Structural features which control folding of homologous proteins in cell-free translation systems. The effect of a mitochondrial-targeting presequence on aspartate aminotransferase.
When the precursor to mitochondrial aspartate aminotransferase (pmAspAT) is synthesized in a rabbit reticulocyte lysate translation system (RRL), its properties are quite unlike those of the purified protein (Mattingly, J.R., Jr., Youssef, J., Iriarte, A., and Martinez-Carrion, M. (1993) J. Biol. Chem. 268, 3925-3937). These results suggest that molecular chaperones present in RRL modulate the folding of pmAspAT. To investigate the structural basis for this, we have used protease resistance to monitor the extent of folding for several related AspATs after synthesis in RRL and in wheat germ extract (WGE). In addition to pmAspAT, the following proteins were examined: the mature form of pmAspAT (delta 2-28 pmAspAT), its cytosolic counterpart (cAspAT), a chimeric protein consisting of the presequence of pmAspAT attached to the amino terminus of cAspAT (pcAspAT), and a pmAspAT variant in which the presequence and the amino-terminal domain of the mature enzyme are deleted (delta 2-57 pmAspAT). In RRL, delta 2-28 pmAspAT folds somewhat faster than intact pmAspAT, whereas the truncated delta 2-57 pmAspAT is unable to fold. In contrast, cAspAT and pcAspAT both fold with extreme rapidity. After synthesis in WGE, pmAspAT and delta 2-28 pmAspAT never acquire a protease-resistant conformation, whereas the folding of cAspAT and pcAspAT still occurs rapidly. We conclude that the presequence has only a minor role in determining the folding rate of the pmAspAT mitochondrial precursor protein in RRL or WGE and has no influence on the folding of the homologous cAspAT. Rather, the primary sequence of the mature part of the protein seems to dictate whether or how molecular chaperones regulate folding events.


[Manual for "Junior Doctors" in medical oncology].
The reform of medical residency introduced in 2017 established the position of Junior Doctor, for its last phase, called the consolidation phase. Its goal is the increasing and supervised autonomy of the resident, in order to better support the transition toward senior practitioner. It appears necessary to define proper guidelines on the status and the specific role of Junior Doctor in medical oncology, in order to help the implementation of the reform of the 3rd cycle. This document is the result of a collaboration between AERIO and CNEC, that respectively represent medical oncology residents and university teachers. It aims to advise and guide local practices, without being compulsory, before the arrival of the first Junior Doctors in November 2021. It explains the Junior Doctors' principal jobs: consultation, multidisciplinary meeting, day clinic, hospitalization, clinical research, quality policy and teaching.


Inhibition of a mouse hepatoma by the alkylating agent Trenimon linked to immunoglobulins.
Trenimon was conjugated in active alkylating form to rabbit anti-mouse H6 hepatoma globulin (AHG) with retention of antibody activity. H6 hepatoma-inoculated mice were given various combinations of conjugates, free Trenimon, and unconjugated immunoglobulins in daily injections for 5 days. Linkage of Trenimon to immunoglobulins reduced systemic toxicity of the drug, with comparative retention of its antitumor activity. The antitumor action of Trenimon was potentiated by AHG irrespective of whether the drug was directly linked to AHG or free AHG was administered along with Trenimon linked to normal rabbit globulin (NRG). In vitro, Trenimon bound to AHG was less inhibitory to hepatoma cells than free Trenimon, but more inhibitory than Trenimon-NRG conjugates. There was no significant endocytosis of conjugates by the hepatoma cells. This suggests that unlike free Trenimon, the target molecules of Trenimon-immunoglobulin conjugates are not intracellular DNA but are located on the surface of the hepatoma cells.


Effects of insulin on steroidogenesis in cultured porcine ovarian theca.
The effects of insulin on porcine thecal steroidogenesis were examined in long-term cultures of hyaluronidase-collagenase dispersed thecal cells. The thecal cultures made significant amounts of progesterone (P) and androstenedione (delta 4 A). Testosterone, dihydrotestosterone, estrone, and estradiol could not be detected in the media. Luteinizing hormone (LH) alone significantly increased P and delta 4 A accumulation. Insulin alone increased P accumulation on days 2 to 4 of culture. Insulin alone did not stimulate delta 4 A accumulation. Insulin plus LH resulted in a significantly greater accumulation of P and delta 4 A than LH alone. These results suggest that insulin may be a regulator of ovarian thecal steroidogenesis.


A randomized study of beef lung and pork mucosal heparin in cardiac surgery.
Beef lung heparin had substantially greater anticoagulant activity than pork mucosal heparin during a preoperative heparin tolerance test and also during cardiopulmonary bypass (CPB) in 100 randomized patients. Supplemental heparin was needed during CPB in many more of the patients receiving pork mucosal heparin. Heparin rebound was detected in 16 patients; this low incidence may result from a relatively high protamine:heparin ratio. There was notably less postoperative bleeding in those who received beef lung heparin. Platelet counts were not altered by either type of heparin. It is surprising that the preoperative tolerance curve only predicted heparin sensitivity during CPB in one-half of the patients. Blood activated coagulation time levels were increased markedly by hemodilution and further raised by hypothermia. These data demonstrate that beef lung heparin is better than pork mucosal heparin for CPB.


Genetic diversity and phylogenetic analysis of the surface layer protein A gene (slpA) among Clostridioides difficile clinical isolates from Tehran, Iran.
Clostridioides difficile is the most common causative agent of healthcare-associated diarrhea. C. difficile strains produce a crystalline surface layer protein (SlpA), encoded by the slpA gene. Previous studies have shown that SlpA varies among C. difficile strains. In this study, we used the SlpA sequence-based typing system (SlpAST) for the molecular genotyping of C. difficile clinical isolates identified in Iran; the PCR ribotypes (RTs) and toxin profiles of the isolates were also characterized. Forty-eight C. difficile isolates were obtained from diarrheal patients, and characterized by capillary electrophoresis (CE) PCR ribotyping and the detection of toxin genes. In addition, the genetic diversity of the slpA gene was investigated by Sanger sequencing. The most common RTs were RT126 (20.8%), followed by RT001 (12.5%) and RT084 (10.4%). The intact PaLoc arrangement representing cdu2+/tcdR+/tcdB+/tcdE+/tcdA+/tcdC+/cdd3+ profile was the predominant pattern and cdtA and cdtB genes were found in one-third of the isolates. Using the SlpA genotyping, 12 main genotypes and 16 subtypes were identified. The SlpA type 078-1 was the most prevalent genotype (20.8%), and identified within the isolates of RT126. The yok-1, gr-1, cr-1 and kr-3 genotypes were detected in 14.5%, 12.5%, 12.5% and 8.3% of isolates, respectively. Almost all the isolates with the same RT were clustered in similar SlpA sequence types. In comparison to PCR ribotyping, SlpAST, as a simple and highly reproducible sequenced-based technique, can discriminate well between C. difficile isolates. This typing method appears to be a valuable tool for the epidemiological study of C. difficile isolates worldwide.


Methanol-precipitated, detergent-treated Newcastle disease virus as an agar-gel precipitation test antigen.
Methanol-precipitated, detergent-treated (Triton X-100) Newcastle disease virus (NDV) was found suitable as an agar gel precipitation test (AGPT) antigen. One hundred and twenty sera were tested by the haemagglutination-inhibition (HI) test and AGPT against NDV. There was a very significant increase in the proportion of AGPT positive samples with increase in HI titre. Hence AGPT can be recommended as a field based test for seromonitoring following vaccinations against ND where laboratory facilities are inadequate.


Criticality-Enhanced Magnetocaloric Effect in Quantum Spin Chain Material Copper Nitrate.
In this work, a systematic study of Cu(NO3)2·2.5 H2O (copper nitrate hemipentahydrate, CN), an alternating Heisenberg antiferromagnetic chain model material, is performed with multi-technique approach including thermal tensor network (TTN) simulations, first-principles calculations, as well as magnetization measurements. Employing a cutting-edge TTN method developed in the present work, we verify the couplings J = 5.13 K, α = 0.23(1) and Landé factors g∥= 2.31, g⊥ = 2.14 in CN, with which the magnetothermal properties have been fitted strikingly well. Based on first-principles calculations, we reveal explicitly the spin chain scenario in CN by displaying the calculated electron density distributions, from which the distinct superexchange paths are visualized. On top of that, we investigated the magnetocaloric effect (MCE) in CN by calculating its isentropes and magnetic Grüneisen parameter. Prominent quantum criticality-enhanced MCE was uncovered near both critical fields of intermediate strengths as 2.87 and 4.08 T, respectively. We propose that CN is potentially a very promising quantum critical coolant.


Growth Hormone Receptor Knockdown Sensitizes Human Melanoma Cells to Chemotherapy by Attenuating Expression of ABC Drug Efflux Pumps.
Melanoma remains one of the most therapy-resistant forms of human cancer despite recent introductions of highly efficacious targeted therapies. The intrinsic therapy resistance of human melanoma is largely due to abundant expression of a repertoire of xenobiotic efflux pumps of the ATP-binding cassette (ABC) transporter family. Here, we report that GH action is a key mediator of chemotherapeutic resistance in human melanoma cells. We investigated multiple ABC efflux pumps (ABCB1, ABCB5, ABCB8, ABCC1, ABCC2, ABCG1, and ABCG2) reportedly associated with melanoma drug resistance in different human melanoma cells and tested the efficacy of five different anti-cancer compounds (cisplatin, doxorubicin, oridonin, paclitaxel, vemurafenib) with decreased GH action. We found that GH treatment of human melanoma cells upregulates expression of multiple ABC transporters and increases the EC50 of melanoma drug vemurafenib. Also, vemurafenib-resistant melanoma cells had upregulated levels of GH receptor (GHR) expression as well as ABC efflux pumps. GHR knockdown (KD) using siRNA in human melanoma cells treated with sub-EC50 doses of anti-tumor compounds resulted in significantly increased drug retention, decreased cell proliferation and increased drug efficacy, compared to mock-transfected controls. Our set of findings identify an unknown mechanism of GH regulation in mediating melanoma drug resistance and validates GHR as a unique therapeutic target for sensitizing highly therapy-resistant human melanoma cells to lower doses of anti-cancer drugs.


