Sequence variability in the putative coding region of TT virus: evidence for two rather than several major types.
Recently a new human virus, TT virus (TTV) was identified in the serum of a patient with post-transfusion hepatitis of unknown aetiology. Comparative sequence analysis of a 222 nt fragment of ORF 1 of TTV was performed to assess the genomic variability of this virus. Phylogenetic analysis of the nucleotide sequences of 76 TTV isolates collected in 17 countries segregated them into two major groups: TTV 1 and TTV 2. The TTV 1 group comprised two distinct subgroups, which corresponded to previously described TTV subtypes 1a and 1b. The TTV 2 group was separated into four main branches, two of which included sequences previously provisionally attributed as TTV types 2 and 3. Bootstrap resampling, however, did not support the reliability of this grouping, suggesting that the isolates in the TTV 2 group should be considered as subtypes of a single type rather than different TTV types.


Immobilization of proline activated lipase within metal organic framework (MOF).
The immobilization of enzyme with enhanced catalytic activity is the major challenge. In this work, we have activated the lipase in the presence of proline and successfully immobilized into zeolitic imidazolate framework (ZIF)-8 by biomineralization method. The prepared lipase-proline MOF exhibited 135% enhanced catalytic activity as compared to free counterpart. Further, it exhibited four-fold improved thermal stability with respect to native enzyme after immobilization. In Michaelis-Menten kinetic studies, Km values for lipase-proline MOF were found to be lower, whereas, it exhibited higher Vmax than lipase-MOF and free lipase. The lipase-MOF and lipase-proline MOF were showed 56% and 72% residual activity, respectively after six cycles of reuse.


A bacteriocin-mediated antagonism by Enterococcus faecium BC25 against ruminal Streptococcus bovis.
A bacteriocin-like activity produced by Enterococcus faecium BC25, isolated from the the rumen of a cow, was partially purified and characterized. The active substance was prepared by ammonium sulfate and chloroform/methanol precipitation of culture supernatant. The bacteriocin was a protein of molecular mass 17.5 kDa. Activity was inactivated by trypsin and proteinase K. The bacteriocin BC25 inhibited growth of amylolytic ruminal strains of Streptococcus bovis, including S. bovis AO 24/85. Results showed that bacteriocine substance BC25 has a bacteriostatic effect when present in concentrations exceeding 125 AU/ml. Agar overlays and batch culture growth experiments proved that E. faecium BC25 was producing bacteriocin that inhibited the growth of S. bovis.


Remarkable tethering effect on DNA cleavage of propargylic sulfone conjugates with intercalating moieties.
A number of novel propargylic sulfone conjugates 3 and 4 with intercalating moieties were synthesized and evaluated for DNA cleavage activity through nucleic base alkylation. A remarkable enhancement in DNA cleaving potency was observed with those conjugates 3 possessing a suitable spacer, a right attachment point at the aromatic ring, and a good intercalator.


Cytological localization of inverted repeated DNA sequences in Vicia faba.
Inverted repeated DNA sequences have been isolated from sheared Vicia faba DNA by hydroxylapatite column chromatography, treated with nuclease S1, tritiated by the nick translation method and hybridized in situ on squashes of Vicia faba root tips. Silver grains appear grouped in a rather limited portion of interphase nuclei and form a sort of band across them. The central regions of metaphase chromosomes are preferentially labeled, labeling being excluded from telomeres, centromeres and secondary constrictions. These results are briefly discussed in relation to those obtained in other species and the functional significance of inverted repeats.


The CDK inhibitor p21 is a novel target gene of ATF4 and contributes to cell survival under ER stress.
Activating transcription factor 4 (ATF4) is well known for its role in the endoplasmic reticulum (ER) stress response. ATF4 also transcriptionally induces multiple effectors that determine cell fate depending on cellular context. In addition, ATF4 can communicate both pro-apoptotic and pro-survival signals. How ATF4 mediates its prosurvival roles, however, requires further investigation. Here, we report that the CDK inhibitor p21 is a novel target gene of ATF4. We identified two ATF4-responsive elements, one of which directly binds ATF4, within the first intron of the p21 gene. Importantly, overexpression of p21 enhances cell survival following ER stress induction, while p21 knockdown increases cell death. These results suggest that p21 induction plays a vital role in the cellular response to ER stress and indicate that p21 is a prosurvival effector of ATF4.


[Topical treatment of peri-implantitis with metronidazole dental gel 25%. Clinical analysis and microbiological control].
Aim of this research is to verify the efficacy of metronidazole dental gel 25%, used as a topical antibiotic for the treatment of peri-implantitis. The efficacy of the medicine in the starting phase of the disease (mucositis), as well as during peri-implantitis involving bone-bearing loss has been evaluated. Twenty patients were chosen as a sample for the study. After careful evaluation of the several protocols concerning peri-implantitis treatment, some diagnostic clinical parameters have been recorded for each patient and metronidazole dental gel 25% (Elyzol Cabon) has been administered in two applications at perimplants pocket (one after a week). For each patient 3 microbiological drawings were made for the evaluation of the bacterial population around the implant site. From the microbiological examination a decrease of Gram- and an increase of Gram+ which returned in almost all the patients to normal levels have been observed; moreover, in all the patients a gradual decrease of PMN, resolution index of the inflammatory process was obtained, confirmed also by a remarkable improvement of all the observed diagnostic parameters, except for the peri-implants bone radiotransparency which was unchanged. The research showed that the peri-implant diseases can be positively resolved by using the metronidazole dental gel 25% topical antibiotic. This drug led to a 60-70% decrease of Gram-, and 40-50% increase for Gram+, bringing these back to normal values in almost all the patients. Moreover, a good recovery of the peri-implants soft tissues has been observed.


Protective effect of supplemental low intensity white light on ultraviolet-B exposure-induced impairment in cyanobacterium Spirulina platensis: formation of air vacuoles as a possible protective measure.
Intact trichomes of Spirulina platensis were exposed to 1-5 h of low (0.2 mW cm(-2)) or high (0.6 mW cm(-2)) intensity UV-B (280-320 nm) radiation, alone or with photosynthetically active radiation (PAR) of supplemental 50 muE m(-2) s(-1) white light (WL). The mitigating effect of supplemental WL on UV-B induced alterations in Spirulina were investigated by monitoring time-dependent change in photosystem (PS) II mediated O(2) evolution, absorption, circular dichroism (CD) spectra, and ultrastructure. At low intensity, UV-B induced loss in PS II-catalyzed O(2) evolution, but caused no change in the absorption spectrum. At high intensity, UV-B caused a decrease in absorption by phycobilisomes (PBsomes), which was only partly prevented by the presence of low-intensity supplemental WL. The CD spectral analysis revealed that UV-B exposure caused time-dependent enhancement of the negative psi-type bands at 452 and 689 nm, reflecting alterations in the macroaggregation of chlorophyll-protein complexes. This enhancement of negative PS II-type bands was substantially arrested by the presence of supplemental WL exposure, even when UV-B exposure was continued for 5 h. These changes in UV-B-induced CD spectrum suggest alterations in the antenna structure of Spirulina involving both PBsomes and Chlorophyll a. Thus, supplemental low intensity WL arrests, to large extent, the macroaggregation of pigment-protein complexes. Furthermore, the electron micrographs of Spirulina revealed that UV-B exposure caused disorganization of the cellular ultrastructure, while the inclusion of supplemental WL enhanced the formation of air vacuoles in Spirulina. We suggest that the formation of vacuoles by supplemental WL is a protective feature against UV-B.


Folie a deux versus genetically driven delusional disorder: case reports and nosological considerations.
Folie a deux is thought to occur in persons who live close together, so that delusions of the primarily ill patient are adopted by the partner. Alternatively, if both patients are related, they may share the same genetically driven psychiatric illness. We present the cases of two psychotic couples (monozygotic twins and a mother and daughter pair, respectively), in which both subjects featured an almost identical psychopathological syndrome with the same delusional content and thus fulfilled the diagnostic criteria of folie a deux. When Leonhard's diagnostic system was applied, all four patients were found to suffer from affect-laden paraphrenia, arguing against the presence of folie a deux in those dyads. Thus, thorough psychopathological examination in suspected folie a deux settings can help to obtain valid diagnoses.


Safety of rituximab in a patient with chronic renal failure and low-grade non-Hodgkin lymphoma.
An 82-year-old man, with known chronic renal failure and a creatinine clearance (CrCl) of 18 mL/min, was diagnosed with low-grade marginal zone non-Hodgkinlymphoma (NHL) of his right orbit. He received rituximab as single treatment (375 mg/m 2 every 3 weeks). After eight cycles, almost complete response was obtained, rituximab was extremely well-tolerated, and his creatinine levels remained stable throughout the whole treatment.


Population genomic analysis reveals domestication of cultivated rye from weedy rye.
Rye (Secale cereale) is an important crop with multiple uses and a valuable genetic resource for wheat breeding. However, due to its complex genome and outcrossing nature, the origin of cultivated rye remains elusive. The geneticist N.I. Vavilov proposed that cultivated rye had been domesticated from weedy rye, rather than directly from wild species like other crops. Unraveling the domestication history of rye will extend our understanding of crop evolution and upend our inherent understanding of agricultural weeds. To this end, in this study we generated the 8.5 Tb of whole-genome resequencing data from 116 worldwide accessions of wild, weedy, and cultivated rye, and demonstrated that cultivated rye was domesticated directly from weedy relatives with a similar but enhanced genomic selection by humans. We found that a repertoire of genes that experienced artificial selection is associated with important agronomic traits, including shattering, grain yield, and disease resistance. Furthermore, we identified a composite introgression in cultivated rye from the wild perennial Secale strictum and detected a 2-Mb introgressed fragment containing a candidate ammonium transporter gene with potential effect on the grain yield and plant growth of rye. Taken together, our findings unravel the domestication history of cultivated rye, suggest that interspecific introgression serves as one of the likely causes of obscure species taxonomy of the genus Secale, and provide an important resource for future rye and wheat breeding.


Cadmium(II)-113 NMR studies of the mechanism of metal ion activation of yeast enolase.
Yeast enolase binds one mole of 113Cd2+ per subunit at a site that consists of all oxyligands in a distorted octahedral environment. This "conformational" metal ion's environment undergoes further distortion on addition of substrate/product or analogs. At pH's below the optimum value the shifted resonance tends to break up into several, suggesting the existence of several slowly exchanging intermediate forms. At acid pH's, on addition of one additional mole/subunit of 113Cd2+, which greatly increases catalysis, "conformational" resonance(s) further broadens, suggesting that the second, "catalytic" metal ion increases the rates of interconversion between "conformational" species. At more alkaline pH's, near the optimum pH, the "conformational" peak is sharpened, which suggests that very fast interconversion is occurring. The position of the "catalytic" metal ion resonance also suggests all oxyligands in a distorted octahedral geometry. The "catalytic" resonance is often broadened to the point where it cannot be seen, suggesting rapid changes in its geometry due to interconversion of substrate and product.


