A pan-cancer analysis of Dyskeratosis congenita 1 (DKC1) as a prognostic biomarker.
Dyskeratosis congenita 1 (DKC1), a critical component of telomerase complex, is highly expressed in a variety of human cancers. However, the association of DKC1 with cancer occurrence and development stages is not clear, making a pan-cancer analysis crucial. We conducted a study using various bioinformatic databases such as TIMER, GEPIA, UALCAN, and KM plotter Analysis to examine the different expressions of DKC1 in multiple tissues and its correlation with pathological stages. Through KEGG analysis, GO enrichment analysis and Venn analysis, we were able to reveal DKC1-associated genes and signaling pathways. In addition, we performed several tests including the CCK, wound healing assay, cell cycle arrest assay, transwell assay and Sa-β-gal staining on DKC1-deleted MDA-231 cells. Our study demonstrates that DKC1 has relatively low expression specificity in different tissues. Furthermore, we found that in ACC, KICH, KIRP and LIHC, the expression level of DKC1 is positively correlated with pathological stages. Conversely, in NHSC, KIRP, LGG, LIHC, MESO and SARC, we observed a negative influence of DKC1 expression level on the overall survival rate. We also found a significant positive correlation between DKC1 expression and Tumor Mutational Burden in 14 tumors. Additionally, we observed a significantly negative impact of DKC1 DNA methylation on gene expression at the promoter region in BRCA. We also identified numerous phosphorylation sites concentrated at the C-terminus of the DKC1 protein. Our GO analysis revealed a correlation between DKC1 and ribosomal biosynthesis pathways, and the common element UTP14A was identified. We also observed decreased rates of cell proliferation, migration and invasion abilities in DKC1-knockout MDA-MB-231 cell lines. Furthermore, DKC1-knockout induced cell cycle arrest and caused cell senescence. Our findings suggest that the precise expression of DKC1 is closely associated with the occurrence and developmental stages of cancer in multiple tissues. Depletion of DKC1 can inhibit the abilities of cancer cells to proliferate, migrate, and invade by arresting the cell cycle and inducing cell senescence. Therefore, DKC1 may be a valuable prognostic biomarker for the diagnosis and treatment of cancer in various tissues.


A novel Cellvibrio mixtus family 10 xylanase that is both intracellular and expressed under non-inducing conditions.
Hydrolysis of the plant cell wall polysaccharides cellulose and xylan requires the synergistic interaction of a repertoire of extracellular enzymes. Recently, evidence has emerged that anaerobic bacteria can synthesize high levels of periplasmic xylanases which may be involved in the hydrolysis of small xylo-oligosaccharides absorbed by the micro-organism. Cellvibrio mixtus, a saprophytic aerobic soil bacterium that is highly active against plant cell wall polysaccharides, was shown to express internal xylanase activity when cultured on media containing xylan or glucose as sole carbon source. A genomic library of C. mixtus DNA, constructed in lambdaZAPII, was screened for xylanase activity. The nucleotide sequence of the genomic insert from a xylanase-positive clone that expressed intracellular xylanase activity in Escherichia coli revealed an ORF of 1137 bp (xynC), encoding a polypeptide with a deduced M(r) of 43413, defined as xylanase C (XylC). Probing a gene library of Pseudomonas fluorescens subsp. cellulosa with C. mixtus xynC identified a xynC homologue (designated xynG) encoding XylG; XylG and xynG were 67% and 63% identical to the corresponding C. mixtus sequences, respectively. Both XylC and XylG exhibit extensive sequence identity with family 10 xylanases, particularly with non-modular enzymes, and gene deletion studies on xynC supported the suggestion that they are single-domain xylanases. Purified recombinant XylC had an M(r) of 41000, and displayed biochemical properties typical of family 10 polysaccharidases. However, unlike previously characterized xylanases, XylC was particularly sensitive to proteolytic inactivation by pancreatic proteinases and was thermolabile. C. mixtus was grown to late-exponential phase in the presence of glucose or xylan and the cytoplasmic, periplasmic and cell envelope fractions were probed with anti-XylC antibodies. The results showed that XylC was absent from the culture media but was predominantly present in the periplasm of C. mixtus cells grown on glucose, xylan, CM-cellulose or Avicel. These data suggest that C. mixtus can express non-modular internal xylanases whose potential roles in the hydrolysis of plant cell wall components are discussed.


Shedding light on surface exposition of poly(ethylene glycol) and folate targeting units on nanoparticles of poly(ε-caprolactone) diblock copolymers: Beyond a paradigm.
Polymeric nanoparticles (NPs) of poly(ε-caprolactone) (PCL) covered with a hydrophilic poly(ethylene glycol) (PEG) shell are usually prepared from diblock PEG-PCL copolymers through different techniques. Furthermore PEG, NPs can be decorated with targeting ligands to accumulate in specific cell lines. However, the density and conformation of PEG on the surface and its impact on the exposition of small targeting ligands has been poorly considered so far although this has a huge impact on biological behaviour. Here, we focus on PEG-PCL NPs and their folate-targeted version to encourage accumulation in cancer cells overexpressing folate receptor α. NPs were prepared with mixtures of PEG-PCL with different PEG length (short 1.0kDa, long 2.0kDa,) and a folate-functionalized PEG-PCL (PEG 1.5kDa) by the widely employed solvent displacement method. In depth characterization of NPs surface by 1H NMR, fluorescence and photon correlation spectroscopy evidenced a PEGylation extent below 7% with PEG in a mushroom conformation and the presence of folate more exposed to water pool in the case of copolymer with short PEG. NPs with short PEG adsorbed HSA forming a soft corona without aggregating. Although limited, PEGylation overall reduced NPs uptake in human macrophages. Uptake of NPs exposing folate prepared with short PEG was higher in KB cells (FR+) than in A549 (FR-), occurred via FR-receptor and involved lipid rafts-dependent endocytosis. In conclusion, the present results demonstrate that PEG length critically affects protein interaction and folate exposition with a logical impact on receptor-mediated cell uptake. Our study highlights that the too simplistic view suggesting that PEG-PCL gives PEG-coated NPs needs to be re-examined in the light of actual surface properties, which should always be considered case-by-case.


Clinical experiences with vecuronium in children.
The neuromuscular blocking effects of vecuronium have been investigated clinically in children regarding its efficacy and safety. The drug is given either of the two doses--0.08 mg/kg body weight or 0.1 mg/kg body weight in 40 children as a single intravenous bolus injection for endotracheal intubation. Vecuronium in doses of 0.1 mg/kg body weight provided satisfactory clinical relaxation for ideal intubating conditions in all children in reference to the ease of intubation, intermediate duration of action, spontaneous or easy reversal of the neuromuscular block, cardiovascular stability and absence of serious side-effects. Thus vecuronium may be regarded as a useful muscle relaxant in paediatric anaesthesia.


Anti-Helicobacter pylori activity of anacardic acids from Amphipterygium adstringens.
Amphipterygium adstringens (Schltdl.) Standl. (Anacardiaceae) is widely used in traditional Mexican medicine for the treatment of gastritis and ulcers. In this work, we studied the anti-Helicobacter pylori activity of its bark, this Gram-negative bacterium is considered the major etiological agent of chronic active gastritis and peptic ulcer disease, and it is linked to gastric carcinoma. From a bio-guided assay of the fractions obtained form a continuous Soxhlet extraction of the bark, we identified that petroleum ether fraction had significant antimicrobial activity against Helicobacter pylori. From this fraction, we isolated an anacardic acids mixture and three known triterpenes: masticadienonic acid; 3alpha-hydroxymasticadienonic acid; 3-epi-oleanolic; as well as the sterol beta-sitosterol. Only the anacardic acids mixture exhibits a potent dose-dependent antibacterial activity (MIC=10 microg/ml in broth cultures). It is enriched in saturated alkyl phenolic acids (C15:0, C16:0, C17:0 C19:0) which represents a novel source of these compounds with potent anti-Helicobacter pylori activity. The promising use of anacardic acids and Amphipterygium adstringens bark in the development of an integral treatment of Helicobacter pylori diseases is discussed.


Adverse drug events associated with the use of analgesics, sedatives, and antipsychotics in the intensive care unit.
As critically ill patients frequently receive analgesics, sedatives, and antipsychotics to optimize patient comfort and facilitate mechanical ventilation, adverse events associated with the use of these agents can affect all organ systems and result in substantial morbidity and mortality. Although many of these adverse effects are common pharmacologic manifestations of the agent, and therefore frequently reversible, others are idiosyncratic and thus unexpected. The critically ill are more susceptible to adverse drug events than nonintensive care unit patients due to the high doses and long periods for which each of these agents are often administered, the frequent use of intravenous formulations that contain adjuvants that may lead to toxicity in some instances, and the high prevalence of end-organ dysfunction that affects the pharmacokinetic and pharmacodynamic response to therapy. This paper will review the most common and serious adverse drug events reported to occur with the use of sedatives, analgesics, and antipsychotics in the intensive care unit; highlight the pharmacokinetic, pharmacodynamic, and pharmacogenetic factors that can influence analgesic, sedative, and antipsychotic response and safety in the critically ill; and identify strategies that can be used to minimize toxicity with these agents.


Updates on Diabetic Foot and Charcot Osteopathic Arthropathy.
Diabetes mellitus affects approximately 30.8 million people currently living in the USA. Chronic diabetes complications, including diabetic foot complications, remain prevalent and challenging to treat. We review clinical diagnosis and challenges providers may encounter when managing diabetic foot ulcers and Charcot neuroarthropathy. Mechanisms controlling these diseases are being elucidated and not fully understood. Offloading is paramount to heal and manage diabetic foot ulcers and Charcot neuroarthropathy. Diabetic foot ulcers recur and the importance of routine surveillance and multidisciplinary approach is essential. Several predictors of failure in Charcot foot include a related diabetic foot ulcer, midfoot or rearfoot location of the Charcot event, and progressive bony changes on interval radiographs. Patients with diabetic foot ulcer and/or Charcot neuroarthropathy are in need of consistent and regular special multidisciplinary care. If not diagnosed early and managed effectively, morbidity and mortality significantly increase.


Versatile low-copy-number plasmid vectors for cloning in Escherichia coli.
Small low-copy-number plasmid vectors were constructed by in vitro and in vivo recombinant DNA techniques. pLG338 and pLG339 are derived from pSC105, have a copy number of six to eight per chromosome, and carry genes conferring resistance to tetracycline and kanamycin. pLG338 (7.3 kb) has unique restriction endonuclease sites for BamHI, SalI, HincII, SmaI, XhoI, EcoRI and KpnI, the first five lying within a drug resistance gene. pLG339 (6.2 kb) lacks the KpnI site, but has unique SphI and PvuII sites. These versatile vectors should be useful for cloning many genes coding for membrane and regulatory proteins which cannot be cloned into high-copy-number plasmids.