Ghrelin, leptin and high-molecular-weight adiponectin in relation to depressive symptoms in older adults: Results from the Longitudinal Aging Study Amsterdam.
Ghrelin, leptin and high-molecular-weight (HMW) adiponectin have been linked to depression in middle-aged adults. Pathophysiological mechanisms of depression change as age progresses and it is unclear whether the same associations exist in older adults. We analyzed the associations between ghrelin, leptin and HMW adiponectin and depressive symptoms (Center for Epidemiologic Studies Depression (CES-D) score ≥ 16) in a community-dwelling cohort of 898 participants in a multivariable logistic regression analysis at baseline and after three years of follow-up, were applicable stratified by sex, age and waist-hip-ratio (WHR). At baseline no significant associations were found. After three years of follow-up ghrelin was associated with higher odds for depressive symptoms (fully adjusted continuous analysis OR 2.27, 95% CI 1.42 - 3.61). There was effect modification for age and WHR, with significant associations in participants younger than 69.7 years (median) and with a WHR below 0.9554 (mean). In the sex-stratified analysis for leptin we found significant associations in men (fully adjusted continuous analysis OR 1.07, 95% CI 1.02 - 1.12). For HMW adiponectin there were no significant associations in the multivariable analysis. As our cohort consisted of relatively healthy participants with intact cognitive function, selection bias may have contributed to lack of significant baseline associations. Our results show significant associations between ghrelin and - for men only - leptin and depressive symptoms after three years of follow up. This may provide a new therapeutic window for treatment of depressive symptoms in older adults, as both ghrelin and leptin are positively influenced by weight loss.


Bioresorbable β-TCP-FeAg nanocomposites for load bearing bone implants: High pressure processing, properties and cell compatibility.
In this paper, the processing and properties of iron-toughened bioresorbable β-tricalcium phosphate (β-TCP) nanocomposites are reported. β-TCP is chemically similar to bone mineral and thus a good candidate material for bioresorbable bone healing devices; however intrinsic brittleness and low bending strength make it unsuitable for use in load-bearing sites. Near fully dense β-TCP-matrix nanocomposites containing 30vol% Fe, with and without addition of silver, were produced employing high energy attrition milling of powders followed by high pressure consolidation/cold sintering at 2.5GPa. In order to increase pure iron's corrosion rate, 10 to 30vol% silver were added to the metal phase. The degradation behavior of the developed composite materials was studied by immersion in Ringer's and saline solutions for up to 1month. The mechanical properties, before and after immersion, were tested in compression and bending. All the compositions exhibited high mechanical strength, the strength in bending being several fold higher than that of polymer toughened β-TCP-30PLA nanocomposites prepared by the similar procedure of attrition milling and cold sintering, and of pure high-temperature sintered β-TCP. Partial substitution of iron with silver led to an increase in both strength and ductility. Furthermore, the galvanic action of silver particles dispersed in the iron phase significantly accelerated in vitro degradation of β-TCP-30(Fe-Ag) nanocomposites. After 1month immersion, the composites retained about 50% of their initial bending strength. In cell culture experiments, β-TCP-27Fe3Ag nanocomposites exhibited no signs of cytotoxicity towards human osteoblasts suggesting that they can be used as an implant material.


Decision-making tools to assist prognosis and treatment choices in early breast cancer: a review.
Breast cancer remains the most common type of cancer affecting women worldwide with an estimated lifetime risk of 1:8. With developments in adjuvant treatment and the identification of breast cancer subtypes, rising expectation of 'personalized' and 'targeted' therapy, decisions on systemic therapy have become increasingly more difficult. In a bid to assist clinicians in correctly selecting patients in whom systemic adjuvant therapy would be of most benefit, a number of decision-making tools have been developed. In this article, the authors will review some of these tools, explore how they were developed and assess the impact they have had on daily clinical practice.


53BP1 oligomerization is independent of its methylation by PRMT1.
p53 binding protein 1 (53BP1) participates in the repair of DNA double stranded breaks (DSBs) where it is recruited to or near sites of DNA damage. Although little is known about the biochemical functions of 53BP1, the protein possesses several motifs that are likely important for its role as a DNA damage response element. This includes two BRCA1 C-terminal repeats, tandem Tudor domains, and a variety of phosphorylation sites. Here we show that a glycine-arginine rich (GAR) stretch of 53BP1 lying upstream of the Tudor motifs is methylated. We demonstrate that arginine residues within this region are important for asymmetric methylation by the PRMT1 methyltransferase. We further show that sequences upstream of the Tudor domains that do not include the GAR stretch are sufficient for 53BP1 oligomerization in vivo. Thus, although Tudor domains bind methylated proteins, 53BP1 homo-oligomerization occurs independently of Tudor function. Lastly, we find that deficiencies in 53BP1 generate a "hyper-rec" phenotype. Collectively, these data provide new insight into 53BP1, an important component in maintaining genomic stability.


Corynebacterium rouxii, a recently described member of the C. diphtheriae group isolated from three dogs with ulcerative skin lesions.
Corynebacterium (C.) diphtheriae is one of the two etiological pathogens for human diphtheria with significant morbidity and mortality. Recently, members of its biovar Belfanti have been described as two novel species, C. belfantii and C. rouxii. The most important virulence factor and also the premise to cause diphtheria is the isolate's capacity to encode and express the diphtheria toxin (DT). In contrast to C. ulcerans, which represents a potentially zoonotic pathogen, C. diphtheriae (incl. the novel deduced species) has almost exclusively been found to comprise a human pathogen. We here report three rare cases of C. rouxii isolation from dogs suffering from disseminated poly-bacterial exsudative to purulent dermatitis and a traumatic labial defect, respectively. The isolates were identified as C. diphtheriae based on commercial biochemistry and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-TOF MS) analysis. However, recently described specific spectral peaks were highly similar to spectra of C. rouxii, which was confirmed by whole genome sequencing. Further investigations of the dog isolates for the presence of DT by tox gene qPCR revealed negative results. The findings from this study point out that skin infections in companion animals can be colonized by uncommon and so believed human specific pathogens, thereby resembling the clinical signs of cutaneous diphtheria.


Clinical course, treatment and visual outcome of an outbreak of Burkholderia contaminans endophthalmitis following cataract surgery.
Postoperative endophthalmitis is a rare but dreaded complication of intraocular surgery and often results in severe visual impairment or blindness. The present study describes the clinical course, treatment and visual outcome of an outbreak of Burkholderia contaminans endophthalmitis following cataract surgery. Among 290 patients who underwent uneventful phacoemulsification cataract surgery at one outpatient clinic between January 4th and 28th 2019, 6 cases developed Burkholderia contaminans endophthalmitis. Clinical data were collected by retrospective review of patient records. Microbiological samples from vitreous aspirates, intraocular lenses (IOL) and lens capsules were cultured, and recA and draft whole genome sequences analysed. The recA sequences of all Burkholderia contaminans isolates and the allelic profile of the isolates were identical. All cases had a similar clinical presentation with rapid development of endophthalmitis symptoms with variable time to onset. The mean time to admission was 34 days (12-112 days). All cases had a seemingly favourable response to intravitreal antibiotics. However, acute recurrences occurred after long time periods (12-71 days). The cases experienced between 0 and 3 recurrences. Due to persistent infection, the cases received between 5 and 15 treatments (mean 7.8) including IOL and lens capsule explantation in 5 of 6 cases. Burkholderia contaminans was detected in all explanted lens capsules. The final corrected distance visual acuity (CDVA, Snellen chart) was between 0.8 and 1.2 and all cases had final CDVA ≥0.8. A persistent and intensive treatment approach including total lens capsule and IOL explantation is recommended for Burkholderia contaminans endophthalmitis following cataract surgery and may lead to a favourable visual result.


PDCD1 and PDCD1LG1 polymorphisms affect the susceptibility to multiple myeloma.
Single-nucleotide polymorphisms (SNPs) of the programmed cell death protein-1 (PDCD1), programmed cell death protein-1 ligand-1 (PDCD1LG1), and cytotoxic T lymphocyte-associated antigen-4 (CTLA4) genes are implicated in the pathogenesis of some cancers. We investigated the role of PDCD1, PDCD1LG1, and CTLA4 SNPs in MM pathogenesis and the susceptibility to and clinical features of multiple myeloma (MM). We obtained genomic DNA from 124 patients with MM and 211 healthy controls and detected PDCD1 (rs36084323, rs41386349, and rs2227982), PDCD1LG1 (rs2297136 and rs4143815), and CTLA4 (rs733618, rs11571316, rs231775, and rs3087243) genotypes using the polymerase chain reaction-restriction fragment length polymorphism method or the TaqMan allelic discrimination real-time PCR method. The patients with MM had a significantly higher frequency of the PDCD1 GCC/GCC haplotype (rs36084323/rs41386349/rs2227982) compared with the healthy controls. PDCD1 rs2227982 CC genotype was associated significantly with a higher frequency of bone lesions. Patients with PDCD1LG1 rs2297136 TT and TC types (high-expression types) showed lower albumin level than those with CC genotype. In addition, the PDCD1LG1 rs4143815 CC and CG types (high-expression types) were associated significantly with higher frequency of patients who were treated with thalidomide and/or bortezomib. However, there was no statistical significance between CTLA4 polymorphisms and clinical variables of patients with MM. There were no significant differences between all the polymorphisms and OS. Our study indicates that the PDCD1 haplotype is associated with a susceptibility to MM. The PDCD1 rs2227982 and PDCD1LG1 rs2297136 affect the clinical features of multiple myeloma patients.


Chemical carcinogen-induced rat mammary carcinogenesis is a potential model of p21-activated kinase positive female breast cancer.
The p21-activated kinase 1 (PAK1) gene encodes a serine/threonine kinase that is overexpressed in a subset of human breast carcinomas with poor prognosis. The laboratory rat (Rattus norvegicus) orthologous gene is located at Mammary carcinoma susceptibility 3 (Mcs3) QTL on rat chromosome 1. We used quantitative PCR to determine effects of Mcs3 genotype and 7,12-dimethylbenz(a)anthracene (DMBA) exposure on Pak1 expression. There was no effect of Mcs3 genotype; however, there was a 3.5-fold higher Pak1 level in DMBA-exposed mammary glands (MGs) than in unexposed glands (P < 0.05). Sequence variants in Pak1 exons did not alter amino acid sequence between Mcs3-susceptible and -resistant strains. Protein expression of PAK1/Pak1 in human breast carcinomas and DMBA-exposed rat mammary glands was detected using immunohistochemistry (IHC). Rat mammary glands from 12-wk-old females unexposed to DMBA were negative for Pak1, whereas 24% of carcinogen-exposed mammary glands from age-matched females stained positive for Pak1. The positive mammary glands exposed to carcinogen had no pathological signs of disease. Human breast carcinomas, used as comparative controls, had a 22% positivity rats. This was consistent with other human breast cancer studies of PAK1 expression. Similar frequencies of human/rat PAK1/Pak1 expression in female breast carcinomas and carcinogen-induced rat mammary glands, showing no visible pathogenesis of disease, suggests aberrant PAK1 expression is an early event in development of some breast cancers. Laboratory rats will be a useful experimental organism for comparative studies of Pak1-mediated mechanisms of breast carcinogenesis. Future studies of PAK1 as a diagnostic marker of early breast disease are warranted.