Correction: Estimation of injury costs: financial damage of English Premier League teams' underachievement due to injuries.
[This corrects the article DOI: 10.1136/bmjsem-2019-000675.].


Purification and characterization of smooth muscle myosin light chain kinase.
Smooth muscle myosin light chain kinase was purified from turkey gizzards. The enzyme was extracted from washed myofibrils and the final step of purification was affinity chromatography using calmodulin coupled to Sepharose 4B. The purified enzyme was characterized with respect to its physical, chemical, and kinetic properties. It has a molecular weight of 130,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and 124,000 by sedimentation equilibrium centrifugation under nondenaturing conditions. It is an asymmetric molecule with a Stokes radius of 75 A, a sedimentation coefficient of 4.45 S, and a frictional coefficient of 1.85. Smooth muscle myosin light chain kinase is dependent on the calcium-binding protein calmodulin for activity. It has an apparent K0.5 for calmodulin of 10(-9) M and binds 1 mol of calmodulin/mol of myosin kinase in the presence of calcium. The binding of calmodulin increases the sedimentation coefficient from 4.45 S to 5.05 S, and the Stokes radius from 75 A to 79 A, and does not alter the frictional coefficient. The enzyme has a Km for ATP and the 20,000-dalton light chain of smooth muscle myosin of 50 microM and 5 microM, respectively. It phosphorylates the 20,000-dalton light chain of smooth muscle myosin more rapidly than the equivalent light chain from cardiac and skeletal muscles. It does not phosphorylate histones, alpha-casein, phosphorylase kinase, or phosphorylase b at a significant rate.


A2A Adenosine Receptor Partial Agonism Related to Structural Rearrangements in an Activation Microswitch.
In drug design, G protein-coupled receptor (GPCR) partial agonists enable one to fine-tune receptor output between basal and maximal signaling levels. Here, we add to the structural basis for rationalizing and monitoring partial agonism. NMR spectroscopy of partial agonist complexes of the A2A adenosine receptor (A2AAR) revealed conformations of the P-I-F activation motif that are distinctly different from full agonist complexes. At the intracellular surface, different conformations of helix VI observed for partial and full agonist complexes manifest a correlation between the efficacy-related structural rearrangement of this activation motif and intracellular signaling to partner proteins. While comparisons of A2AAR in complexes with partial and full agonists with different methods showed close similarity of the global folds, this NMR study now reveals subtle but distinct local structural differences related to partial agonism.


Bactericidal effect of intense pulsed light on seeds without loss of viability.
This study investigated the microbial inactivation effects of intense pulsed light (IPL) treatment as an alternative to chemical treatment for decontaminating the radish and pak choi seeds. The f R values (which indicate the resistance to IPL treatment) for radish and pak choi seeds were 24.50, 20.81 J/cm2, respectively. This resistance exhibited by seeds to IPL treatment is related to their surface roughness. Their Rq (the root-mean-square roughness), average surface roughness (Ra), and 10-point height roughness (Rz) values indicate that each crevice on a rough surface could shelter microorganisms from IPL. Viability tests of seeds exposed to IPL treatment indicated that the average germination rates of treated seeds exceeded 85% on day 3 of germination, which is considered as an acceptable criterion for germination. Also, on day 5 of germination the average shoot lengths of sprouts exposed to IPL did not differ significantly from those of untreated seeds.


Human Placental Mesenchymal Stem Cell-derived Exosomes in Combination with Hyperbaric Oxygen Synergistically Promote Recovery after Spinal Cord Injury in Rats.
Spinal cord injury (SCI) is a critical medical condition during which sensorimotor function is lost. Current treatments are still unable to effectively improve these conditions, so it is important to pay attention to other effective approaches. Currently, we investigated the combined effects of human placenta mesenchymal stem cells (hPMSCs)-derived exosomes along with hyperbaric oxygen (HBO) in the recovery of SCI in rats. Ninety male mature Sprague-Dawley (SD) rats were allocated into five equal groups, including; sham group, SCI group, Exo group (underwent SCI and received hPMSCs-derived exosomes), HBO group (underwent SCI and received HBO), and Exo+HBO group (underwent SCI and received hPMSCs-derived exosomes plus HBO). Tissue samples at the lesion site were obtained for the evaluation of stereological, immunohistochemical, biochemical, molecular, and behavioral characteristics. Findings showed a significant increase in stereological parameters, biochemical factors (GSH, SOD, and CAT), IL-10 gene expression and behavioral functions (BBB and EMG Latency) in treatment groups, especially Exo+HBO group, compared to SCI group. In addition, MDA levels, the density of apoptotic cells and gliosis, as well as expression of inflammatory genes (TNF-α and IL-1β) were considerably reduced in treatment groups, especially Exo+HBO group, compared to SCI group. We conclude that co-administration of hPMSCs-derived exosomes and HBO has synergistic neuroprotective effects in animals undergoing SCI.


An Integrative Informatics Approach to Explain the Mechanism of Action of N1-(Anthraquinon-2-yl) Amidrazones as BCR/ABL Inhibitors.
Drugs incorporating heterocyclic chemical skeletons possess a plethora of therapeutic activities such as anticancer, antimicrobial, antihypertensive, and antipsychiatric effects. It is becoming routine, nowadays, to use cheminformatics and bioinformatics methods to elucidate the mechanism(s) of action of such drugs. This study aimed to probe the activity of a recently published series of N1- (anthraquinon-2-yl) amidrazone piperazine derivatives employing computational strategies[1], identify their structural basis of binding to BCR/ABL kinase domain, and explain their anticancer activities in human breast adenocarcinoma (MCF-7) and chronic myelogenous leukemia (K562) cell lines. We applied an in silico integrative informatics approach integrating molecular descriptors, docking studies, cheminformatics, and network analysis. Our results highlighted the possible involvement of the BCR/ABL and DRD2 pathways in the anticancer activity of the studied compounds, and induced fit docking (IFD) indicated that the BCR/ABL kinase domain is a putative drug target. Additionally, high-scoring docking poses identified a unique network of hydrogen bonding with amino acids Y253, K271, E286, V299, L301, T315, M318, I360, R362, V379, and D3810. Using an integrative informatics approach to characterize our anticancer compounds, we were able to explain the biological differences between synthesized and biologically validated amidrazone piperazine anticancer agents. We were also able to postulate a mechanism of action of this novel group of anticancer agents.


Transitions of Care Among Patients Undergoing Percutaneous Coronary Intervention for Stable Angina: Insights From the Veterans Affairs Clinical Assessment Reporting and Tracking Program.
Background Effective transitions from the procedural to outpatient setting are essential to ensure high-quality cardiovascular care across health care systems, particularly among patients undergoing invasive cardiac procedures. We evaluated the association of postprocedural follow-up visits and antiplatelet prescriptions with clinical outcomes among patients undergoing percutaneous coronary intervention for stable angina at community or Veterans Affairs (VA) hospitals. Methods and Results Patients who actively received care within the VA Healthcare System and underwent percutaneous coronary intervention for stable angina at a community or VA hospital between October 1, 2015, and September 30, 2019, were identified. We compared mortality for patients receiving community or VA care, and among subgroups of community-treated patients by the presence of a postprocedural follow-up visit within 30 days or prescription for antiplatelet (P2Y12) medication within 120 days of the procedure. Among 12 837 patients who survived the first 30 days, 5133 were treated at community hospitals, and 7704 were treated in the VA. Prescriptions for antiplatelet therapy were less common for those treated in the community (85%) compared with the VA at 1 year (95%; hazard ratio [HR], 0.46; 95% CI, 0.44-47). Compared with VA-treated patients, the hazards for death were similar for patients treated in the community with a follow-up visit (HR, 1.17; 95% CI, 0.97-1.40) or with a fill for an antiplatelet therapy (HR, 1.08; 95% CI, 0.90-1.30). However, patients treated in the community without a follow-up visit had an 86% (HR, 1.86; 95% CI, 1.40-2.48) increased hazard of death, and those without antiplatelet prescription fill had a 144% increased hazard of death (HR, 2.44; 95% CI, 1.85-3.21) compared with all VA-treated patients. Conclusions Patients treated at community facilities have a decreased chance of receiving antiplatelet prescriptions after percutaneous coronary intervention with a concordant increased hazard of mortality, emphasizing the importance of transitions of care across health care systems when assessing cardiovascular quality.


Association of gut microbiota with post-operative clinical course in Crohn's disease.
The gut microbiome is altered in Crohn's disease. Although individual taxa have been correlated with post-operative clinical course, global trends in microbial diversity have not been described in this context. We collected mucosal biopsies from the terminal ileum and ascending colon during surgery and post-operative colonoscopy in 6 Crohn's patients undergoing ileocolic resection (and 40 additional Crohn's and healthy control patients undergoing either surgery or colonoscopy). Using next-generation sequencing technology, we profiled the gut microbiota in order to identify changes associated with remission or recurrence of inflammation. We performed 16S ribosomal profiling using 101 base-pair single-end sequencing on the Illumina GAIIx platform with deep coverage, at an average depth of 1.3 million high quality reads per sample. At the time of surgery, Crohn's patients who would remain in remission were more similar to controls and more species-rich than Crohn's patients with subsequent recurrence. Patients remaining in remission also exhibited greater stability of the microbiota through time. These observations permitted an association of gut microbial profiles with probability of recurrence in this limited single-center study. These results suggest that profiling the gut microbiota may be useful in guiding treatment of Crohn's patients undergoing surgery.


Molecular cloning and characterization of a phenylalanine ammonia-lyase gene (LrPAL) from Lycoris radiata.
LrPAL is a novel full-length cDNA isolated from Lycoris radiata by degenerate oligonucleotide primer PCR (DOP-PCR), 3'- and 5'-RACE approaches, harbours an open reading frame (ORF) encoding a 708 amino acid product. Sequence alignment showed that the deduced amino acid sequence of LrPAL shared more than 80% identity with other PAL sequences reported in Arabidopsis thaliana and other plants. RT-PCR revealed that LrPAL transcripts were higher in bud flowers and wilting flowers (5 days after blooming) than in blooming flowers. The transcript levels of LrPAL in leaves were significantly induced by methyl jasmonate (MJ) and nitric oxide (NO), and salicylic acid (SA). Similarly, HPLC analysis showed that galantamine (GAL) content was also higher in bud flowers and wilting flowers than in blooming flowers. The GAL content in leaves was significantly induced by MJ and NO, and inhibited by SA. This study enables us to further elucidate the role of LrPAL in the biosynthesis of GAL in Lycoris radiata at a molecular level.


