[Complanatoside A and complanatoside B in rats administered with raw and salt-processed Astragali Complanati Semen: a pharmacokinetics study].
An UPLC-MS/MS method was developed to simultaneously determine complanatoside A and complanatoside B in rat plasma with rutin as the internal standard and applied to examine the effect of salt-processing on pharmacokinetics of these two flavonoid glycosides. The pharmacokinetic parameters were estimated using DAS 3.2.6 and subjected to independent sample t-test with SPSS 23.0. No significant difference in T_(max) of complanatoside B was observed between the raw and processed groups; however, in the processed group, C_(max) and AUC_(0-12 h) of complanatoside B increased obviously(P<0.05), while MRT_(0-12 h) decreased from(3.34±0.44) h to(1.81±0.36) h(P<0.05). C_(max) [(14.72±11.13) μg·L~(-1)] and MRT_(0-24) [(3.93±0.26) h] of complanatoside A in the raw group were statistically different from those [(35.64±21.99) μg·L~(-1),(1.43±0.24) h] in the processed group(P<0.05). As a result, salt-processing can facilitate the in vivo adsorption and accelerate the excretion of complanatoside A and complanatoside B.


A biologically active lipid, thromboxane, as a regulator of angiogenesis and lymphangiogenesis.
Thromboxane (TX) and prostaglandins are metabolites of arachidonic acid, a twenty-carbon unsaturated fatty acid, and have a variety of actions that are exerted via specific receptors. Angiogenesis is defined as the formation of new blood vessels from pre-existing vascular beds and is a critical component of pathological conditions, including inflammation and cancer. Lymphatic vessels play crucial roles in the regulation of interstitial fluid, immune surveillance, and the absorption of dietary fat from the intestine; and they are also involved in the pathogenesis of various diseases. Similar to angiogenesis, lymphangiogenesis, the formation of new lymphatic vessels, is a critical component of pathological conditions. The TP-dependent accumulation of platelets in microvessels has been reported to enhance angiogenesis under pathological conditions. Although the roles of some growth factors and cytokines in angiogenesis and lymphangiogenesis have been well characterized, accumulating evidence suggests that TX induces the production of proangiogenic and prolymphangiogenic factors through the activation of adenylate cyclase, and upregulates angiogenesis and lymphangiogenesis under disease conditions. In this review, we discuss the role of TX as a regulator of angiogenesis and lymphangiogenesis, and its emerging importance as a therapeutic target.


[Isolation of imazalil and prochloraz resistant Penicillium variable strains from a commercial citrus warehouse].
"In vitro" sensitivity of two strains of Penicillium variabile to different concentrations of Imazalil and Prochloraz has been studied. Both strains were isolated from a citrus packing-house in which Imazalil was used. These strains were resistant to both compounds showing a ED50 higher than 100 ppm.


Anti-proliferative effects of the novel ceramide analog (S)-2-(benzylideneamino)-3-hydroxy-N-tetrade-cylpropanamide in chemoresistant cancer.
The ceramide-sphingosine-1-phosphate rheostat is a promising therapeutic target. Here, the novel ceramide analog (S)-2-(benzylideneamino)-3-hydroxy-N-tetrade-cylpropanamide is shown to block proliferation and enhance the efficacy of the clinical chemotherapeutics, etoposide and doxorubicin. These results demonstrate the therapeutic potential of this compound in treating both endocrine resistant and chemoresistant breast cancer.


Screening of peripheral arterial disease in primary health care.
The screening tool for diagnosing lower extremity arterial disease is the assessment of the ankle-brachial index (ABI), which is widely used in general practice. However, resting ABI can easily produce a false negative result. In light of this, our goal was to determine the proportion of definitive diagnoses (peripheral arterial disease [PAD] confirmed or refuted) among patients screened in general practice, and the rate of cases in which the need for further specialized examination is necessary, with special attention to groups having non-compressible arteries and ABI negative symptomatic status. The aim of our work is to improve the efficiency of primary health care screening in PAD and reduce the extremely high domestic amputation ratio. Eight hundred and sixteen patients were screened. We used the Edinburgh Questionnaire and recorded medical histories, major risk factors, current complaints, and medication. Physical examinations were performed, including ABI testing. Thirty-three percent complained about lower extremity claudication; 23% had abnormal ABI values; 13% of the patients within the normal ABI range had complaints of dysbasia; and 12% were in the non-compressible artery group. The ABI-negative symptomatic group's risk factor profile showed a close similarity to the clear PAD-positive and non-compressible artery groups. The percentage of PAD could be higher than the number of patients diagnosed by ABI screening. Nearly a quarter of the population fell into the non-compressible artery and ABI-negative symptomatic groups, together defined as the "murky zone". When screening purposely for PAD, these patients deserve special attention due to the insufficient selectivity and sensitivity of measurements. If there is high clinical suspicion of PAD in spite of normal ABI values, further assessment may be considered.


Chlamydia trachomatis diversity viewed as a tissue-specific coevolutionary arms race.
The genomes of pathogens are thought to have evolved under selective pressure provided by the host in a coevolutionary arms race (the 'Red Queen's Hypothesis'). Traditionally, adaptation by pathogens is thought to rely not on whole chromosome dynamics but on gain/loss of specific genes, yielding differential abilities to infect distinct tissues. Thus, it is not known whether distinct host organs differently shape the genome of the same pathogen. We tested this hypothesis using Chlamydia trachomatis as model species, looking at 15 serovars that infect different organs: eyes, genitalia and lymph nodes. We analyzed over 51,000 base pairs from all serovars using various phylogenetic approaches and a non-phylogenetic indel-based algorithm to study the evolution of individual and concatenated loci. This survey comprised about 33% of all single nucleotide polymorphisms in C. trachomatis chromosomes. We present a model in which genome evolution indeed correlates with the cell type (epithelial versus lymph cells) and organ (eyes versus genitalia) that a serovar infects, illustrating an adaptation to physiologically distinct niches, and discarding genetic drift as the dominant evolutionary driving force. We show that radiation of serovars occurred primarily by accumulation of single nucleotide polymorphisms in intergenomic regions, housekeeping genes, and genes encoding hypothetical and cell envelope proteins. Furthermore, serovar evolution also correlates with ecological success, as the two most successful serovars showed a parallel evolution. We identified a single nucleotide polymorphism-based tissue-specific arms race for strains in the same species, reflecting global chromosomal dynamics. Studying such tissue-specific arms race scenarios is crucial for understanding pathogen-host interactions during the course of infectious diseases, in order to dissect pathogen biology and develop preventive and therapeutic strategies.


Chances of late surgery in relation to length of wait lists.
The proportion of patients who undergo surgery within a clinically safe time is an important performance indicator in health systems that use wait lists to manage access to care. However, little is known about chances of on-time surgery according to variations in existing demand. We sought to determine what proportion of patients have had late coronary bypass surgery after registration on wait lists of different size in a network of hospitals with uniform standards for timing of surgery. Using records from a population-based registry, we studied wait-list times prospectively collected in a cohort of patients registered on wait lists for coronary artery bypass grafting procedures. We compared the number of weeks from registration to surgery against target access times established for three urgency groups. The chances of undergoing surgery within target time have been evaluated in relation to wait-list size at registration and the number of surgeries performed without registration on a wait list. In 1991-2001, two in three patients were at risk of late surgery when registered on wait lists for isolated coronary bypass procedures in British Columbia, Canada. Although urgent patients had never seen a wait list with clearance time exceeding one week, the odds of on-time surgery were reduced by 25%, odds ratio [OR] = 0.75 (95% confidence interval [CI] 0.65-0.87) for every additional operation performed without registration on a list. When the wait list at registration required a clearance time of over one month, semi-urgent patients had 51% lower odds of on-time surgery as compared to lists with clearance time less than one week, OR = 0.49 (95%CI 0.41-0.60), after adjustment for age, sex, comorbidity, calendar period, hospital and week on the list. In the non-urgent group, the odds were 69% lower, OR = 0.31 (95%CI 0.20-0.47). Every time an operation in the same hospital was performed without registration on a wait list, the odds of on-time surgery for listed patients were reduced by 7%, OR = 0.93 (95%CI 0.91-0.95) in the semi-urgent group, and by 10%, OR = 0.90 (95%CI 0.87-0.94), in the non-urgent group. Chances of late surgery increase with the wait-list size for semi-urgent and non-urgent patients needing coronary bypass surgery. The weekly number of patients who move immediately from angiography to the operation without registration on a wait list reduced chances of surgery within target time in all urgency groups of listed patients. When advising patients who will be placed on the wait list about the expected time to treatment, hospital managers should take into account the current list size as well as the weekly number of patients who require CABG immediately after undergoing coronary angiography.


Evaluation of branched-chain amino acid intake in children with maple syrup urine disease and methylmalonic aciduria.
The biochemical and growth responses to dietary branched-chain amino acid (BCAA) intake were studied in two children; one with a disorder of branched-chain amino acid metabolism, maple syrup urine disease (MSUD) (McKusick 24860), and another with methylmalonic aciduria (MMA) (McKusick 25100). Biochemical control of MSUD focussed on plasma leucine levels while measurement of plasma ammonia levels was used in MMA. From 0 to 2.75 years both patients exhibited five episodes of toxicity. In each case toxicity was associated with dietary indiscretion or infection. The quantity of protein tolerated was always less in the MMA patient and was approximately 1 g/kg/day. From 1 to 2.75 years each patient's growth velocity approximated their predicted growth channel except during periods of toxicity. In both cases leucine intake, which gave normal growth without toxicity, was always judged to be less than the FAO/WHO recommendations. The BCAA intake of the MMA patient was remarkably stable from 0.5 to 2.75 years and at 2 years of age isoleucine and valine intake approximated the FAO/WHO recommendations. From 2 to 2.75 years BCAA intake (mg/kg/day) of the MSUD patient was less than that of the MMA patient and well below FAO/WHO recommendations. Neuromotor development in both patients is normal.


Polychondritis and rheumatoid arthritis. Case report and review of the literature.
We present the case history of a 50-year-old man with seropositive erosive rheumatoid arthritis of 30-years standing who developed polychondritis simultaneously with several extra-articular rheumatoid manifestations, such as anaemia, subcutaneous nodules, pericarditis and episcleritis. The relevant literature is reviewed. Gradually, all symptoms and signs disappeared after start of treatment with 30 mg prednisone and 100 mg azathioprine daily. We suggest that the polychondritis in this patient was also an extra-articular manifestation of rheumatoid arthritis.