Brain-derived neurotrophic factor precursor in the immune system is a novel target for treating multiple sclerosis.
Rationale: Brain-derived neurotrophic factor precursor (proBDNF) is expressed in the central nervous system (CNS) and the immune system. However, the role of proBDNF in the pathogenesis of multiple sclerosis (MS) is unknown. Methods: Peripheral blood and post-mortem brain and spinal cord specimens were obtained from multiple sclerosis patients to analyze proBDNF expression in peripheral lymphocytes and infiltrating immune cells in the lesion site. The proBDNF expression profile was also examined in the experimental autoimmune encephalomyelitis (EAE) mouse model, and polyclonal and monoclonal anti-proBDNF antibodies were used to explore their therapeutic effect in EAE. Finally, the role of proBDNF in the inflammatory immune activity of peripheral blood mononuclear cells (PBMCs) was verified in vitro experiments. Results: High proBDNF expression was detected in the circulating lymphocytes and infiltrated inflammatory cells at the lesion sites of the brain and spinal cord in MS patients. In the EAE mouse model, proBDNF was upregulated in CNS and in circulating and splenic lymphocytes. Systemic but not intracranial administration of anti-proBDNF blocking antibodies attenuated clinical scores, limited demyelination, and inhibited proinflammatory cytokines in EAE mice. Immuno-stimulants treatment increased the proBDNF release and upregulated the expression of p75 neurotrophic receptors (p75NTR) in lymphocytes. The monoclonal antibody against proBDNF inhibited the inflammatory response of PBMCs upon stimulations. Conclusion: The findings suggest that proBDNF from immune cells promotes the immunopathogenesis of MS. Monoclonal Ab-proB may be a promising therapeutic agent for treating MS.


Inactivation of prolyl endopeptidase by a peptidylchloromethane. Kinetics of inactivation and identification of sites of modification.
The kinetics of inactivation of prolyl endopeptidase by acetyl-Ala-Ala-Pro-CH2Cl were studied by progress-curve methods in the presence of substrate. The kinetic mechanism was found to involve the formation of an initial complex between the enzyme and the chloromethane followed by an inactivation step. The substrate was shown to compete for the formation of the initial complex, indicating that binding at the active site was a prerequisite for inactivation. After reaction of the enzyme with [3H]acetyl-Ala-Ala-Pro-CH2Cl, it was possible to isolate five labelled peptides. Four of these peptides contained a cysteine residue as the site of modification, whereas the fifth peptide contained no cysteine and a histidine residue was identified as the site of modification. This residue (His-680) probably represents the active-site histidine of prolyl endopeptidase.


Endoscopic thyroidectomy via areola approach for stage T1 papillary thyroid carcinoma: feasibility, safety, and oncologic outcomes.
To evaluate the feasibility, safety, and oncologic outcomes associated with endoscopic thyroidectomy via the areolar approach (ETAA), compared with conventional open thyroidectomy (COT) for the treatment of stage T1 papillary thyroid carcinoma (PTC). Between January 2021 and June 2022, a total of 1204 patients diagnosed with PTC underwent screening, out of which 138 patients were selected for inclusion in the study population after propensity score matching (92 patients in the ETAA group and 46 patients in the COT group). The study included the collection and analysis of clinicopathologic characteristics, intraoperative outcomes, postoperative outcomes, complications, and follow-up data using R software. The operative time for the ETAA group was longer than that for the COT group (160.42 ± 32.21 min vs. 121.93 ± 29.78 min, p < 0.0001). However, there were no significant differences between the two groups in terms of intraoperative blood loss, the extent of surgical resection, the number of dissected lymph nodes, the number of metastatic lymph nodes, and the rate of parathyroid autotransplantation. Postoperative drainage and C-reactive protein levels were higher in the ETAA group than in the COT group, but there were no significant differences in 24-hour visual analogue scale scores, white blood cell counts, drainage duration, or postoperative hospital stay. Complication rates were similar between the two groups, and no permanent recurrent laryngeal nerve palsy or hypoparathyroidism was observed. Patients who underwent ETAA reported greater cosmetic satisfaction and quality of life than those who underwent COT. During the follow-up phase, only one patient in the COT group developed lateral cervical lymph node involvement requiring reoperation. ETAA is a safe and feasible surgical method for patients with stage T1 PTC, providing results similar to COT in terms of oncologic completeness, while avoiding neck scars, with excellent cosmetic effects. Chinese Clinical Trial Registry center, identifier ChiCTR2300077109.


Readiness of managers and health care workers for e-Health: a cross-sectional study in Khartoum primary health care centers, Sudan.
e-Health is defined as "the use, in the health sector, of digital data- transmitted, stored and retrieved electronically-for clinical, educational and administrative purposes, both at the local site and at a distance". In Primary health care (PHC), the role of e-Health in promoting PHC systems defines its need to achieve the PHC aims. This literary work aims to study the readiness of managers and healthcare workers for e-Health at Khartoum state PHC centers. This was a facility-based cross-sectional study that took place between February and August 2022. A sample size of 327 was calculated, and multistage cluster sampling was used. A validated questionnaire was used, and the generated data were analyzed using the Statistical Package for Social Sciences (SPSS). Variables were described as n (%) and mean ± SD. Non-parametric tests and Spearman's correlation were used to investigate the association of readiness scores with different categorical and numerical variables, respectively. A standard multiple regression model was used to model the associations. A total of 262 forms were completed. The overall readiness percentages were low for both managers (52.8%) and healthcare workers (55.3%). Factors associated with e-Health readiness included occupation, doctors' level of expertise, PHC center, and PHC center type. This study reports low levels of e-Health readiness as reflected by managers and HCWs in Khartoum State PHC. Project planners need to be aware of the obstacles and threats faced by e-Health initiatives if they are not carefully planned, executed, and monitored. Special attention must be given to addressing health inequities and inequalities to ensure that these projects will contribute to fostering accessibility to health services and narrowing the digital divide.


Diet-induced increases in adiposity, but not plasma lipids, promote macrophage infiltration into white adipose tissue.
Obesity, hyperlipidemia, and insulin resistance are cardinal features of the metabolic syndrome and individually increase the risk for developing diabetes and cardiovascular disease, a risk that is amplified when they are simultaneously present. It is becoming increasingly clear that macrophages can infiltrate white adipose tissue (WAT) in the obese state, and their presence is associated with pathophysiological consequences of obesity, such as inflammation and insulin resistance. To determine whether hyperlipidemia could potentiate macrophage infiltration into WAT in the presence of obesity, obesity-prone agouti yellow mice (A(y)/a) on a hyperlipidemia-prone LDL receptor (LDLR)-deficient (LDLR(-/-)) background were placed on chow or Western diet. In addition, A(y)/a mice that were LDLR sufficient were also placed on Western diet. Both genetics and diet increased the degree of adiposity; however, plasma lipids were elevated only in the Western diet-fed LDLR(-/-) mice. The extent of macrophage accumulation in WAT correlated with the degree of adiposity. However, hyperlipidemia did not impact macrophage recruitment to WAT or the downstream metabolic consequences of macrophage accumulation in WAT, such as inflammation and insulin resistance. These data have important implications for the pathogenesis of diet-induced obesity in humans, even when plasma lipid abnormalities are not present.


Rat mandibular distraction osteogenesis: part III. Gradual distraction versus acute lengthening.
Distraction osteogenesis is a well-established method of endogenous tissue engineering. This technique has significantly augmented our armamentarium of reconstructive craniofacial procedures. Although the histologic and ultrastructural changes associated with distraction osteogenesis have been extensively described, the molecular mechanisms governing successful membranous distraction remain unknown. Using an established rat model, the molecular differences between successful (i.e., osseous union with gradual distraction) and ineffective (i.e., fibrous union with acute lengthening) membranous bone lengthening was analyzed. Herein, the first insight into the molecular mechanisms of successful membranous bone distraction is provided. In addition, these data provide the foundation for future targeted therapeutic manipulations designed to improve osseous regeneration. Vertical mandibular osteotomies were created in 52 adult male Sprague-Dawley rats, and the animals were fitted with customized distraction devices. Twenty-six animals underwent immediate acute lengthening (3 mm; a length previously shown to result in fibrous union) and 26 animals were gradually distracted (after a 3-day latency period, animals were distracted 0.25 mm twice daily for 6 days; total = 3 mm). Four mandibular regenerates were harvested from each group for RNA analysis on 5, 7, 9, 23, and 37 days postoperatively (n = 40). Two mandibular regenerates were also harvested from each group and prepared for immunohistochemistry on postoperative days 5, 7, and 37 (n = 12). In addition to the 52 experimental animals, 4 control rats underwent sham operations (skin incision only) and mandibular RNA was immediately collected. Control and experimental specimens were analyzed for collagen I, osteocalcin, tissue inhibitor of metalloproteinase-1, and vascular endothelial growth factor mRNA and protein expression. In this study, marked elevation of critical extracellular matrix molecules (osteocalcin and collagen I) during the consolidation phase of gradual distraction compared with acute lengthening is demonstrated. In addition, the expression of an inhibitor of extracellular matrix turnover, tissue inhibitor of metalloproteinase-1, remained strikingly elevated in gradually distracted animals. Finally, this study demonstrated that neither gradual distraction nor acute lengthening appreciably alters vascular endothelial growth factor expression. These results suggest that gradual distraction osteogenesis promotes successful osseous bone repair by regulating the expression of bone-specific extracellular matrix molecules. In contrast, decreased production or increased turnover of bone scaffolding proteins (i.e., collagen) or regulators of mineralization (i.e., osteocalcin) may lead to fibrous union during acute lengthening.


Dental student skill retention eight months after simulator-supported training in oral radiology.
The purpose of this project was to investigate the long-term effects on skill to interpret spatial information in radiographs after conventional and simulator-supported training. The study was a follow-up of a previously reported randomized experimental study. The original study population was comprised of fifty-seven dental students. Forty-five individuals agreed to participate in a follow-up study eight months after completion of the original study. During the time interval between completion of the previous study and the follow-up study, the participants underwent an examination in oral radiology and had theoretical and clinical training in other topics than oral radiology. Skill at interpreting spatial information in radiographs was assessed with a previously used test instrument. The test instrument was identical with the instrument used for baseline assessment in the original study. The results showed that the skill to interpret spatial relations in radiographs eight months after completion of simulator-supported training was significantly better (p=0.01) than before training. The conventional training showed almost the same pattern, but the difference was smaller and not statistically significant (p=0.11). It is concluded that simulator-supported training is a valuable adjunct to conventional educational methods in oral radiology.