Detection of Complex Sounds in Quiet Conditions by Seals and Sea Lions.
To test how accurately baseline audiometric data predict detection of complex stimuli, absolute detection thresholds for frequency-modulated (FM), amplitude-modulated (AM), and harmonic stimuli were obtained for one Phoca vitulina (harbor seal) and one Zalophus californianus (California sea lion) at frequencies spanning the functional range of hearing. These thresholds were then compared with a priori predictions based on the tonal audiograms of these subjects. Predicted thresholds were accurate for most FM signals and for AM signals for the California sea lion. Predictions were unreliable for harmonic signals for both species and for AM signals for the harbor seal.


Effect of thoracoscopic splanchnic denervation on pain processing in chronic pancreatitis patients.
Central sensitisation due to visceral pancreatic nociceptive input may play an important role in chronic pancreatitis pain. Using quantitative sensory testing (QST), this first study investigates whether thoracoscopic splanchnic denervation (TSD), performed to reduce nociceptive visceral input, affects central sensitisation in chronic pancreatitis patients. We studied 19 chronic pancreatitis patients (11 men, 8 women on stable opioid medication) and 18 healthy volunteers as preoperative controls. Preoperatively and 6 weeks after TSD, pain numeric rating scores, opioid medication, and thresholds to electric skin stimulation and pressure pain (measured in dermatomes T10 (pancreas), C5, T4, L1, L4) were documented. Treatment success was defined as cessation of opioids 6 weeks after TSD. Six weeks after TSD, there was a trend towards lower pain scores, only 10 patients were still on opioids (P<0.05 vs. preoperatively) and thresholds overall were significantly higher than preoperatively (pressure pain: +25%, P<0.001; electric: sensation +55%, pain detection +34%, pain tolerance +21%, P<0.05). Gender-specific differences in hypoalgesia patterns were seen. Preoperatively, TSD treatment successes consumed significantly less opioids than failures, without significant differences in preoperative patterns of neuroplasticity. TSD for chronic pancreatitis pain resulted in fewer patients on opioids and overall increases in pain thresholds. Our results suggest that TSD for reducing visceral nociceptive input may be effective in reducing resulting central sensitisation. Although patients benefiting from TSD consume less opioids preoperatively, we were unable to clearly link treatment success with specific perioperative patterns of neuroplasticity such as the presence or absence of hyperalgesia.


Cross-correlation between spine and hip joint kinematics differs in healthy individuals and subgroups of ankylosing spondylitis patients during trunk lateral flexion.
The effects of sacroiliitis and syndesmophyte formation on the cross-correlation between spine and hip joint kinematics in ankylosing spondylitis (AS) are poorly understood. To investigate the cross-correlation between spine and hip joint kinematics differs in healthy individuals and ankylosing spondylitis patients during trunk lateral flexion. Fifty AS patients and thirty-nine healthy adults (controls) were recruited from a medical center. The patients were divided into two subgroups, namely the sacroiliitis (n = 28) and syndesmophyte (n = 22) subgroups. An inertial motion system was used to record kinematic data of spine, pelvic and hip joints during lateral trunk flexion. The maximal cross-correlation coefficient (CCF) and time lag of motion between the spine and hip joint were analyzed. The syndesmophyte group had the smallest range of motion in all recorded motion. The sacroiliitis group exhibited higher thoracic flexion, pelvic pitch, and pelvic rotation than the other two groups. In the syndesmophyte group, the CCF between lumbar lateral flexion (LLF) and hip abduction were weakly and LLF and hip rotation were strongly correlated. Considering in time sequence, LLF occurred earlier than hip abduction and hip rotation during trunk lateral flexion; however, both AS subgroups exhibited longer time lags than in the control group. The cross-correlation between spine and hip joint kinematics differs in healthy individuals and AS patients during trunk lateral flexion. The motion pattern changes in patients with AS of differing severity may also alter the loads on the spine and hip joints.


Anchored Minimal Clinically Important Difference Metrics: Considerations for Bias and Regression to the Mean.
Minimal clinically important differences (MCIDs) are used to understand clinical relevance. However, repeated observations produce biased analyses unless one accounts for baseline observation, known as regression to the mean (RTM). Using an International Knee Documentation Committee (IKDC) survey dataset, we can demonstrate the effect of RTM on MCID values by (1) MCID-estimate dependence on baseline observation and (2) MCID-estimate bias being higher when the posttest-pretest data correlation is lower. We created 10 IKDC datasets with 5000 patients and a specific correlation under both equal and unequal variances. For each 10-point increase in baseline IKDC, MCID decreased by 3.5, 2.7, 1.9, 1.2, and 0.7 points when posttest-pretest correlations were 0.10, 0.25, 0.50, 0.75, and 0.90, respectively, under equal variances. Not accounting for RTM resulted in a static 20-point MCID. Minimal clinically important difference estimates may be unreliable. Minimal clinically important difference calculations should include the correlation and variances between posttest and pretest data, and researchers should consider using a baseline covariate-adjusted receiver operating characteristic curve analysis to calculate MCID.


Acetaminophen differentially enhances social behavior and cortical cannabinoid levels in inbred mice.
Supratherapeutic doses of the analgesic acetaminophen (paracetomol) are reported to promote social behavior in Swiss mice. However, we hypothesized that it might not promote sociability in other strains due to cannabinoid CB(1) receptor-mediated inhibition of serotonin (5-HT) transmission in the frontal cortex. We examined the effects of acetaminophen on social and repetitive behaviors in comparison to a cannabinoid agonist, WIN 55,212-2, in two strains of socially-deficient mice, BTBR and 129S1/SvImJ (129S). Acetaminophen (100mg/kg) enhanced social interactions in BTBR, and social novelty preference and marble burying in 129S at serum levels of ≥70 ng/ml. Following acetaminophen injection or sociability testing, anandamide (AEA) increased in BTBR frontal cortex, while behavior testing increased 2-arachidonyl glycerol (2-AG) levels in 129S frontal cortex. In contrast, WIN 55,212-2 (0.1mg/kg) did not enhance sociability. Further, we expected CB(1)-deficient (+/-) mice to be less social than wild-type, but instead found similar sociability. Given strain differences in endocannabinoid response to acetaminophen, we compared cortical CB(1) and 5-HT(1A) receptor density and function relative to sociable C57BL/6 mice. CB(1) receptor saturation binding (Bmax=958±117 fmol/mg protein), and affinity for [(3)H] CP55,940 (K(D)=3±0.8 nM) was similar in frontal cortex among strains. CP55,940-stimulated [(35)S] GTPγS binding in cingulate cortex was 136±12, 156±22, and 75±9% above basal in BTBR, 129S and C57BL/6 mice. The acetaminophen metabolite para-aminophenol (1 μM) failed to stimulate [(35)S] GTPγS binding. Hence, it appears that other indirect actions of acetaminophen, including 5-HT receptor agonism, may underlie its sociability promoting properties outweighing any CB(1) mediated suppression by locally-elevated endocannabinoids in these mice.


Genetic profile characterization and segregation analysis of 10 X-STRs in a sample from Santander, Colombia.
Ten X-chromosome short tandem repeats (X-STRs: DXS8378, DXS7132, DXS9898, DXS6809, DXS6789, DXS101, GATA172D05, HPRTB, DXS8377, and DXS7423) were analyzed in a sample of unrelated individuals (108 males and 110 females) from the Santander Department in Colombia. In this sample, gene diversities varied between 63.56%, for DXS8378, and 91.41%, for DXS8377. For this set of 10 X-STRs, a high discrimination power was obtained for both male (1 in 3 x 10(6)) and female (1 in 9 x 10(10)) samples and a high mean exclusion chance in father/daughter duos (99.993%) and in father/mother/daughter trios (99.9999%), demonstrating the usefulness of this set of markers in forensic and kinship analysis. Hardy-Weinberg equilibrium was tested in the female sample and no significant deviations were found. Pairwise analysis showed significant differences in the comparison with samples from Spain, Peru, and Argentina and with African American and Hispanic samples from New York. This same set of X-STRs was also typed in 51 mother/father/daughter trios, 43 mother/son duos, and in a single father/daughter pair. In total, four mutations were observed; one at DXS7132 and at DXS6809, and two at DXS8377. Two mutations were paternal and one maternal; and to a fourth mutation, it was not possible to define its origin.


Cancer and Covid-19: A preliminary study on the trauma aspects of coronavirus in cancer patients.
Because of Covid 19, it has become necessary to revise the treatment of cancer patients ("how" and "when"). That has had important psychological repercussions. The purpose of this study is the evaluation of the impact of Covid19 in terms of Post-Traumatic Stress Disorder and Depression and the potential association with coping strategies. We conducted an exploratory study with 106 patients undergoing treatment, using following questionnaires: Screening Questionnaire for Disaster Mental Health and Mini-Mental Adjustment to Cancer. Only 25.5% of our sample showed symptoms of posttraumatic stress disorder (PTSD) and 6.6% revealed a probable presence of depression. In addition, it came up a significant correlation between SQD_P and the coping styles "Hopelessness" (r = 0.41 p < 0.001) and "Anxious Preoccupation" (r = 0.45, p < 0.001). A strong correlation also emerged between non-Covid 19 patients and PTSD (r = 0.29, p = 0.002). Our preliminary data did not reveal a prevalence of PTSD, but the persistence of the health emergency requires to focus future research on protective and risk factors related to PTSD and psychological distress in cancer patients, in order to reduce the mental health burden of Covid19.


Is hyperuricemia an independent prognostic factor for IgA nephropathy: a systematic review and meta-analysis of observational cohort studies.
Hyperuricemia has been reported to be correlated with IgA nephropathy (IgAN). However, whether hyperuricemia or elevated serum uric acid (SUA) is an independent prognostic factor of IgAN remains unknown. Therefore, this systematic review and meta-analysis evaluated the prognostic value of hyperuricemia and elevated SUA in IgAN. Databases including PubMed, EMBASE, the Cochrane Central Register of Controlled Trials (CENTRAL), and Open Gray were reviewed systematically. The kidney failure events of IgAN were defined as a doubling of serum creatinine, halving of eGFR, end-stage renal disease (ESRD), or death. The risk ratio (RR) between hyperuricemia and IgAN-caused kidney failure was evaluated before and after adjustment for relevant covariates. The RR between elevated SUA and IgAN-caused kidney failure was evaluated after adjustment for relevant covariates. A total of 11 548 patients from 14 studies were included in this meta-analysis. Hyperuricemia was found to be an independent prognostic factor of IgAN (unadjusted RR = 2.79, 95% CI = 1.93-4.03, p for heterogeneity <0.00001, I2 = 91%; adjusted RR = 2.12, 95% CI = 1.64-2.73, p for heterogeneity = 0.86, I2 = 0%). Subgroup and sensitivity analyses confirmed the stability of these results. Similarly, elevated SUA was positively correlated with kidney failure events of IgAN (adjusted RR = 1.25, 95% CI = 1.19-1.31, p for heterogeneity = 0.6, I2 = 0%). Our meta-analysis showed that hyperuricemia and elevated SUA were both independently associated with an increased incidence of kidney failure events in IgAN patients.