Enzymes and enzymatic mechanisms in enzymatic degradation of lignocellulosic biomass: A mini-review.
Enzymatic hydrolysis is the key step limiting the efficiency of the biorefinery of lignocellulosic biomass. Enzymes involved in enzymatic hydrolysis and their interactions with biomass should be comprehended to form the basis for looking for strategies to improve process efficiency. This article updates the contemporary research on the properties of key enzymes in the lignocellulose biorefinery and their interactions with biomass, adsorption, and hydrolysis. The advanced analytical techniques to track the interactions for exploiting mechanisms are discussed. The challenges and prospects for future research are outlined.


Temperature-mediated acquisition of rare heterologous symbionts promotes survival of coral larvae under ocean warming.
Reef-building corals form nutritional symbioses with endosymbiotic dinoflagellates (Symbiodiniaceae), a relationship that facilitates the ecological success of coral reefs. These symbionts are mostly acquired anew each generation from the environment during early life stages ("horizontal transmission"). Symbiodiniaceae species exhibit trait variation that directly impacts the health and performance of the coral host under ocean warming. Here, we test the capacity for larvae of a horizontally transmitting coral, Acropora tenuis, to establish symbioses with Symbiodiniaceae species in four genera that have varying thermal thresholds (the common symbiont genera, Cladocopium and Durusdinium, and the less common Fugacium and Gerakladium). Over a 2-week period in January 2018, a series of both no-choice and four-way choice experiments were conducted at three temperatures (27, 30, and 31°C). Symbiont acquisition success and cell proliferation were measured in individual larvae. Larvae successfully acquired and maintained symbionts of all four genera in no-choice experiments, and >80% of larvae were infected with at least three genera when offered a four-way choice. Unexpectedly, Gerakladium symbionts increased in dominance over time, and at high temperatures outcompeted Durusdinium, which is regarded as thermally tolerant. Although Fugacium displayed the highest thermal tolerance in culture and reached similar cell densities to the other three symbionts at 31°C, it remained a background symbiont in choice experiments, suggesting host preference for other symbiont species. Larval survivorship at 1 week was highest in larvae associated with Gerakladium and Fugacium symbionts at 27 and 30°C, however at 31°C, mortality was similar for all treatments. We hypothesize that symbionts that are currently rare in corals (e.g., Gerakladium) may become more common and widespread in early life stages under climate warming. Uptake of such symbionts may function as a survival strategy in the wild, and has implications for reef restoration practices that use sexually produced coral stock.


Kinetic Values, Mobility (in)equalities, and Ageing in Smart Urban Environments.
The idea of the right to mobility has been fundamental to modern Western citizenship and is expressed in many legal and government documents. Although there is widespread acceptance regarding the importance of mobility in older adults, there have been few attempts to develop ethical and theoretical tools to portray mobility (in)equalities in old age. This paper develops a novel conceptualisation of kinetic values focusing on older adults whose ability to move has been restricted for internal and external reasons. Informed by the phenomenological theory of kinaesthesia, I suggest that kinetic values are related to four principal dimensions: self-motion, being-moved, co-motion, and forced movement. I assume that these dimensions can address the key dilemma of human dignity among older adults who suffer from losing their autonomy and agency through their mobility impairments and who are at risk of being confined to their homes. To concretise the formulation of kinetic values, I study movement as part of technological equipment and urban infrastructure to examine what kinds of kinetic values mobility services and assistive robots convey for older adults in smart urban environments. By bridging disciplines, the phenomenological approach provides a novel understanding of mobility and the interplay among assistive technologies, kinaesthesia, and urban technological infrastructure. The approach suggests that kinetic values should be interpreted more comprehensively so that kinaesthesia can become better identified as a positive life-promoting practice.


School start times and academic achievement - A systematic review on grades and test scores.
Early school times clash with the late sleep of adolescents, leading to wide-spread sleep restriction in students. Evidence suggests that delaying school starts is beneficial for sleep and recent studies investigated whether this also translates into improved academic achievement. We thus conducted a systematic review of the literature on school start times, grades and test scores in middle and high-school students. We reviewed 21 studies following the PRISMA guidelines and assessed the evidence quality using a pre-defined risk of bias tool. Nine studies reported no association of later starts with achievement, while the remaining reported mixed (5), positive (5), negative (1) or unclear (1) results. Considering the heterogeneity in academic outcomes, study types, amount of delay and exposure, and the substantial risk of bias, a meta-analysis was not warranted - instead we provide grouped reviews and discussion. Overall, no generalisable improvements in achievement with later starts emerge beyond the level of single studies. This does not necessarily preclude improvements in students' learning but highlights shortcomings of the literature and the challenges of using grades and test scores to operationalise academic achievement. Given other previously reported positive outcomes, our results suggest that schools could start later while achievement is likely maintained.


Delayed-interval delivery in a quadruplet pregnancy after intrauterine death of a partial molar pregnancy and preterm delivery. A case report.
Delayed-interval delivery is infrequent in twin gestation and more rare in triplet and quadruplet gestation. Coexistence of a triploid pregnancy with a normal fetus has not previously been reported to have resulted in survival of the normal fetus. A 26-year-old woman, gravida 2, para 0-0-1-0, was diagnosed with a quadruplet pregnancy. At 16 1/2 weeks' gestation she developed preeclampsia and severe hyperemesis. Ultrasound was consistent with partial molar pregnancy in quadruplet D. Quadruplet D died in utero, and the preeclampsia and hyperemesis resolved. At 19 5/7 weeks, spontaneous rupture of the membranes and preterm labor occurred, and quadruplet A, stillborn female weighing 260 g, was delivered. With the use of antibiotic therapy, tocolysis and bed rest, the remaining two fetuses were maintained in utero until 32 6/7 weeks' gestation, when quadruplet B, a 1,470-g female, and quadruplet C, a 1,700-g female, were delivered. This was the first reported case of surviving fetuses coexisting with a partial molar pregnancy. This case was also complicated by preterm delivery and successful delayed-interval birth in a quadruplet pregnancy.


Tumor relevant protein functional interactions identified using bipartite graph analyses.
An increased surge of -omics data for the diseases such as cancer allows for deriving insights into the affiliated protein interactions. We used bipartite network principles to build protein functional associations of the differentially regulated genes in 18 cancer types. This approach allowed us to combine expression data to functional associations in many cancers simultaneously. Further, graph centrality measures suggested the importance of upregulated genes such as BIRC5, UBE2C, BUB1B, KIF20A and PTH1R in cancer. Pathway analysis of the high centrality network nodes suggested the importance of the upregulation of cell cycle and replication associated proteins in cancer. Some of the downregulated high centrality proteins include actins, myosins and ATPase subunits. Among the transcription factors, mini-chromosome maintenance proteins (MCMs) and E2F family proteins appeared prominently in regulating many differentially regulated genes. The projected unipartite networks of the up and downregulated genes were comprised of 37,411 and 41,756 interactions, respectively. The conclusions obtained by collating these interactions revealed pan-cancer as well as subtype specific protein complexes and clusters. Therefore, we demonstrate that incorporating expression data from multiple cancers into bipartite graphs validates existing cancer associated mechanisms as well as directs to novel interactions and pathways.


Tumor size determines the efficacy of percutaneous ethanol injection for the treatment of small hepatocellular carcinoma.
This study was aimed at defining the therapeutic value of percutaneous ethanol injection in patients with solitary hepatocellular carcinoma less than 4 cm. Ultrasound-guided ethanol injection was performed in 24 cirrhotic patients (9 Child A, 10 Child B and 5 Child C), with hepatocellular carcinoma not suitable for surgical treatment. Its efficacy was assessed by repeated ultrasound, computed tomography and tumor biopsy during a follow-up ranging between 4 and 41 mo. Ethanol injection did not achieve a complete tumor necrosis in five cases after a minimum of 12 injections. Seven of the remaining 19 cases, with initial success, have shown recurrence during follow-up, thus resulting in 50% success rate, which was significantly related to baseline tumor size. The six patients with nodules less than 2 cm achieved a complete response, whereas this was recorded in 2 of the 7 with tumor size between 2 and 3 cm, and in only 1 of the 11 cases between 3 and 4 cm. The 1- and 2-yr survival of Child's A and B patients was 87% and 70%, respectively. These results indicate that percutaneous ethanol injection is a useful treatment for hepatocellular carcinoma, especially in tumors less than 3 cm. The high survival rate among patients with nonadvanced liver disease suggests that this therapeutic approach can be considered an alternative approach to surgical resection for tumors smaller than 3 cm.


The NCI-MATCH trial: lessons for precision oncology.
The NCI-MATCH (Molecular Analysis for Therapy Choice) trial ( NCT02465060 ) was launched in 2015 as a genomically driven, signal-seeking precision medicine platform trial-largely for patients with treatment-refractory, malignant solid tumors. Having completed in 2023, it remains one of the largest tumor-agnostic, precision oncology trials undertaken to date. Nearly 6,000 patients underwent screening and molecular testing, with a total of 1,593 patients (inclusive of continued accrual from standard next-generation sequencing) being assigned to one of 38 substudies. Each substudy was a phase 2 trial of a therapy matched to a genomic alteration, with a primary endpoint of objective tumor response by RECIST criteria. In this Perspective, we summarize the outcomes of the initial 27 substudies in NCI-MATCH, which met its signal-seeking objective with 7/27 positive substudies (25.9%). We discuss key aspects of the design and operational conduct of the trial, highlighting important lessons for future precision medicine studies.


Dosimetric verification using monte carlo calculations for tissue heterogeneity-corrected conformal treatment plans following RTOG 0813 dosimetric criteria for lung cancer stereotactic body radiotherapy.
The recently activated Radiation Therapy Oncology Group (RTOG) studies of stereotactic body radiation therapy (SBRT) for non-small-cell lung cancer (NSCLC) require tissue density heterogeneity correction, where the high and intermediate dose compliance criteria were established based on superposition algorithm dose calculations. The study was aimed at comparing superposition algorithm dose calculations with Monte Carlo (MC) dose calculations for SBRT for NSCLC and to evaluate whether compliance criteria need to be adjusted for MC dose calculations. Fifteen RTOG 0236 study sets were used. The planning tumor volumes (PTV) ranged from 10.7 to 117.1 cm(3). SBRT conformal treatment plans were generated using XiO (CMS Inc.) treatment planning software with superposition algorithm to meet the dosimetric high and intermediate compliance criteria recommended by the RTOG 0813 protocol. Plans were recalculated using the MC algorithm of a Monaco (CMS, Inc.) treatment planning system. Tissue density heterogeneity correction was applied in both calculations. Overall, the dosimetric quantities of the MC calculations have larger magnitudes than those of the superposition calculations. On average, R(100%) (ratio of prescription isodose volume to PTV), R(50%) (ratio of 50% prescription isodose volume to PTV), D(2 cm) (maximal dose 2 cm from PTV in any direction as a percentage of prescription dose), and V(20) (percentage of lung receiving dose equal to or larger than 20 Gy) increased by 9%, 12%, 7%, and 18%, respectively. In the superposition plans, 3 cases did not meet criteria for R(50%) or D(2 cm). In the MC-recalculated plans, 8 cases did not meet criteria for R(100%), R(50%), or D(2 cm). After reoptimization with MC calculations, 5 cases did not meet the criteria for R(50%) or D(2 cm). Results indicate that the dosimetric criteria, e.g., the criteria for R(50%) recommended by RTOG 0813 protocol, may need to be adjusted when the MC dose calculation algorithm is used.


