conformational substates. The fits to the intensity ratios of the vFe-His subbands yield a freezing temperature of Tf = 117 K and a transition region of delta T = 55 K. In comparison we have utilized the above thermodynamic model to reanalyze earlier data on the temperature dependence of the ratio Ao/A1 of two subbands underlying the infrared absorption band of the CO stretching vibration in CO-ligated myoglobin (A. Ansari, J. Berendzen, D. Braunstein, B. R. Cowen, H. Frauenfelder, M. K. Kong, I. E. T. Iben, J. Johnson, P. Ormos, T. B. Sauke, R. Scholl, A. Schulte, P. J. Steinbach, R. D. Vittitow, and R. D. Young, 1987, Biophys. Chem. 26:237-335). This yields thermodynamic parameters, in particular the freezing temperature (Tf = 231 K) and the width of the transition region (AT =8 K), which are significantly different from the corresponding parameters obtained from the above vFe-His data, but very close to values describing the transition of protein bound water from a liquid into an amorphous state. These findings and earlier reported data on the temperature dependence exhibited by the Soret absorption bands of various deoxy and carbonmonoxymyoglobins led us to the conclusion that the fluctuations between conformational substates of the heme environment in carbonmonoxymyoglobin are strongly coupled to motions within the hydration shell, whereas the thermal motions between the substates of the Fe-HisF8 linkage in deoxymyoglobin proceed on an energy landscape that is mainly determined by the intrinsic properties of the protein. The latter differ from protein fluctuations monitored by Mossbauer experiments ondeoxymyoglobin crystals which exhibit a strong coupling to the protein bound water and most probably reflect a higher tier in the hierarchical arrangement of substates and equilibrium fluctuations.


Computational modeling of oligonucleotide positional densities for human promoter prediction.
The gene promoter region controls transcriptional initiation of a gene, which is the most important step in gene regulation. In-silico detection of promoter region in genomic sequences has a number of applications in gene discovery and understanding gene expression regulation. However, computational prediction of eukaryotic poly-II promoters has remained a difficult task. This paper introduces a novel statistical technique for detecting promoter regions in long genomic sequences. A number of existing techniques analyze the occurrence frequencies of oligonucleotides in promoter sequences as compared to other genomic regions. In contrast, the present work studies the positional densities of oligonucleotides in promoter sequences. The analysis does not require any non-promoter sequence dataset or any model of the background oligonucleotide content of the genome. The statistical model learnt from a dataset of promoter sequences automatically recognizes a number of transcription factor binding sites simultaneously with their occurrence positions relative to the transcription start site. Based on this model, a continuous naïve Bayes classifier is developed for the detection of human promoters and transcription start sites in genomic sequences. The present study extends the scope of statistical models in general promoter modeling and prediction. Promoter sequence features learnt by the model correlate well with known biological facts. Results of human transcription start site prediction compare favorably with existing 2nd generation promoter prediction tools.


Cruciform DNA Sequences in Gene Promoters Can Impact Transcription upon Oxidative Modification of 2'-Deoxyguanosine.
Sequences of DNA typically adopt B-form duplexes in genomes, although noncanonical structures such as G-quadruplexes, i-motifs, Z-DNA, and cruciform structures can occur. A challenge is to determine the functions of these various structures in cellular processes. We and others have hypothesized that G-rich G-quadruplex-forming sequences in human genome promoters serve to sense oxidative damage generated during oxidative stress impacting gene regulation. Herein, chemical tools and a cell-based assay were used to study the oxidation of guanine to 8-oxo-7,8-dihydroguanine (OG) in the context of a cruciform-forming sequence in a gene promoter to determine the impact on transcription. We found that OG in the nontemplate strand in the loop of a cruciform-forming sequence could induce gene expression; conversely when OG was in the same sequence on the template strand, gene expression was inhibited. A model for the transcriptional changes observed is proposed in which OG focuses the DNA repair process on the promoter to impact expression. Our cellular and biophysical studies and literature sources support the idea that removal of OG from duplex DNA by OGG1 yields an abasic site (AP) that triggers a structural shift to the cruciform fold. The AP-bearing cruciform structure is presented to APE1, which functions as a conduit between DNA repair and gene regulation. The significance is enhanced by a bioinformatic study of all human gene promoters and transcription termination sites for inverted repeats (IRs). Comparison of the two regions showed that promoters have stable and G-rich IRs at a low frequency and termination sites have many AT-rich IRs with low stability.