Dual channel ion imprinted fluorescent polymers for dual mode simultaneous chromium speciation analysis.
A simple core shell structured fluorescent sensor was constructed to realize simultaneous detection of hexavalent and trivalent chromium ions. Briefly, blue-carbon dots (b-CDs) were embedded into a silica sphere, then a Cr(iii) imprinted silica layer doped with red-CDs (r-CDs) was coated onto the b-CDs@SiO2. Cr(vi) can selectively quench b-CDs based on the inner filter effect and Cr(iii) can selectively quench r-CDs based on electron transfer with the aid of the ion imprinting technique. In this strategy, it was not necessary to reduce Cr(vi) to Cr(iii) or oxidize Cr(iii) to Cr(vi), the chromium speciation of both can be detected simultaneously. When Cr(vi) was detected in the blue channel, the fluorescence intensity quenching effect was seen at 440 nm, and was linear from 0.01 to 10.0 μM, with a detection limit of 3.8 nM. For the detection of Cr(iii) in the red channel, the fluorescence intensity quenching effect was seen at 605 nm, and was linear from 0.1 to 15.0 μM, with a detection limit of 46 nM. This strategy enjoyed the advantages of simple construction, convenient detection, good selectivity and high sensitivity.


Readers select a comprehension mode independent of pronoun: Evidence from fMRI during narrative comprehension.
Perspective is a crucial feature for communicating about events. Yet it is unclear how linguistically encoded perspective relates to cognitive perspective taking. Here, we tested the effect of perspective taking with short literary stories. Participants listened to stories with 1st or 3rd person pronouns referring to the protagonist, while undergoing fMRI. When comparing action events with 1st and 3rd person pronouns, we found no evidence for a neural dissociation depending on the pronoun. A split sample approach based on the self-reported experience of perspective taking revealed 3 comprehension preferences. One group showed a strong 1st person preference, another a strong 3rd person preference, while a third group engaged in 1st and 3rd person perspective taking simultaneously. Comparing brain activations of the groups revealed different neural networks. Our results suggest that comprehension is perspective dependent, but not on the perspective suggested by the text, but on the reader's (situational) preference.


[Malaria in Norway--diagnosis, treatment and prevention].
The paper describes and discusses procedures and problems related to diagnostics, therapy and prophylaxis of malaria in Norway. A high degree of suspicion in physicians evaluating febrile travellers is of the utmost importance. The clinical symptoms and findings are initially rather unspecific, with fever and fever related symptoms. A definite diagnosis is made by the demonstration of malaria parasites in thin and thick blood smears, which is best performed by an infectious disease physician. Before treatment is started, it is important to determine whether the patient is infected with Plasmodium falciparum or with one of the more benign malaria species. If this is difficult microscopically, a test that detects P. falciparum antigen in blood may be useful in some cases. The therapy of benign malaria is still chloroquine plus primaquine; most patients with malaria falciparum can be routinely treated with mefloquine. Cases of complicated falciparum malaria, usually due to delayed diagnosis and start of treatment, require extensive and sophisticated treatment, usually including parenteral treatment with quinine. Prophylaxis consists of prevention of mosquito bites and chemoprophylaxis; the importance of avoiding bites should never be underestimated. A more widespread use of mefloquine among travellers to Africa is the most important change in chemoprophylaxis in recent years. Finally the article discusses possible improvements in diagnostic procedure, therapy and chemoprophylaxis.


Does facet joint inflammation induce radiculopathy?: an investigation using a rat model of lumbar facet joint inflammation.
The association between lumbar facet joint inflammation and radiculopathy was investigated using behavioral, histologic, and immunohistochemical testing in rats. To develop a rat model of lumbar facet joint inflammation and ascertain whether facet joint inflammation induces radiculopathy using this model. Both mechanical and chemical factors have been identified as important for inducing radiculopathy. In lumbar spondylosis, facet joint osteophytes may contribute to nerve root compression, which may induce radiculopathy. Furthermore, inflammation may occur in the facet joint, as in other synovial joints. Inflamed synovium may thus release inflammatory cytokines and induce nerve root injury with subsequent radiculopathy. A piece of gelatin sponge containing complete adjuvant was inserted into the L5-L6 facet joint in rats (arthritis group). Saline was used in the control group. Mechanical allodynia was determined using the von Frey test. Inflammatory cells infiltrating the epidural space were counted, and changes in cartilage were assessed histologically. Tumor necrosis factor (TNF)-alpha-immunoreactive cells in the L5 dorsal root ganglion were counted. Mechanical allodynia was observed in the arthritis group from day 3, gradually recovering during the observation period. Significantly larger numbers of inflammatory cells had infiltrated the epidural space by days 3 and 7 in the arthritis group than in controls. Numbers of TNF-alpha-immunoreactive cells were significantly increased at days 1 and 3 in the arthritis group compared with controls. Predominantly small nociceptive neurons were stained. When inflammation was induced in a facet joint, inflammatory reactions spread to nerve roots, and leg symptoms were induced by chemical factors. These results support the possibility that facet joint inflammation induces radiculopathy.


Brain tumor vessel response to synchrotron microbeam radiation therapy: a short-term in vivo study.
The aim of this work focuses on the description of the short-term response of a 9L brain tumor model and its vasculature to microbeam radiation therapy (MRT) using magnetic resonance imaging (MRI). Rat 9L gliosarcomas implanted in nude mice brains were irradiated by MRT 13 days after tumor inoculation using two orthogonal arrays of equally spaced 28 planar microbeams (25 microm width, 211 microm spacing and dose 500 Gy). At 1, 7 and 14 days after MRT, apparent diffusion coefficient, blood volume and vessel size index were mapped by MRI. Mean survival time after tumor inoculation increased significantly between MRT-treated and untreated groups (23 and 28 days respectively, log-rank test, p < 0.0001). A significant increase of apparent diffusion coefficient was observed 24 h after MRT in irradiated tumors versus non-irradiated ones. In the untreated group, both tumor size and vessel size index increased significantly (from 7.6 +/- 2.2 to 19.2 +/- 4.0 mm(2) and +23%, respectively) between the 14th and the 21st day after tumor cell inoculation. During the same period, in the MRT-treated group, no difference in tumor size was observed. The vessel size index measured in the MRT-treated group increased significantly (+26%) between 14 and 28 days of tumor growth. We did not observe the significant difference in blood volume between the MRT-treated and untreated groups. MRT slows 9L tumor growth in a mouse brain but MRI results suggest that the increase in survival time after our MRT approach may be rather due to a cytoreduction than to early direct effects of ionizing radiation on tumor vessels. These results suggest that MRT parameters need to be optimized to further damage tumor vessels.


African swine fever virus serotype-specific proteins are significant protective antigens for African swine fever.
African swine fever (ASF) is an emerging disease threat for the swine industry worldwide. No ASF vaccine is available and progress is hindered by lack of knowledge concerning the extent of ASFV strain diversity and the viral antigens conferring type-specific protective immunity in pigs. Available data from vaccination/challenge experiments in pigs indicate that ASF protective immunity may be haemadsorption inhibition (HAI) serotype-specific. Recently, we have shown that two ASFV proteins, CD2v (EP402R) and C-type lectin (EP153R), are necessary and sufficient for mediating HAI serological specificity (Malogolovkin et al., 2015).. Here, using ASFV inter-serotypic chimeric viruses and vaccination/challenge experiments in pigs, we demonstrate that serotype-specific CD2v and/or C-type lectin proteins are important for protection against homologous ASFV infection. Thus, these viral proteins represent significant protective antigens for ASFV that should be targeted in future vaccine design and development. Additionally, these data support the concept of HAI serotype-specific protective immunity.


Efficacy and pitfalls of digital technologies in healthcare services: A systematic review of two decades.
The use of technology in the healthcare sector and its medical practices, from patient record maintenance to diagnostics, has significantly improved the health care emergency management system. At that backdrop, it is crucial to explore the role and challenges of these technologies in the healthcare sector. Therefore, this study provides a systematic review of the literature on technological developments in the healthcare sector and deduces its pros and cons. We curate the published studies from the Web of Science and Scopus databases by using PRISMA 2015 guidelines. After mining the data, we selected only 55 studies for the systematic literature review and bibliometric analysis. The study explores four significant classifications of technological development in healthcare: (a) digital technologies, (b) artificial intelligence, (c) blockchain, and (d) the Internet of Things. The novel contribution of current study indicate that digital technologies have significantly influenced the healthcare services such as the beginning of electronic health record, a new era of digital healthcare, while robotic surgeries and machine learning algorithms may replace practitioners as future technologies. However, a considerable number of studies have criticized these technologies in the health sector based on trust, security, privacy, and accuracy. The study suggests that future studies, on technological development in healthcare services, may take into account these issues for sustainable development of the healthcare sector.


The mechanobiology of tendon fibroblasts under static and uniaxial cyclic load in a 3D tissue engineered model mimicking native extracellular matrix.
Tendon mechanobiology plays a vital role in tendon repair and regeneration; however, this mechanism is currently poorly understood. We tested the role of different mechanical loads on extracellular matrix (ECM) remodelling gene expression and the morphology of tendon fibroblasts in collagen hydrogels, designed to mimic native tissue. Hydrogels were subjected to precise static or uniaxial loading patterns of known magnitudes and sampled to analyse gene expression of known mechano-responsive ECM-associated genes (Collagen I, Collagen III, Tenomodulin, and TGF-β). Tendon fibroblast cytomechanics was studied under load by using a tension culture force monitor, with immunofluorescence and immunohistological staining used to examine cell morphology. Tendon fibroblasts subjected to cyclic load showed that endogenous matrix tension was maintained, with significant concomitant upregulation of ECM remodelling genes, Collagen I, Collagen III, Tenomodulin, and TGF-β when compared with static load and control samples. These data indicate that tendon fibroblasts acutely adapt to the mechanical forces placed upon them, transmitting forces across the ECM without losing mechanical dynamism. This model demonstrates cell-material (ECM) interaction and remodelling in preclinical a platform, which can be used as a screening tool to understand tendon regeneration.