[Elective single embryo transfer - the Scandinavian experiment].
Limiting the number of embryos transferred from three to two does not reduce the high risk of twin pregnancy (between 21 and 40%). Scandinavian centers have proposed in the 2000s the elective single embryo transfer (eSET) as the only means to reduce maternal, neonatal and psychological consequences related to multiple births. Pooled results from prospective randomized controlled trials and prospective cohort studies comparing eSET and transfer of two embryos (DET) in a selected population have confirmed the almost complete disappearance of twins when eSET was effective but the compromising effect of eSET upon live birth rates was discussed. Optimizing the eSET overall pregnancy rate need to associate a freezing policy and to define risk factors for increased chance of multiple birth (patient age, diagnosis, number of top-embryos or unsuccessful attempts). The extension of eSET practice to an unselected population irrespective of embryo quality is still controversial. The choice between offering one cycle of SET or DET in an unselected patient population depends on the society's willingness to optimize the in vitro fertilization results according to a defined health care policy: the first one is the twins disappearance with reduced overall pregnancy rate and the second one is a reduced twin birth rate with maintain of the total pregnancy percentage. The real question is to define what percentage of twin pregnancy could be considered as acceptable.


Atypical Teratoid/Rhabdoid Tumor of the Cerebellum in an Adult: Case Report and Literature Review.
Atypical teratoid/rhabdoid tumor (AT/RT) is a rare tumor that is most frequently encountered in the pediatric patient population. AT/RT accounts for approximately 1%-2% of all pediatric central nervous system tumors and roughly 10%-20% of tumors in patients younger than 3 years of age. While AT/RT has been encountered in the adult population, the vast majority of the cases reported occur in the supratentorial space. In the existing literature, only 3 adult cases that arise from the cerebellum have ever been reported. A 38-year-old female presented with 6 months of worsening nausea, emesis, vertigo, diplopia, and coordination difficulty. Magnetic resonance imaging revealed a T1 avidly contrast-enhancing mass, composed of both cystic and solid areas, extending from the cerebellum into the fourth ventricle. Following a gross total resection, surgical pathology was consistent with AT/RT, with tumor cell loss of integrase interactor-1 (INI-1) observed via immunohistochemical staining. This case represents just the fourth ever reported case of AT/RT arising from the cerebellum in an adult and the oldest reported age to date of a cerebellar AT/RT occurring in a female. Due to the paucity of reported adult AT/RT cases, little is known about adults with AT/RT. Further reports will function to improve the general understanding of AT/RT in the adult population.


Factors Which Predict Adverse Outcomes in Anterior Cervical Discectomy and Fusion Procedures in the Nonelderly Adult Population.
Retrospective cohort. The largest published cohort of anterior cervical discectomy and fusion (ACDF) patients was queried to better characterize demographic and operative factors that predict 90-day complication and 2-year reoperation risk. The MarketScan Database was queried from 2007 to 2016 to identify adult patients until 65 years, who underwent an ACDF procedure using International Classification of Diseases 9th Version (ICD-9) and Current Procedural Terminology (CPT) codes. MarketScan is a national insurance claims database that contains millions of patient records across all 50 states. Multivariate logistic regression was used to identify factors associated with complications until 90 days and reoperations until 2 years. Of 138,839 ACDF procedures, 8500 patients (6.1%) experienced a complication within 90 days of the ACDF, and 7433 (5.4%) underwent surgical revision by 2 years. While the use of anterior cervical plating did not predict 2-year reoperation, it was associated with dramatically reduced 90-day complication risk (adjusted odds ratio [aOR]: 0.32; 95% confidence interval [CI]: 0.30-0.34; P <0.001). Upon multivariate analysis, female sex (aOR: 0.83; 95% CI: 0.79-0.87; P <0.001) was associated with decreased risk of 2-year reoperation, while depression predicted a 50% increase in reoperation risk (aOR: 1.51; 95% CI: 1.43-1.59; P <0.001). The single largest factor associated with reoperation risk, however, was the presence of a 90-day postoperative complication (aOR: 1.79; 95% CI: 1.66-1.94; P <0.001). Increased patient comorbidities and the use of bone morphogenic protein were found to increase the risk for postoperative complications, while cervical plating was associated with a strong decline in this risk. In addition, poor patient mental health outweighed the adverse of impact of other comorbidities on 2-year revision risk. The presence of a postoperative complication was the key modifiable risk factor associated with reoperation risk. Conclusions from this study may help surgeons better identify high-risk ACDF patients for more careful patient selection, counseling, informed consent, and management.


Exposure to phenanthrene and depuration: Changes on gene transcription, enzymatic activity and lipid peroxidation in gill of scallops Nodipecten nodosus.
Understanding the mechanism of phenanthrene (PHE) biotransformation and related cellular responses in bivalves can be an important tool to elucidate the risks of polycyclic aromatic hydrocarbons (PAHs) to aquatic organisms. In the present study it was analyzed the transcriptional levels of 13 biotransformation genes related to cytochrome P450 (CYP), glutathione S-transferase (GST), sulfotransferase (SULT), flavin-containing monooxygenase and fatty acid-binding proteins by qPCR in gill of scallops Nodipecten nodosus exposed for 24 or 96h to 50 or 200μgL(-1) PHE (equivalent to 0.28 and 1.12μM, respectively), followed by depuration in clean water for 96h (DEP). Likewise, it was quantified the activity of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione reductase (GR), glucose 6-phosphate dehydrogenase (G6PDH), GST and levels of lipid peroxidation. Increased transcriptional levels of CYP2UI-like, CYP2D20-like, CYP3A11-like, GSTomega-like, SULT1B1-like genes were detected in organisms exposed to PHE for 24 or 96h. In parallel, GR and GPX activities increased after 96h exposure to 200μgL(-1) PHE and G6PDH activity increased after 24h exposure to 50μgL(-1) PHE. This enhancement of antioxidant and phase I and II biotransformation systems may be related to the 2.7 and 12.5 fold increases in PHE bioaccumulation after 96h exposure to 50 and 200μgL(-1) PHE, respectively. Interestingly, DEP caused reestablishment of GPX and GR activity, as well as to the transcript levels of all upregulated biotransformation genes (except for SULT1B1-like). Bioaccumulated PHE levels decreased 2.5-2.9 fold after depuration, although some biochemical and molecular modifications were still present. Lipid peroxidation levels remained lower in animals exposed to 200μgL(-1) PHE for 24h and DEP. These data indicate that N. nodosus is able to induce an antioxidant and biotransformation-related response to PHE exposure, counteracting its toxicity, and DEP can be an effective protocol for bivalve depuration after PHE exposure.


Thiolated chitosan: development and in vivo evaluation of an oral delivery system for leuprolide.
The aim of the present study was to develop an oral delivery system for the peptide drug leuprolide. Gel formulations based on unmodified chitosan/reduced glutathione (GSH) and chitosan-thioglycolic acid (chitosan-TGA)/GSH were prepared, and their effect on the absorption of leuprolide was evaluated in vitro and in vivo in male Sprague Dawley rats. Transport studies were performed with freshly excised rat intestinal mucosa mounted in Ussing-type chambers. Due to the addition of gel formulations comprising 0.5% (m/v) unmodified chitosan/0.5% (m/v) GSH and 0.5% (m/v) chitosan-TGA/0.5% (m/v) GSH, the transport of leuprolide across excised mucosa was improved up to 2.06-fold and 3.79-fold, respectively, in comparison with leuprolide applied in buffer (P(app)=2.87 ± 0.77 × 10⁻⁶ cm/s). In vivo, the addition of oral gel formulation comprising 8 mg of unmodified chitosan, 1mg of GSH and 1mg of leuprolide increased the area under the plasma concentration-time curve (AUC₀₋₈) of leuprolide 1.39-fold in comparison with leuprolide having been administered just in saline. Moreover, the administration of oral gel formulation comprising 8 mg of chitosan-TGA, 1mg of GSH and 1mg of leuprolide resulted in a further enhanced leuprolide plasma concentration, and the area under the plasma concentration-time curve (AUC₀₋₈) of leuprolide was increased 3.72-fold in comparison with the control. With the oral gel formulation comprising 8 mg of chitosan-TGA, a relative bioavailability (versus s.c. injection) of 4.5% was achieved in contrast to the control displaying a relative bioavailability of 1.2%. Thus, according to the achieved results, it is suggested that chitosan-TGA in combination with GSH is a valuable tool for improving the oral bioavailability of the peptide drug leuprolide.


Influence of different therapeutic strategies on exhaled NO and lung inflammation in asthma and COPD.
Nitric oxide (NO), a simple free radical gas, elicits a diverse range of physiological and pathophysiological effects, and plays an important role in pulmonary diseases. Nitrosative stress and nitration of proteins in airway epithelium maybe responsible for steroid resistance in asthma and their ineffectiveness in chronic obstructive pulmonary disease (COPD), supporting the potential role of future therapeutic strategies aimed at regulating NO synthesis in asthma and COPD. Here, we have reviewed the potential role of NO modulators (NO synthase inhibitors and NO donors), which if given on a regular basis may have clinical benefit in asthma and COPD.


Preparation of water soluble chitosan by hydrolysis using hydrogen peroxide.
Chitosan is not soluble in water, which limits its wide application particularly in the medicine and food industry. In the present study, water soluble chitosan (WSC) was prepared by hydrolyzing chitosan using hydrogen peroxide under the catalysis of phosphotungstic acid in homogeneous phase. Factors affecting hydrolysis were investigated and the optimal hydrolysis conditions were determined. The WSC structure was characterized by Fourier transform infrared spectroscopy. The resulting products were composed of chitooligosaccharides of DP 2-9. The WSC content of the product and the yield were 94.7% and 92.3% (w/w), respectively. The results indicate that WSC can be effectively prepared by hydrolysis of chitosan using hydrogen peroxide under the catalysis of phosphotungstic acid.