Effect of ceramic layering on the fit of cobalt-chromium alloy 3-unit fixed dental prostheses fabricated by additive, soft milling, and casting technologies.
The change in fit after ceramic layering of additively manufactured cobalt-chromium alloy frameworks has not been evaluated extensively. The purpose of this in vitro study was to compare the fit of cobalt-chromium alloy fixed dental prostheses fabricated by different techniques before and after ceramic layering. A cobalt-chromium alloy master model was prepared to receive a 3-unit fixed dental prosthesis. Sixty cobalt-chromium alloy frameworks (N=60) were manufactured by using 3 manufacturing techniques (n=20): selective laser melting (Mediloy S-Co), soft milling (Ceramill Sintron), and conventional casting as the control group (Girobond NB). The replica technique was used to measure the marginal and internal discrepancies. The frameworks were then layered with ceramic, and the same fit measuring procedure was repeated. The results were compared before and after ceramic layering within each group. The data were analyzed using the Levene, ANOVA, and paired-samples t tests (α=.05). A significant difference was found within groups before and after ceramic layering for selective laser melting (P=.006) and soft milling (P=.009) but not for conventional casting (P>.05). No statistical difference was reported in the marginal region for conventional casting group (P=.155) in contrast with the soft milling and selective laser melting groups (P=.003). Soft milling showed increased gap values in the marginal (P=.006) and occlusal regions (P=.004). Ceramic layering increased the discrepancy of the laser-sintered and milled frameworks, increasing the marginal discrepancy.


miR-186 induces tetraploidy in arsenic exposed human keratinocytes.
Chronic inorganic arsenic (iAs) exposure in drinking water is a global issue affecting >225 million people. Skin is a major target organ for iAs. miRNA dysregulation and chromosomal instability (CIN) are proposed mechanisms of iAs-induced carcinogenesis. CIN is a cancer hallmark and tetraploid cells can better tolerate increase in chromosome number and aberration, contributing to the evolution of CIN. miR-186 is overexpressed in iAs-induced squamous cell carcinoma relative to iAs-induced hyperkeratosis. Bioinformatic analysis indicated that miR-186 targets mRNAs of important cell cycle regulators including mitotic checkpoint serine/threonine kinase B (BUB1) and cell division cycle 27 (CDC27). We hypothesized that miR-186 overexpression contributes to iAs-induced transformation of keratinocytes by targeting mitotic regulators leading to induction of CIN. Ker-CT cells, a near diploid human keratinocyte cell line, were transduced with miR-186 overexpressing or scrambled control lentivirus. Stable clones were isolated after puromycin selection. Clones transduced with lentivirus expressing either a scrambled control miRNA or miR-186 were maintained with 0 or 100 nM iAs for 4 weeks. Unexposed scrambled control clones were considered as passage matched controls. Chronic iAs exposure increased miR-186 expression in miR-186 clones. miR-186 overexpression significantly reduced CDC27 levels irrespective of iAs exposure. The percentage of tetraploid or aneuploid cells was increased in iAs exposed miR-186 clones. Aneuploidy can arise from a tetraploid intermediate. Suppression of CDC27 by miR-186 may lead to impairment of mitotic checkpoint complex formation and its ability to maintain cell cycle arrest leading to chromosome misalignment. As a result, cells overexpressing miR-186 and chronically exposed to iAs may have incorrect chromosome segregation and CIN. These data suggest that dysregulation of miRNA by iAs mediates tetraploidy, aneuploidy and chromosomal instability contributing to iAs-induced carcinogenesis.


Increase in outpatient knee arthroscopy in the United States: a comparison of National Surveys of Ambulatory Surgery, 1996 and 2006.
This study was proposed to investigate the changes in the utilization of knee arthroscopy in an ambulatory setting over the past decade in the United States as well as its implications. The National Survey of Ambulatory Surgery, last carried out in 1996, was conducted again in 2006 by the Centers for Disease Control and Prevention. We analyzed the cases with procedure coding indicative of knee arthroscopy or anterior cruciate ligament reconstruction. To produce estimates for all arthroscopic procedures on the knee in an ambulatory setting in the United States for each year, we performed a design-based statistical analysis. The number of arthroscopic procedures on the knee increased 49% between 1996 and 2006. While the number of arthroscopic procedures for knee injury had dramatically increased, arthroscopic procedures for knee osteoarthritis had decreased. In 1996, knee arthroscopies performed in freestanding ambulatory surgery centers comprised only 15% of all orthopaedic procedures, but the proportion increased to 51% in 2006. There was a large increase in knee arthroscopy among middle-aged patients regardless of sex. In 2006, >99% of arthroscopic procedures on the knee were in an outpatient setting. Approximately 984,607 arthroscopic procedures on the knee (95% confidence interval, 895,999 to 1,073,215) were performed in an outpatient setting in 2006. Among those, 127,446 procedures (95% confidence interval, 95,124 to 159,768) were for anterior cruciate ligament reconstruction. Nearly 500,000 arthroscopic procedures were performed for medial or lateral meniscal tears. This study revealed that the knee arthroscopy rate in the United States was more than twofold higher than in England or Ontario, Canada, in 2006. Our study found that nearly half of the knee arthroscopic procedures were performed for meniscal tears. Meniscal damage, detected by magnetic resonance imaging, is commonly assumed to be the source of pain and symptoms. Further study is imperative to better define the symptoms, physical findings, and radiographic findings that are predictive of successful arthroscopic treatment.


Recombinant human erythropoietin antagonizes trastuzumab treatment of breast cancer cells via Jak2-mediated Src activation and PTEN inactivation.
We found that the receptor for erythropoietin (EpoR) is coexpressed with human epidermal growth factor receptor-2 (HER2) in a significant percentage of human breast tumor specimens and breast cancer cell lines. Exposure of HER2 and EpoR dual-positive breast cancer cells to recombinant human erythropoietin (rHuEPO) activated cell signaling. Concurrent treatment of the cells with rHuEPO and trastuzumab reduced the cells' response to trastuzumab both in vitro and in vivo. We identified Jak2-mediated activation of Src and inactivation of PTEN as underlying mechanisms through which rHuEPO antagonizes trastuzumab-induced therapeutic effects. Furthermore, we found that compared with administration of trastuzumab alone, concurrent administration of rHuEPO and trastuzumab correlated with shorter progression-free and overall survival in patients with HER2-positive metastatic breast cancer.


Structural Dependent Eu3+ Luminescence, Photoelectric and Hysteresis Effects in Porous Strontium Titanate.
Eu3+ doped porous nanostructured SrTiO3 films and powder fabricated by sol-gel route without using any precursor template are characterized by different morphology and phase composition. The films show red and yellow luminescence with the most intensive photoluminescence (PL) bands at 612 nm and 588 nm, respectively. Raman, secondary ion mass spectrometry (SIMS), and X-ray diffraction (XRD) analysis of undoped nanostructured porous SrTiO3 films showed the presence of TiO2, SrO, and SrTiO3 phases and their components. The undoped porous SrTiO3 films are photosensitive and demonstrate resistive switching. The capacitance-voltage hysteresis loops with the width of about 6 V in the frequency range of 2 kHz-2 MHz were observed.


Detection of T cells reactive to intestinal alkaline phosphatase, an autoantigen in acute bacterial infections, and discrimination between autoantigen-specific CD5+ and CD5- B cells.
Autoantibodies of the IgG isotype, specifically directed against intestinal alkaline phosphatase (IAP), occur transiently in the majority of sera from patients with acute bacterial infections. Sometimes they are observed in autoimmune diseases. Using a T cell proliferation assay, it was found that isolated peripheral blood mononuclear cells (PBMC) from IAP autoantibody (IAPA)-positive patients (n = 18) responded significantly to IAP, whereas proliferation could not be induced in PBMC from healthy donors (n = 11). Significant stimulation of PBMC from patients (n = 11) was not obtained by use of transferrin, a common autoantigen in humans, indicating the specificity of stimulation of IAP-reactive T cells. Furthermore, T cell proliferation was observed when a highly purified IAP fragment (CNBr 21) spanning amino acids 334-462 of the primary structure of IAP was used as antigen. Thus, it was shown that an immunodominant T cell epitope resides within the CNBr 21 fragment which also contains a discontinuous B cell epitope as evaluated previously. Double immunocytochemical staining of T cell-depleted PBMC with IAP and an anti-human CD5 antibody allowed the detection of CD5+ B lymphocytes, which probably produce natural IAPA (nIAPA). These nIAPA-specific CD5+ B cells occurred with approximately the same frequency among T cell-depleted PBMC from healthy donors and those from patients. In contrast, IAPA-producing CD5- B cells were found in B cell-enriched preparations from patients, but not in those from healthy individuals.


The magnitude and associated factors of post-operative pain among adult patients.
Postoperative pain continues to be a serious consequence of surgical intervention. Several factors may contribute to the development of postoperative pain; these could be preoperative factors, demographic factors, anesthetic factors, and surgical factors. The aim of this study was to assess the magnitude and factors associated with postoperative pain among surgical patients. An institutional-based prospective longitudinal study included 265 postoperative patients from the surgical wards of Gambella General Hospital from April 15, 2021, to June 30, 2021. A consecutive sampling technique was used to recruit study participants. The patients were followed up for 24 h postoperatively. A numerical rating scale (NRS-11) is used for the assessment of pain. Data analysis was done using the Statistical Package for Social Science (SPSS) 25. Logistic regression analysis was used to calculate the association between dependent and independent variables with a 95% confidence interval and a p-value<0.05 was considered statistically significant. A total of 270 data points were collected. Of these, a total of 265 with a 98.1% response rate were analyzed. The incidence of postoperative pain was 69%, 74%, and 77.0% at 2 h, 12 h, and 24 h, respectively. The following factors were strongly associated with the dependent variable: patient age, 18-45 years old [AOR = 2.8; (95%CI: 1.13, 6.74, p = 0.026)], skin incision length, 10 cm [AOR = 2.5; (95%CI: 1.30, 5.13, p = 0.007)], preoperative pain [AOR = 2.4, (95%CI: 1.02, 5.60, p = 0.045)], and surgeon experience [AOR = 2.1, (95%CI. and Recommendation: In the current study the magnitude of postoperative pain was high, 220 patients were complaining POP (83%). Age of the patient, length of skin incision, preoperative pain, and experience of surgeons were the independent associated factors for the experience of postoperative pain. Preoperative pain management should exercise among adult surgical patients in order to reduce the incidence of postoperative pain, and the length of surgical skin incisions should be minimized.


A previously unknown way of heme detoxification in the digestive tract of cats.
Free heme is a highly toxic molecule for a living organism and its detoxification is a very important process, especially for carnivorous animals. Here we report the discovery of a previously unknown process for neutralizing free heme in the digestive tract of domestic cats. The cornerstone of this process is the encapsulation of heme into carbonated hydroxyapatite nanoparticles, followed by their excretion with faeces. This way of heme neutralization resembles the formation of insoluble heme-containing particles in the digestive tracts of other hematophagous species (for example, the formation of insoluble hemozoin crystals in malaria-causing Plasmodium parasites). Our findings suggest that the encapsulation of heme molecules into a hydroxyapatite matrix occurs during the transition from the acidic gastric juice to the small intestine with neutral conditions. The formation of these particles and their efficiency to include heme depends on the bone content in a cat's diet. In vitro experiments with heme-hydroxyapatite nanoparticles confirm the proposed scenario.