The Impact of Social Media on Applicants' Perceptions of Plastic Surgery Training Programs.
Given recent COVID-19 restrictions on in-person visiting subinternships and interviews, this study sought to evaluate the program information that was most influential to future plastic surgery applicants as they researched residency programs on social media. An electronic survey targeting medical students interested in plastic surgery was deployed to assess the importance of various information sources in forming perceptions of residency programs. Applicants were invited to participate through an Instagram "Story" (where the survey was embedded) and through an electronic survey link sent via email to interested program applicants and interviewees. There were 83 respondents, among which 92% were current medical students planning to apply to Plastic Surgery. The most utilized resources that informed program interest were: mentors (86%), peers/partners (60%), and geographic location preference (55%). Among social media content, applicants most desired posts about resident life (66%) and team bonding activities (61%). Overall, 72% of respondents agreed/strongly agreed that social media played a role in informing their interest to apply to a specific residency program. The study demonstrated that prospective plastic surgery applicants expect programs to have a social media presence, and thus, programs should invest time and thought in their social media strategy. While electronic sources are not the most important sources of information rated among applicants, social media plays an influential role in guiding interest in specific programs. To best inform applicant perspectives during the recruiting process, programs should prioritize content that gives a picture of "resident life" and team dynamics.


Decreased duration and altered topography of electroencephalographic microstates in patients with panic disorder.
The topography and temporal sequence of scalp electrical fields were analyzed by adaptive segmentation of the continuous electroencephalogram (EEG) in 27 patients with panic disorder and 28 control subjects during rest phases and during the viewing of a neutral (mushroom) or an emotionally relevant (casualty) picture. The results indicate decreased duration of brain microstates in panic patients during all conditions. Comparison of the resting phases with the viewing conditions revealed a significant acceleration of EEG microstates in both the patients and the control subjects. Patients and control subjects differed in the topography of the fields during rest: control subjects showed a left-anterior/right-posterior orientation, while panic patients showed a predominantly right-anterior/left-posterior orientation. Neither group displayed any topographic changes when viewing the mushroom picture. However, when viewing the anxiety-specific casualty picture, panic patients shifted fields in a different way than did control subjects. Centroid topography does not permit clear localization of the cortical generators. It is concluded that panic patients show a generally increased cortical activation compared with healthy control subjects, and activate different neuronal arrays when viewing an anxiety-specific stimulus.


Effect of amiodarone on electric induction, morphology, and rate of ventricular tachycardia and its relation to clinical efficacy.
Using His bundle electrograms and programmed ventricular stimulation, the effects of chronic amiodarone treatment on induction, morphology, and the rate of ventricular tachycardia (VT) were studied in 17 consecutive patients treated with amiodarone for control of recurrent sustained VT or ventricular fibrillation. Studies were done before and after treatment with amiodarone for an average duration of 5.3 (range 2 to 18) months. During the control study, sustained VT could be induced in 16 patients. VT was initiated by single or double right ventricular (RV) extrastimuli in 14 patients, by double left ventricular (LV) extrastimuli in 1 patient, and by RV burst pacing in 1 patient. Only one pattern (morphology) of VT similar to that of spontaneous VT was induced in 12 patients and two patterns of VT in 4 patients. The average cycle length (CL) (mean +/- SD) of induced VT was 325.8 +/- 61.2 ms. After amiodarone, VT could be induced in 7 of 17 patients and was initiated by single RV extrastimuli in 5 patients, double RV extrastimuli in 1 patient, and RV burst pacing in 1 patient. In 3 of 5 patients in whom VT could be initiated by single RV extrastimuli, initiation of VT required double RV or double LV extrastimuli in the control study; in 1 of 5 patients VT could not be induced in the control study. Amiodarone induced nonclinical, polymorphic VT in 4 patients in whom only clinical VT could be induced during the control study. Compared to control, the CL of induced VT was significantly longer (322 +/- 65.7 vs 416 +/- 41.5 ms; P less than 0.001).+


Tracing intracellular proteolytic pathways. Proteolysis of fatty acid synthase and other cytoplasmic proteins in the yeast Saccharomyces cerevisiae.
Yeast fatty acid synthase consists of two independent polypeptide strains, alpha and beta. The functional multienzyme complex, composed of six alpha- and six beta-subunits, is rather stable against proteolysis in vivo. Mutations in one of the subunits or deletion of one subunit lead to degradation of the nonmutated remaining fatty acid synthase protein. We show that the unassembled alpha-subunit of this enzyme is short-lived, and degradation depends on the presence of active cytoplasmic proteinase yscE, the yeast proteasome. The unassembled beta-subunit is degraded by a nonvacuolar proteolytic system under vegetative growth conditions. However, starvation of a vacuolar proteinase mutant strain, which lacks the alpha-subunit of fatty acid synthase, leads to appearance of the unassembled beta-subunit is isolated vacuoles. This indicates that the major vacuolar peptidases proteinase yscA and yscB are at least partly involved in degradation of the beta-subunit of fatty acid synthase. In a proteinase yscA and yscB double mutant strain wild type for fatty acid synthase both subunits of fatty acid synthase, alpha and beta, are detectable in vacuoles. In addition, under the same starvation conditions other cytoplasmic proteins are found in the vacuole of a proteinase yscA and yscB double mutant strain. The experiments in conjunction with the previous finding of the appearance of vesicles in vacuoles of starved cells (Simeon, A., van der Klei, I.J., Veenhuis, M., and Wolf, D. H. (1992) FEBS Lett. 301, 231-235) indicate that transport of these tested cytoplasmic proteins into the vacuole is an unselective bulk process induced by nutritional stress.


Analysis of Novel Mutations and Methylmalonyl-CoA Mutase Levels in Thai Patients with Isolated Methylmalonic Acidemia.
Isolated methylmalonic acidemia (MMA) is an autosomal recessive, inherited disorder that results from either a mut defect of the methylmalonyl-CoA mutase apoenzyme (MCM, the product of the MUT gene) or a cbl defect in the synthesis of its cofactor, adenosylcobalamin (AdoCbl). In this study, biochemical and mutational analyses of three patients clinically diagnosed with MMA were performed. No MCM activity was detected in leukocyte extracts of two patients, while high MCM activity was found in the other, suggesting mut (0) and cbl defects, respectively. A novel (c.IVS6 -3 to -8delCTTTTT, p.K444_L445insFC*) and two known mutations in the MUT gene and one novel (c.227_36delGACCCAAAGA, p.R76Mfs*14) mutation in the MMAB gene were identified. In addition, MCM immunoblot analysis of leukocyte extract samples of these three patients and eight patients previously reported by our group, as well as their parents, showed a good correlation between the MCM protein and activity levels. Patients with mut (0) defective subtypes lacked MCM activity and had no MCM band, while patients carrying the cbl defects had high MCM activity levels and an intense MCM band at about 83 kDa, in comparison to those in their parents. These data expand the mutation spectrum of MMA deficiency. In addition, the examination of MCM protein level may be used as an alternative technique to determine the mut (0) and cbl defective subgroups.


GABA-mimetic action of etomidate.
A comparison of antagonism by bicuculline or strychnine of the effects of GABA or etomidate on rat isolated superior cervical ganglia, frog isolated hemisected spinal cords and rat central neurones in vivo indicates that etomidate has GABA-mimetic actions.


Involvement of thioredoxin-interacting protein (TXNIP) in glucocorticoid-mediated beta cell death.
Glucocorticoid hormones (GCs) are widely used to treat a variety of inflammatory and immune diseases. However, their long-term administration is associated with adverse metabolic effects, including glucose intolerance and diabetes. Our objective was to elucidate the mechanisms by which GCs affect beta cell survival with a specific emphasis on the role of the thioredoxin-interacting protein (TXNIP) in beta cell apoptosis. Human and mouse islets, together with MIN6 beta cells, were exposed to dexamethasone (Dex) and apoptosis was assessed by measuring the percentage of sub-G1 cells, the appearance of cleaved caspase-3 or by using a TUNEL assay. Dex-upregulated expression of Txnip mRNA was analysed by real-time PCR, and GC-modulated production and modification of proteins were determined by western blotting. We provide evidence that TXNIP, a negative regulator of the antioxidant thioredoxin (TRX), is strongly induced in beta cells by GCs and that its induction is dependent on p38 mitogen-activated protein kinase (MAPK) activation. TXNIP downregulation by RNA interference, overexpression of the radical scavenger TRX1 or elevation of intracellular cAMP levels attenuated the Dex-mediated apoptosis. Dex-induced Txnip expression and beta cell apoptosis are mediated by the glucocorticoid receptor (GR), as the GR antagonist RU486 fully abolishes these effects. Altogether, our data suggest TXNIP as a novel mediator of GC-induced apoptosis in beta cells and further contribute to our understanding of beta cell death pathways.


Metastasis on a Schuchardt incision after Schauta-Amreich operation for cervical carcinoma.
Laparoscopic pelvic lymph node dissection in combination with a vaginal radical Schauta-Amreich operation has become an alternative for women with early-stage cervical cancer. We described the first case of metastasis on Schuchardt's incision after Schauta-Amreich operation for neuroendocrine cervical cancer. Metastasis on Schuchardt's incision after Schauta-Amreich operation can rarely be the metastatic localization of the cervical carcinoma.