Premenopausal and postmenopausal women differ in their cardiovascular and neuroendocrine responses to behavioral stressors.
Middle-aged (45-51 years) women performed four tasks while their heart rate, blood pressure, and plasma catecholamines were measured. The tasks were serial subtraction, mirror image tracing, speech, and postural tilt. The speech task was considered to be particularly relevant to women because of its emphasis on social skills. Fifteen premenopausal women reported menstruating regularly and were tested in the early follicular phase. Sixteen postmenopausal women reported not menstruating for at least 12 months and their hormonal status was verified by serum levels of follicle-stimulating hormone. Results showed that postmenopausal women exhibited greater increases from baseline in heart rate during all tasks, relative to premenopausal women, with a particularly pronounced increase during the speech task. Postmenopausal women exhibited greater increases from baseline in systolic blood pressure and epinephrine, relative to premenopausal women, during the speech task only. Explanations for the stressor-specific effect of menopausal status were discussed. The results suggest that reproductive hormones may interact with stressor characteristics to determine middle-aged women's physiological responses to stress.


Pencycuron dissipation in soil: effect of application rate and soil conditions.
Dissipation of the fungicide pencycuron was examined under controlled laboratory conditions in clay loam soils from rice cultivated fields of alluvial soil (Typic udifluvent) and coastal saline soil (Typic endoaquept) at field rate, twice the field rate and 10 times the field rate with and without decomposed cow manure maintained at 60% of maximum water-holding capacity (WHC) and waterlogged soil condition. The half-life values depended on the initial concentrations of pencycuron. Pencycuron, in general, degraded fastest in coastal saline soil and in soil amended with decomposed cow manure at 60% of maximum WHC of soil.


Genetic and quantum chemical basis of the mutagenicity of nitroarenes for adenine-thymine base pairs.
The mutagenicity of nitroarenes for Salmonella typhimurium strains with adenine-thymine base pairs at the mutational site is dependent upon enzymic reduction of the nitro function. Although the electrophilic metabolites of nitroarenes are capable of mutating adenine-thymine base pairs, they show a marked preference for guanine-cytosine pairs when given a choice. Quantum chemical calculations indicate the reactivity order for nucleophilic sites in an AT run of base pairs to be the N-7 of adenine (N7(A)) first, followed by an approximately equal reactivity for C-8 of adenine (C8(A)) and O4 of thymine (O4(T)). Given the low probability of reaction of electrophilic metabolites of nitroarenes with adenine-thymine base pairs, the mutagenic potency of nitroarenes for strains with adenine-thymine base pairs at the mutational site is remarkable.


Effect of heat treatments on stability of altemariol, alternariol monomethyl ether and tenuazonic acid in sunflower flour.
A study was carried out to evaluate the effect of heat treatment on the stability of alternariol (AOH), alternariol monomethyl ether (AME) and tenuazonic acid (TeA) in sunflower flour and the effectiveness of this treatment by a biological assay in rats. The concentrations of AOH and AME remained constant during heating at 100°C for up to 90 minutes while TeA concentration decreased with time to 50% after 90 minutes. The most effective treatment in reducing AOH and AME levels was heating at 121°C for 60 minutes. Histopathological evaluation in the biological assay in rats fed withAlternaria toxins showed marked atrophy and fusion of villi in the intestines and liver cell damage; these lesions were less severe in rats fed heat-treated sunflower flour in line with the reduced toxin content. However, a lower weight gain and a noticeable renal damage in rats were produced when they fed decontaminated flour.


Cu(II) and dopamine bind to α-synuclein and cause large conformational changes.
α-Synuclein (AS) is an intrinsically disordered protein that can misfold and aggregate to form Lewy bodies in dopaminergic neurons, a classic hallmark of Parkinson's disease. The binding of Cu(II) and dopamine to AS was evaluated by nanopore analysis with α-hemolysin. In the absence of Cu(II), wild-type AS (1 μM) readily translocated through the pore with a blockade current of--85 pA, but mostly bumping events were observed in the presence of 25 μM Cu(II). A binding site in the N-terminus was confirmed, because Cu(II) had no effect on the event profile of a peptide consisting of the C-terminal 96-140 residues. In the presence of dopamine (25 μM), the translocation events at--85 pA shifted to--80 pA, which also represents translocation events, because the event time decreases with increasing voltage. Events at--80 pA were also observed for the mutant A30P AS in the presence of dopamine. Event profiles for an N-terminal 1-60-residue peptide and a C-terminal 96-140-residue peptide were both altered in the presence of 25 μM dopamine. In contrast, dopamine had little effect on the CD spectrum of AS, and a single binding site with a Ka of 3.5 × 10(3) m(-1) was estimated by isothermal titration calorimetry. Thus, dopamine can interact with both the N-terminus and the C-terminus. Two-dimensional NMR spectroscopy of AS in the presence of dopamine showed that there were significant changes in the spectra in all regions of the protein. According to these findings, a model is presented in which dopamine induces folding between the N-terminus and C-terminus of AS. Partially folding conformations such as this may represent important intermediates in the misfolding of AS that leads to fibrillization.


Analysis of the mobility of DNA double-strand break-containing chromosome domains in living mammalian cells.
DNA double-strand breaks (DSBs) are among the most dangerous types of DNA damage. Unrepaired, DSBs may lead to cell death, and when misrejoined, they can result in potentially carcinogenic chromosome rearrangements. The induction of DSBs and their repair take place in a chromatin microenvironment. Therefore, understanding and describing the dynamics of DSB-containing chromatin is of crucial importance for understanding interactions among DSBs and their repair. Recent developments have made it possible to study ionizing radiation-induced foci of DSB repair proteins in vivo. In this chapter, we describe techniques that can be applied to visualize and analyze the spatio-temporal dynamics of DSB-containing chromatin domains in mammalian cell nuclei. Analogous procedures may also be applied to the analysis of mobility of other intranuclear structures in living cells.


Evaluation of arterial grafts prior to coronary bypass in pseudoxanthoma elasticum.
Pseudoxanthoma elasticum is a rare, inherited connective tissue disorder associated with coronary and peripheral arterial disease and accelerated atherosclerosis in medium sized arteries. We describe 110-month symptom-free survival in a patient with pseudoxanthoma elasticum who underwent coronary bypass using the left internal mammary artery at 56 years of age. The vessel was evaluated pre-operatively with angiography to determine suitability, and he has not required further investigation or intervention due to lack of symptoms.


Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b.
The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). These ER-retained Kv3.1b proteins were susceptible to degradation, when co-expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression.


Applicability of cloud point extraction coupled with microwave-assisted back-extraction to the determination of organophosphorous pesticides in human urine by gas chromatography with flame photometry detection.
A procedure for the determination of organophosphorous pesticides (OPPs) - phorate, diazinon, parathion-methyl, fenthion and quinalphos - in human urine was developed using the cloud point extraction of nonionic surfactant (Triton X-114) coupled with microwave-assisted back-extraction prior to gas chromatography with flame photometry detection (GC-FPD) analysis. The upper organic solution obtained from back-extraction was centrifugated simply for further cleanup for the sake of automatic injection. A preconcentration factor of 50 was obtained for these five OPPs extracted from only 10 mL of a sample. The limits of detection (LODs) were 0.07 ng mL(-1) for phorate, fenthion and quinalphos, 0.04 ng mL(-1) for diazinon and 0.08 ng mL(-1) for parathion-methyl. The limits of quantification (LOQs) were 0.21, 0.12, 0.24, 0.21 and 0.21 ng mL(-1), respectively. Accuracy of the method was evaluated by bias, which ranged from +6.85 to -14.68%. Precision was also good; the relative standard deviations (R.S.D.s) were less than 9%. The method showed to be potential for biological monitoring.


Correlation among cytokines, bronchopulmonary dysplasia and modality of ventilation in preterm newborns: improvement with melatonin treatment.
Improved survival because of advances in neonatal care has resulted in an increased number of infants at risk for chronic lung disease. Even though the etiology of lung injury is multifactorial, recent animal and clinical data indicate that pulmonary damage depends in large part on the ventilatory strategies used. Ventilator-associated lung injury was believed to result from the use of high pressure, thus, the term barotraumas. This trauma is believed to involve free-radical damage. Oxidant injury is a serious cause of lung injury. In the present study, 110 newborns with respiratory distress syndrome were studied; 55 were treated with melatonin and the other 55 with placebo. All the subjects were mechanically ventilated with or without guaranteed volume. Proinflammatory cytokines [interleukin (IL)-6, IL-8 and tumor necrosis factor (TNF)-alpha] were measured in tracheobronchial aspirate and the clinical outcome was evaluated. Melatonin treatment reduced the proinflammatory cytokines and improved the clinical outcome. The beneficial action of melatonin presumably related to its antioxidative actions.


Managing the multifaceted nature of type 2 diabetes using once-weekly injectable GLP-1 receptor agonist therapy.
As a highly prevalent chronic condition associated with complications and high mortality rates, it is important for pharmacists to have a comprehensive understanding of the impact of type 2 diabetes (T2D) and available treatment options. The use of injectable glucagon-like peptide 1 receptor agonists (GLP-1 RAs) is recommended as an effective and convenient treatment regimen for improving glycaemic control in individuals with T2D, with a good safety profile; however, the wider extent of its potential benefits often are unknown to clinical pharmacists. The objective of this article is to provide an overview of the impact of T2D on individuals and to discuss the multifaceted role of once-weekly (QW) GLP-1 RAs in addressing these challenges. This is a narrative review of the published literature regarding the use of injectable GLP-1 RAs in managing health complications in people with T2D. Recent findings reveal additional benefits of GLP-1 RAs in managing T2D complications, including atherosclerotic cardiovascular (CV) disease, retinopathy, neuropathy, and nephropathy. Dulaglutide and semaglutide have been shown to provide additional CV benefit in patients at high risk of CV events compared with standard of care/placebo and may offer renal protection in patients with chronic kidney disease. Cost-effectiveness studies, taking into consideration these different complications, have shown that QW GLP-1 RAs were cost-effective compared with other therapies. GLP-1 RAs may also help to improve overall health-related quality of life, reducing the risk of depression and 'diabetes distress', and limiting the risk of hypoglycaemia. From the literature, this appears to be the first review of the evidence supporting the multifaceted role of QW GLP-1 RAs in T2D, with particular emphasis on their use in comorbid conditions, as well as associated potential financial and well-being benefits. The results suggest that QW GLP-1 RAs may be an attractive treatment option for improving glycaemic control in T2D, especially in individuals with (or at risk of) additional comorbidities or health complications.