Controversy: Noninvasive and invasive cortical stimulation show efficacy in treating stroke patients.
Stroke is the leading cause of disability in the adult population of western industrialized countries. Despite significant improvements of acute stroke care, two thirds of stroke survivors have to cope with persisting neurologic deficits. Adjuvant brain stimulation is a novel approach to improving the treatment of residual deficits after stroke. Transcranial magnetic stimulation (TMS), transcranial direct current stimulation (tDCS), and epidural electrical stimulation have been used in first trials on small cohorts of stroke patients. Effect sizes in the order of 8% to 30% of functional improvement have been reported, but a publication bias toward presenting "promising" but not negative results is likely. Many questions regarding underlying mechanisms, optimal stimulation parameters, combination with other types of interventions, among others, are open. This review addresses six controversies related to the experimental application of brain stimulation techniques to stroke patients. Cortical stimulation after stroke will need to be individually tailored and a thorough patient stratification according to type and extent of clinical deficit, lesion location, lesion size, comorbidities, time in the recovery process, and perhaps also age and gender will be necessary. There is consensus that cortical stimulation in stroke patients is still experimental and should only be applied in the frame of scientific studies.


A Deep-Learning Enabled Automatic Fetal Thalamus Diameter Measurement Algorithm.
The analysis of maternal factors that impact the normal development of the fetal thalamus is an emerging field of research and requires the retrospective measurement of fetal thalamus diameter (FTD). Unfortunately, FTD is not measured in routine 2D ultrasound (2D-US) screenings of fetuses. Manual measurement of FTD is a laborious, difficult, and error-prone process because the thalamus lacks well-defined boundaries in 2D-US images of the fetal brain as it has a similar echogenicity to the surrounding brain tissue. Traditional methods based on statistical shape models (SSMs) perform poorly in measuring FTD due to the noisy textures and fuzzy edges of the fetal thalamus in 2D-US images of the fetal brain. To overcome these difficulties, we propose a deep learning-based automatic FTD measurement algorithm, FTDNet. FTDNet measures FTD by learning to directly detect the measurement landmarks through supervised learning. The algorithm first detects the region of the brain that contains the thalamus structure, and then focuses on processing that region for FTD landmark detection. Our FTD dataset, developed through a consensus between two ultrasonographers, contains 1,111 pairs of landmark coordinates for measuring FTD and verified bounding boxes surrounding the fetal thalamus. To assess FTDNet's measurement consistency compared to the ground truth, we used the intraclass correlation coefficient (ICC). FTDNet achieved an ICC score of 0.734, significantly outperforming the prior SSM method and other baseline comparison methods. Our findings are an important step forward in understanding the maternal factors which influence fetal brain development.Clinical relevance- This work proposes an end-to-end thalamus detection and measurement algorithm for measuring fetal thalamus diameter. Our work represents a significant step in the research of how maternal factors can impact fetal thalamus development. The development of an automatic and accurate method for measuring FTD through deep learning has the potential to greatly advance this field of study.


Atomic Substitution Enabled Synthesis of Vacancy-Rich Two-Dimensional Black TiO2- x Nanoflakes for High-Performance Rechargeable Magnesium Batteries.
Rechargeable magnesium (Mg) batteries assembled with dendrite-free, safe, and earth-abundant metal Mg anodes potentially have the advantages of high theoretical specific capacity and energy density. Nevertheless, owing to the large polarity of divalent Mg2+ ions, the insertion of Mg2+ into electrode materials suffers from sluggish kinetics, which seriously limit the performance of Mg batteries. Herein, we demonstrate an atomic substitution strategy for the controlled preparation of ultrathin black TiO2- x (B-TiO2- x) nanoflakes with rich oxygen vacancies (OVs) and porosity by utilizing ultrathin 2D TiS2 nanoflakes as precursors. We find out that the presence of OVs in B-TiO2- x electrode material can greatly improve the electrochemical performances of rechargeable Mg batteries. Both experimental results and density functional theory simulations confirm that the introduction of OVs can remarkably enhance the electrical conductivity and increase the number of active sites for Mg2+ ion storage. The vacancy-rich B-TiO2- x nanoflakes exhibit high reversible capacity and good capacity retention after long-term cycling at large current densities. It is hoped that this work can provide valuable insights and inspirations on the defect engineering of electrode materials for rechargeable magnesium batteries.


[The discourse of managers on equity: a challenge for Brazil's Unified Health System (SUS)].
Equity is one of the pillars of Brazil's Unified Health System (SUS). However, analysis of the scientific literature shows that the concept of equity is polysemic and open to many interpretations. The scope of this article is to identify the meanings conferred upon equity in the discourse of SUS managers. Semi-structured interviews were conducted with individuals in three municipalities in the state of Sao Paulo, namely Marília, São Carlos and Santos. The interviews were recorded, transcribed and organized using the Discourse of the Collective Subject method. In the analysis of the results, three core categories were identified: Treat everyone in an egalitarian manner; treat the unequal in an unequal manner, prioritizing the most needy in accordance with their social and economic status; and treating the unequal in an unequal manner, prioritizing specific groups in accordance with risk criteria. The managers acknowledge equity as being a SUS principle and seek prudent clinical and epidemiological criteria to justify their choices in allocating resources. There is a need for further research into the concept and operationalization of equity and the need to hear the other actors of SUS who are involved in the actions and are jointly responsible for decisions on healthcare.


Analysis of a recombinant mouse hepatitis virus expressing a foreign gene reveals a novel aspect of coronavirus transcription.
We have inserted heterologous genetic material into the nonessential gene 4 of the coronavirus mouse hepatitis virus (MHV) in order to test the applicability of targeted RNA recombination for site-directed mutagenesis of the MHV genome upstream of the nucleocapsid (N) gene and to develop further genetic tools for site-directed mutagenesis of structural genes other than N. Initially, a 19-nucleotide tag was inserted into the start of gene 4a of MHV strain A59 with the N gene deletion mutant Alb4 as the recipient virus. In further work, the entire gene for the green fluorescent protein (GFP) was inserted in place of gene 4, creating the currently largest known RNA virus. The expression of GFP was demonstrated by Western blot analysis of infected cell lysates; however, the level of GFP expression was not sufficient to allow detection of fluorescence of viral plaques. Northern blot analysis of transcripts of GFP recombinants showed the expected alteration of the pattern of the nested MHV subgenomic mRNAs. Surprisingly, though, GFP recombinants also produced an RNA species that was the same size as wild-type mRNA4. Analysis of the 5' end of this species revealed that it was actually a collection of mRNAs originating from 10 different genomic fusion sites, none possessing a canonical intergenic sequence. The finding of these aberrant mRNAs suggests that long-range RNA or the ribonucleoprotein structure of the MHV genome can sometimes be the sole determinant of the site of initiation of transcription.


Membrane-induced conformational change in human apolipoprotein H.
The interaction of apolipoprotein H (Apo H) with lipid membrane has been considered to be a basic mechanism for the biological function of the protein. Previous reports have demonstrated that Apo H can interact only with membranes containing anionic phospholipids. Here we study the membrane-induced conformational change of Apo H by CD spectroscopy with two different model systems: anionic-phospholipid-containing liposomes [such as 1, 2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) and cardiolipin], and the water/methanol mixtures at moderately low pH, which mimic the micro-physicochemical environment near the membrane surface. It is found that Apo H undergoes a remarkable conformational change on interaction with liposomes containing anionic phospholipid. To interact with liposomes containing DMPG, there is a 6.8% increase in alpha-helix in the secondary structures; in liposomes containing cardiolipin, however, there is a 12.6% increase in alpha-helix and a 9% decrease in beta-sheet. The similar conformation change in Apo H can be induced by treatment with an appropriate mixture of water/methanol. The results indicate that the association of Apo H with membrane is correlated with a certain conformational change in the secondary structure of the protein.


Robotic versus laparoscopic gastrectomy for adenocarcinoma in the US: a propensity score-matching analysis of 11,173 patients on oncological adequacy.
Surgery remains the cornerstone treatment for gastric cancer. Previous studies have reported better lymphadenectomy with minimally invasive approaches. There is a paucity of data comparing robotic and laparoscopic gastrectomy in the US. Herein, we examined whether oncological adequacy differs between laparoscopic and robotic approaches. The National Cancer Database was utilized to identify patients who underwent gastrectomy for adenocarcinoma between 2010 and 2019. A propensity score-matching analysis between robotic gastrectomy (RG) versus laparoscopic gastrectomy (LG) was performed. The primary outcomes were lymphadenectomy ≥ 16 nodes and surgical margins. A total of 11,173 patients underwent minimally invasive surgery for gastric adenocarcinoma between 2010 and 2019. Of those 8320 underwent LG and 2853 RG. Comparing the unmatched cohorts, RG was associated with a higher rate of adequate lymphadenectomy (63.5% vs 57.1%, p < .0.0001), higher rate of negative margins (93.8% vs 91.9%, p < 0.001), lower rate of prolonged length of stay (26.0% vs 29.6%, p < .0.001), lower 90-day mortality (3.7% vs 5.0%, p < 0.0001), and a better 5-year overall survival (OS) (56% vs 54%, p = 0.03). A propensity score-matching cohort with a 1:1 ratio was created utilizing the variables associated with lymphadenectomy ≥ 16 nodes. The matched analysis revealed that the rate of adequate lymphadenectomy was significantly higher for RG compared to LG, 63.5% vs 60.4% (p = 0.01), respectively. There was no longer a significant difference between RG and LG regarding the rate of negative margins, prolonged length of stay, 90-day mortality, rate of receipt of postoperative chemotherapy, and OS. This propensity score-matching analysis with a large US cohort shows that RG was associated with a higher rate of adequate lymphadenectomy compared to LR. RG and LG had a similar rate of negative margins, prolonged length of stay, receipt of postoperative chemotherapy, 90-day mortality, and OS, suggesting that RG is a comparable surgical approach, if not superior to LG.


Neural cell adhesion molecule mediates contact-dependent inhibition of growth of near-diploid mouse fibroblast cell line m5S/1M.
A near-diploid mouse fibroblast cell line m5S/1M used in this study shows a high sensitivity to contact-dependent inhibition of growth, and the addition of EGF causes both morphological change and loss of contact-dependent inhibition of growth. The m5S/1M cell and its transformants obtained by x-ray irradiation have been used to search for the cell surface glycoproteins that are responsible for the growth regulation via cell-cell interactions. Lectin blotting analyses showed that the expression of the cell surface glycoprotein of 140 kD (140KGP) is highly sensitive to the transformation induced either by x-ray irradiation or by the EGF stimulation. We purified the 140KGP and found that it was composed of two glycoproteins. The major component of 140KGP was identified as neural cell adhesion molecule (NCAM) by amino acid sequence analyses of the peptide fragments and by the cross-reactivity with anti-NCAM mAb, clone H22.214.171.124. Monoclonal antibody against 140KGP (clone LN-10) recognizes all three isoforms of NCAM expressed on m5S/1M cell and showed that the expression of NCAM was highly sensitive to the transformation. Furthermore, the immobilized LN-10 strongly inhibited the growth of actively proliferating m5S/1M cells and the LN-10 in a soluble form showed a significant growth-stimulating effect on the confluent quiescent cultures of m5S/1M cells. The results show that NCAM plays a major role in the contact-dependent inhibition of growth of m5S/1M, and that NCAM might be involved in the regulation of cell growth during embryogenesis and formation of nervous systems.