The Blalock-Taussig shunt in infants: standard versus modified.
In recent years, the modified Blalock-Taussig shunt--a polytetrafluoroethylene graft from the subclavian artery to the pulmonary artery--has been preferred over the standard shunt by some surgeons because (1) it requires less dissection and (2) length of native vessels is not critical. From January, 1979, to June, 1985, we operated on 51 infants less than 1 year of age, including 26 less than 1 week of age, to palliate severe complex cyanotic congenital cardiac malformations. Twenty-four modified Blalock-Taussig shunts and 29 standard Blalock-Taussig shunts were created. The groups were concurrent. We reviewed all available cineangiograms and measured branch pulmonary and subclavian arteries. Pulmonary artery index was not different preoperatively in patients given a modified versus a standard Blalock-Taussig shunt (144 +/- 118 and 118 +/- 59 mm2/m2, respectively), but it was greater postoperatively in patients with a modified shunt (431 +/- 188 and 189 +/- 106 mm2/m2) (p = 0.07). Distortion of the pulmonary artery occurred less often after a modified Blalock-Taussig shunt (4/11) than a standard Blalock-Taussig shunt (6/8) (p = 0.06), though none of the distortions was severe. Early and late shunt failure occurred less often with a modified shunt (5/24) than with a standard shunt (15/29) (p less than 0.05). The modified Blalock-Taussig shunt had advantages over the standard Blalock-Taussig shunt in our series: pulmonary artery growth was greater, distortion of pulmonary arteries was less commonly seen, and shunt failure occurred less often. Thus, in infants, we believe the modified Blalock-Taussig shunt should be considered a reasonable alternative to the standard Blalock-Taussig shunt.


Why do males emerge before females? protandry as a mating strategy in male and female butterflies.
The reproductive strategy of butterfly males can be defined as being to maximize the number of females mated. We have earlier shown that, if the eclosion period of females is regarded as given, males should emerge before females to achieve maximal reproductive success. However, females may also be considered to have a reproductive strategy with respect to the issue "when to emerge". In this paper we assume that females are selected to minimize the time spent unmated (to minimize prereproductive death), and analyze when females should optimally emerge in relation to males to achieve this end. We show that there is no conflict between the sexes with respect to the timing of eclosion when the length of the eclosion period is approximately equal for males and females. Thus, protandry should be considered a reproductive strategy of both males and females.


Echocardiographic parameters for the assessment of congestive heart failure in dogs with myxomatous mitral valve disease and moderate to severe mitral regurgitation.
The objective of this study was to evaluate the influence of congestive heart failure (CHF) on echocardiographic variables in dogs with moderate or severe mitral valve regurgitation (MR) secondary to myxomatous mitral valve disease (MMVD). The secondary objective was to investigate the preliminary use of left ventricular early inflow-outflow index (LVEIO) and L-waves as indicators of CHF. Forty-five dogs with moderate or severe MR without (n = 23) or with (n = 22) CHF were retrospectively included. Echocardiographic variables of elevated left ventricular (LV) filling pressures and cardiac function, LVEIO and the presence of L-waves were obtained and statistically compared between the two groups. On general linear model analysis, mitral inflow peak E-wave velocity, E:A ratio, ratio of E to isovolumic relaxation time (IVRT) and LVEIO were increased, while left atrium (LA) fractional shortening was reduced, in dogs with CHF. Peak E-wave velocity, peak A-wave velocity, E:A ratio, LA minimum diameter, LA diameter before the P-wave, LA fractional shortening and LVEIO were found to be predictive of CHF. The absence of L-waves indicated a reduced risk of CHF. Several echocardiographic variables may be useful in identifying elevated LV filling pressures, consistent with CHF, in dogs with moderate and severe MR. Left ventricular early inflow-outflow index and L-waves might be of interest as clinical and prognostic markers in dogs with MMVD and require further investigation.


Classification of large vessel vasculitis: Can we separate giant cell arteritis from Takayasu arteritis?
The two main variants of large vessel vasculitis include Takayasu arteritis and giant cell arteritis. While these two conditions have historically been considered different conditions, recent evidence questions whether they are a spectrum of the same disease. Classification criteria are limited in distinguishing between cases with phenotypic overlap. The limitations of the current criteria and directions of future research are reviewed.


De-Intensification Of Blood Glucose Lowering Medication In People Identified As Being Over-Treated: A Mixed Methods Study.
To evaluate if, one year after notification of possible overtreatment, diabetes care providers de-intensified glucose-lowering medications and to gain insight into the opinions and beliefs of both care providers and patients regarding de-intensification. Mixed methods using routine care data from five health-care centres in the Netherlands. Patient characteristics and medication prescription of patients, previously identified as possibly over-treated, were extracted from patients' medical records. Opinions of care providers were obtained through interviews. Patients received questionnaires about their diabetes treatment and were asked to participate in focus groups. A total of 64 elderly patients with type 2 diabetes were previously identified as possibly over-treated and included; 57.8% male, median age 75 years (IQR=72-82), median diabetes duration 12 years (IQR=8-18). De-intensification was implemented in more than half (n=36) of them. Care providers preferred person-centred care above just setting general HbA1c target values, considering patient characteristics (such as comorbidity) and patient's preference. Patients valued glucose levels as most important in determining their treatment. Both patients and care providers felt that de-intensification should occur gradually. Treatment had been de-intensified in more than half of the patients (56.3%). Insight in reasons for not de-intensifying elderly patients is important since treatment for them can be "person-centred care". De-intensification is an iterative and time-intensive process.


Positive practice routines in overcoming resistance to the treatment of school phobia: a case study with follow-up.
The treatment is described of a 6-year-old first grade girl who refused to attend her classroom without adult accompaniment. For eight weeks an attempt was made, using different adult escorts, to prompt/shape classroom attendence through classroom exposure. However, the outcome was the development of an unwavering preference to remain in the principal's office after the child would not tolerate separation from these adults. Rather then continue to take her to the classroom, an intervention was initiated designed to structure her daily experiences and provide instruction. In this she was expected to practice several educational activities at designated times in the principal's office. This intervention resulted in a complete elimination of the child's classroom avoidance within five weeks and concomitant improvements in both social and academic functioning. Although the child's mother suddenly died during intervention, follow-up at one and 12 months in a different school indicated the maintenance of all therapeutic gains. Unique aspects of the procedures pertaining to refractory problems and some tentative behavioral guidelines to overcoming resistance to treatment are discussed.


Intravitreal Ziv-Aflibercept: Clinical Effects and Economic Impact.
During the past decade, drugs that inhibit the actions of vascular endothelial growth factor (VEGF) have become standard-of-care treatment for a variety of chorioretinal vascular conditions. The off-label, intravitreal use of ziv-aflibercept (Zaltrap) has provided clinicians with an additional cost-effective drug. The commercial preparation of ziv-aflibercept contains the same aflibercept (VEGF-trap) molecule as Eylea but has a much higher osmolarity (1000 mOsm/kg vs 300 mOsm/kg). Initial concerns regarding cytotoxicity and long-term safety of intravitreal ziv-aflibercept have been largely negated after a series of publications failed to identify adverse ocular and systemic side effects. Both treatment-naive and anti-VEGF‒resistant cases of neovascular age-related macular degeneration (nAMD), diabetic macular edema (DME), retinal vein occlusion (RVO), and choroidal neovascular membrane (CNVM) may respond as well to ziv-aflibercept as to aflibercept. A higher dose of ziv-aflibercept (2 mg in 0.08 mL) does not cause any adverse effects during short-term follow-up period (1 month). Data from various sources suggest that ziv-aflibercept may be as cost effective as bevacizumab, thereby making it an attractive treatment option in low- and middle-income countries. However, problems with off-label use, compounding, and counterfeiting limit its availability in many countries. Data from prospective, randomized, multicenter clinical trials are still required to convince physicians and regulatory bodies of its clinical efficacy and potential as early therapy.


Medallion-like dermal dendrocyte hamartoma: A rare congenital CD34-positive dermal lesion clinically and pathologically overlapping with fibroblastic connective tissue nevus.
Both medallion-like dermal dendrocyte hamartoma and fibroblastic connective tissue nevus are rare benign dermal lesions composed of CD34-positive spindle cells. Although regarded as different diseases, it is sometimes difficult to distinguish between them due to their clinical and pathological similarities. We present a case of medallion-like dermal dendrocyte hamartoma that could also be diagnosed as fibroblastic connective tissue nevus and propose the possibility of overlap in these diseases.


Molecular Genetics of Prostate Cancer and Role of Genomic Testing.
Prostate cancer (PCa) is characterized by profound genomic heterogeneity. Recent advances in personalized treatment entail an increasing need of genomic profiling. For localized PCa, gene expression assays can support clinical decisions regarding active surveillance and adjuvant treatment. In metastatic PCa, homologous recombination deficiency, microsatellite instability-high (MSI-H), and CDK12 deficiency constitute main actionable alterations. Alterations in DNA repair genes confer variable sensitivities to poly(ADP-ribose)polymerase inhibitors, and the use of genomic instability assays as predictive biomarker is still incipient. MSI can be assessed by immunohistochemistry To date there is a lack of consensus as to testing standards.


Adolescents' nonmedical use and excessive medical use of prescription medications and the identification of substance use subgroups.
The purpose of this study was to identify subgroups of adolescents based on their past 12 months use of tobacco, alcohol, marijuana, illicit drugs, and nonmedical use and excessive medical use of prescription medications. A cross-sectional Web-based survey of adolescents from two middle and high school districts in Southeastern Michigan was conducted. The sample included 2,744 middle school (7th and 8th grade) and high school (9th through 12th grade) students. Participants had a mean age of 14.8 years (SD = 1.9 years); 50.4% were female, 64.1% were Caucasian, and 30.6% were African American. Participants completed measures of the past 12 months of substance use, parental monitoring, parental substance use, and internalizing and externalizing problems. Exploratory latent class analysis (LCA) indicated four classes. The largest class was composed of participants with low probabilities of using any substances (low/no use class), and the smallest class was composed of participants with relatively high probabilities of using all substances (multiple substances class). A third class included participants with high probabilities of using tobacco, alcohol, and marijuana (TAM). The fourth class consisted of participants with relatively high probabilities of alcohol use, nonmedical prescription drug use, and excessive medical use of prescription drugs (ANM). Female gender predicted membership in the ANM and multiple substance classes, and parental monitoring, parental substance use problems, internalizing, and externalizing problems uniquely predicted membership in all three high-risk risk classes. Results indicated three high-risk subgroups of adolescents, each characterized by a different pattern of substance use. Two risk groups are characterized by relatively high probabilities of nonmedical use and excessive medical use of prescription medications.


Fluorescence anisotropy reveals order and disorder of protein domains in the nuclear pore complex.
We present a new approach for studying individual protein domains within the nuclear pore complex (NPC) using fluorescence polarization microscopy. The NPC is a large macromolecular complex, the size and complexity of which presents experimental challenges. Using fluorescence anisotropy and exploiting the symmetry of the NPC and its organization in the nuclear envelope, we have resolved order and disorder of individual protein domains. Fluorescently tagging specific domains of individual nucleoporins revealed both rigid and flexible domains: the tips of the FG domains are disordered, whereas the NPC-anchored domains are ordered. Our technique allows the collection of structural information in vivo, providing the ability to probe the organization of protein domains within the NPC. This has particular relevance for the FG domain nucleoporins, which are crucial for nucleocytoplasmic transport.