Lumacaftor/ivacaftor therapy fails to increase insulin secretion in F508del/F508del CF patients.
Glucose tolerance abnormalities including cystic fibrosis related diabetes (CFRD) are common in patients with cystic fibrosis (CF). The underlying pathophysiology is not fully understood. Emerging evidence suggests that CFTR dysfunction may directly or indirectly impact β-cell function, offering the potential for improvement with CFTR modulator therapy. In small pilot studies, treatment with ivacaftor improved insulin secretion in patients with the G551D CFTR mutation. In the current study, we examined the impact of lumacaftor/ivacaftor therapy on glucose tolerance and insulin secretion in patients with CF who were homozygous for the F508del mutation. 39 subjects from the PROSPECT Part B study who had been prescribed lumacaftor/ivacaftor by their CF care team at a CF Foundation's Therapeutic Development Network center were recruited. Subjects underwent 2-hour oral glucose tolerance tests (OGTTs) at baseline prior to first dose of lumacaftor/ivacaftor, and at 3, 6 and 12 months on therapy. OGTT glucose, insulin and c-peptide parameters were compared. Compared to baseline, OGTT fasting and 2 hour glucose levels, glucose area under the curve, insulin area under the curve and time to peak insulin level were not significantly different at 3, 6 and 12 months on lumacaftor/ivacaftor therapy. Similarly, C-peptide levels were no different. Lumacaftor/ivacaftor therapy did not improve insulin secretion or glucose tolerance in patients with CF who were homozygous for the F508del mutation.


Concomitant adenocarcinoma and colonic non-Hodgkin's lymphoma in a patient with ulcerative colitis: a case report and molecular analysis.
Ulcerative colitis (UC) complicated by colonic lymphoma is rare, although UC is often accompanied by adenocarcinoma of the colon. A concurrent existence of adenocarcinoma and lymphoma in a patient with UC is extremely rare, and has not yet been analyzed at the molecular level. We report a 64-year-old female patient with concomitant adenocarcinoma and diffuse large B-cell lymphoma (DLBCL) in the colon of UC. The genetic changes in these two neoplasms were analyzed. The colon adenocarcinomas had a mutation in MSH6 gene, DNA methylation in CDKN2A gene, and increased microsatellite instability (MSI), although these genetic changes were not recognized in either DLBCL or non-neoplastic UC mucosa. The DLBCL was diagnosed as primary colonic lymphoma, and confirmed Epstein-Barr virus (EBV) infection. The adenocarcinomas and the non-neoplastic UC mucosa were EBV-negative. Our case presented here clearly shows that the development of adenocarcinoma and lymphoma in the colon with UC was caused by individual mechanisms.


Solvent-temperature perturbations of ionizable groups as a tool for the investigation of the active site of enzymes.
Co-solvent and temperature effects on the pK of histidine (imidazolium) residue 46 of trypsin, as well as of weak electrolytes (buffers), which have been reported in two preceding papers, can be satisfactorily explained in terms of enthalpy-entropy compensation patterns. Such patterns have been generated for various mixed solvents between 20 degrees and minus 20 degrees and minus 50 degrees. Under these conditions compensation temperature, T-c, is strongly dependent on the nature of the ionizable group studied: 240 plus or minus 10 K for neutral acids and 310 plus or minus 5 K for cationic acids. This work focuses on the possibilities offered and on the problems raised by the use of this methodology as a tool in the investigation of the active site of enzymes. Furthermore, it is shown in the case of histidine residue 46 of trypsin that the co-solvent effect vanishes at the compensation temperature, a result of great practical significance if applicable to any ionizable group at the active site of enzymes.


An introduction to therapeutic approaches to vascular cognitive impairment.
Vascular cognitive impairment (VCI), encompassing vascular dementia, has been claimed as the "second-most common dementia" after Alzheimer Disease. Whether or not this is true, the clinical picture of most dementia in older people includes vascular disease. There are no validated pharmacological targets for prevention or treatment of VCI. This has inspired a multitude of potential treatment approaches, reflected by the articles in this Special Issue. These include in vitro testing of the novel oral anticoagulant dabigatran for protection against β-amyloid neurotoxicity, and an overview of neuroinflammation in VCI and the role of circulating markers (PIGF, VEGF-D) identified by the MarkVCID study. There are reviews of potential therapeutics, including adrenomedullin and nootropic preparations (exemplified by cerebrolysin). The role of sleep is reviewed, with possible therapeutic targets (5HT2A receptors). There is a clinical study protocol (INVESTIGATE-SVD) and a feasibility analysis for a secondary prevention trial in small vessel disease. Clinical data include secondary analyses of blood pressure and cerebral blood flow from a longitudinal clinical trial (NILVAD), differences between methylphenidate and galantamine responders and non-responders (STREAM-VCI), appraisal of treatment approaches in India, and primary outcomes from a randomised trial of Argentine tango dancing to preserve cognition in African American women (ACT). Treating vascular disease has great potential to improve global cognitive health, with public health impacts alongside individual benefit. Vascular disease burden varies across populations, offering the possibility of proactively addressing health inequity in dementia using vascular interventions. The next 5-10 years will witness cost-effective lifestyle interventions, repurposed drugs and novel therapeutics.


Plasma kallikrein-bradykinin pathway promotes circulatory nitric oxide metabolite availability during hypoxia.
Nitric oxide (NO) is an indispensible signalling molecule under hypoxic environment for both ethnic high altitude natives as well as lowland residents at high altitude. Several studies have reported higher levels of NO and bioactive NO products for both high altitude natives as well as healthy high altitude sojourners. But the metabolic pathways regulating the formation of NO and associated metabolites during hypoxia still remain elusive. In the present study, we profiled plasma proteomes of Ladakhi natives (3520 m) and lowland residents (post 1, 4 and 7 days stay) at the same altitude. This has resulted in the identification of 208 hypoxia responsive proteins (p < 0.05) and kininogen-plasma kallikrein-bradykinin as a major pathway regulating eNOS activity during hypoxia. In corroboration, we have also observed significant higher levels of plasma biomarkers for NO production (l-citrulline, nitrite, nitrate) for Ladakhi natives as compared to both lowland individuals healthy high altitude sojourners indicating higher NO availability. Since hypoxia-induced free radicals reduce NO availability, we also measured plasma levels of 8-isoprostanes, protein carbonyls and protein oxidation products in both Ladakhi natives and high altitude sojourners. Interestingly Ladakhi natives had significant lower levels of oxidative stress in comparison to high altitude sojourners but higher than lowland controls. These results suggest that plasma kallikrein-bradykinin-eNOS pathway along with moderate oxidative stress contributes to high altitude adaptation of Ladakhi natives.


Gamma delta T cells provide an early source of interferon gamma in tumor immunity.
Interferon (IFN)-gamma is necessary for tumor immunity, however, its initial cellular source is unknown. Because gammadelta T cells primarily produce this cytokine upon activation, we hypothesized that they would provide an important early source of IFN-gamma in tumor immunosurveillance. To address this hypothesis, we first demonstrated that gammadelta T cell-deficient mice had a significantly higher incidence of tumor development after challenge with a chemical carcinogen methylcholanthrene (MCA) or inoculation with the melanoma cell line B16. In wild-type mice, gammadelta T cells were recruited to the site of tumor as early as day 3 after inoculation, followed by alphabeta T cells at day 5. We then used bone marrow chimeras and fetal liver reconstitutions to create mice with an intact gammadelta T cell repertoire but one that was specifically deficient in the capacity to produce IFN-gamma. Such mice had a higher incidence of tumor development, induced either with MCA or by inoculation of B16 melanoma cells, compared with mice with IFN-gamma-competent gammadelta T cells. Moreover, genetic deficiency of gammadelta T cells resulted in impaired IFN-gamma production by tumor antigen-triggered alphabeta T cell upon immunization with tumor lysate. These results demonstrate that gammadelta T cells can play a necessary role in tumor immunity through provision of an early source of IFN-gamma that in turn may regulate the function of tumor-triggered alphabeta T cells.


Increased dimeric IgA-producing B cells in tonsils in IgA nephropathy determined by in situ hybridization for J chain mRNA.
The origin of mesangial IgA deposits in IgA nephropathy (IgAN) remains obscure. A significant proportion of deposited immunoglobulin is dimeric (J chain-positive). Previous studies of J chain expression within lymphoid tissue in IgAN have utilized antibodies which other investigators have found to be non-specific. To address this problem, we have developed and in situ hybridization (ISH) method for the detection of J chain mRNA within IgA plasma cells. Tonsils from 12 patients with IgAN and 12 controls were studied using (i) non-isotopic ISH for J chain mRNA, and (ii) combined immunofluorescence (IF) and fluorescent ISH. J chain mRNA-positive cells were identified in germinal centres, and within the subepithelial and interfollicular zones. A greater number of J chain mRNA-positive cells were found in the germinal centres of patients (mean 57.7 +/- 4.6 cells/10(5) micron2) compared with controls (mean 36.9 +/- 3.5 cells/10(5) micron2) (P < 0.001). Combined IF and fluorescent ISH showed a greater proportion of J chain mRNA-positive interfollicular IgA cells in patient tonsils (32 +/- 3.4%) compared with controls (21 +/- 2.3%; P < 0.02). These results indicate a shift towards dimeric IgA production in the tonsils in IgAN. In addition, the finding of excess numbers of J chain-positive Iga-negative cells within germinal centres suggests that an abnormality may be present at the B cell differentiation stage before IgA switching. These results further highlight immune abnormalities within the tonsil as a central feature of abnormal polymeric IgA biology in this common form of glomerulonephritis.


Group II intron mobility occurs by target DNA-primed reverse transcription.
Mobile group II introns encode reverse transcriptases and insert site specifically into intronless alleles (homing). Here, in vitro experiments show that homing of the yeast mtDNA group II intron aI2 occurs by reverse transcription at a double-strand break in the recipient DNA. A site-specific endonuclease cleaves the antisense strand of recipient DNA at position +10 of exon 3 and the sense strand at the intron insertion site. Reverse transcription of aI2-containing pre-mRNA is primed by the antisense strand cleaved in exon 3 and results in cotransfer of the intron and flanking exon sequences. Remarkably, the DNA endonuclease that initiates homing requires both the aI2 reverse transcriptase protein and aI2 RNA. Parallels in their reverse transcription mechanisms raise the possibility that mobile group II introns were ancestors of nuclear non-long terminal repeat retrotransposons and telomerases.