Systemic inhibition of miR-451 increases fibrotic signaling and diminishes autophagic response to exacerbate renal damage in Tallyho/Jng mice.
miRNAs provide fine tuning of gene expression via inhibition of translation. miR-451 has a modulatory role in cell cycling via downregulation of mechanistic target of rapamycin. We aimed to test whether chronic systemic inhibition of miR-451 would enhance renal fibrosis (associated with deranged autophagy). Adult TallyHo/Jng mice (obese insulin resistant) were randomized to two treatment groups to receive either miR-451 inhibition [via a locked nucleic acid construct] or a similar scrambled locked nucleic acid control for 8 wk. All mice were fed a high-fat diet (60% kcal from fat) ad libitum and humanely euthanized after 12 wk. Kidneys and blood were collected for analysis. Renal expression of miR-451 was sixfold lower in inhibitor-treated mice compared with control mice. miR-451 inhibition increased kidney weight and collagen and glycogen deposition. Blood chemistry revealed significantly higher Na+ and anion gap (relative metabolic acidosis) in inhibitor-treated mice. Western blot analysis and immunohistochemistry of the kidney revealed that the inhibitor increased markers of renal injury and fibrosis, e.g., kidney injury molecule 1, neutrophil gelatinase-associated lipocalin, transforming growth factor-β, 14-3-3 protein-ζ, mechanistic target of rapamycin, AMP-activated protein kinase-α, calcium-binding protein 39, matrix metallopeptidase-9, and the autophagy receptor sequestosome 1. In contrast, the inhibitor reduced the epithelial cell integrity marker collagen type IV and the autophagy markers microtubule-associated protein 1A/1B light chain 3B and beclin-1. Taken together, these results support a protective role for miR-451 in reducing renal fibrosis by enhancing autophagy in obese mice.


Detection and staging of chronic obstructive pulmonary disease using a computed tomography-based weakly supervised deep learning approach.
Chronic obstructive pulmonary disease (COPD) is underdiagnosed globally. The present study aimed to develop weakly supervised deep learning (DL) models that utilize computed tomography (CT) image data for the automated detection and staging of spirometry-defined COPD. A large, highly heterogeneous dataset was established, consisting of 1393 participants retrospectively recruited from outpatient, inpatient, and physical examination center settings of four large public hospitals in China. All participants underwent both inspiratory chest CT scans and pulmonary function tests. CT images, spirometry data, demographic information, and clinical information of each participant were collected. An attention-based multi-instance learning (MIL) model for COPD detection was trained using CT scans from 837 participants. External validation of the COPD detection was performed with 620 low-dose CT (LDCT) scans acquired from the National Lung Screening Trial (NLST) cohort. A multi-channel 3D residual network was further developed to categorize GOLD stages among confirmed COPD patients. The attention-based MIL model used for COPD detection achieved an area under the receiver operating characteristic curve (AUC) of 0.934 (95% CI: 0.903, 0.961) on the internal test set and 0.866 (95% CI: 0.805, 0.928) on the LDCT subset acquired from the NLST. The multi-channel 3D residual network was able to correctly grade 76.4% of COPD patients in the test set (423/553) using the GOLD scale. The proposed chest CT-DL approach can automatically identify spirometry-defined COPD and categorize patients according to the GOLD scale. As such, this approach may be an effective case-finding tool for COPD diagnosis and staging. • Chronic obstructive pulmonary disease is underdiagnosed globally, particularly in developing countries. • The proposed chest computed tomography (CT)-based deep learning (DL) approaches could accurately identify spirometry-defined COPD and categorize patients according to the GOLD scale. • The chest CT-DL approach may be an alternative case-finding tool for COPD identification and evaluation.


Association of ambient air pollution with depressive and anxiety symptoms in pregnant women: A prospective cohort study.
Air pollution is associated with depressive and anxiety symptoms in the general population. However, this relationship among pregnant women remains largely unknown. To evaluate the association between pregnancy air pollution exposure and maternal depressive and anxiety symptoms during the third trimester assessed using the Center for Epidemiologic Studies-Depression and State-Trait Anxiety Inventory scales, respectively. We analyzed 1481 pregnant women from a cohort study in Seoul. Maternal exposure to particulate matter with an aerodynamic diameter <2.5 μm (PM2.5) and <10 μm (PM10), as well as to nitrogen dioxide (NO2) and ozone (O3) for each trimester and the entire pregnancy was assessed at participant's residential address by land use regression models. We estimated the relative risk (RR) and corresponding confidence interval (CI) of the depressive and anxiety symptoms associated with an interquartile range (IQR) increase in PM2.5, PM10, NO2, and O3 using modified Poisson regression. In single-pollutant models, an IQR increase in PM2.5, PM10, and NO2 during the second trimester was associated with an increased risk of depressive symptoms (PM2.5 RR = 1.15, 95% CI: 1.04, 1.27; PM10 RR = 1.13, 95% CI: 1.04, 1.23; NO2 RR = 1.15, 95% CI: 1.03, 1.29) after adjusting for relevant covariates. Similarly, an IQR increase in O3 during the third trimester was associated with an increased risk of depressive symptoms (RR = 1.09, 95% CI: 1.01, 1.18), while the IQR increase in O3 during the first trimester was associated with a decreased risk (RR = 0.89, 95% CI: 0.82, 0.96). Exposure to PM2.5, PM10, and NO2 during the second trimester was significantly associated with anxiety symptoms. The associations with PM2.5 and O3 in single-and multi-pollutant models were consistent. Our findings indicate that increased levels of particulate matter, NO2, and O3 during pregnancy may elevate the risk of depression or anxiety in pregnant women.


Tailoring the oral sensation and digestive behavior of konjac glucomannan-gelatin binary hydrogel based bigel: Effects of composition and ratio.
In the food industry, there is a growing demand for bigels that offer both adaptable oral sensations and versatile delivery properties. Herein, we developed bigels using a binary hydrogel of konjac glucomannan (KGM) and gelatin (G) combined with a stearic acid oleogel. We closely examined how the oleogel/hydrogel volume ratio (φ) and the KGM/G mass ratio (γ) influenced various characteristics of the bigels, including their microstructure, texture, rheological properties, thermal-sensitivity, oral tribology, digestive stability, and nutraceutical delivery efficiency. A noteworthy observation was the structural evolution of the bigels with increasing φ values: transitioning from oleogel-in-hydrogel to a bicontinuous structure, and eventually to hydrogel-in-oleogel. Lower γ values yielded a softer, thermally-responsive bigel, whereas higher γ values imparted enhanced viscosity, stickiness, and spreadability to the bigel. Oral tribology assessments demonstrated that φ primarily influenced the friction sensations at lower chewing intensities. In contrast, γ played a significant role in augmenting oral friction perceptions during more intense chewing. Additionally, φ dictated the controlled release and bioaccessibility of curcumin, while γ determined digestive stability. This study provides valuable insights, emphasizing that through meticulous selection and adjustment of the hydrogel matrix composition, bigels can be custom-fabricated to achieve specific oral sensations and regulated digestive behaviors.


Structure-guided design and synthesis of P1' position 1-phenylcycloalkylamine-derived pentapeptidic BACE1 inhibitors.
Previously, we reported potent pentapeptidic BACE1 inhibitors with the hydroxymethylcarbonyl isostere as a substrate transition-state mimic. To improve the in vitro potency, we further reported pentapeptidic inhibitors with carboxylic acid bioisosteres at the P(4) and P1' positions. In the current study, we screened new P1' position 1-phenylcycloalkylamine analogs to find non-acidic inhibitors that possess double-digit nanomolar range IC(50) values. An extensive structure-activity relationship study was performed with various amine derivatives at the P1' position. The most potent inhibitor of this pentapeptide series, KMI-1830, possessing 1-phenylcyclopentylamine at the P1' position had an IC(50) value of 11.6 nM against BACE1 in vitro enzymatic assay.


Prevalence of myopia: A large-scale population-based study among children and adolescents in weifang, china.
This study aimed to determine the prevalence of myopia among school-aged children and adolescents at the whole city level of Weifang, China. This study was a large scale school-based cross-sectional study among children and adolescents aged 5-20 years old. Participants were selected by the school-based registration system in 2020. All the subjects underwent spherical equivalent (SE) error with non-cycloplegic autorefraction measurement. Myopia was defined as an SE refraction of ≤-0.75 diopters (D) and graded into low myopia (-0.75 to -3.00 D), moderate myopia (-3.01 to -5.99 D), and high myopia (≤-6.00 D). A total of 1,059,838 participants were eligible for this survey and 1,013,206 (95.6% participation rate) were selected through data quality control, which comprised 17 districts/counties and 1,254 schools, including 861 elementary schools, 313 middle schools, and 80 high schools. The mean age of participants was 11.57 ± 3.36 years (5-20 years), and the male-to-female ratio was 1.11. The whole city-level prevalence of total myopia was 75.35%. The prevalence of total myopia among the students in the Hanting District was 45.47%, but ≈82.37% of students living in Changyi have myopia. The overall prevalence of low myopia in elementary, middle, and high school students was 48.56, 47.30, and 31.62%, respectively, while high myopia (SE ≤ -6.00 D) prevalence was 1.12, 8.89, and 20.12%, respectively. The overall prevalence of myopia increased fastest in children aged 7-9 years old. The prevalence of high myopia was 7.59% for girls and 6.43% for boys, respectively (p < 0.001). The prevalence of myopia increased with increasing age and grade, but SE decreased with increasing age and grade. The current investigation demonstrated a high proportion of myopia among school students in the city of Weifang, and gradually increased with age, and the prevalence of myopia was the highest in Changyi areas. The high myopia prevalence for girls was higher than that in boys.


Detecting nonhemolytic Pasteurella haemolytica infections in healthy Rocky Mountain bighorn sheep (Ovis canadensis canadensis): influences of sample site and handling.
Effects of sampling procedures on ability to culture Pasteurella spp. from Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were examined experimentally. Sample site influenced (P less than 0.0001) recovery of P. haemolytica in adult bighorn sheep. We isolated nonhemolytic P. haemolytica from 18 of 19 tonsillar swabs and 18 of 19 tonsillar biopsies from adult sheep, yet only four of 19 nasal swabs yielded isolates. Sample handling also affected (P less than 0.0001) recovery of P. haemolytica. Nonhemolytic P. haemolytica was cultured from 14 of 19 tonsillar swabs plated directly onto blood agar, but from only two of 19 swabs stored for 24 hr in modified Stuart's medium. We detected nonhemolytic P. haemolytica at least once in bronchial aspirates from four and in nasal swabs from three of six bighorn lambs. Based on direct cultures of tonsillar swabs and/or biopsies, all 26 bighorn sheep (seven lambs, 19 adults) sampled were infected with nonhemolytic P. haemolytica; only two lambs developed pneumonia during the study period. Thirty-four of 37 nonhemolytic P. haemolytica isolates tested were biotype T; three were biotype A. Serotypes 3; 4; 3, 4 and 3, 4, 10 were identified in a subsample of 17 isolates. Our data suggest tonsillar swabs or biopsies plated directly onto blood agar and incubated immediately offer the greatest probability of recovering nonhemolytic P. haemolytica from health bighorn sheep.