Benzodioxane-benzamides as promising inhibitors of Escherichia coli FtsZ.
The conserved process of cell division in bacteria has been a long-standing target for antimicrobials, although there are few examples of potent broad-spectrum compounds that inhibit this process. Most currently available compounds acting on division are directed towards the FtsZ protein, a self-assembling GTPase that is a central element of the division machinery in most bacteria. Benzodioxane-benzamides are promising candidates, but poorly explored in Gram-negatives. We have tested a number of these compounds on E. coli FtsZ and found that many of them significantly stabilized the polymers against disassembly and reduced the GTPase activity. Reconstitution in crowded cell-like conditions showed that FtsZ bundles were also susceptible to these compounds, including some compounds that were inactive on protofilaments in dilute conditions. They efficiently killed E. coli cells defective in the AcrAB efflux pump. The activity of the compounds on cell growth and division generally showed a good correlation with their effect in vitro, and our experiments are consistent with FtsZ being the target in vivo. Our results uncover the detrimental effects of benzodioxane-benzamides on permeable E. coli cells via its central division protein, implying that lead compounds may be found within this class for the development of antibiotics against Gram-negative bacteria.


MFSD2A potentiates gastric cancer response to anti-PD-1 immunotherapy by reprogramming the tumor microenvironment to activate T cell response.
The efficacy of anti-programmed cell death protein 1 (PD-1) immunotherapy in various cancers, including gastric cancer (GC), needs to be potentiated by more effective targeting to enhance therapeutic efficacy or identifying accurate biomarkers to predict clinical responses. Here, we attempted to identify molecules predicting or/and promoting anti-PD-1 therapeutic response in advanced GC (AGC). The transcriptome of AGC tissues from patients with different clinical responses to anti-PD-1 immunotherapy and GC cells was analyzed by RNA sequencing. The protein and mRNA levels of the major facilitator superfamily domain containing 2A (MFSD2A) in GC cells were assessed via quantitative real-time polymerase chain reaction, Western blotting, and immunohistochemistry. Additionally, the regulation of anti-PD-1 response by MFSD2A was studied in tumor-bearing mice. Cytometry by Time-of-Flight, multiple immunohistochemistry, and flow cytometry assays were used to explore immunological responses. The effects of MFSD2A on lipid metabolism in mice cancer tissue and GC cells was detected by metabolomics. Higher expression of MFSD2A in tumor tissues of AGC patients was associated with better response to anti-PD-1 immunotherapy. Moreover, MFSD2A expression was lower in GC tissues compared to adjacent normal tissues, and its expression was inversely correlated with GC stage. The overexpression of MFSD2A in GC cells enhanced the efficacy of anti-PD-1 immunotherapy in vivo by reprogramming the tumor microenvironment (TME), characterized by increased CD8+ T cell activation and reduced its exhaustion. MFSD2A inhibited transforming growth factor β1 (TGFβ1) release from GC cells by suppressing cyclooxygenase 2 (COX2)-prostaglandin synthesis, which consequently reprogrammed TME to promote anti-tumor T cell activation. MFSD2A potentially serves as a predictive biomarker for anti-PD-1 immunotherapy response in AGC patients. MFSD2A may be a promising therapeutic target to potentiate the efficacy of anti-PD-1 immunotherapy by reprogramming the TME to promote T cells activation.


Effect of micro- and nanoparticle shape on biological processes.
In the drug delivery field, there is beyond doubt that the shape of micro- and nanoparticles (M&NPs) critically affects their biological fate. Herein, following an introduction describing recent technological advances for designing nonspherical M&NPs, we highlight the role of particle shape in cell capture, subcellular distribution, intracellular drug delivery, and cytotoxicity. Then, we discuss theoretical approaches for understanding the effect of particle shape on internalization by the cell membrane. Subsequently, recent advances on shape-dependent behaviors of M&NPs in the systemic circulation are detailed. In particular, the interaction of M&NPs with blood proteins, biodistribution, and circulation under flow conditions are analyzed. Finally, the hurdles and future directions for developing nonspherical M&NPs are underscored.


Generation of zero-valent sulfur from dissimilatory sulfate reduction in sulfate-reducing microorganisms.
Dissimilatory sulfate reduction (DSR) mediated by sulfate-reducing microorganisms (SRMs) plays a pivotal role in global sulfur, carbon, oxygen, and iron cycles since at least 3.5 billion y ago. The canonical DSR pathway is believed to be sulfate reduction to sulfide. Herein, we report a DSR pathway in phylogenetically diverse SRMs through which zero-valent sulfur (ZVS) is directly generated. We identified that approximately 9% of sulfate reduction was directed toward ZVS with S8 as a predominant product, and the ratio of sulfate-to-ZVS could be changed with SRMs' growth conditions, particularly the medium salinity. Further coculturing experiments and metadata analyses revealed that DSR-derived ZVS supported the growth of various ZVS-metabolizing microorganisms, highlighting this pathway as an essential component of the sulfur biogeochemical cycle.


Is elective neck treatment indicated for T2N0 squamous cell carcinoma of the glottic larynx?
This is an analysis of 98 patients with T2N0 squamous cell carcinoma of the glottic larynx treated with radiation therapy. Patients received irradiation to the primary lesion alone; the neck was not treated electively. All patients had at least 2 years of follow-up; patients who died within 2 years from treatment with the neck continuously disease-free were excluded from the analysis. The rate of control of neck disease following irradiation was as follows: primary site continuously disease-free, 73/75 (97%); primary tumor recurrence, 18/23 (78%). Salvage treatment was successful in 4 of 7 patients who developed recurrent disease in the neck. We conclude that elective treatment to the clinically negative neck is not indicated for patients with T2N0 squamous cell carcinoma of the glottic larynx. However, patients who develop a local recurrence following irradiation have a substantial risk of harboring disease in the neck and should undergo a neck dissection in conjunction with the surgical procedure selected to resect the recurrent disease at the primary site.


The current status of S-100B as a biomarker in melanoma.
Melanoma is the most malignant type of all skin cancer types. It causes over 75% of all skin cancer related mortality. In the Netherlands, the total number of new diagnosed melanoma patients is expected to increase from 2400 patients in 2000 to 4800 patients in 2015. After surgical treatment, 20-28% of melanoma patients present with loco-regional recurrence, 26-60% with regional recurrences, and 15-50% with distant metastases. Early detection of lymph node (micro) metastases by means of a sentinel lymph node biopsy (SLNB) is therefore of crucial importance since early lymph node dissection decrease treatment morbidity and improve overall survival. However when patients present with palpable nodes, given the heterogeneity in survival, the suspicion rises that numerous patients have a form of subclinical dissemination, which can remain undetected by current modern imaging methods. Biomarkers could illuminate on this matter, although there is very little understanding of their biological significance. It can be expected that the strongest biological markers are surrogates of key biological events. The protein S-100B seems to be the best analyzed biomarker in melanoma. It has the potential to identify high-risk stage III melanoma patients who may benefit from adjuvant systematic treatment. In the stratification of new adjuvant therapeutic trials in patients with loco-regional recurrences, we therefore recommend the use of S-100B in the stratification. Since an effective (adjuvant) therapy for loco-regional metastatic and disseminated melanoma is recently introduced, the use of S-100B seems to alter dramatically in the near future.


Leonurine attenuates angiotensin II-induced cardiac injury and dysfunction via inhibiting MAPK and NF-κB pathway.
Hypertension is a common risk factor for heart failure, and excessive angiotensin II (Ang II) leads to hypertensive cardiac alterations such as hypertrophy, cardiac fibrosis, remodeling, and dysfunction. Leonurine is the major active alkaloid compound obtained from the traditional Chinese herbal medicine, Leonurus japonicus Houtt. The effects of leonurine on Ang II-induced hypertensive cardiac injury remain unknown. In the present study, we investigated the cardioprotective effects of leonurine in Ang II-infused mice and explored the underlying mechanisms in cardiomyocytes. Cardiac injury was induced by Ang II infusion in experimental mice with or without leonurine (at 10 or 20 mg/kg) treatment. H9c2 cells and neonatal rat primary cardiomyocytes were used to investigate the mechanisms through which leonurine exerts its protection effects. The results showed that leonurine significantly alleviated Ang II-induced cardiac hypertrophy, fibrosis, and inflammation in both mice and cultured cardiomyocytes. Echocardiography revealed that leonurine preserved cardiac function in mice. Further investigations revealed that leonurine inhibited the activation of the mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) pathways to reduce inflammatory response and injuries in Ang II-challenged cardiomyocytes. Inhibition of MAPKs and NF-κB in cardiomyocytes abolished the anti-inflammatory effects of leonurine. Our study provides evidence that leonurine exerts protective effects against Ang II-induced hypertensive cardiac remodeling and dysfunction by inhibiting the MAPK and NF-κB pathways. Leonurine may be a promising agent for treating hypertensive heart failure.


Relationship between level of milk production and estrous behavior of lactating dairy cows.
The objective of this study was to determine if there is an association between level of milk production and duration of estrus as determined by standing activity recorded by a radiotelemetry system. Holstein cows (n = 267; 50 DIM) were fitted with a transmitter that allowed continuous recording of standing activity. Cows were housed in a free-stall barn and milked twice daily. Ovulation was confirmed for all estruses (n = 380). Average milk production for the 10 days before the day of estrus was used to classify cows as lower (< 39.5 kg/day) or higher (>/= 39.5 kg/day) producers at the time of estrous expression. Follicle size and serum estradiol (E(2)) concentrations were determined in a subset of cows (n = 71) on the day of estrus. Duration (6.2 +/- 0.5 h versus 10.9 +/- 0.7 h; P < 0.0001), standing events (6.3 +/- 0.4 versus 8.8 +/- 0.6; P = 0.001), and standing time (21.7 +/- 1.3 s versus 28.2 +/- 1.9 s; P = 0.007) were shorter for estruses from higher (46.4 +/- 0.4 kg/day; n = 146) than lower producers (33.5 +/- 0.3 kg/day; n = 177). Milk production was correlated with the duration of estrus (r = -0.51; P < 0.0001; n = 323). Higher producers had lower E(2) concentrations than lower producers (6.8 +/- 0.5; n = 31 versus 8.6 +/- 0.5 pg/ml; n = 40; P = 0.01) in spite of larger pre-ovulatory follicle diameter (18.6 +/- 0.3; n = 31 versus 17.4 +/- 0.2 mm; n = 40; P 0.004). Interestingly, E(2) concentrations were not correlated with diameter of the pre-ovulatory follicle (r = -0.17; P = 0.15) but milk production was correlated with both E(2) concentrations (r = -0.57; P < 0.0001) and diameter of the pre-ovulatory follicle (r = 0.45; P < 0.0001). Thus, high milk production decreases duration of estrus probably due to decreased circulating concentrations of E(2).