African swine fever virus MGF360-9L promotes viral replication by degrading the host protein HAX1.
African swine fever virus (ASFV) infection causes African swine fever (ASF), a virulent infectious disease that threatens the safety of livestock worldwide. Studies have shown that MGF360-9 L is important for the virulence of ASFV and the host protein HS1-associated protein X-1 (HAX1) plays an important role in viral pathogenesis. This study aimed to clarify the mechanism by which HAX1 mediates ASFV replication through interactions with MGF360-9 L. The regions of interaction between MGF360-9 L and HAX1 were predicted and validated. HAX1 overexpression and RNA interference studies revealed that HAX1 is a host restriction factor that suppresses ASFV replication. Moreover, HAX1 expression was inhibited in ASFV-infected mature bone marrow-derived macrophages, and infection with the virulent MGF360-9 L gene deletion strain (∆MGF360-9 L) attenuated the inhibitory effect of the wild-type strain (WT) on HAX1 expression, suggesting a complex regulatory relationship between MGF360-9 L and HAX1. Furthermore, the E3 ubiquitin ligase RNF114 interacted with MGF360-9 L and HAX1, MGF360-9 L degraded HAX1 through the ubiquitin-proteasome pathway, and RNF114 facilitated the degradation of HAX1 by MGF360-9L-linked K48 ubiquitin chains through the ubiquitin-proteasome pathway, thereby facilitating ASFV replication. In conclusion, this study has enriched our understanding of the regulatory networks between ASFV proteins and host proteins and provided a reference for investigation into the pathogenesis and immune escape mechanism of ASFV.


The arthroscopic square knot: a biomechanical comparison with open and arthroscopic knots.
To compare the loop and knot security of arthroscopic square knots with other arthroscopic and open knots with the use of 2 commonly used suture types. Five knot configurations were tested 12 times each. The arthroscopic square knot was compared with the open square knot, arthroscopic and open half hitches with alternating posts, and the Duncan loop. Load-to-failure testing was performed at a rate of 1.25 mm/sec, and cyclic testing was performed between 7 N and 30 N at a rate of 1 N/sec for 50 cycles. Two suture types were tested: No. 2 Ethibond suture and No. 2 FiberWire. Among the No. 2 Ethibond sutures, the arthroscopic Duncan loop had the highest load-to-failure at 165 N. The remaining knot types failed at between 142 N and 148 N. The load at 3 mm of lengthening was between 100 N and 120 N for all knots. The No. 2 FiberWire failed at higher loads than the No. 2 Ethibond suture for all knot types except the Duncan loop. The arthroscopic half hitches, arthroscopic square knots, and open half hitches all failed at between 220 N and 264 N. The open square knot failed at 188 N, and the Duncan loop failed at 147 N. The load at 3 mm of lengthening was between 130 N and 165 N for all knots except the Duncan loop, which failed at 95 N. With cyclic testing, arthroscopic square knots performed better than all knot types, but the differences were clinically insignificant. Arthroscopic square knots have the same or greater strength when compared with other arthroscopic or open knots tied with the same suture type, and they perform as well or better in the face of cyclic loads. Good knot security can be attained with all of the knots tested, regardless of suture type. Equivalent knots tied with No. 2 FiberWire fail at higher loads, except for those tied with the Duncan loop. Arthroscopic square knots can be used in the clinical setting with no compromise in function when compared with open square knots.


Predictors of postnatal depression in the slums Nairobi, Kenya: a cross-sectional study.
Postnatal depression (PND) is a universal mental health problem that prevents mothers' optimal existence and mothering. Although research has shown high PND prevalence rates in Africa, including Kenya, little research has been conducted to determine the contributing factors, especially in low-resource communities. This study aimed to investigate the PND risk factors among mothers attending Lang'ata and Riruta Maternal and Child Health Clinics (MCH) in the slums, Nairobi. This study was cross-sectional. It is part of a large study that investigated the effectiveness of a brief psychoeducational intervention on PND. Postnatal mothers (567) of 6-10 weeks postanatal formed the study population. Depression rate was measured using the original 1961 Beck's Depression Inventory (BDI). In addition, a sociodemographic questionnaire (SDQ) was used to collect hypothesized risk variables. Multivariable logistic regression analysis was used to explore predictors of PND. The overall prevalence of PND in the sample of women was 27.1%. Women aged 18-24 (β = 2.04 95% C.I.[0.02; 4.05], p = 0.047), dissatisfied with body image (β = 4.33 95% C.I.[2.26; 6.41], p < 0.001), had an unplanned pregnancy (β = 2.31 95% C.I.[0.81; 3.80], p = 0.003 and felt fatigued (β = - 1.85 95% C.I.[- 3.50; 0.20], p = 0.028) had higher odds of developing PND. Participants who had no stressful life events had significantly lower depression scores as compared to those who had stressful life events (β = - 1.71 95% C.I.[- 3.30; - 0.11], p = 0.036) when depression was treated as a continuous outcome. Sensitivity analysis showed that mothers who had secondary and tertiary level of education had 51 and 73% had lower likelihood of having depression as compared to those with a primary level of education (A.O.R = 0.49 95% C.I.[0.31-0.78], p = 0.002) and (A.O.R = 0.27 95% C.I.[0.09-0.75], p = 0.013) respectively. This study reveals key predictors/risk factors for PND in low-income settings building upon the scanty data. Identifying risk factors for PND may help in devising focused preventive and treatment strategies.


[Biological active substances of plant origin. Flavanones: dietary sources, biovailability, the influence on xenobiotic metabolizing enzymes].
This review summarizes data concerning distribution, as well as the main food sources and daily consumption of flavanones, a relatively small group of flavonoids exhibiting wide range of biological activities, which are mainly specific for citrus fruits. The questions on their bioavailability and metabolism are discussed. The results of the in vitro and in vivo studies on the effects of flavanones on the activity of I and II phase drug metabolizing enzymes are shown.


Effect of salt stress on the production and properties of extracellular polysaccharides produced by Cryptococcus laurentii.
The composition, main structural features and molecular properties of exopolysaccharides (EP) produced by Cryptococcus laurentii var. laurentii CCY 17-3-16 under optimal (EPo) and NaCI-stress conditions (EPs) as well as their subfractions isolated by gel chromatography were studied using chemical, FT-IR and NMR spectroscopy methods. The results showed that under stress conditions the yeast produced EP with a lower content of protein and phosphorus. In comparison to EPo, the EPs exhibited a substantially larger proportion of high molecular mass populations. NMR analysis of EPs revealed a higher degree of branching with single xylose side chains of the heteromannan components. The increase of the molecular mass and degree of branching of the macromolecular chains of the heteromannan components might in part be related to the function of EPs to protect the yeast cells from water loss and maintain growth conditions under the salt stress.


Modulation of airway responses to influenza A/PR/8/34 by Delta9-tetrahydrocannabinol in C57BL/6 mice.
Delta(9)-tetrahydrocannabinol (Delta(9)-THC) has been widely established as a modulator of host immune responses. Accordingly, the objective of the present study was to examine the effects of Delta(9)-THC on the immune response within the lungs and associated changes in the morphology of the bronchiolar epithelium after one challenge with a nonlethal dose of the influenza virus A/PR/8 (PR8). C57BL/6 mice were treated by oral gavage with Delta(9)-THC and/or vehicle (corn oil) for 5 consecutive days. On day 3, mice were instilled intranasally with 50 plaque-forming units of PR8 and/or vehicle (saline) 4 h before Delta(9)-THC exposure. Mice were subsequently killed 7 and 10 days postinfection (dpi). Viral hemagglutinin 1 (H1) mRNA levels in the lungs were increased in a dose-dependent manner with Delta(9)-THC treatment. Enumeration of inflammatory cell types in bronchoalveolar lavage fluid showed an attenuation of macrophages and CD4(+) and CD8(+) T cells in Delta(9)-THC-treated mice compared with controls. Likewise, the magnitude of inflammation and virus-induced mucous cell metaplasia, as assessed by histopathology, was reduced in Delta(9)-THC-treated mice by 10 dpi. Collectively, these results suggest that Delta(9)-THC treatment increased viral load, as assessed by H1 mRNA levels, through a decrease in recruitment of macrophages and lymphocytes, particularly CD4(+) and CD8(+) T cells, to the lung.


Fractography of human intact long bone by bending.
Human intact tibiae were tested using the static bending method to learn about the relationship between the fracture surface and the failure mode. The bending test was applied to test pieces and to whole bones. The fracture surface was observed by scanning electron microscopy. The bone fracture is closely related to the architecture of the bone substance, especially to the direction of the Haversian canals and the lamellae. The failure mode and the sequence of the break line of the bone can be found out by the observation on the fracture surface. Hardly any crushing effects caused by the compressive force is seen. The mechanical properties of the fractured bone can be estimated to some extend by considering the direction of the break line and the failure mode. The strength calculated by the simple beam formula for elastic materials can not be obtained directly because of the plastic deformation of the bone. The results of the tensile test may be applied to the fracture using the static bending moment.


Data and methods to assess occupational exposure to personnel involved in cardiac catheterization procedures.
To provide normalized scatter exposure data and methods for reliable estimation of cumulative effective dose and eye-lens equivalent dose to personnel involved in fluoroscopically guided cardiac catheterization (FGCC) procedures. An anthropomorphic phantom was placed supine on the table of a modern digital C-arm angiographic system and 17 different fluoroscopic projections commonly employed during FGCC procedures were represented. Scatter exposure rates at the waist and eye level were measured for varying exposure parameters and position in the operating room. The effect of beam field size, patient size, use of radioprotective garments and small variations in projection angulation and table height on scatter radiation was investigated. Apart from the position and use of radio-protective garments, radiation burden to operators during fluoroscopic guidance was found to remarkably depend beam field size (>45% reduction if a 10 × 10 cm(2) instead of 15 × 15 cm(2) fluoroscopy beam is used) and patient size (>25% increased scatter for obese patients). In contrast, the variation of measured scatter exposure from a given projection was found to be <10% when the source to skin distance was altered by ±10 cm or beam angulation of a specific projection was altered by ±10°. Presented scatter exposure data charts and methods allow for prospective and retrospective estimation of effective dose and eye-lens equivalent dose to personnel involved in any FGCC procedure. Projection specific maps of scatter exposure produced may enhance familiarization of involved medical staff to good radiation protection practice and optimization of working habits in the cardiac catheterization lab.


Spectrophotometric studies and application of imipramine-eriochrome cyanine R system for determination of imipramine in pharmaceuticals.
Eriochrome cyanine R (ECR) has been tested as reagent for the determination of imipramine. It reacts in neutral medium with imipramine forming reddish compound, which can be quantitatively extracted into n-butanol. This property has been successfully used for the extractive-spectrophotometric determination of imipramine. Beer's law is obeyed in concentration range of 10-80 microg ml(-1) of imipramine. The method was applied to the determination of imipramine in its pharmaceutical.