Effects of atrazine and chlorpyrifos on acetylcholinesterase and Carboxylesterase in brain and muscle of common carp.
The aim of this study was to investigate the effects of chlorpyrifos (CPF), atrazine (ATR) and the mixture of them on acetylcholinesterase (AChE) and carboxylesterase (CbE) in brain and muscle of common carp, respectively. 220 carps were averagely divided into 11 groups according to the different treatments and concentration, including the exposure and recovery experiments. The activities of AChE and CbE of the brain and muscle were determined at the end of the exposure and the recovery. The results showed that in the control group, the specific enzymatic activities in the brain were higher than that in the muscle. The activities of AChE and CbE in the exposure groups were significantly lower than that in the control group except for the CbE activity in the ATR low-dose group. There was a negative dose-response relationship between the activities of AChE and CbE and the pesticides concentration. The activities of AChE and CbE in the recovery groups were significantly higher than that in the exposure group except for the CbE activity in the ATR low-dose group, AChE activity in the high-dose group of the mixture of ATR and CPF, and AChE activity of the brain in the CPF high-dose group. The results suggested that: (1) brain AChE may be considered as a very sensitive and early biomarker of exposure to CPF, ATR, or the mixture of ATR and CPF, (2) brain CbE may be used as a secondary biomarker for evaluating the exposure to CPF, ATR, or the mixture of ATR and CPF and (3) the change of the AChE and CbE activities caused by the mixture of ATR and CPF was more sensitive than that caused by the ATR or CPF alone.


Database of ab initio L-edge X-ray absorption near edge structure.
The L-edge X-ray Absorption Near Edge Structure (XANES) is widely used in the characterization of transition metal compounds. Here, we report the development of a database of computed L-edge XANES using the multiple scattering theory-based FEFF9 code. The initial release of the database contains more than 140,000 L-edge spectra for more than 22,000 structures generated using a high-throughput computational workflow. The data is disseminated through the Materials Project and addresses a critical need for L-edge XANES spectra among the research community.


The cascade hypothesis of keratoconus.
Keratoconus, a non-inflammatory thinning of the cornea, is a leading indication for corneal transplantation. For its causation, we propose a "Cascade Hypothesis" stating that keratoconus corneas have abnormal or defective enzymes in the lipid peroxidation and/or nitric oxide pathways leading to oxidative damage. The accumulation of oxidative, cytotoxic by-products causes an alteration of various corneal proteins, triggering a cascade of events, (i.e. apoptosis, altered signaling pathways, increased enzyme activities, fibrosis). This hypothesis is supported by biochemical, immunohistochemical and molecular data presented in this review. Based upon this evidence, one can speculate that keratoconus patients should minimize their exposure to oxidative stress. Protective steps should include wearing ultraviolet (UV) protection (in the contact lenses and/or sunglasses), minimizing the mechanical trauma (eye rubbing, poorly fit contact lenses) and keeping eyes comfortable with artificial tears, non-steroidal anti-inflammatory drugs and/or allergy medications.


Analysis of Differential Expression of Synaptic Vesicle Protein 2A in the Adult Rat Brain.
Synaptic vesicle protein 2A (SV2A), which plays an important role in the pathophysiology of epilepsy, is a unique vesicular protein recognized as a pharmacological target of anticonvulsant drugs. Furthermore, SV2A is a potential synaptic density marker, as it is ubiquitously expressed throughout the brain in all nerve terminals independently of their neurotransmitter content. Due to the growing interest in this protein, we thoroughly analyzed SV2A levels, expression patterns and colocalization in both excitatory and inhibitory synapses among different brain structures in healthy rats. In addition, we discuss the main semiquantitative methodologies used to study SV2A because these techniques might represent powerful tools for evaluating synaptic changes associated with brain disorders. Our results showed that the SV2A expression levels differed among the analyzed structures, and a positive correlation between the SV2A mRNA copy number and protein level was observed by Western blot. In addition, immunohistochemistry demonstrated slight but consistent asymmetrical SV2A levels in different laminated structures, and SV2A expression was increased by up to 40% in some specific layers compared to that in others. Finally, triple immunofluorescence revealed strong SV2A colocalization with GABAergic terminals, mainly around the principal cells, suggesting that SV2A primarily participates in this inhibitory system in different rat brain structures. Although the SV2A protein is considered a good candidate marker of synaptic density, our data show that changes in its expression in pathological processes must be viewed as not only increased or decreased synapse numbers but also in light of the type of neurotransmission being affected.


Protein phosphatase type-1 and type-2 catalytic subunits both bind inhibitor-2 and monoclonal immunoglobulins.
Protein phosphatases involved in cellular regulation have been categorized functionally into two major types by substrate specificity and sensitivity to protein inhibitors. In this classification type-1 phosphatases are inhibited by the heat-stable protein inhibitor-2 (I-2), whereas type-2 phosphatases are considered insensitive to inhibition by this protein. This study demonstrates that the phosphorylase phosphatase activity of both purified type-1 and type-2 catalytic subunits can be blocked by micromolar concentrations of I-2. Heparin also was more effective at inhibiting the type-1 compared to type-2 phosphatase but required thousandfold higher concentrations than I-2. The specificity of the interaction with I-2 indicates that the tertiary structures of the two phosphatase catalytic subunits closely resemble one another. However, only the type-1, not the type-2, protein phosphatase activity was neutralized by immunoglobulins affinity-purified against the Mr = 33,000 catalytic fragment of the type-1 phosphatase. Preparations of rabbit skeletal muscle type-1 phosphatase catalytic fragment and of bovine cardiac type-2 phosphatase catalytic subunit were compared by "Western" immunoblotting with sheep polyclonal and mouse monoclonal immunoglobulins raised against the respective proteins. Monoclonal anti-type-2 immunoglobulins preferentially stained the type-2 phosphatase catalytic subunit used as antigen, but displayed cross-reaction with 10-50 times more of the type-1 phosphatase. In contrast, as found with their effects on activity, sheep anti-type-1 immunoglobulins were specific; immunoblotting detected the type-1, not the type-2, catalytic protein. We conclude that the two catalytic proteins have at least one common primary structural epitope recognized by the monoclonal immunoglobulins. These data, taken together with other recent immunochemical results, support a hypothesis that this family of enzymes was derived from a common ancestral protein phosphatase catalytic subunit.


The KSHV viral IL-6 homolog is sufficient to induce blood to lymphatic endothelial cell differentiation.
The predominant tumor cell of Kaposi's Sarcoma (KS) is the spindle cell, a cell of endothelial origin that expresses markers of lymphatic endothelium. In culture, Kaposi's Sarcoma-associated herpesvirus (KSHV) infection of blood endothelial cells drives expression of lymphatic endothelial cell specific markers, in a process that requires activation of the gp130 receptor and the JAK2/STAT3 and PI3K/AKT signaling pathways. While expression of each of the KSHV major latent genes in endothelial cells failed to increase expression of lymphatic markers, the viral homolog of human IL-6 (vIL-6) was sufficient for induction and requires the JAK2/STAT3 and PI3K/AKT pathways. Therefore, activation of gp130 and downstream signaling by vIL-6 is sufficient to drive blood to lymphatic endothelial cell differentiation. While sufficient, vIL-6 is not necessary for lymphatic reprogramming in the context of viral infection. This indicates that multiple viral genes are involved and suggests a central importance of this pathway to KSHV pathogenesis.


Treatment response after palliative radiotherapy for bleeding gastric cancer: a multicenter prospective observational study (JROSG 17-3).
Palliative radiotherapy seems to be rarely performed for incurable gastric cancer. In this first multicenter study, we examined the effectiveness of palliative radiotherapy and investigated whether biologically effective dose (BED) is associated with survival, response, or re-bleeding. Eligibility criteria included blood transfusion or hemoglobin levels < 8.0 g/dL. The primary endpoint was the intention-to-treat (ITT) bleeding response rate at 4 weeks. Response entailed all of the following criteria: (i) hemoglobin levels ≥ 8.0 g/dL; (ii) 7 consecutive days without blood transfusion anytime between enrollment and blood sampling; and (iii) no salvage treatment (surgery, endoscopic treatment, transcatheter embolization, or re-irradiation) for bleeding gastric cancer. Re-bleeding was defined as the need for blood transfusion or salvage treatment. We enrolled 55 patients from 15 institutions. The ITT response rates were 47%, 53%, and 49% at 2, 4, and 8 weeks, respectively. The per-protocol response rates were 56%, 78%, and 90% at 2, 4, and 8 weeks, respectively. Neither response nor BED (α/β = 10) predicted overall survival. Multivariable Fine-Gray model showed that BED was not a significant predictor of response. Univariable Cox model showed that BED was not significantly associated with re-bleeding. Grades 1, 2, 3, and, ≥ 4 radiation-related adverse events were reported in 11, 9, 1, and 0 patients, respectively. The per-protocol response rate increased to 90% during the 8-week follow-up. The frequent occurrence of death starting shortly after enrollment lowered the ITT response rate. BED was not associated with survival, bleeding response, or re-bleeding.


Application of engineered extracellular vesicles for targeted tumor therapy.
All cells, including prokaryotes and eukaryotes, could release extracellular vesicles (EVs). EVs contain many cellular components, including RNA, and surface proteins, and are essential for maintaining normal intercellular communication and homeostasis of the internal environment. EVs released from different tissues and cells exhibit excellent properties and functions (e.g., targeting specificity, regulatory ability, physical durability, and immunogenicity), rendering them a potential new option for drug delivery and precision therapy. EVs have been demonstrated to transport antitumor drugs for tumor therapy; additionally, EVs' contents and surface substance can be altered to improve their therapeutic efficacy in the clinic by boosting targeting potential and drug delivery effectiveness. EVs can regulate immune system function by affecting the tumor microenvironment, thereby inhibiting tumor progression. Co-delivery systems for EVs can be utilized to further improve the drug delivery efficiency of EVs, including hydrogels and liposomes. In this review, we discuss the isolation technologies of EVs, as well as engineering approaches to their modification. Moreover, we evaluate the therapeutic potential of EVs in tumors, including engineered extracellular vesicles and EVs' co-delivery systems.