Subunit interactions in rabbit-muscle glyceraldehyde-phosphate dehydrogenase, as measured by NAD+ and NADH binding.
1. The binding parameters for NADH and NAD+ to rabbit-muscle glyceraldehyde-phosphate dehydrogenase (D-glyceraldehyde-3-phosphate:NAD+ oxidoreductase (phosphorylating), EC 220.127.116.11) have been measured by quenching of the flourescence of the protein and the NADH. 2. The fact that the degree of protein fluorescence quenching by bound NAD+ or NADH, excited at 285 nm and measured at 340 nm ('blue' tryptophans), is not linearly related to the saturation functions of these nucleotides, leads to a slight overestimation of the interaction energy and an underestimation of the concentration of sites, if linearity is assumed. 3. This is also the case for NADH, but not for NAD+, when the protein fluorescence is excited at 305 nm and measured at 390 nm ('red' tryptophans). 4. The binding of NAD+ can be described by a model in which the binding of NAD+, via negative interactions within the dimer, induces weaker binding sites, with the result that the microscopic dissociation constant is 0.08 microM at low saturation and 0.18 microM for the holoenzyme. 5. The binding of NADH can be described on the basis of the same model, the dissociation constant at low saturation being 0.5 microM and of the holoenzyme 1.0 microM. 6. The fluorescence of bound NADH is not sensitive to the conformational changes that cause the decrease in affinity of bound NAD+ or NADH. 7. The binding of NAD+ to the 3-phosphoglyceroyl enzyme can be described by a dissociation constant that is at least two orders of magnitude greater than the dissociation constants of the unacylated enzyme. The affinity of NAD+ to this form of the enzyme is in agreement with the Ki calculated from product inhibition by NAD+ of the reductive dephosphorylation of 1,3-diphosphoglycerate.


Transforming growth factor beta 1 in ductal carcinoma in situ and invasive carcinomas of the breast.
Transforming growth factor beta (TGF-beta) is a multi-functional regulatory protein which can affect growth, immune responses, angiogenesis and the formation of extracellular matrix. Its role in breast carcinomas has been investigated using an antiserum to TGF-beta 1 and immunohistochemistry. 27 ductal carcinomas in situ and 54 invasive carcinomas were examined, employing formalin-fixed, paraffin-embedded material. There was no reactivity in 55.5% of in situ carcinomas in comparison with the invasive tumour where only a third were negative. Prominent reactivity was seen in 11% of in situ tumours, and 20% of invasive carcinomas. There was no correlation between detection of transforming growth factor beta 1, and histological grade, oestrogen receptor status, epidermal growth factor receptor status and Ki-67 labelling for the invasive carcinomas. There was a significant relationship between prominent reactivity and node status, all carcinomas with this degree of staining having metastasised. This, along with the differences between in situ and invasive carcinomas, suggests that TGF-beta 1 may be a determining factor for invasion and metastasis.


Development and validation of a carotid atherosclerosis risk prediction model based on a Chinese population.
This study aimed to identify independent risk factors for carotid atherosclerosis (CAS) and construct and validate a CAS risk prediction model based on the Chinese population. This retrospective study included 4,570 Chinese adults who underwent health checkups (including carotid ultrasound) at the Zhenhai Lianhua Hospital, Ningbo, China, in 2020. All the participants were randomly assigned to the training and validation sets at a ratio of 7:3. Independent risk factors associated with CAS were identified using multivariate logistic regression analysis. The least absolute shrinkage and selection operator combined with 10-fold cross-validation were screened for characteristic variables, and nomograms were plotted to demonstrate the risk prediction model. C-index and receiver operating characteristic curves, calibration plots, and decision curve analysis (DCA) were used to evaluate the risk model's discrimination, calibration, and clinical applicability. Age, body mass index, diastolic blood pressure, white blood cell count, mean platelet volume, alanine transaminase, aspartate transaminase, and gamma-glutamyl transferase were identified as independent risk factors for CAS. In the training, internal validation, and external validation sets, the risk model showed good discriminatory power with C-indices of 0.961 (0.953-0.969), 0.953 (0.939-0.967), and 0.930 (0.920-0.940), respectively, and excellent calibration. The results of DCA showed that the prediction model could be beneficial when the risk threshold probabilities were 1-100% in all sets. Finally, a network computer (dynamic nomogram) was developed to facilitate the physicians' clinical operations. The website is https://nbuhgq.shinyapps.io/DynNomapp/. The development of risk models contributes to the early identification and prevention of CAS, which is important for preventing and reducing adverse cardiovascular and cerebrovascular events.


Improvement of tuberous sclerosis complex (TSC) skin tumors during long-term treatment with oral sirolimus.
Oral mechanistic target of rapamycin inhibitors have been shown to reduce visceral tumor volume in patients with tuberous sclerosis complex (TSC). We sought to evaluate the cutaneous response to oral sirolimus in patients with TSC and an indication for systemic treatment, including long-term effects. A retrospective analysis of 14 adult patients with TSC prescribed sirolimus to treat lymphangioleiomyomatosis was performed. Serial photographs of angiofibromas, shagreen patches, and ungual fibromas taken before, during, and after the treatment period were blinded, then assessed using the Physician Global Assessment of Clinical Condition (PGA). Microscopic and molecular studies were performed on skin tumors harvested before and during treatment. Sirolimus significantly improved angiofibromas (median treatment duration 12 months; median PGA score 4.5 [range 1.5-5]; Wilcoxon signed rank test, P = .018) and shagreen patches (median treatment duration 10 months; median PGA score 4.5 [range 3.5-5]; Wilcoxon signed rank test, P = .039), whereas ungual fibromas improved in some patients (median treatment duration 6.5 months; median PGA score 4.66 [range 2.75-5]; Wilcoxon signed rank test, P = .109). Clinical, immunohistochemical, or molecular evidence of resistance was not observed (range 5-64 months of treatment). This was a retrospective analysis limited to adult women with lymphangioleiomyomatosis. Oral sirolimus is an effective long-term therapy for TSC skin tumors, particularly angiofibromas, in patients for whom systemic treatment is indicated.


Primary diagnosis of pulmonary embolism with unenhanced MRI for patients not eligible for CTPA: Clinical outcome.
To follow up the clinical outcome of patients with suspected pulmonary embolism (PE), in those only imaged using unenhanced, free-breathing magnetic resonance imaging (MRI). Fifty-seven patients aged 29-99 years (mean 70, SD 18) that could not undergo Computed Tomography Pulmonary Angiography (CTPA) were offered alternative imaging diagnostics in parallel with ongoing methodological studies validating MRI vs CTPA. Contraindications included renal failure (n = 44), severe iodine contrast allergy (n = 10), pregnancy (n = 2) and radioactive iodine therapy (n = 1). The unenhanced MRI protocol was based on free-breathing, steady-state free precession with no cardiac or respiratory gating. Retrospective review of the electronic medical record (EMR) was made of 0-12 months post-imaging and was collected during 2012-2018. All 57 MRIs were of diagnostic quality and 12 pulmonary embolisms were diagnosed. Of the 57 patients, 44 were already on, or had started anticoagulation therapy due to clinical suspicion of PE. Four of the patients were put on anticoagulation after the positive MRI and 13 were taken off anticoagulation after a negative MRI report. Other diagnoses reported (considering dyspnea) were pleural effusion (n = 24), consolidation (n = 12) and pericardial effusion (n = 2). One patient had a deep vein thrombosis (DVT) within three months of our negative MRI result and then had a stroke within one year. Another patient suffered a stroke within three months of being diagnosed (by MRI) with PE and given anticoagulation as treatment. Our method supported or altered clinical decision-making and treatment in this cohort. A diagnostic tool for PE without intravenous contrast agent or radiation is of great benefit for certain patients.


The Role of New Technologies in the Diagnosis and Surveillance of Non-Muscle Invasive Bladder Carcinoma: A Prospective, Double-Blinded, Monocentric Study of the XPERT© Bladder Cancer Monitor and Narrow Band Imaging© Cystoscopy.
Follow-up is essential for the early detection of recurrent non-muscle invasive bladder cancers (NMIBC). This study investigates the clinical relevance of new diagnostic tools such as an mRNA-based urine test (XPERT© Bladder Cancer Monitor, XBCM) and Narrow Band Imaging© (NBI) and compares them with the established follow-up diagnostics (white-light cystoscopy (WLC) and urine cytology). This was a prospective, double-blind, single-center study that involved patients undergoing NMIBC screening at a tertiary care center. Enrollment occurred between January 2018 and March 2020. In addition to standard care (WLC, cytology, and ultrasound), patients underwent XBCM urine testing and NBI cystoscopy. In total, 301 WLCs were performed; through this, 49 patients demonstrated NMIBC recurrence. NBI cystoscopy was congruent with WLC in all patients. Cytology showed a sensitivity (SE) and specificity (SP) of 27% and 97% (PPV: 65%; NPV 87%), respectively, whereas XBCM showed SE and SP of 58% and 89%, respectively (PPV: 51%; NPV: 92%; AUC: 0.79 (0.716-0.871)). Subgroup analysis showed improved SE and similar SP (PPV, NPV) for high grade (HG) recurrence, with a SE of 74% and SP of 89% (39%, 97%). NBI cystoscopy does not necessarily provide additional benefit over standard WLC. However, the XBCM may provide better SE and a diagnostic advantage in instances of HG disease recurrence.


Adoption of water reuse technologies: An assessment under different regulatory and operational scenarios.
Water reuse technologies may alleviate the water scarcity problems that affect many world regions, but their adoption is still limited. In particular, key actors in the adoption of water reuse technologies are water utilities, that provide both urban water and wastewater treatment services. Water utilities are embedded in the urban water system, which includes several stakeholders (urban water users, citizens at large, the environment) that may drive or pose barriers to water reuse adoption. Therefore, to ensure a smooth introduction of water reuse technologies, it is fundamental to understand how water reuse interacts with the existing urban water system and impacts its stakeholders. This paper contributes to the ongoing debate on water reuse by conceptualizing the interaction between water reuse technologies and the urban water system and its stakeholders, and addressing the adoption decision of water utilities by assessing its economic and environmental consequences. Based on a review of literature, policy and other secondary documents, and on primary data coming from interviews with experts from a water utility operating in Southern Italy, the study models the utility's response to a shift from urban to reuse water. It then simulates how reuse water introduction impacts on the utility and other stakeholders of the water system, under various regulatory and operational scenarios defined through a thorough analysis of policy documents and literature. Results show that the adoption of water reuse reduces the utility's margin by cannibalizing urban water demand, but appropriate policy measures may enhance the economic sustainability of reuse. System-level performances, such as impact on freshwater savings, costs for users, effects on the public budget, are also assessed, showing how different regulatory options moderate the intensity of impacts for the different stakeholders of the water system. Furthermore, the adoption of reuse water by the most distant users is found to enhance the economic sustainability of reuse and positively impact the utility's margin.