Cognitive impairments and mental health of patients with post-COVID-19: A cross-sectional study.
Persistent cognitive complaints belong to the most frequent symptoms after COVID-19. This study explored the neuropsychological profile, mental health and risk factors for cognitive impairment in post-COVID-19 patients. The patients were recruited consecutively in the Post COVID Center of the University Hospital of Erlangen between 12/2022 and 05/2023. They underwent an extensive neuropsychological assessment including the Verbal Learning Memory Test (VLMT), the digit span backwards from the Wechsler Memory Scale-Revised (WMS-R), the Trail Making Test (TMT) Part A and B, the d2 Test of Attention and the Regensburger Verbal Fluency Test (RWT). For each cognitive domain we calculated the frequency of age-adjusted scores below the measure-specific norms. Depressive symptoms were measured with the Patient-Health-Questionnaire-9 (PHQ-9). Logistic regression analyses were computed. In 110 patients (mean age: 42.5 ± 11.9 years; 68.2% women), the most frequent cognitive deficits were observed for verbal fluency, working speed, delayed recall and attention. In almost every cognitive domain high education levels were associated with a decreased risk for cognitive impairment. Higher age was a risk factor for working speed and delayed recall and a protective factor for verbal fluency. Clinically relevant depressive symptoms were associated with an elevated risk for an impairment regarding some cognitive functions. Cognitive dysfunctions were common among the post-COVID-19 patients. Differentiated exploration of cognitive impairments is crucial for a proper characterization of the post-COVID syndrome. In future research parameters of cognitive impairment should be correlated to alterations in biological markers of the disease like markers of immunological and microcirculation change.


Topography of somatosensory evoked potentials after stimulation of the median nerve.
We studied topography of major negative-positive peaks, NI, PI, NII, PII and NIII, of scalp recorded somatosensory evoked potentials (SEP) after stimulation of the median nerve. Unlike the diffusely distributed P14, NI, PI and NII recorded from contralateral hemisphere after unilateral stimulation normally showed statistically significant latency increase from frontal (N17, P20, N29) to central (N19, P23, N32) and parietal (N20, P26, N34) electrodes. However, NIII (N60) had considerable inter- and intra-individual variations with no consistent antero-posterior latency shift. In contrast to well localized N19 and N32 peaks at the contralateral central electrode, the N17, P20 and N29 peaks were registered over the bifrontal and ipsilateral central regions as well as in the vertex. The parietal N20 peak was also present at the occipital electrodes bilaterally. In patients with localized cerebral lesions, types of SEP abnormalities varied considerably, presumably reflecting complex somatosensory afferent pathways. A small lesion in posterolateral thalamus may totally eliminate NI, NII and NIII components over both hemispheres, sparing only P14 whereas a sizable lesion in the frontal or parietal lobe may affect only NII or NIII. Capsular lesions spare P14 and frontal N17 but may alter all the subsequent SEP components, or NII or NIII selectively. In some cases, the corresponding peaks at the central and parietal electrodes may be affected independently. The complex relationships between the type of SEP abnormalities and the location of cerebral lesions can best be explained by postulating the presence of multiple, at least partially independent, thalamocortical projections mediating regionally specific somatosensory inputs.


Evaluation of a research awareness training programme to support research involvement of older people with dementia and their care partners.
Best-practice guidelines recommend that appropriate support be provided to public contributors to facilitate their involvement in research. One form of support is research awareness training. Older people with dementia and care partners were involved in four Research User Groups (RUGs) in the UK, France, Cyprus and Greece. We delivered research awareness training (RAT) to the RUGs. The aim of this study was to evaluate the acceptability and perceived outcomes of the training from the perspective of RUG members. At the end of each research training session, participants completed the Training Acceptability Rating Scale-section 2, which records the respondent's impressions of the training process and the outcomes of training. Participants were also invited to take part in semi-structured interviews at the end of the programme. Thirty-four RUG members completed the TARS-section 2 with 23 completing semi-structured interviews. Over two-thirds (67%) of participants rated their overall satisfaction with the RAT 'a great deal'. Qualitative responses indicated that participants found group work to be beneficial for learning, the structure of training activities and topics covered appropriate. The type and format of the training materials were viewed as helpful, and they valued the new knowledge gained. The training contents were applicable, useful and relevant to the participants' role within the research. We highlight the importance of facilitating participation by (a) fostering awareness of relevant research issues and (b) tailoring delivery of training according to the needs of the participants.


Evaluation of early precut with needle-knife in difficult biliary cannulation during ERCP.
There is scarce information on whether performing the precut procedure early rather than after several cannulation attempts is associated with different success and complication rates. The aim of this retrospective study was is to compare the early precut technique with the standard one in terms of the results and complications. The contemporary success rate and postoperative complications in 792 endoscopic retrograde cholangiopancreatography cases were frequently observed during the period from June 2007 to May 2011, and 56 of these cases were carried out with precut biliary sphincterotomy after the standard sphincterotomy had failed. The success rate for standard sphincterotomy was 89.8%: 51 out of 56 cases were carried out with precut biliary sphincterotomy and succeeded. The total success rate was 96.3%. The difference was significant (χ2=25.62, p<0.01) compared to the success rate of first cannulation, while the difference in complication rates between precut and standard sphincterotomy was minor (9.9 vs. 12.5%, p>0.05). Early precut with a needle-knife in a difficult biliary cannulation was safe and effective if performed by experienced endoscopists.


Be Careful When Using Peer-Influence on Nudging Solicitation: Evidence of Potential Negative Effect from a Sample of Chinese University Students.
Peer information is now commonly used in solicitation. However, scholars have long focused on testing its effectiveness on increasing the donation amount without paying attention to its potential negative effects on donors. Thus, the current study employs high vs low peer donation amount (HPDA vs LPDA) information to explore its effect on "how-much-to-donate" decisions and the corresponding neural and psychological reactions at the same time. Student samples from a Chinese university and behavioral experiments with the event-related potential (ERP) method were used in this study. The behavioral results are consistent with previous research in which HPDA was positively associated with higher donation levels. ERP results show the mechanisms behind decision-making can be summarized into a cognitive approach represented by cost-benefit analysis and an affective approach represented by reward perception. More surprisingly, in contrast to the behavioral results, LPDA elicits higher level of reward perception than HPDA. The results indicate that although HPDA leads to higher levels of donation, donors do not show higher levels of reward anticipation at the neurological level, indicating the increment of donation may come at the cost of donors. Theoretical and practical implications are also discussed.


Chiari 1 Formation Redefined-Clinical and Radiographic Observations in 388 Surgically Treated Patients.
The subject of Chiari formation is revisited and redefined. Results of surgical treatment of patients with Chiari formation by atlantoaxial fixation are presented. Results were analyzed of 388 patients with Chiari formation surgically treated during 2010 to June 2019. Two hundred and sixty-six patients had syringomyelia. Two hundred and three patients had no craniovertebral bone abnormality and 74 patients had group A and 111 patients had group B basilar invagination. Twenty-nine patients had been earlier treated by foramen magnum decompression surgery with or without duroplasty. Clinical parameters, analysis of video recordings both before and after surgery, and patient self-assessment were included in the analysis of outcome. Immediate postoperative and sustained clinical improvement was observed in 385 patients (99.4%). Satisfactory clinical outcome in most patients after atlantoaxial fixation and without any manipulation of neural structures, dura, or bone in the region of foramen magnum consolidates the viewpoint that atlantoaxial instability is the nodal point of pathogenesis of Chiari 1 formation. The study suggests that Chiari 1 formation may be a secondary natural neural alteration in the face of atlantoaxial instability. The role of foramen magnum decompression surgery needs to be reassessed.


Isolation of nucleolar DNA-binding proteins by simultaneous, competitive DNA--sephadex affinity chromatography.
A new method is described for the isolation of sequence-specific DNA-binding proteins, based on simultaneous, competitive binding to homologous or heterologous DNA. Since both types of DNA are bound to a solid matrix (Sephadex beads), the method may be used for the preparative isolation of proteins which bind preferentially to specific DNA sequences. Fractionation of nucleolar proteins from Novikoff hepatoma ascites cells by this technique yields two protein fractions which show much greater apparent specificity of binding to homologus DNA, in a nitrocellulose filter-binding assay, than unfractionated nucleolar proteins.


Internal decompression for spinal stenosis (IDSS) for decompression and use of interlaminar dynamic device (CoflexTM) for stabilization in the surgical management of degenerative lumbar canal stenosis with or without mild segmental instability: our initial results.
Internal decompression of spinal stenosis (IDSS) and Posterior dynamic stabilization (PDS) form a bridge between decompression laminectomy alone and rigid fusion, by attempting to sustain beneficial effects of decompression and stabilization in an attempt to prevent bad effects of relentless degeneration. To evaluate the clinical outcome in operated patient of posterior dynamic stabilization. Data were collected over 1 year in prospective, nonrandomized follow-up study using outcome scales. Single surgeon, tertiary care centre in Mumbai, India. Described here is the operative technique of posterior dynamic stabilization using CoFlexTM and clinical outcome of 67 consecutive patients using Visual Analogue Scale and Oswestry Disability Score.


Regulation of C-myc protooncogene expression in osteoblastic cells by arachidonic acid metabolites: relationship to proliferation.
Prostaglandin E2 and leukotriene B4 are metabolites of arachidonic acid with well-characterized effects on osteoblastic cells. Prostaglandin E2 has been shown to be a potent bone-resorbing agent and to stimulate as well as inhibit osteoblastic cell proliferation. Leukotriene B4 has also been demonstrated to stimulate or inhibit osteoblastic cell proliferation, depending on the cell type tested. In the present study, the potential relationship of the effects of prostaglandin E2 and leukotriene B4 on osteoblastic cell proliferation to c-myc protooncogene expression was investigated. Prostaglandin E2 has been shown previously to inhibit normal rat osteoblastic cell proliferation. The present studies show that prostaglandin E2 at 10(-6) M decreased c-myc expression in these cells. In the human osteoblastic osteosarcoma cell line, G292, prostaglandin E2 increased c-myc expression and inhibited proliferation. In contrast, epidermal growth factor increased DNA synthesis as well as c-myc expression. Prostaglandin E2 also inhibited proliferation of another human osteoblastic osteosarcoma cell line, Saos-2, but it did not produce any changes in c-myc expression. In these cells, epidermal growth factor did not affect either DNA synthesis or c-myc expression. Leukotriene B4 did not show any effects on c-myc expression in any of the osteoblastic cells tested.