Isolated fallopian tube torsion: a rare but important event for women of reproductive age.
To report two cases of isolated fallopian tubal torsion that emphasize the rarity and difficult of diagnosis. Case report. A district general hospital. We report two cases of hematosalpinx due to torsion of the fallopian tube. The symptoms were not specific; both patients had acute lower abdominal pain. The first diagnosis in both cases was a torsion of a pelvic complex mass like a tumor. Laparoscopy is necessary, but often surgery is performed too late to preserve the tube. Resolution of symptoms with preservation of fertility. Both patients had the tube removed. Early diagnosis and surgical intervention is mandatory when isolated fallopian tube torsion is suspected, particularly in a nulliparous young woman.


Five-Year Glycemic Trajectories Among Healthy African-American and European-American Offspring of Parents With Type 2 Diabetes.
Cross-sectional surveys report a higher prevalence of diagnosed type 2 diabetes mellitus (T2DM) in African Americans (AA) than European Americans (EA). We studied 5-year glycemic excursions among AA and EA in the Pathobiology of Prediabetes in A Biracial Cohort study, to assess ethnic disparities. Pathobiology of Prediabetes in A Biracial Cohort followed normoglycemic offspring of parents with T2DM for 5 years, with serial assessments of oral glucose tolerance test , anthropometry, body fat, insulin sensitivity and beta-cell function. The primary outcome was progression to prediabetes (impaired fasting glucose and/or impaired glucose tolerance). We further analyzed 5-year changes in fasting (FPG) and 2-hour plasma glucose (2hrPG). One hundred and one (52 AA, 49 EA) out of 343 subjects developed prediabetes during follow-up. The change in FPG ranged from -24 mg/dl to +38 mg/dl. The FPG remained stable (± 5 mg/dl from baseline) in 50% of EA and 46.8% of AA and the 2hrPG remained stable (± 25 mg/dl from baseline) in 73.7% of EA and 71.0 % of AA during follow-up. The proportions with change in FPG of 5mg/dl to >25 mg/dl and 2hrPG of 25 mg/dl to >50 mg/dl were similar in EA and AA offspring, as were the 10th - 90th percentiles of the distribution of 5-year changes in FPG and 2hrPG. During 5 years of follow-up, black and white offspring of parents with T2DM exhibited remarkable phenotypic concordance of glycemic trajectories. Thus, parental history of T2DM may be a stronger factor than race/ethnicity in the prediction of longitudinal glycemic trends.


Effect of neonatal body temperature on postanoxic, potentially neurotoxic iron accumulation in the rat brain.
In asphyxiated newborns iron, released from heme and ferritin and deposited in the brain, contributes to neurodegeneration. Because hypothermia provides neuroprotection, newborn mammals, showing spontaneously reduced body temperature, might avoid the iron-mediated neurotoxicity. Therefore, we decided to study the effects of body temperature and chelation of iron with deferoxamine on iron accumulation in the brain of three weeks old rats exposed neonatally to a critical anoxia. At the age of two days, newborn rats were exposed to anoxia in 100% nitrogen atmosphere. Rectal temperature was kept at 33 degrees C (typical of the rat neonates), or elevated to a level typical of febrile (39 degrees C) adults. Control rats were exposed to atmospheric air in the respective thermal conditions. Half of the rats exposed to anoxia under hyperthermic conditions were injected with deferoxamine (DF), immediately after anoxia and 24 h later. Regional changes in cerebral iron deposition were examined in the frontal cortex, the hippocampus and the striatum, using iron histochemistry, when the rats reached the age of three weeks. Increased iron staining was found in neurons of each of the three cerebral regions in rats exposed to neonatal anoxia under hyperthermic conditions and the iron accumulation was prevented by postanoxic DF injection. In conclusion, febrile body temperature amplifies cerebral hyperferremia, which might induce neurodegenerative disturbances in the brain. On the other hand, a protection against the brain hyperferremia can be achieved by both the reduced physiological neonatal body temperature and by postasphyxic DF administration.


Visible transient expression of EGFP requires intranuclear injection of large copy numbers.
For the development of human artificial chromosomes (HACs) as a gene transfer vehicle we need to assess the efficiency of de novo chromosome formation depending on the type and the copy number of transferred DNA constructs. In order to check transient EGFP expression as a reporter to immediately detect presence of transfected DNA, we microinjected approximately 1 to 10(5) copies of pEGFP-N1 plasmid into the nucleus of various cell types. Whether using primary, immortalized, or tumor cells, at least 10(3)-10(4) copies were required to generate a visible green signal in the majority of the 50-90% of cells surviving injection. Generally, the cells showed relatively constant, copy number-dependent signals. 10(3) copies resulted in faint and 10(5) in bright fluorescence under the microscope. In addition, the different copy number groups contained a small fraction of cells showing much stronger fluorescence, indicating activation or lack of suppression which facilitates detection of as few as 10(2) transferred copies in rare instances. Thus, transient expression from single copies is not sufficient to reliably detect presence of DNA in the nucleus. The result is relevant for the development of low copy HAC transfer protocols.


Identification of novel prognostic risk signature of breast cancer based on ferroptosis-related genes.
Ferroptosis is a type of cell regulated necrosis triggered by intracellular phospholipid peroxidation, which is more immunogenic than apoptosis. Therefore, genes controlling ferroptosis may be promising candidate biomarkers for tumor therapy. In this study, we investigate the function of genes associated with ferroptosis in breast cancer (BC) and systematically evaluate the relationship between ferroptosis-related gene expression and prognosis of BC patients from the Cancer Genome Atlas database. By using the consensus clustering method, 1203 breast cancer samples were clustered into two clearly divided subgroups based on the expression of 237 ferroptosis-related genes. Then differentially expressed analysis and least absolute shrinkage and selection operator were used to identify the prognosis-related genes. Furthermore, the genetic risk signature was constructed using the expression of prognosis-related genes. Our results showed that the genetic risk signature can identify patient subgroups with distinct prognosis in either training cohort or validation, and the genetic risk signature was associated with the tumor immune microenvironment. Finally, the Cox regression analysis indicated that our risk signature was an independent prognostic factor for BC patients and this signature was verified by the polymerase chain reaction and western blot. Within this study, we identified a novel prognostic classifier based on five ferroptosis-related genes which may provide a new reference for the treatment of BRCA patients.


Direct Anterior Approach and Perioperative Fracture With a Single-Taper Wedge Femoral Component.
Despite growing interest in direct anterior approach total hip arthroplasty, perioperative femoral fracture and early aseptic loosening are increasingly recognized complications. Previous research has documented the role of surgeon experience in association with these femoral complications. The purpose of this study was to explore the relationship between femoral component design and early periprosthetic femoral complications. This was an extension of previous work with an updated patient cohort of 5090 consecutive direct anterior primary total hip arthroplasties at a single institution with a single-taper, wedge femoral stem comprising 4 variants involving length and geometry: group 1, full-length, standard profile; group 2, full-length, reduced distal profile; group 3, short-length, standard profile; and group 4, short-length, reduced distal profile. Records were reviewed retrospectively for the incidence of early postoperative periprosthetic fracture or aseptic loosening and analyzed with regard to patient demographics and femoral stem type. There were 42 (0.83%) periprosthetic femur complications observed in the early postoperative period. Increased age (P < .001) and female gender (P = .023) were significantly associated with incidence of femoral complications in univariate analysis, while age maintained this significant relationship in multivariate analysis (P < .001). There was a trend toward increased complication rate in patients receiving a short stem with full profile taper (1.27%, P = .0539). Despite an overall low rate of femoral complications after direct anterior total hip arthroplasty, the risk is increased in elderly patients and females. Furthermore, femoral stem design may portend an elevated risk of these complications.


Effect of substrate on Ca2(+)-concentration required for activity of the Ca2(+)-dependent proteinases, mu- and m-calpain.
The Ca2+ concentrations required for half-maximal activity of mu- and m-calpain purified from bovine skeletal muscle were tested using four different protein substrates and three different synthetic peptide substrates. Hammersten casein, the commonly used substrate for measuring mu- and m-calpain activity, required 2.5 microM Ca2+ for half-maximal activity of mu-calpain and 290 microM Ca2+ for half-maximal activity of m-calpain. When Hammersten casein was dialyzed against 8 M urea and 10 mM EDTA to remove all endogenous Ca2+, it required 1.9 and 290 microM Ca2+ for half-maximal activity of mu- and m-calpain, respectively. Rabbit skeletal muscle myofibrils and rabbit skeletal muscle troponin required 65 microM and 24 microM Ca2+ for half-maximal activity of mu-calpain and 380 microM and 580 microM Ca2+ for half-maximal activity of m-calpain, respectively. The three synthetic substrates tested, Suc-Leu-Tyr-MCA, Boc-Leu-Thr-Arg-MCA, and Suc-Leu-Leu-Val-Tyr-MCA, required 1.6 microM to 3.7 microM Ca2+ for half-maximal activity of mu-calpain and 200 to 560 microM Ca2+ for half-maximal activity of m-calpain.


A Potential Filling Material for Wound Healing and Shaping: Acellular Dermal Matrix Combined with Autologous Dermis.
The currently used subcutaneous fillers are not effective enough. The use of autologous tissues or cells will reduce immune rejection and increase the stability of the fillers, while increasing the absorbability of filling materials. To establish a durable and safe filling material, we herein combined acellular dermal matrix (ADM) with autologous dermal cells and tested the performance of this matrix in wound healing and shaping. To prepare the ADM, stratified skin was obtained from the back of two New Zealand rabbits and decellularized to obtain ADM. The ADM was ultrasonically mixed with as-prepared autologous dermis in vitro. The mixture was injected as a subcutaneous filler into the back area of 20 New Zealand rabbits. In this procedure, different concentration ratios of the mixture were injected, and the volume change of the filler was measured and categorized into five groups. At 4 months and 8 months after filling, samples were obtained for quantitative evaluation of the thickness and vessel density, as well as qualitative evaluation of colonization. Examination of the ADM conformed that the matrix had no cells. ADM and the prepared autologous dermis were evenly mixed, and the filler was gradually absorbed. The comprehensive evaluation of ADM and autologous dermis mixture showed that group C had the best filling effect with the least infiltration of inflammatory cells and the highest vascular density compared to A, B, D and E groups in the study. When combined at the ratio of 50%:50%, the autologous dermis combined and acellular dermal matrix showed better performance compared to the other four different ratios in this study. This implies that this combination is potentially safer, effective and stable as a filling material compared to ADM or dermal matrix alone. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Real-time imaging of pulvinus bending in Mimosa pudica.
Mimosa pudica is a plant that rapidly shrinks its body in response to external stimuli. M. pudica does not perform merely simple movements, but exhibits a variety of movements that quickly change depending on the type of stimuli. Previous studies have investigated the motile mechanism of the plants from a biochemical perspective. However, an interdisciplinary study on the structural characteristics of M. pudica should be accompanied by biophysical research to explain the principles underlying such movements. In this study, the structural characteristics and seismonastic reactions of M. pudica were experimentally investigated using advanced bio-imaging techniques. The results show that the key factors for the flexible movements by the pulvinus are the following: bendable xylem bundle, expandable/shrinkable epidermis, tiny wrinkles for surface modification, and a xylem vessel network for efficient water transport. This study provides new insight for better understanding the M. pudica motile mechanism through structural modification.