Executive dyscontrol: an important factor affecting the level of care received by older retirees.
To examine the relative contributions of Executive Control Function (ECF), general cognition, mood, problem behavior, physical disability, demographic variables, and the number of prescribed medications to the level of care received by older retirees. Multivariate regression and discriminant modeling. A single Continuing Care Retirement Community (CCRC) in San Antonio, Texas. A total of 107 older retirees (mean age = 83.7+/-7.2 years), including 17 community-dwelling, well, older controls and 90 CCRC residents. CCRC subjects represented a convenience sample of consecutive referrals for geropsychiatric assessment. Sixty-one subjects resided at a noninstitutionalized level of care, and 46 were institutionalized. Tests of ECF (the Executive Interview (EXIT25)), general cognition (the Mini-Mental State Examination (MMSE)), mood (the Geriatric Depression Scale short-form (sGDS)), problem behavior (the Nursing Home Behavior Problem Scale (NHBPS)), physical disability (the Cumulative Illness Rating Scale (CIRS)), age, gender, years of education, and the number of prescribed medications were studied. All variables except gender and education varied significantly across level of care. Four variables made significant independent contributions; EXIT25 score (r2 = .48, P< .001), medication usage (partial r2 = .11, P<.001), sGDS score (partial r2 = .06, P = .001), and problem behavior (partial NHBPS r2 = .04, P<.04). These variables accounted for 69% of the total variance in level of care (R2 = .69; F (df 7,99) = 32.1, P<.001). A discriminant model based on the number of prescribed medications, EXIT25, sGDS, and NHBPS scores classified 83.2% of cases correctly (Wilke's lambda = .50, F(5,101) = 20.1; P<.001). The MMSE enters but fails to contribute significantly, independent of the other variables. Age and CIRS scores fail to enter. Cognitive (particularly ECF) impairment contributes most to the observed variance in level of care received by older retirees in this CCRC. In contrast, markers of general cognition, depression, and physical illness contributed relatively little additional variance. ECF is not detected well by traditional cognitive measures and must be sought by specific tests. Further study is needed to replicate these findings in other populations.


Targeting of mesenchymal stem cells to ovarian tumors via an artificial receptor.
Mesenchymal Progenitor/Stem Cells (MSC) respond to homing cues providing an important mechanism to deliver therapeutics to sites of injury and tumors. This property has been confirmed by many investigators, however, the efficiency of tumor homing needs to be improved for effective therapeutic delivery. We investigated the feasibility of enhancing MSC tumor targeting by expressing an artificial tumor-binding receptor on the MSC surface. Human MSC expressing an artificial receptor that binds to erbB2, a tumor cell marker, were obtained by transduction with genetically modified adenoviral vectors encoding an artificial receptor (MSC-AR). MSC-AR properties were tested in vitro in cell binding assays and in vivo using two model systems: transient transgenic mice that express human erbB2 in the lungs and ovarian xenograft tumor model. The levels of luciferase-labeled MSCs in erbB2-expressing targeted sites were evaluated by measuring luciferase activity using luciferase assay and imaging. The expression of AR enhanced binding of MSC-AR to erbB2-expressing cells in vitro, compared to unmodified MSCs. Furthermore, we have tested the properties of erbB2-targeted MSCs in vivo and demonstrated an increased retention of MSC-AR in lungs expressing erbB2. We have also confirmed increased numbers of erbB2-targeted MSCs in ovarian tumors, compared to unmodified MSC. The kinetic of tumor targeting by ip injected MSC was also investigated. These data demonstrate that targeting abilities of MSCs can be enhanced via introduction of artificial receptors. The application of this strategy for tumor cell-based delivery could increase a number of cell carriers in tumors and enhance efficacy of cell-based therapy.


emo-1, a Caenorhabditis elegans Sec61p gamma homologue, is required for oocyte development and ovulation.
emo-1(oz1) is a member of a class of hermaphrodite sterile mutations in Caenorhabditis elegans that produce endomitotic oocytes in the gonad arm. Oocytes in emo-1(oz1) mutants exhibit multiple defects during oogenesis. After meiotic maturation, ovulation fails, trapping oocytes in the gonad arm where they become endomitotic. emo-1 encodes a homologue of the Sec61p gamma subunit, a protein necessary for translocation of secretory and transmembrane proteins into the endoplasmic reticulum of yeast and mammalian cells. A putative emo-1 null mutation, oz151, displays embryonic lethality. The oz1 sterile mutation is a transposable element insertion into the emo-1 3' untranslated region that almost completely eliminates germline mRNA accumulation. Genetic mosaic analysis using the oz1 allele indicates that emo-1(+) expression in germ cells is required for fertility. The J67 monoclonal antibody, which recognizes an oocyte surface antigen (Strome, S. 1986. In Gametogenesis and the Early Embryo. J.G. Gall, editor. Alan R. Liss, Inc., New York. 77-95.), does not stain oz1 oocytes, a finding consistent with defective protein transport in the mutant. We propose that the emo-1 gene product acts in the transport of secreted and transmembrane proteins in C. elegans oocytes, and is necessary for both oogenesis and the coupling of ovulation with meiotic maturation.


Research on structural strengthening technology for regenerative denitration catalysts.
The cost of replacing failed selective catalytic reduction (SCR) catalysts and their disposal as hazardous solid waste is high. If failed catalysts are recovered and regenerated into new SCR denitration catalysts, the cost of flue gas denitration can be effectively reduced. However, regenerated SCR catalysts have relatively low structural strength and activity and cannot yet form an effective replacement. In this study, aluminum dihydrogen phosphate, aluminum nitrate, and aluminum sulfate were used as structural strengthening agents in the regeneration of SCR catalysts, and over-impregnation, drumming-assisted impregnation, and ultrasonic-assisted preparation techniques were compared. The corresponding regenerated SCR catalysts were then prepared and analyzed for compressive strength, wear strength, H2-TPR, NH3-TPD, and in situ IR. Factors influencing the structural strength, physical properties, and catalytic activity of the regenerated catalysts were investigated. The best results were obtained as follows: compressive strength of 4.57 MPa, wear rate of 0.088% kg-1, and denitration of 58% after 10 min of drumming-assisted impregnation in an aluminum sulfate solution with a concentration of 16%. Based on this, a synergistic method for catalyst activity and structural strengthening was explored to support the design of better SCR catalysts for regeneration.


Second metatarsal stress fracture in sport: comparative risk factors between proximal and non-proximal locations.
Stress fractures of the second metatarsal are common injuries in athletes and military recruits. There are two distinct areas in the second metatarsal where stress fractures develop: one proximal (at the base) and the other non-proximal (distal). Diagnosis can be difficult, and there is a difference in prognosis and treatment of the two types of stress fracture. Therefore differentiation of fracture location is warranted. Differences in risk factors and clinical outcomes between proximal and non-proximal stress fractures have not been studied. To determine whether different risk factors and/or clinical outcomes associated with proximal and non-proximal stress fractures of the second metatarsal exist. Patients diagnosed with proximal stress fractures of the second metatarsal were included in the study. Retrospectively, an age-matched control group with a non-proximal stress fracture was selected for comparison. Statistical analysis involved bivariate comparisons of demographic variables and clinical measurement between the two groups. Patients with proximal stress fractures were more likely to be chronically affected, usually exhibited an Achilles contracture, showed differences in length of first compared with second metatarsal, were more likely to experience multiple stress fractures, and exhibited low bone mass. In addition, a high degree of training slightly increased the risk of a non-proximal fracture, whereas low training volume was associated with a proximal stress fracture. The signs, symptoms and clinical findings associated with proximal metatarsal stress fractures are different from those of non-proximal stress fractures.


Ionic conductance pathways in the mouse medullary thick ascending limb of Henle. The paracellular pathway and electrogenic Cl- absorption.
Net Cl- absorption in the mouse medullary thick ascending limb of Henle (mTALH) involves a furosemide-sensitive Na+:K+:2 Cl- apical membrane symport mechanism for salt entry into cells, which occurs in parallel with a Ba++-sensitive apical K+ conductance. The present studies, using the in vitro microperfused mouse mTALH, assessed the concentration dependence of blockade of this apical membrane K+-conductive pathway by Ba++ to provide estimates of the magnitudes of the transcellular (Gc) and paracellular (Gs) electrical conductances (millisiemens per square centimeter). These studies also evaluated the effects of luminal hypertonicity produced by urea on the paracellular electrical conductance, the electrical Na+/Cl- permselectivity ratio, and the morphology of in vitro mTALH segments exposed to peritubular antidiuretic hormone (ADH). Increasing luminal Ba++ concentrations, in the absence of luminal K+, produced a progressive reduction in the transcellular conductance that was maximal at 20 mM Ba++. The Ba++-sensitive transcellular conductance in the presence of ADH was 61.8 +/- 1.7 mS/cm2, or approximately 65% of the total transepithelial conductance. In phenomenological terms, the luminal Ba++-dependent blockade of the transcellular conductance exhibited negative cooperativity. The transepithelial osmotic gradient produced by luminal urea produced blebs on apical surfaces, a striking increase in shunt conductance, and a decrease in the shunt Na+/Cl- permselectivity (PNa/PCl), which approached that of free solution. The transepithelial conductance obtained with luminal 800 mM urea, 20 mM Ba++, and 0 K+ was 950 +/- 150 mS/cm2 and provided an estimate of the maximal diffusion resistance of intercellular spaces, exclusive of junctional complexes. The calculated range for junctional dilution voltages owing to interspace salt accumulation during ADH-dependent net NaCl absorption was 0.7-1.1 mV. Since the Ve accompanying ADH-dependent net NaCl absorption is 10 mV, lumen positive, virtually all of the spontaneous transepithelial voltage in the mouse mTALH is due to transcellular transport processes. Finally, we developed a series of expressions in which the ratio of net Cl- absorption to paracellular Na+ absorption could be expressed in terms of a series of electrical variables. Specifically, an analysis of paired measurement of PNa/PCl and Gs was in agreement with an electroneutral Na+:K+:2 Cl- apical entry step. Thus, for net NaCl absorption, approximately 50% of Na+ was absorbed via a paracellular route.