Effect of selected alcohol dehydrogenase inhibitors on human hepatic lactate dehydrogenase activity - an in vitro study.
Metabolic acidosis severely complicates methanol and ethylene glycol intoxications. Acidosis is caused by acid metabolites and can be intensified by lactate elevation. Lactate concentration depends on the NADH(2)/NAD ratio. Lactate dehydrogenase (LDH, E.C.126.96.36.199.) supplies more lactate when the level of NADH(2) is elevated. The aim of the study was to evaluate the effect of alcohol dehydrogenase (ADH) inhibitors and substrates: cimetidine, EDTA, 4-methylpyrazole (4-MP), Ukrain and ethanol on LDH activity. The activity of LDH was determined spectrophotometrically in human liver homogenates incubated with cimetidine, EDTA, 4-MP and Ukrain at concentrations of 2 x 10(-6), 10(-5) and 5 x 10(-5) m as well as ethanol at concentrations of 12.50, 25.00, 50.00 mm. The LDH activity was significantly increased by 10(-5) and 5 x 10(-5) m concentrations of cimetidine and 4-MP, and by all concentrations of ethanol. The most effective change of LDH activity of about 26% (P<0.01) was observed at the highest concentration of ethanol. Ukrain inhibited LDH activity at both concentrations, i.e. 10(-5) and 5 x 10(-5) m (P<0.05). However, EDTA did not significantly influence LDH activity. The data showed that ethanol and 4-MP, the main antidotes in methanol or ethylene glycol poisoning, may increase liver LDH activity - an undesirable effect during the therapy of patients intoxicated with these alcohols. On the other hand, the decrease of LDH activity in the presence of Ukrain is a promising finding but definitely requires further investigation.


Tears and aqueous humor from horses inoculated with Leptospira contain antibodies which bind to cornea.
An antigenic relationship between Leptospira interrogans and equine cornea was previously described by us. An enzyme-linked immunosorbent assay was employed in the present work to investigate the existence of anti-leptospira and anti-cornea antibodies in tears, aqueous humor and serum from horses inoculated i.m. with those antigens. Ten days after a booster by the same route, antibodies that bind to microtiter plates, coated with an homogenate of either equine cornea or leptospira, were detected in those fluids and in the sera. At the same time, the corneas of the horses began to develop a diffuse opacity. This finding of anti-leptospira antibodies in equine tears and aqueous humor shows the pathway along which they arrive at the cornea and bind to it.


RNA-binding protein RBM28 can translocate from the nucleolus to the nucleoplasm to inhibit the transcriptional activity of p53.
RNA-binding protein RBM28 (RBM28), as a nucleolar component of spliceosomal small nuclear ribonucleoproteins, is involved in the nucleolar stress response. Whether and how RBM28 regulates tumor progression remains unclear. Here, we report that RBM28 is frequently overexpressed in various types of cancer and that its upregulation is associated with a poor prognosis. Functional and mechanistic assays revealed that RBM28 promotes the survival and growth of cancer cells by interacting with the DNA-binding domain of tumor suppressor p53 to inhibit p53 transcriptional activity. Upon treatment with chemotherapeutic drugs (e.g., adriamycin), RBM28 is translocated from the nucleolus to the nucleoplasm, which is likely mediated via phosphorylation of RBM28 at Ser122 by DNA checkpoint kinases 1 and 2 (Chk1/2), indicating that RBM28 may act as a nucleolar stress sensor in response to DNA damage stress. Our findings not only reveal RBM28 as a potential biomarker and therapeutic target for cancers but also provide mechanistic insights into how cancer cells convert stress signals into a cellular response linking the nucleolus to regulation of the tumor suppressor p53.


Chronic allopurinol and adenine therapy in Duchenne muscular dystrophy: effects on muscle function, nucleotide degradation, and muscle ATP and ADP content.
Prompted by the controversy on the efficacy of allopurinol in Duchenne muscular dystrophy and by our observations of an abnormal adenine nucleotide turnover in this disease, we conducted an 18 month, double-blind clinical trial with allopurinol and adenine in 14 Duchenne boys paired according to age and functional activity. Detailed clinical evaluation was performed trimonthly. Muscle ATP and ADP content was measured before and after 1 year of treatment. The effect of therapy on adenine nucleotide turnover was determined. No significant difference was observed between the treated and placebo groups, but both showed a significant deterioration (p less than 0.05) in most clinical parameters. Muscle ATP was reduced in Duchenne dystrophy (p less than 0.02) but did not change with therapy, and no correction of the abnormal adenine nucleotide degradation was observed.


Dependence of the deactivation reactions of photosystem II on the redox state of plastoquinone pool A varied under anaerobic conditions; Equilibria on the acceptor side of photosystem II.
Dark adapted spinach chloroplasts and Chlorella with variably reduced plastoquinone pools were given 1 or 2 saturating flashes. Under these conditions the rate of deactivation of state S2 of the oxygen evolving site of Photosystem II (B, Kok, B, Forbush, M. McGloin (1970) Photochem. Photobiol. 11, 457-475) is highly dependent on the pool redox state, undergoing a nearly 10-fold acceleration upon transforming the plastoquinone pool (A) from 100% oxidized to 90% reduced. Deactivation of state S3 is unaffected by the same variation of the pool redox state. These observations are attributed to a high concentration of Photosystem II reduced primary electron acceptor, Q-, coincident with the formation of S2 and a low concentration coincident with the formation of S3, under the conditions of highly reduced plastoquinone pool. Simultaneous determination of Q- and A2- result in an estimated equilibrium constant of 15-20 for reaction Q-B in equilibrium QB- and a value greater than 50 for equilibrium Q-B- in equilibrium QB2-, where B is the secondary electron acceptor described by B. Bouges-Bocquet ((1973) Biochim. Biophys. Acta. 314, 250-256) and B.R. Velthuys and J. Amesz ((1974) Biochim. Biophys. Acta. 333, 85-94). It is proposed that doubly reduced B becomes protonated in the last reaction.


Complex role experiences and personal differentiation: a real-life experiment.
This paper analyses the relationship between the complexity of role experiences and personal differentiation. "Complex role experiences" are defined by high levels of voluntarism, multiplexity, quasi-responsibility and trial-and-error behaviour. The ability to differentiate between diverse principles of role enactment and various criteria of interpersonal relations is conceptualized as "personal differentiation". Our hypothesis is that a pre-designed educational programme providing complex role experiences enhances personal differentiation. This hypothesis was tested in an experimental study on Israeli adolescents participating in summer camps. The results show that the active exposure to and assumption of variegated roles in different settings on a voluntaristic basis, while performing meaningful and quasi-responsible actions by means of trial-and-error behaviour, enhances role and interpersonal differentiation, and hence personal differentiation, among adolescents.


Combined SEM/AVS and attenuation of concentration models for the assessment of bioavailability and mobility of metals in sediments of Sepetiba Bay (SE Brazil).
This study proposes a new methodology to study contamination, bioavailability and mobility of metals (Cd, Cu, Ni, Pb, and Zn) using chemical and geostatistics approaches in marine sediments of Sepetiba Bay (SE Brazil). The chemical model of SEM (simultaneously extracted metals)/AVS (acid volatile sulfides) ratio uses a technique of cold acid extraction of metals to evaluate their bioavailability, and the geostatistical model of attenuation of concentrations estimates the mobility of metals. By coupling the two it was observed that Sepetiba Port, the urban area of Sepetiba and the riverine discharges may constitute potential sources of metals to Sepetiba Bay. The metals are concentrated in the NE area of the bay, where they tend to have their lowest mobility, as shown by the attenuation model, and are not bioavailable, as they tend to associate with sulfide and organic matter originated in the mangrove forests of nearby Guaratiba area.


SARS-CoV-2 Nsp1 binds the ribosomal mRNA channel to inhibit translation.
The SARS-CoV-2 non-structural protein 1 (Nsp1), also referred to as the host shutoff factor, suppresses host innate immune functions. By combining cryo-electron microscopy and biochemistry, we show that SARS-CoV-2 Nsp1 binds to the human 40S subunit in ribosomal complexes, including the 43S pre-initiation complex and the non-translating 80S ribosome. The protein inserts its C-terminal domain into the mRNA channel, where it interferes with mRNA binding. We observe translation inhibition in the presence of Nsp1 in an in vitro translation system and in human cells. Based on the high-resolution structure of the 40S-Nsp1 complex, we identify residues of Nsp1 crucial for mediating translation inhibition. We further show that the full-length 5' untranslated region of the genomic viral mRNA stimulates translation in vitro, suggesting that SARS-CoV-2 combines global inhibition of translation by Nsp1 with efficient translation of the viral mRNA to allow expression of viral genes.


Environmental stimulation promotes changes in the distribution of phorbol ester receptors.
The translocation of protein kinase C (PKC) from the cytosol to the membrane might be functionally involved in learning and memory. Using [3H]-phorbol 12,13-dibutyrate (3H-PDBu) binding three pools of binding sites could be distinguished in tissue preparations: Pool a comprised the soluble receptors which bound phorbol ester with low affinity in the absence of calcium. Pool b was composed of high-affinity phorbol ester binding sites identified in the soluble fraction upon addition of calcium. Pool c represented stably membrane-bound receptors binding phorbol ester independently of calcium. 3H-PDBu binding was then measured in the cortices and hippocampi of rats trained in an eight-arm radial maze. A progressive training-dependent increase of membrane-bound binding activity with a concomitant decrease in the soluble fraction was detected independent of learning the maze task. These results suggest that it is the experience of an enriched environment by the repeated behavioral stimulation in a maze rather than the acquisition of a memory task that leads to enhanced incorporation of phorbol ester receptors (PKC) into the cell membrane.


Skin irritants of Euphorbia fortissima.
By means of a combination of partition and chromatographic methods six irritant constituents were isolated from the fresh latex of Euphorbia fortissima. Compounds A-D were di-esters of the common parent diterpene 12-deoxyphorbol, and compounds E and F were mono-esters of the same diterpene. The fresh latex had an irritant dose 50% (ID50) on mice of 0-64 mug mul- minus 1. Compounds A-D are short-acting irritants reaching a maximum activity within 4 h of application to the skin, whilst the monoesters maintained potent irritant effects for up to 24 h. Selective hydrolysis of the di-esters at the C-20 primary ester group also produced mono-esters of greater potency after 24 h. An increase in the length of the fatty acid located at C-13 produced greater biological activity in both the mono- and di-ester groups.