Development and characterization of mucoadhesive patches of salbutamol sulfate for unidirectional buccal drug delivery.
Buccal patches of salbutamol sulfate were prepared using five different water soluble polymers in various proportions and combinations using PEG-400/PG as plasticizers. A 32 full factorial design was used to design the experiments for each polymer combination. Patches were laminated on one side with a water impermeable backing layer for unidirectional drug release. The thickness of medicated patches ranged between 0.2 and 0.4 mm and showed an increase in mass whenever PEG-400 was used as plasticizer. The surface pH of all patches approached neutral. Eight formulations which had shown high folding endurance (> 300) were selected for evaluation. Patches prepared with PEG-400 showed a high swelling index. The residence time of the tested patches ranged between 105 and 130 min. Formulations A10, A32, B10 and B32 fitted the Higuchi model best, whereas formulations A19 and B19 showed super case II transport drug release. Stability studies indicated that there was no change in the chemical and physical characteristics during the test period of 6 months.


The Role of Religion and Spirituality in Conceptualizing Reproductive Loss: Clinical Implications of the Narratives of Gestational Surrogates in the USA.
Reproductive losses experienced by gestational surrogates, who do not share a genetic connection to the child they are carrying, may not be acknowledged or treated as compassionately as women who lose their "own" pregnancies. In-depth, semi-structured interviews were conducted with 17 gestational surrogates from diverse religious backgrounds. Grounded theory analysis of data included line-by-line coding, and emergent themes were identified. The results suggest that understanding and utilizing the religious/spiritual beliefs of a gestational surrogate may be one way for health professionals to provide implications counseling prior to surrogacy and also as a means to help process and grieve losses that may occur within surrogacy.


MR Template-Based Individual Brain PET Volumes-of-Interest Generation Neither Using MR nor Using Spatial Normalization.
For more anatomically precise quantitation of mouse brain PET, spatial normalization (SN) of PET onto MR template and subsequent template volumes-of-interest (VOIs)-based analysis are commonly used. Although this leads to dependency on the corresponding MR and the process of SN, routine preclinical/clinical PET images cannot always afford corresponding MR and relevant VOIs. To resolve this issue, we propose a deep learning (DL)-based individual-brain-specific VOIs (i.e., cortex, hippocampus, striatum, thalamus, and cerebellum) directly generated from PET images using the inverse-spatial-normalization (iSN)-based VOI labels and deep convolutional neural network model (deep CNN). Our technique was applied to mutated amyloid precursor protein and presenilin-1 mouse model of Alzheimer's disease. Eighteen mice underwent T2-weighted MRI and 18F FDG PET scans before and after the administration of human immunoglobin or antibody-based treatments. To train the CNN, PET images were used as inputs and MR iSN-based target VOIs as labels. Our devised methods achieved decent performance in terms of not only VOI agreements (i.e., Dice similarity coefficient) but also the correlation of mean counts and SUVR, and CNN-based VOIs was highly accordant with ground-truth (the corresponding MR and MR template-based VOIs). Moreover, the performance metrics were comparable to that of VOI generated by MR-based deep CNN. In conclusion, we established a novel quantitative analysis method both MR-less and SN-less fashion to generate individual brain space VOIs using MR template-based VOIs for PET image quantification. The online version contains supplementary material available at 10.1007/s13139-022-00772-4.


Investigation of a Microemulsion Containing Clotrimazole and Itraconazole for Transdermal Delivery for the Treatment of Sporotrichosis.
The aim of this study was to develop a microemulsion (ME) formulation containing an association of itraconazole (ITC) and clotrimazole (CLT) as a transdermal delivery system for the treatment of sporotrichosis. Pseudoternary phase diagrams were constructed to optimize the ME formulation. The ME formulation selected contained 1% (w/w) ITC and 1% (w/w) CLT and was composed of 23.07% Tween® 60 (surfactant), 23.07% propylene glycol (cosurfactant/cosolvent), 30.77% benzyl alcohol (oil), and 21.09% water. The ITC/CLT-loaded ME (ITC/CLT-ME) had a droplet size value of 217 ± 0.9 nm, with a polydispersity index of 0.5 ± 0.1. Permeation experiments on pig ear skin were conducted for ITC/CLT-ME, and the results indicated that the drug permeation performance was influenced by CLT, indicating that CLT acts as a promoter enhancer. In the in vitro antifungal activity assay using Sporothrix brasiliensis yeast, the inhibition halo produced by ITC/CLT-ME exhibited a mean diameter of 43.67 ± 2.31 mm. The ITC/CLT-ME formulation did not cause skin irritation in mice. The results suggest that ITC/CLT-ME is a promising tool for the transdermal treatment of sporotrichosis.


Bovine model of reproductive aging: response to ovarian synchronization and superstimulation.
The responsiveness of the hypothalamo-pituitary axis to steroid treatments for ovarian synchronization and the ovarian superstimulatory response to exogenous FSH was compared in 13-14 year old cows and their 1-4 year old young daughters. We tested the hypotheses that aging in cattle is associated with: (1) decreased follicular wave synchrony after estradiol and progesterone treatment; (2) delayed LH surge and ovulation in response to exogenous preovulatory estradiol treatment; (3) reduced superstimulatory response to exogenous FSH. Higher plasma FSH concentrations (P<0.01), and a tendency (P=0.07) for fewer 4-5 mm follicles at wave emergence were observed in old cows (n=10) than in young cows (n=9). The suppressive effect of estradiol/progesterone treatment on FSH was similar between old and young cows. Although the preovulatory LH surge in response to estradiol treatment was delayed in old than young cows (P=0.01), detected ovulation times were not different. No difference in ovarian superstimulatory response was detected between age groups, but old cows (n=8) tended (P=0.10) to have fewer large follicles (>or=9 mm) 12 h after last FSH treatment than in young cows (n=7). We concluded that pituitary and ovarian responsiveness to estradiol/progesterone synchronization treatment was similar between old and young cows, but aging was associated with a delayed preovulatory LH surge subsequent to estradiol treatment. Old cows tended to have fewer large follicles after superstimulatory treatment than young cows.


Associated risk factors with disease severity and antiviral drug therapy in patients with COVID-19.
Due to the latent onset of novel coronavirus disease 2019 (COVID-19), it is important to identify patients with increased probabilities for disease progression early in order to implement timely medical strategies. This study aimed to identify the factors associated with increased COVID-19 severity and evaluate the current antiviral drugs, especially in severe patients. This was a retrospective observational study performed at the No. 7 Hospital of Wuhan (Wuhan, China) with hospitalized patients confirmed with COVID-19 from January 11 to March 13, 2020. Multivariable logistic regression analysis was used to identify the associated factors of severe COVID. Treatments of antivirus drugs were collected and evaluated. Of the 550 patients, 292 (53.1%) were female and 277 (50.4%) were > 60 years old. The most common symptom was fever (n = 372, 67.7%), followed by dry cough (n = 257, 46.7%), and dyspnea (n = 237, 43.1%), and fatigue (n = 224, 40.7%). Among the severe patients, 20.2% required invasive ventilator support and 18.0% required non-invasive ventilator. The identified risk factors for severe cases were: age ≥ 60 years (odds ratio (OR) =3.02, 95% confidence interval (CI): 1.13-8.08, P = 0.028), D-dimer > 0.243 μg/ml (OR = 2.734, 95%CI: 1.012-7.387, P = 0.047), and low oxygenation index (OR = 0.984, 95%CI: 0.980-0.989, P < 0.001). In severe cases, the benefits (relief of clinical symptoms, clinical outcome, and discharge rate) of arbidol alone was 73.3%, which was better than ribavirin (7/17, 41.2%, P = 0.029). Age > 60 years, D-dimer > 0.243 μg/ml, and lower oxygenation index were associated with severe COVID-19. Arbidol might provide more clinical benefits in treating patients with severe COVID-19 compared with ribavirin.


Anisotropy of photosynthetic membranes and the degree of fluorescence polarization.
The degree of fluorescence polarization, P, of unoriented and magnetically oriented spinach chloroplasts as a function of excitation (400-680 nm) and emission wavelengths (675-750 nm) is reported. For unoriented chloroplasts P can be divided into two contributions, PIN and PAN. The latter arises from the optical anisotropy of the membranes which is due to the orientation with respect to the membrane plane of pigment molecules in vivo. The intrinsic polarization PIN, which reflects the energy transfer between different pigment molecules and their degree of mutual orientation, can be measured unambiguously only if (1) oriented membranes are used and the fluorescence is viewed along a direction normal to the membrane planes, and (2) the excitation is confined to the Qy (approximately 660-680 nm) absorption band of chlorophyll in vivo. With 670-680 nm excitation, values of P using unoriented chloroplasts can be as high as +14%, mostly reflecting the orientational anisotropy of the pigments. Using oriented chloroplasts PIN is shown to be +5+/-1%. The excitation wavelength dependence studies of PIN indicate that the carotenoid and chlorophyll Qy transition moments tends to be partially oriented with respect to each other on a local level (within a given photosynthetic unit or its immediate neighbors).


Phaeohyphomycosis infection leading to flexor tendon rupture: a case report.
A rare previously unreported cause of flexor tendon rupture is described. A 66-year-old man presented with a fully extended left middle finger, accompanied by swelling and purulent drainage. Prior to presentation, he had received a steroid injection for left middle finger stenosing tenosynovitis and subsequently developed culture-proven phaeohyphomycosis fungal infection and secondary enterococcal bacterial infection, requiring pharmacotherapy and incision, drainage, and debridement for abscess formation. Clinical and magnetic resonance imaging findings were consistent with the diagnosis of closed flexor tendon rupture of the left middle finger. Antifungal and antibiotic therapy followed by two-stage flexor tendon reconstruction was performed. Six months postoperatively, full passive range of motion was achieved and the proximal interphalangeal and distal interphalangeal joints of the left middle finger actively flexed to 125 degrees and 90 degrees, respectively.


Lipids from nerve tissues of the horseshoe crab, Limulus polyphemus.
1. The predominant lipids of nerve cords, ganglion and brain from horseshoe crabs were cholesterol (11% of lipid) and phospholipid (81% of lipid). 2. Major phospholipids were phosphatidyl ethanolamine and phosphatidyl choline with lesser amounts of phosphatidyl serine and phosphatidyl inositol and sphingomyelin. 3. The phospholipid fraction was characterized by a high content of plasmalogen, i.e. alk-1-enyl acyl phosphatides, so that 42% of the ethanolamine phosphatides were the plasmalogen, phosphatidal ethanolamine. 4. Phosphatidyl choline and phosphatidyl ethanolamine were high in polyunsaturation with 20:4 and 20:5 major fatty acids. Sphingomyelin had predominantly long chain saturated fatty acids. 5. Cerebrosides and gangliosides, which are associated with vertebrate nerve tissues, were absent from nerves of horseshoe crabs.