Multivariate classification of schizophrenia and its familial risk based on load-dependent attentional control brain functional connectivity.
Patients with schizophrenia (SCZ), as well as their unaffected siblings (SIB), show functional connectivity (FC) alterations during performance of tasks involving attention. As compared with SCZ, these alterations are present in SIB to a lesser extent and are more pronounced during high cognitive demand, thus possibly representing one of the pathways in which familial risk is translated into the SCZ phenotype. Our aim is to measure the separability of SCZ and SIB from healthy controls (HC) using attentional control-dependent FC patterns, and to test to which extent these patterns span a continuum of neurofunctional alterations between HC and SCZ. 65 SCZ with 65 age and gender-matched HC and 39 SIB with 39 matched HC underwent the Variable Attentional Control (VAC) task. Load-dependent connectivity matrices were generated according to correct responses in each VAC load. Classification performances of high, intermediate and low VAC load FC on HC-SCZ and HC-SIB cohorts were tested through machine learning techniques within a repeated nested cross-validation framework. HC-SCZ classification models were applied to the HC-SIB cohort, and vice-versa. A high load-related decreased FC pattern discriminated between HC and SCZ with 66.9% accuracy and with 57.7% accuracy between HC and SIB. A high load-related increased FC network separated SIB from HC (69.6% accuracy), but not SCZ from HC (48.5% accuracy). Our findings revealed signatures of attentional FC abnormalities shared by SCZ and SIB individuals. We also found evidence for potential, SIB-specific FC signature, which may point to compensatory neurofunctional mechanisms in persons at familial risk for schizophrenia.


2. Contact lens care and ocular surface homeostasis.
The early focus of contact lens wear and ocular health was on oxygen delivery. However, as we learn more about how the eye works, and investigate how the contact lens interacts with the cornea, the role of the tear film has risen in prominence. A healthy tear film is critical for normal ocular homeostasis, and abnormalities of the tear film are the primary cause of dry eye. In order to improve patient eye health and comfort during lens wear, we need to further elucidate the relationship among contact lenses, contact lens solutions, the tear film, and the corneal epithelium, and find ways to maintain homeostasis of the ocular surface. In this section, we review the latest data and opinions on this complex relationship between contact lenses and lens care solutions.


[Prominent ears. Correction using a modified Chongchet technique].
The authors describe their experience with the correction of prominent ears using a modified Chongchet technique. This method is based on the investigations of GIBSON and DAVIS (1958), who showed the tendency of cartilage to warp when one surface is cut. The prominent ear is folded back to produce the desired antihelix fold, which is outlined with methylene blue. An elliptical piece of retroauricular skin is excised. An incision is made through the whole thickness of the cartilage, following the posterior marked linea of the new antihelical fold, from the superior crus to the tail of antihelix. The anterior surface of the cartilage is dissected from the skin. A series of parallel incisions are made through the taut outer layer of the future fold. The tension on that side is released, and therefore it will bend towards the opposite side. The antihelical fold is corrected. This method was used in 192 patients with prominent ears. The results were satisfactory, with a normal-looking fold of antihelix. No recurrence of the deformity was seen.


A specific hybridisation internalisation probe (SHIP) enables precise live-cell and super-resolution imaging of internalized cargo.
Facilitated by the advancements in microscopy, our understanding of the complexity of intracellular vesicle traffic has dramatically increased in recent years. However, distinguishing between plasma membrane-bound or internalised ligands remains a major challenge for the studies of cargo sorting to endosomal compartments, especially in small and round cells such as lymphocytes. The specific hybridization internalisation probe (SHIP) assay, developed for flow cytometry studies, employs a ssDNA fluorescence internalisation probe and a complementary ssDNA quenching probe to unambiguously detect the internalized receptors/cargo. Here, we adopted the SHIP assay to study the trafficking of receptor/ligand complexes using B lymphocytes and B cell receptor-mediated antigen internalization as a model system. Our study demonstrates the potential of the SHIP assay for improving the imaging of internalized receptor/ligand complexes and establishes the compatibility of this assay with multiple imaging modalities, including live-cell imaging and super-resolution microscopy.


Neighborhood Environment and Self-Rated Health Among Urban Older Adults.
Objective: This study examines associations between neighborhood environment and self-rated health (SRH) among urban older adults. Method: We selected 217 individuals aged 65+ living in a de-industrialized Midwestern city who answered questions on the 2009 Speak to Your Health survey. The relationship between neighborhood environment and SRH was analyzed using regression models. Neighborhood variables included social support and participation, perceived racism, and crime. Additional models included actual crime indices to compare differences between perceived and actual crime. Results: Seniors who have poor SRH are 21% more likely to report fear of crime than seniors with excellent SRH (p = .01). Additional analyses revealed Black seniors are 7% less likely to participate in social activities (p = .005) and 4% more likely to report experiencing racism (p < .001). Discussion: More than 80% of older adults live in urban areas. By 2030, older adults will account for 20% of the U.S. Given the increasing numbers of older adults living in urban neighborhoods, studies such as this one are important. Mitigating environmental influences in the neighborhood that are associated with poor SRH may allow urban older adults to maintain health and reduce disability.


Functional relationship between cyclic AMP-dependent protein phosphorylation and platelet inhibition.
Exposure of human platelets to prostacyclin (PGI2), iloprost or prostaglandin E1 (PGE1) elicits the cyclic AMP-dependent phosphorylation of proteins of 22, 24, 30, 39, 50, 60 and 250 kDa (P22, P24 etc.). P22 was recently identified as rap 1B, a ras-like protein, and P24 was shown to be the beta-chain of glycoprotein Ib. We found that cyclic AMP-dependent phosphorylation of all proteins except P22 was maximal 1 min after exposure of platelets to PGI2, iloprost or PGE1; maximal phosphorylation of P22 occurred after 45 min of incubation. Inhibition of thrombin-induced platelet activation required only a 30 s incubation with PGI2 or iloprost; at this time phosphorylation of P22 was only slightly increased. Although at maximal concentrations PGI2 was more potent than PGE1 in inhibiting thrombin-induced platelet activation, no difference in the degree and the kinetics of cyclic AMP-dependent protein phosphorylation was found. Platelets that had been preincubated and washed in the presence of PGE1 and later resuspended in the absence of PGE1 responded fully to activation by thrombin despite maximal phosphorylation of P22 and P24. Furthermore, addition of PGI2 to PGE1-washed platelets prevented thrombin-induced platelet activation, but did not evoke further phosphorylation of P22 or P24. Phosphorylation of P39 and P50 correlated better with PGI2-induced inhibition of platelet activation. In experiments in which PGE1-induced inhibition of platelet activation was overcome by the addition of thrombin, no dephosphorylation of proteins phosphorylated by cyclic AMP-dependent kinases was observed. These experiments indicate that: (a) phosphorylation of rap 1B and glycoprotein Ib is not related to platelet inhibition by cyclic AMP; (b) phosphorylation of other proteins such as P39 and P50 probably plays a role in mediating cyclic AMP-dependent platelet inhibition; (c) reactions other than cyclic AMP-dependent protein phosphorylation may participate in platelet inhibition by cyclic AMP.


Histidine decarboxylase, a pyridoxal phosphate-dependent enzyme, is an autoantigen of gastric enterochromaffin-like cells.
Patients with autoimmune polyendocrine syndrome type 1 often have autoantibodies against neurotransmitter synthesizing enzymes, including the pyridoxal phosphate-dependent enzymes glutamic acid decarboxylase and aromatic L-amino acid decarboxylase. Using a candidate approach, we have identified the histamine-synthesizing enzyme histidine decarboxylase, also pyridoxal phosphate dependent, as an autoantigen in this disorder. Anti-histidine decarboxylase antibodies reacting with in vitro translated antigen were found in 36/97 (37%) of autoimmune polyendocrine syndrome type 1 patients studied. The antibodies also reacted with the native enzyme in HMC-1 cell lysates and did not cross-react with the highly homologous aromatic L-amino acid decarboxylase. Anti-histidine decarboxylase antibodies were associated with a history of intestinal dysfunction (P = 0.017). Gastric and duodenal biopsies from a patient with anti-histidine decarboxylase antibodies were studied by immunohistochemistry. The oxyntic mucosa was found to lack the histamine producing enterochromaffin-like cells, suggestive of an autoimmune destruction. To our knowledge, this is the first report of autoantibodies against histidine decarboxylase and absence of gastric enterochromaffin-like cells.


The laser: an alternative for palliative treatment of obstructing intraluminal lesions.
Twenty nine patients with carcinoma obstructing the colon, six obstructing the gastroesophageal junction, six obstructing the tracheobronchial tree, and one patient with recurrent transitional cell cancer of the bladder invading the rectum, not amenable to surgical resection, were treated by laser photocoagulation. Forty were treated with the Nd:YAG laser and two with the CO2 laser. There were 26 men and 16 women with an average age of 73.8 years. The locations of the tumors were 17 rectal, six rectosigmoid, one transverse colon, two descending colon, four ascending colon, six gastroesophageal, and six in the tracheobronchial tree. Symptoms consisted of obstruction and/or bleeding. Seventeen patients were poor surgical candidates, 25 were unresectable due to widespread disease, two had recurrence at the anastomosis, and three patients had another primary site not controlled by their previous surgical procedure. There were seven complications--three perforations, a rectovaginal fistula, two failures in therapy, and lower GI bleeding. Nineteen patients have died of their disease. Tumors were eradicated in two patients. Forty patients had remained free of referring symptoms of obstruction and bleeding.


Judicious Arrangement of Symbols on a Simulated Augmentative and Alternative Communication Display Optimizes Visual Attention by Individuals With Down Syndrome.
Aided augmentative and alternative communication (AAC) displays are often designed as symmetrical row-column grids, with each square in the grid containing a symbol. To maximize vocabulary on displays, symbols are often placed close to one another, and background color cuing is used to signal/differentiate symbols across different grammatical categories. However, from a visual and developmental standpoint, these display features (close-set symbols and use of background color cues) may not be optimal. In particular, placing symbols quite close together may result in visual crowding, in which individual symbols cannot be distinguished due to the presence of many neighbors, or flankers. This research sought to examine the role of display arrangement and background color cuing on the efficiency of visual attention during search. Ten adolescents and adults with Down syndrome underwent a visual search task while a research-based eye tracking system recorded their patterns of visual attention. Participants searched for symbol targets on displays with varying levels of visual crowding and background color cuing. Spatial arrangements that reduced visual crowding and that used the spatial organization to cue the grammatical category of symbols resulted in significantly fewer fixations to nonrelevant distracters during search and reduced the likelihood of fixations away from the target once it was located. Background color was helpful in reducing the latency to find the target. Spatial cues may offer a powerful means to maximize the efficiency of search within AAC displays. Background color cuing may facilitate speed to locate targets in these older individuals. Implications for AAC design, as well as future avenues for maximizing (growing) vocabulary, are discussed.