Retinal and circumpapillary nerve fiber layer thickness and associated factors in children.
To evaluate the distribution of macula and circumpapillary retina nerve fiber layer (cpRNFL) thickness and other associated factors among grade-1 primary school children in Lhasa using spectral-domain optical coherence tomography (SD-OCT). OCT assessment was conducted on 1856 grade-1 students from 7 primary schools in Lhasa, Tibet following a successful random stratified sampling of the students. Each child underwent comprehensive general and ocular examinations as well as an SD-OCT detection (12 × 9 mm, 3D wide scan mode, Topcon 3D OCT-1) to assess the thickness of the macula, ganglion cell-inner plexiform layer (GCIPL), ganglion cell complex (GCC), and cpRNFL. Multivariate and correlation analyses were performed to evaluate the association of the demographic and ocular variables. The average age of the 1762 (94.43%) students who underwent OCT assessment was 6.83 ± 0.46 years. Among them, 984 (53.02%) were boys. The number of students who had macular, cpRNFL, and optic disc scans completed and with adequate image quality were 1412 (82.2%), 1277 (74.4%), and 1243 (72.4%), respectively. The average macula full retinal thickness (FRT), GCIPL, GCC, and cpRNFL thickness of the students was 279.19 ± 10.61 μm, 76.41 ± 4.70 μm, 108.15 ± 6.15 μm, and 112.33 ± 13.5 μm, respectively. Multivariate regression and correlation analysis further revealed that boys and girls had significant differences in their average cpRNFL thickness. Moreover, GCC and GCIPL thickness was negatively correlated with IOP but positively correlated with the body mass index. The thickness of all the layers of the macula and cpRNFL were positively correlated with spherical equivalent. Further to this, the average macular FRT, GCIPL, and GCC thicknesses were positively correlated with cpRNFL global thickness. This study describes the normal distribution of macular retina, cpRNFL, and optic disc parameters in grade-1 Tibetan children in Lhasa. It contributes to the establishment of a normative ophthalmology database of Tibetan children, and advances the ability of OCT in ophthalmic disorder diagnosis during long-term monitoring in plateau.


An improved system for exposure of cultured mammalian cells to gaseous compounds in the chromosomal aberration assay.
A gas exposure system using rotating vessels was improved for exposure of cultured mammalian cells to gaseous compounds in the chromosomal aberration assay. This system was composed of 12 square culture vessels, a device for preparation of air containing test gas, and positive and negative control gases at target concentrations and for supplying these gases to the culture vessels, and a roller apparatus in an incubator. Chinese hamster lung cells (CHL/IU) were grown on one side of the inner surface of the square culture vessel in the MEM medium. Immediately prior to exposure, the medium was changed to the modified MEM. Air in the culture vessel was replaced with air containing test gas, positive or negative control gas. Then, the culture vessels were rotated at 1.0 rpm. The monolayered culture cells were exposed to test gas during about 3/4 rotation at upper positions and alternatively immersed into the culture medium during about 1/4 rotation at lower positions. This system allowed the chromosomal aberration assay simultaneously at least at three different concentrations of a test gas together with positive and negative control gases with and without metabolic activations, and duplicate culture at each exposure concentration. Seven gaseous compounds, 1,3-butadiene, chlorodifluoromethane, ethyl chloride, methyl bromide, methyl chloride, propyne, and vinyl chloride, none of which has been tested to date, were tested on CHL/IU for the chromosomal aberration assay using this gas exposure system. All the compounds except chlorodifluoromethane showed positive responses of the structural chromosomal aberrations, whereas polyploidy was not induced by any of these gases. This improved gas exposure system proved to be useful for detecting chromosomal aberrations of gaseous compounds.


Peritonitis in peritoneal dialysis: bacterial colonization by biofilm spread along the catheter surface.
We have used modern techniques of direct microscopic examination and quantitative bacterial recovery to show the existence of a route of bacterial colonization along the external and internal surfaces of Tenckhoff catheters implanted in experimental animals. The external route of progressive bacterial colonization extends from the cutaneous exit site through the dacron cuff and into the peritoneum. Bacterial growth along this route consists primarily of glycocalyx enclosed bacterial biofilms adherent to catheter and tissue surfaces, and this surface colonization may or may not give rise to peritoneal infection in which free-living bacteria are found in the peritoneal fluid. The rate of this progressive bacterial colonization depends on the degree of bacterial contamination of the exit site at the time of implantation. Exit site sterilization (hibitane) delays the process while inoculation with rabbit skin strains of Staphylococcus epidermidis accelerates it. Even with optimal implantation techniques, bacterial colonization proceeds via this subcutaneous route so that most Tenckhoff catheter surfaces are covered with a bacterial biofilm, consisting predominantly of gram positive cocci, within three weeks after the implantation of these devices. The rate of bacterial biofilm development on both surfaces of these Tenckhoff catheters, the bacterial colonization of peritoneal tissues, and the dissemination of bacteria into the peritoneal fluid are all significantly accelerated by dialysis in this experimental animal model of continuous ambulatory peritoneal dialysis (CAPD).


Chaperone-mediated ordered assembly of the SAGA and NuA4 transcription co-activator complexes in yeast.
Transcription initiation involves the coordinated activities of large multimeric complexes, but little is known about their biogenesis. Here we report several principles underlying the assembly and topological organization of the highly conserved SAGA and NuA4 co-activator complexes, which share the Tra1 subunit. We show that Tra1 contributes to the overall integrity of NuA4, whereas, within SAGA, it specifically controls the incorporation of the de-ubiquitination module (DUB), as part of an ordered assembly pathway. Biochemical and functional analyses reveal the mechanism by which Tra1 specifically interacts with either SAGA or NuA4. Finally, we demonstrate that Hsp90 and its cochaperone TTT promote Tra1 de novo incorporation into both complexes, indicating that Tra1, the sole pseudokinase of the PIKK family, shares a dedicated chaperone machinery with its cognate kinases. Overall, our work brings mechanistic insights into the assembly of transcriptional complexes and reveals the contribution of dedicated chaperones to this process.


Hyperthermic intra-peritoneal chemotherapy and anticancer immune response.
Peritoneal carcinomatosis (PC) is a metastatic disease of primary tumors localized in the abdomen. Although this disease is considered a terminal condition, recent treatments combining surgery with heated intra-peritoneal chemotherapy (HIPEC) significantly increase patient survival. We have determined that the protective effect of HIPEC is partially linked to the induction of an efficient anticancer immune response.


Efficacy of a high-dose proton pump inhibitor in patients with gastroesophageal reflux disease: a single center, randomized, open-label trial.
The extraesophageal manifestations of gastroesophageal reflux disease (GERD) are more difficult to manage than the typical symptoms. The efficacy of high-dose and standard-dose proton pump inhibitors against these atypical symptoms is not yet established. In this single center, randomized, open-label study, patients with GERD received rabeprazole for 8 weeks, either 20 mg once daily (standard-dose group) or 20 mg twice daily (high-dose group). Patients were assessed before treatment and at weeks 4 and 8 with a 5-graded scale questionnaire consisting of 2 typical symptoms (heartburn and acid regurgitation) and 8 atypical symptoms (chest pain, cough, globus, wheezing, laryngopharyngitis, hoarseness, belching, and dysphagia). Sufficient improvement of reflux symptoms was defined as ≥50% reduction from the initial questionnaire score. Final analyses included 35 patients in the standard-dose group and 38 patients in the high-dose group. The rate of sufficient improvement for typical symptoms was significantly higher in the high-dose group than in the standard-dose group (100.0% vs. 84.0%, P = 0.040). For atypical symptoms, the rate of sufficient improvement tended to be higher in the high-dose group than in the standard-dose group (82.4% vs. 63.0%, P = 0.087). Scores of typical and some atypical symptoms (cough and globus) improved after treatment, with significant inter-group differences in time-course changes. High-dose rabeprazole is more effective for relieving typical GERD symptoms and some atypical symptoms such as cough and globus than a standard-dose regimen. This research was enrolled in a registry of clinical trials run by United States National Library of Medicine at the National Institutes of Health ( ClinicalTrials.gov Protocol Registration and Results system ID: NCT04001400 ). This study was registered on June 26, 2019 - Retrospectively registered.


Feeding ecology of monk sakis (Pithecia monachus) in a seasonally flooded forest in western Amazonia.
Sakis (genus Pithecia) are frugivorous primates with a preference for seeds that complete their diet with leaves and insects. Fruit pulp and seeds are known to have different nutritional characteristics that change during the process of ripening. The consumption of seeds can be an adaptation to changes in resource availability, as unripe seeds are a more steadily available resource than ripe pulp or young leaves. Here, we present the first study of the feeding ecology of monk sakis (Pithecia monachus). We investigated dietary composition and identified important feeding plants in a seasonally flooded forest within the Área de Conservación Regional Comunal Tamshiyacu-Tahuayo in Peruvian Amazonia. Throughout 20 months, we followed groups of monk sakis by foot and canoe and recorded 459 feeding events. Seeds were the most frequently consumed food item (49%), followed by pulp (mesocarp, pericarp or aril; 25%) and arthropods (22%). Leaves, bark, and flowers were ingested only sporadically. The importance of ripe seeds and arthropods in the diet of the monk sakis differed from other studies: we recorded the consumption of mostly ripe seeds and the share of arthropods was relatively high.


Influence of Land Cover and Soil Moisture based Brown Ocean Effect on an Extreme Rainfall Event from a Louisiana Gulf Coast Tropical System.
Extreme flooding over southern Louisiana in mid-August of 2016 resulted from an unusual tropical low that formed and intensified over land. We used numerical experiments to highlight the role of the 'Brown Ocean' effect (where saturated soils function similar to a warm ocean surface) on intensification and it's modulation by land cover change. A numerical modeling experiment that successfully captured the flood event (control) was modified to alter moisture availability by converting wetlands to open water, wet croplands, and dry croplands. Storm evolution in the control experiment with wet antecedent soils most resembles tropical lows that form and intensify over oceans. Irrespective of soil moisture conditions, conversion of wetlands to croplands reduced storm intensity, and also, non-saturated soils reduced rain by 20% and caused shorter durations of high intensity wind conditions. Developing agricultural croplands and more so restoring wetlands and not converting them into open water can impede intensification of tropical systems that affect the area.