Electrocatalysis of nitrogen pollution: transforming nitrogen waste into high-value chemicals.
On 16 June 2023, the United Nations Environment Programme highlighted the severity of nitrogen pollution faced by humans and called for joint action for sustainable nitrogen use. Excess nitrogenous waste (NW: NO, NO2, NO2-, NO3-, etc.) mainly arises from the use of synthetic fertilisers, wastewater discharge, and fossil fuel combustion. Although the amount of NW produced can be minimised by reducing the use of nitrogen fertilisers and fossil fuels, the necessity to feed seven billion people on Earth limits the utility of this approach. Compared to current industrial processes, electrocatalytic NW reduction or CO2-NW co-reduction offers a potentially greener alternative for recycling NW and producing high-value chemicals. However, upgrading this technology to connect upstream and downstream industrial chains is challenging. This viewpoint focuses on electrocatalytic NW reduction, a cutting-edge technology, and highlights the challenges in its practical application. It also discusses future directions to meet the requirements of upstream and downstream industries by optimising production processes, including the pretreatment and supply of nitrogenous raw materials (e.g. flue gas and sewage), design and macroscopic preparation of electrocatalysts, and upscaling of reactors and other auxiliary equipment.


The role of plastidic cytochrome c in algal electron transport and photophosphorylation.
By an improved isolation procedure chloroplasts could be obtained from the alga Bumilleriopsis filiformis (Xanthophyceae) which exhibited high electron transport rates tightly coupled to ATP formation. Uncouplers both stimulate electron transport and inhibit photophosphorylation. These chloroplasts retain almost all soluble cytochrome c-553 besides a membrane-bound cytochrome c-554.5 (=f-554.5). Sonification or iron deficiency removed the soluble cytochrome only with a concurrent decrease of electron transport from water to methyl viologen or to NADP and decreased non-cyclic and cyclic photophosphorylation. However, photosynthetic control and the P/2e ratios remain unaltered. In Bumilleriopsis, which apparently has no plastocyanin, the soluble cytochrome c-553 seemingly links electron transport between the bound cytochrome c and P-700.


The influence of telehealth-based cancer rehabilitation interventions on disability: a systematic review.
To characterize delivery features and explore effectiveness of telehealth-based cancer rehabilitation interventions that address disability in adult cancer survivors. A systematic review of electronic databases (CINAHL Plus, Cochrane Library: Database of Systematic Reviews, Embase, National Health Service's Health Technology Assessment, PubMed, Scopus, Web of Science) was conducted in December 2019 and updated in April 2021. Searches identified 3,499 unique studies. Sixty-eight studies met inclusion criteria. There were 81 unique interventions across included studies. Interventions were primarily delivered post-treatment and lasted an average of 16.5 weeks (SD = 13.1). They were most frequently delivered using telephone calls (59%), administered delivered by nursing professionals (35%), and delivered in a one-on-one format (88%). Risk of bias of included studies was primarily moderate to high. Included studies captured 55 measures of disability. Only 54% of reported outcomes had data that allowed calculation of effect sizes ranging -3.58 to 15.66. The analyses suggest small effects of telehealth-based cancer interventions on disability, though the heterogeneity seen in the measurement of disability makes it hard to draw firm conclusions. Further research using more diverse samples, common measures of disability, and pragmatic study designs is needed to advance telehealth in cancer rehabilitation. Telehealth-based cancer rehabilitation interventions have the potential to increase access to care designed to reduce disability across the cancer care continuum.


Translation Comes First: Ancient and Convergent Selection of Codon Usage Bias Across Prokaryotic Genomes.
Codon usage is the outcome of different evolutionary processes and can inform us about the conditions in which organisms live and evolve. Here, we present R_ENC', which is an improvement to the original S index developed by dos Reis et al. (2004). Our index is less sensitive to G+C content, which greatly affects synonymous codon usage in prokaryotes, making it better suited to detect selection acting on codon usage. We used R_ENC' to estimate the extent of selected codon usage bias in 1800 genomes representing 26 prokaryotic phyla. We found that Gammaproteobacteria, Betaproteobacteria, Actinobacteria, and Firmicutes are the phyla/subphyla showing more genomes with selected codon usage bias. In particular, we found that several lineages within Gammaproteobacteria and Firmicutes show a similar set of functional terms enriched in genes under selected codon usage bias, indicating convergent evolution. We also show that selected codon usage bias tends to evolve in genes coding for the translation machinery before other functional GO terms. Finally, we discuss the possibility to use R_ENC' to predict whether lineages evolved in copiotrophic or oligotrophic environments.


Further researches on the action of hepatectomized rat serum on epithelial differentiation in chick embryo skin.
Skin explants obtained from 12-day old chick embryos, maintained in organ culture containing adult rat serum, show a lower epidermal growth in comparison with skin explants cultured with other adult animal serum; nevertheless the keratogeneous differentiation of the epidermal cell is evident. When the cultures are incubated with medium containing partially hepatectomized rat serum epidermal-cell differentiation and proliferation are absent.


Traumatically intruded permanent incisors: a study of treatment and outcome.
Twenty-nine traumatically intruded permanent maxillary incisors from 20 patients were examined, treated and monitored for 2 years. There was a higher proportion of males than females. The teeth intruded furthest were more frequently extracted. Those which were surgically repositioned were more frequently retained and were associated with marginal bone loss significantly less frequently than those which were passively observed, while external root resorption occurred in similar proportions in both groups. A significantly higher proportion of those teeth that were delayed in presentation were subsequently extracted, and a similar trend was found for those with immature roots at the time of the initial examination.


A modified host-cell reactivation assay to quantify DNA repair capacity in cryopreserved peripheral lymphocytes.
The host-cell reactivation assay (HCRA) is a functional assay that allows the identification of the genes responsible for DNA repair-deficient syndromes, such as Xeroderma pigmentosum, by cross-complementation experiments. It has also been used in molecular epidemiology studies to correlate the low nucleotide excision repair pathway function in peripheral blood lymphocytes with an increased risk of bladder, head and neck, skin and lung cancers. Herein, we present the technical validation of a newly modified HCRA, where nucleofection is used for the transfection of the pmaxGFP plasmid into cryopreserved peripheral blood lymphocytes (PBLs) or lymphoblastoid cell lines. In each sample, 20-24h after transfection, the relative DNA repair capacity (DRC) was quantified by flow cytometry, comparing the transfection efficiency of nucleoporated cells with undamaged plasmid to those transfected with UV-light damaged plasmid in the seven cell lines that were characterized by different DNA repair phenotypes. Dead cells were excluded from the analysis. We observed a high reproducibility of the relative DRC, transfection efficiency and cell viability. The inter-experimental normalization of the flow cytometry resulted in an increased data accuracy and reproducibility. The amount of cells required for each transfection reaction was reduced fourfold, without affecting the final relative DRC. Furthermore, our HCRA demonstrated strong discrimination power in the UV-light dose-response, both in lymphoblastoid cell lines and cryopreserved PBLs. We also observed a strong correlation of the relative DRC data, when samples were measured against two independent batches of both damaged and undamaged plasmid DNA. The relative DRC variable shows a normal distribution when analyzed in the cryopreserved PBLs from a cohort of 35 lung cancer patients and a 5.59-fold variation in the relative DRC is identified among our patients. The mitotic dynamic was discarded as a confounding factor for the relative DRC measurement in this cohort of patients. The results indicate that our method is highly sensitive, reliable and reproducible, and thus, it suitable for population-based studies to quantify in vitro DNA-repair deficiencies.


The H93G Myoglobin Cavity Mutant as a Versatile Scaffold for Modeling Heme Iron Coordination Structures in Protein Active Sites and Their Characterization with Magnetic Circular Dichroism Spectroscopy.
Preparation of heme model complexes is a challenging subject of long-standing interest for inorganic chemists. His93Gly sperm whale myoglobin (H93G Mb) has the proximal His replaced with the much smaller non-coordinating Gly. This leaves a cavity on the proximal side of the heme into which a wide variety of exogenous ligands can be delivered. The end result is a remarkably versatile scaffold for the preparation of model heme adducts to mimic the heme iron coordination structure of native heme proteins. In this review, we first summarize the quantitative evidence for differential ligand binding affinities of the proximal and distal pockets of the H93G Mb cavity mutant that facilitates the preparation of mixed-ligand derivatives. Then we review our use of magnetic circular dichroism and electronic absorption spectroscopy to characterize nitrogen-, oxygen-, and sulfur-donor-ligated H93G Mb adducts with an emphasis on species not easily prepared by other heme model system approaches and those that serve as spectroscopic models for native heme proteins.


First record of a parasitic septate gregarines (Apicomplexa: Sporozoea) in the shrimp Peneaus monodon in Sundarbans of West Bengal.
Investigations on the incidence of septate gregarines in shrimp have immense importance because of severe pathogenicity of the parasite. The septate gregarines infect the midgut of shrimp Peneaus monodon and severe infection disturbs the intestinal tissues. Mostly gregarines of the genus Nematopsis have been identified from cultured peneaid shrimp. It has worldwide in distribution. In India, gregarine parasites have so far been reported from penaeid shrimps of Bombay and Kerala. The species which was isolated from the midgut of shrimp Peneaus monodon collected from Kharibari area of Sunderbans. 9 out of 20 i.e. 45% of the randomly sampled hosts were found to be infected with a species of the genus Nematopsis. Different developmental stages including trophozoites, sporadins, and gametocysts of the Nematopsis sp. infecting the shrimp have been isolated. No correlations have been established between incidence of infection and environmental parameters.


Oral lichenoid lesions, mercury hypersensitivity and combined hypersensitivity to mercury and other metals: histologically-proven reproduction of the reaction by patch testing with metal salts.
We report 11 patients seen between 1991 and 1994 with oral lichenoid lesions (OLL). In 10 cases, there was contact with dental amalgam fillings, and in patient no. 10 with both amalgam restorations and a gold crown. The last patient had, in addition to her OLL, lichen planus of the skin and genital mucosa. In 5 cases, combined sensitization to mercury and other metal salts, particularly gold sodium thiosulfate (GST) and palladium chloride (PDC), was observed. In 10 patients, the lesions considerably improved or totally cleared within 1 to 9 months of replacement of restoration materials. Histological examination of biopsies from the test sites of amalgam, mercuric chloride, GST and PDC, taken 10 or 17 days after application of patch tests, showed lichenoid changes in 7 patients with at least 1 of the allergens. As at least 2 patients had inflammatory lesions of the oral mucosa related to both amalgam and gold restorations, combined sensitization to inorganic and organic mercury derivatives, GST and, in 1 case, PDC, a "dental restoration metal intolerance syndrome" is proposed.