Comparison of three different A1 adenosine receptor antagonists on infarct size and multiple cycle ischemic preconditioning in anesthetized dogs.
A(1) adenosine receptor (AR) antagonists are effective diuretic agents that may be useful for treating fluid retention disorders including congestive heart failure. However, antagonism of A(1)ARs is potentially a concern when using these agents in patients with ischemic heart disease. To address this concern, the present study was designed to compare the actions of the A(1)AR antagonists CPX (1,3-dipropyl-8-cyclopentylxanthine), BG 9719 (1,3-dipropyl-8-[2-(5,6-epoxynorbornyl)]xanthine), and BG 9928 (1,3-dipropyl-8-[1-(4-propionate)-bicyclo-[2,2,2]octyl]xanthine) on acute myocardial ischemia/reperfusion injury and ischemic preconditioning (IPC) in an in vivo dog model of infarction. Barbital-anesthetized dogs were subjected to 60 min of left anterior descending coronary artery occlusion followed by 3 h of reperfusion, after which infarct size was assessed by staining with triphenyltetrazolium chloride. IPC was elicited by four 5-min occlusion/5-min reperfusion cycles produced 10 min before the 60-min occlusion. Multiple-cycle IPC produced a robust reduction ( approximately 65%) in infarct size; this effect of IPC on infarct size was not abrogated in dogs pretreated with any of the three AR antagonists. Surprisingly, in the absence of IPC, pretreatment with CPX or BG 9928 before occlusion or immediately before reperfusion resulted in significant reductions ( approximately 40-50%) in myocardial infarct size. However, treatment with an equivalent dose of BG 9719 had no similar effect. We conclude that the A(1)AR antagonists BG 9719, BG 9928, and CPX do not exacerbate cardiac injury and do not interfere with IPC induced by multiple ischemia/reperfusion cycles. We discuss the possibility that the cardioprotective actions of CPX and BG 9928 may be related to antagonism of A(2B)ARs.


Cognitive Impairment Correlates Linearly with Mean Flow Velocity by Transcranial Doppler below a Definable Threshold.
Low cerebral blood flow can affect cognition in patients with high-grade asymptomatic internal carotid artery stenosis. Current clinical algorithms use stroke risk to determine which patients should undergo revascularization without considering cognitive decline. Although correlations between low-flow and cognitive impairment have been reported, it is not known whether a threshold exists below which such a correlation expresses itself. Such information would be critical in treatment decisions about whether to intervene in patients with high-grade carotid artery stenosis who are at risk for cognitive decline. To determine how reduced blood flow correlates with lower cognitive scores. Patients with ≥80% unilateral internal carotid artery stenosis with no history of stroke were recruited from inpatient and outpatient practices at a single, large, comprehensive stroke center. Patients underwent bilateral insonation of middle cerebral arteries with standard 2-Hz probes over the temporal windows with transcranial Doppler. Cognitive assessments were performed by an experienced neuropsychologist using a cognitive battery comprising 14 standardized tests with normative samples grouped by age. Z-scores were generated for each test and averaged to obtain a composite Z-score for each patient. Multivariable linear regression examined associations between mean flow velocity (MFV) and composite Z-score, adjusting for age, education, and depression. The Davies test was used to determine if there was a breakpoint for a non-zero difference in slope of a segmented relationship over the range of composite Z-score values. Forty-two patients with unilateral high-grade internal carotid artery stenosis without stroke were enrolled (26 males, age = 74 ± 9 years, education = 16 ± 3 years). Average composite Z-score was -0.31 SD below the age-specific normative mean (range -2.8 to +1.2 SD). In linear regression adjusted for age, education, and depression, MFV correlated with cognitive Z-score (β = 0.308, p = 0.043). A single breakpoint in the range of composite Z-scores was identified at 45 cm/s. For MFV <45 cm/s, Z-score decreased 0.05 SD per cm/s MFV (95% CI: 0.01-0.10). For MFV >45 cm/s, Z-score change was nonsignificant (95% CI: -0.07 to 0.05). In high-grade, asymptomatic carotid artery stenosis, cognitive impairment correlated linearly with lower flow in the hemisphere fed by the occluded internal carotid artery, but only below a threshold of MFV = 45 cm/s. Identifying a hemodynamic threshold for cognitive decline using a simple, noninvasive method may influence revascularization decision-making in otherwise "asymptomatic" carotid disease.


Shorter time since inflammatory bowel disease diagnosis in children is associated with lower mental health in parents.
This study assessed the mental health of parents of children with inflammatory bowel disease (IBD), compared their mental health with age-matched and gender-matched references and examined parental and child predictors for mental health problems. A total of 125 mothers and 106 fathers of 125 children with active and inactive IBD from the Swiss IBD multicentre cohort study were included. Parental mental health was assessed by the Symptom Checklist 27 and child behaviour problems by the Strengths and Difficulties Questionnaire. Child medical data were extracted from hospital records. While the mothers reported lower mental health, the fathers' mental health was similar, or even better, than in age-matched and gender-matched community controls. In both parents, shorter time since the child's diagnosis was associated with poorer mental health. In addition, the presence of their own IBD diagnosis and child behaviour problems predicted maternal mental health problems. Parents of children with IBD may need professional support when their child is diagnosed, to mitigate distress. This, in turn, may help the child to adjust better to IBD. Particular attention should be paid to mothers who have their own IBD diagnosis and whose children display behaviour problems.


Isotopic tracing reveals single-cell assimilation of a macroalgal polysaccharide by a few marine Flavobacteria and Gammaproteobacteria.
Algal polysaccharides constitute a diverse and abundant reservoir of organic matter for marine heterotrophic bacteria, central to the oceanic carbon cycle. We investigated the uptake of alginate, a major brown macroalgal polysaccharide, by microbial communities from kelp-dominated coastal habitats. Congruent with cell growth and rapid substrate utilization, alginate amendments induced a decrease in bacterial diversity and a marked compositional shift towards copiotrophic bacteria. We traced 13C derived from alginate into specific bacterial incorporators and quantified the uptake activity at the single-cell level, using halogen in situ hybridization coupled to nanoscale secondary ion mass spectrometry (HISH-SIMS) and DNA stable isotope probing (DNA-SIP). Cell-specific alginate uptake was observed for Gammaproteobacteria and Flavobacteriales, with carbon assimilation rates ranging from 0.14 to 27.50 fg C µm-3 h-1. DNA-SIP revealed that only a few initially rare Flavobacteriaceae and Alteromonadales taxa incorporated 13C from alginate into their biomass, accounting for most of the carbon assimilation based on bulk isotopic measurements. Functional screening of metagenomic libraries gave insights into the genes of alginolytic Alteromonadales active in situ. These results highlight the high degree of niche specialization in heterotrophic communities and help constraining the quantitative role of polysaccharide-degrading bacteria in coastal ecosystems.


Analysis of vitelline envelope synthesis and composition during early oocyte development in gilthead seabream (Sparus aurata).
The oocyte vitelline envelope (VE) of gilthead seabream is composed of four known zona pellucida (ZP) proteins, ZPBa, ZPBb, ZPC, and ZPX. We have previously shown that the gilthead seabream ZP proteins are differentially transcribed in liver and ovary, with the expression in liver being under estrogenic control. However, although mRNA was found in both liver and ovary, only low ZPBa protein levels were detected in liver and plasma. Using isoform-specific ZP antibodies we show that ZPBa and ZPX translation products are present in the cytosol of stage I and II oocytes. In addition, the zpBa and zpX mRNAs were detected in early developing oocytes. During oocyte growth (vitellogenesis), the VE increased in thickness (>10 microm), and we show that the four ZP isoforms are present in different regions of the VE. ZPX was detected closest to the oocyte plasma membrane while the intermediate region was composed of ZPBa, ZPBb, and ZPC. At the outer layer, only ZPC was detected. When oocytes reach the fully grown stage they resume meiosis and hydration. As the oocyte expands, thinning to 4 microm, the VE acquire a striped and compact appearance at the electron microscopy level. This study provides further evidence for the oocyte origin of some ZP proteins in the gilthead seabream and suggests that the ZP proteins are differentially distributed within the VE.


Comparative analysis of Basiliximab and Alemtuzumab induction therapies in blood type A2-to-B kidney transplantation: Impact on kidney function and de novo donor-specific HLA antibodies.
Blood group B kidney transplant candidates have lower transplantation rates and longer waiting times compared to other blood groups. Kidney transplantation from blood group A2-to-B has offered a solution for these patients. This study aimed to investigate the impact of Basiliximab and Alemtuzumab induction therapies on kidney function and de novo donor-specific antibodies (DSA) in blood type A2-to-B kidney transplant recipients within the first 12 months of post-transplant. A retrospective analysis was conducted on 110 consecutive A2-to-B kidney transplant recipients between January 2015 and December 2022. Of these, 46 (41.8%) received Basiliximab, while 64 (58.2%) received Alemtuzumab as induction therapy. Demographics and comorbidities data were collected and compared between the two groups. Serum samples collected at 4- and 12-month intervals post-transplant were used to assess the presence of de novo DSA. Kidney allograft function was evaluated by monitoring serum creatinine levels and assessing Creatinine Clearance based on 24-h urine collection at various time points. During the follow-up period, 20.00% of patients who received Alemtuzumab developed de novo DSA, whereas none of the patients induced with Basiliximab developed de novo DSA (p = 0.038). Recipients who received Basiliximab were older (mean age = 72.00) and received higher Kidney Donor Profile Index (KDPI) kidneys (mean = 75) compared to those induced with Alemtuzumab (mean age = 58.00, mean KDPI = 49) (p < 0.001), with no significant difference observed in the last follow-up creatinine clearance or creatinine levels between the two groups (p = 0.28). The use of Basiliximab as induction immunosuppression in A2-to-B kidney transplant recipients is associated with a lower incidence of de novo HLA DSA formation without significant differences in overall renal function compared to Alemtuzumab.