[Expression of ICAM-1/LFA-1 in the pocket area of adult periodontitis].
To study the expression of ICAM-1/LFA-1 in the pocket area of adult periodontitis. Expressions of ICAM-1/LFA-1 in adult gingival of periodontitis and healthy subjects were studied by alkaline phosphorylase-antialkaline phosphorylase technique. Junctional epithelium and apical part of sulcus epithelium expressed ICAM-1 in both adult periodontitis and healthy gingiva, showing an ICAM-1 gradient change with maximal staining at tooth aspect and weaker staining in the basal layer of keratinocytes. Significantly more LFA-1 positive leukocytes were observed in connective tissues and within pocket epithelium in adult periodontitis than those in healthy gingiva. ICAM-1/LFA-1 may provide important adhesion pathway for leukocytes migration into gingiva sulcus in adult periodontitis lesions.


A retrospective analysis of risk factors for massive hemoptysis in patients with bronchiectasis.
Massive hemoptysis is a common and fatal complication of bronchiectasis. However, the risk factors for massive hemoptysis in patients with bronchiectasis have not yet been reported. This study investigated the potential risk factors for massive hemoptysis in patients with bronchiectasis. This retrospective study included patients with bronchiectasis and their data were obtained from medical records. The risk factors for massive hemoptysis were evaluated by multivariate analysis of patient characteristics, medical history, and computed tomography imaging data, including the number of lesions, lesion location, and laboratory findings. Among 379 patients, 61 (16.09%) experienced severe hemoptysis. Multivariate analysis revealed that diabetes (odds ratio (OR) 2.885; 95% confidence interval (CI) 1.009-8.247), lesions involving two lobes (OR 4.347; 95% CI 1.960-9.638) and three lobes (OR 2.787; 95% CI 1.055-7.363) were significant predictors of severe hemoptysis. However, a disease course between 1 and 5 years (OR 0.300; 95% CI 0.112-0.801) and involvement of the left lower lobe (OR 0.394; 95% CI 0.196-0.793) were protective factors for the prevention of massive hemoptysis. Lesions in the right upper lobe were more likely to cause massive hemoptysis (OR 1.458) than involvement of other lobes. Diabetes and lesions involving two and three lobes, were risk factors for massive hemoptysis in patients with bronchiectasis. Disease duration between 1 and 5 years and involvement of the left lower lobe were protective factors, while lesions in the right upper lobe had a stronger relationship with massive hemoptysis in patients with bronchiectasis.


The Saccharomyces cerevisiae MSI4 gene encodes the yeast counterpart of component A of Rab geranylgeranyltransferase.
We describe the isolation of MSI4 as a multicopy suppressor of ira1 (inhibitory regulator of Ras). We find it encodes a 66-kDa protein with three regions similar to component A of Rab geranylgeranyltransferase and Rab guanine nucleotide dissociation inhibitor. A gene disruption experiment showed that MSI4 is essential for cell growth. To investigate its functions further, we constructed a strain whose MSI4 is driven by the GAL1 promoter. This strain accumulated the endoplasmic reticulum precursor form of a vacuolar enzyme, carboxypeptidase Y, under the restrictive conditions. Moreover, we showed that the activity of geranylgeranyl-transferase for Ypt1p was very low in the mutant shifted to glucose medium. We propose that the MSI4 gene product is a yeast counterpart of component A of Rab geranylgeranyltransferase that is essential for Ypt1p to localize on membranes.