Novel bayes factors that capture expert uncertainty in prior density specification in genetic association studies.
Bayes factors (BFs) are becoming increasingly important tools in genetic association studies, partly because they provide a natural framework for including prior information. The Wakefield BF (WBF) approximation is easy to calculate and assumes a normal prior on the log odds ratio (logOR) with a mean of zero. However, the prior variance (W) must be specified. Because of the potentially high sensitivity of the WBF to the choice of W, we propose several new BF approximations with logOR ∼N(0,W), but allow W to take a probability distribution rather than a fixed value. We provide several prior distributions for W which lead to BFs that can be calculated easily in freely available software packages. These priors allow a wide range of densities for W and provide considerable flexibility. We examine some properties of the priors and BFs and show how to determine the most appropriate prior based on elicited quantiles of the prior odds ratio (OR). We show by simulation that our novel BFs have superior true-positive rates at low false-positive rates compared to those from both P-value and WBF analyses across a range of sample sizes and ORs. We give an example of utilizing our BFs to fine-map the CASP8 region using genotype data on approximately 46,000 breast cancer case and 43,000 healthy control samples from the Collaborative Oncological Gene-environment Study (COGS) Consortium, and compare the single-nucleotide polymorphism ranks to those obtained using WBFs and P-values from univariate logistic regression.


Mitigation of nitrofurantoin-induced toxicity in the perfused rat lung.
1. Nitrofurantoin is an antimicrobial agent which produces pulmonary toxicity via the redox cycling of the nitro group and its radical anion. This futile cycling triggers a complex series of events known collectively as oxidative stress. 2. In the isolated perfused rat lung, nitrofurantoin induced a decrease in tissue levels of glutathione but not protein thiols by the end of the 180 min experiment. There was no decline in tissue levels of angiotensin converting enzyme (a marker of cell disruption). However, edema was extensive as monitored in real time by weight gain (2.71 +/- 0.56 g vs 0.63 +/- 0.53 g in control, P < 0.05, n = 4) and lung mechanical functioning. The edema was matched by an increase in lavage proteins (85 +/- 15 mg vs 16 +/- 9 mg in controls, P < 0.05, n = 4). Electron microscopic examination of tissue indicated that the endothelial cells were detached from the basement membrane which would account for the edema. 3. Co-infusion of penicillamine, N-acetylcysteine or N-(2-mercaptopropionyl)-glycine which can protect tissue from oxidative stress failed to mitigate NFT-induced edema. Allopurinol, an inhibitor of xanthine oxidase and a metal chelator, significantly decreased weight gain but did not prevent the loss of glutathione. These results suggested that allopurinol was not blocking metabolic activation of NFT by xanthine oxidase but scavenging metal cations which can initiate and/or propagate the oxidative stress cascade. 4. We concluded that, in the isolated perfused rat lung, the classic pathway of oxidative stress induced by NFT is interrupted at the stage of GSH loss. These experiments demonstrated that organ function was compromised more than the individual cells. They also suggested that allopurinol may prove beneficial in modulating NFT pulmonary toxicity.


Generalization of SPIHT: Set Partition Coding System.
This paper constructs a set partition coding system (SPACS) to combine the advantages of different types of set partition coding algorithms. General tree (GT) is an important conception introduced in this paper, which can represent tree set and square set simultaneously. With the help of GT, SPIHT is generalized to construct degree- k SPIHT based on the analysis of two kinds of set partition operations. Using the same coding mechanism, SPACS (k,p) is constructed, aided with virtual subbands that are generated by recursive division on the LL band. SPACS belongs to tree-set partition coding algorithms if k and p take smaller values. In particular, SPACS(2,1) is the classical SPIHT. SPACS tends toward a block-set partition coding algorithm as k,p increases. Location bit, amplitude bit, and unnecessary bit are presented, which can be used to analyze the coding efficiency of SPACS. We compress 256 images with 512×512 using SPACS. The numerical results show SPACS achieves some improvements in coding efficiency over SPIHT, especially at very low bitrate. On average, to code every image, SPACS(3,1) (at an average of 3.93 bpp) needs 7792 more location bits but saves 10 218 unnecessary bits, compared with SPIHT (3.94 bpp).


A new member of the alkaline phosphatase superfamily with a formylglycine nucleophile: structural and kinetic characterisation of a phosphonate monoester hydrolase/phosphodiesterase from Rhizobium leguminosarum.
The alkaline phosphatase superfamily comprises a large number of hydrolytic metalloenzymes such as phosphatases and sulfatases. We have characterised a new member of this superfamily, a phosphonate monoester hydrolase/phosphodiesterase from Rhizobium leguminosarum (R/PMH) both structurally and kinetically. The 1.42 A crystal structure shows structural homology to arylsulfatases with conservation of the core alpha/beta-fold, the mononuclear active site and most of the active-site residues. Sulfatases use a unique formylglycine nucleophile, formed by posttranslational modification of a cysteine/serine embedded in a signature sequence (C/S)XPXR. We provide mass spectrometric and mutational evidence that R/PMH is the first non-sulfatase enzyme shown to use a formylglycine as the catalytic nucleophile. R/PMH hydrolyses phosphonate monoesters and phosphate diesters with similar efficiency. Burst kinetics suggest that substrate hydrolysis proceeds via a double-displacement mechanism. Kinetic characterisation of active-site mutations establishes the catalytic contributions of individual residues. A mechanism for substrate hydrolysis is proposed on the basis of the kinetic data and structural comparisons with E. coli alkaline phosphatase and Pseudomonas aeruginosa arylsulfatase. R/PMH represents a further example of conservation of the overall structure and mechanism within the alkaline phosphatase superfamily.


An insult-inducible vector system activated by hypoxia and oxidative stress for neuronal gene therapy.
Gene therapy has demonstrated the protective potential of a variety of genes against stroke. However, conventional gene therapy vectors are limited due to the inability to temporally control their expression, which can sometimes lead to deleterious side effects. Thus, an inducible vector that can be temporally controlled and activated by the insult itself would be advantageous. Using hypoxia responsive elements (HRE) and antioxidant responsive elements (ARE), we have constructed an insult-inducible vector activated by hypoxia and reactive oxygen species (ROS). In COS7 cells, the inducible ARE-HRE-luciferase vectors are highly activated by oxygen deprivation, hydrogen peroxide treatment, and the ROS-induced transcription factor NF-E2-related factor 2 (Nrf2). Using a defective herpes virus, the neuroprotective potential of this inducible vector was tested by over-expressing the transcription factor Nrf2. In primary cortical cultures, expression of the inducible ARE-HRE-Nrf2 protects against oxygen glucose deprivation, similar to that afforded by the constitutively expressed Nrf2. This ARE+HRE vector system is advantageous in that it allows the expression of a transgene to be activated not only during hypoxia but also maintained after reperfusion, thus prolonging the transgene expression during an ischemic insult. This insult-inducible vector system will be a valuable gene therapy tool for activating therapeutic/protective genes in cerebrovascular diseases.


Tremor and Klinefelter's Syndrome.
Klinefelter's syndrome (KS) has been associated with tremor, but reports on tremor phenomenology and treatment are limited. Patient 1 is a 17-year-old male with a dystonic tremor treated with deep brain stimulation (DBS). Patient 2 is a 57-year-old male with a predominant left hand resting tremor and dystonic features. Our cases suggest that the tremor in patients with KS may be dystonic in nature. Patient 1 is also the third reported case of successful treatment with DBS. These cases have implications for elucidating the underlying neurobiological mechanism of tremor and identifying treatment options.


Non-rigid surface shape registration to monitor change in back surface topography.
Measured back surface topographies can be useful to monitor the external manifestation of scoliosis to avoid exposure to large doses of radiation. Manual shape fitting of back surface topographies from successive clinical visits can then be used to detect differences. Automated matching of the measured topographies has been seen as a possible improvement on manual comparisons. Recognizing that two changed surface cannot be expected to be simple rigid replicas of each other, the goal of this research has been to develop a new algorithm based on a non-rigid surface matching algorithm, suited to matching the surfaces into same reference frame while also estimating the parameters of the scoliosis deformities, and eliminating noise due to normal body change caused by growth. Back surface topography data from laser optical scanning have been automatically matched by a least squares non-rigid matching algorithm. The algorithm includes new parameters able to model shape changes caused by normal growth and scoliosis deformation. This non-rigid matching algorithm returned r.m.s. values for surface closeness which were improved by at least 10% over rigid matching. Experiments on various scoliosis data demonstrate that the non-rigid matching algorithm is able to accurately match the surfaces while simultaneously extracting parameters representing patient shape change. The non-rigid algorithm has proven to be an improvement on the classical rigid surface matching approach which allows positional fit rather than shape fit. Measured back surface topographies can be closely matched to monitor the external manifestation of scoliosis.


Indirect Impact of PD-1/PD-L1 Blockade on a Murine Model of NK Cell Exhaustion.
The induction of exhaustion on effector immune cells is an important limiting factor for cancer immunotherapy efficacy as these cells undergo a hierarchical loss of proliferation and cytolytic activity due to chronic stimulation. Targeting PD-1 has shown unprecedented clinical benefits for many cancers, which have been attributed to the prevention of immune suppression and exhaustion with enhanced anti-tumor responses. In this study, we sought to evaluate the role of the PD-1/PD-L1 pathway in murine natural killer (NK) cell activation, function, and exhaustion. In an in vivo IL-2-dependent exhaustion mouse model, neutralization of the PD-1/PD-L1 pathway improved NK cell activation after chronic stimulation when compared to control-treated mice. These cells displayed higher proliferative capabilities and enhanced granzyme B production. However, the blockade of these molecules during long-term in vitro IL-2 stimulation did not alter the progression of NK cell exhaustion (NCE), suggesting an indirect involvement of PD-1/PD-L1 on NCE. Given the expansion of CD8 T cells and regulatory T cells (Tregs) observed upon acute and chronic stimulation with IL-2, either of these two populations could influence NK cell homeostasis after PD-L1/PD-1 therapy. Importantly, CD8 T cell activation and functional phenotype were indeed enhanced by PD-1/PD-L1 therapy, particularly with anti-PD-1 treatment that resulted in the highest upregulation of CD25 during chronic stimulation and granted an advantage for IL-2 over NK cells. These results indicate a competition for resources between NK and CD8 T cells that arguably delays the onset of NCE rather than improving its activation during chronic stimulation. Supporting this notion, the depletion of CD8 T cells reversed the benefits of PD-1 therapy on chronically stimulated NK cells. These data suggest a bystander effect of anti-PD1 on NK cells, resulting from the global competition that exists between NK and CD8 T cells for IL-2 as a key regulator of these cells' activation. Thus, achieving an equilibrium between these immune cells might be important to accomplish long-term efficacy during anti-PD-1/IL-2 therapy.