Identification of host miRNAs that may limit human rhinovirus replication.
To test whether the replication of human rhinovirus (HRV) is regulated by microRNAs in human bronchial epithelial cells. For the present study, the human cell line BEAS-2B (derived from normal human bronchial epithelial cells) was adopted. DICER knock-down, by siRNA transfection in BEAS-2B cells, was performed in order to inhibit microRNA maturation globally. Alternatively, antisense oligonucleotides (anti-miRs) were transfected to inhibit the activity of specific microRNAs. Cells were infected with HRV-1B. Viral replication was assessed by measuring the genomic viral RNA by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Association between microRNA-induced-silencing-complex and viral RNA was detected by Ago2 co-immunoprecipitation followed by RT-qPCR. Targetscan v.6 was used to predict microRNA target sites on several HRV strains. Here, we show that microRNAs affect replication of HRV-1B. DICER knock-down significantly reduced the expression of mature microRNAs in a bronchial epithelial cell line (BEAS-2B) and in turn, increased the synthesis of HRV-1B RNA. Additionally, HRV-1B RNA co-immunoprecipitated with argonaute 2 protein, an important effector for microRNA activity suggesting that microRNAs bind to viral RNA during infection. In order to identify specific microRNAs involved in this interaction, we employed bioinformatics analysis, and selected a group of microRNAs that have been reported to be under-expressed in asthmatic bronchial epithelial cells and were predicted to target different strains of rhinoviruses (HRV-1B, -16, -14, -27). Our results suggest that, out of this group of microRNAs, miR-128 and miR-155 contribute to the innate defense against HRV-1B: transfection of specific anti-miRs increased viral replication, as anticipated in-silico. Taken together, our results suggest that pathological changes in microRNA expression, as already reported for asthma or chronic obstructive pulmonary disease have the potential to affect Rhinovirus replication and therefore may play a role in virus-induced exacerbations.


Entomopathogenic nematodes for the control of the codling moth (Cydia pomonella L.) in field and laboratory trials.
Three commercially available entomopathogenic nematode (EPN) strains (Steinernema feltiae and Heterorhabditis bacteriophora Hb1 and Hb2) and two local species (S. jeffreyense and S. yirgalemense) were evaluated for the control of the codling moth (Cydia pomonella). In field spray trials, the use of S. jeffreyense resulted in the most effective control (67%), followed by H. bacteriophora (Hb1) (42%) and S. yirgalemense (41%). Laboratory bioassays using spray application in simulated field conditions indicate S. feltiae to be the most virulent (67%), followed by S. yirgalemense (58%). A laboratory comparison of the infection and penetration rate of the different strains showed that, at 14°C, all EPN strains resulted in slower codling moth mortality than they did at 25°C. After 48 h, 98% mortality was recorded for all species involved. However, the washed codling moth larvae, cool-treated (at 14°C) with S. feltiae or S. yirgalemense, resulted in 100% mortality 24 h later at room temperature, whereas codling moth larvae treated with the two H. bacteriophora strains resulted in 68% and 54% control, respectively. At 14°C, S. feltiae had the highest average penetration rate of 20 IJs/larva, followed by S. yirgalemense, with 14 IJs/larva. At 25°C, S. yirgalemense had the highest penetration rate, with 39 IJs/larva, followed by S. feltiae, with 9 IJs/larva. This study highlights the biocontrol potential of S. jeffreyense, as well as confirming that S. feltiae is a cold-active nematode, whereas the other three EPN isolates tested prefer warmer temperatures.


Genetic mapping of the powdery mildew resistance gene in soybean PI 567301B.
Powdery mildew (PMD) of soybean [Glycine max (L.) Merr.] is caused by the fungus Microsphaera diffusa. Severe infection of PMD on susceptible varieties often causes premature defoliation and chlorosis of the leaves, which can result in considerable yield losses under favorable environmental conditions for disease development in the field. A total of 334 F(7)-derived recombinant inbred lines (RILs) from a cross of a PMD susceptible soybean cultivar Wyandot and PMD-resistant PI 567301B were used for genetic mapping of PMD resistance in PI 567301B and for development of molecular markers tightly linked to the gene. The result of the PMD screening for each line in the field was in agreement with that in the greenhouse test. The genetic map containing the PMD resistance gene was constructed in a 3.3 cM interval flanked by two simple sequence repeat (SSR) markers on chromosome 16. The PMD resistance gene was mapped at the same location with SSR marker BARCSOYSSR_16_1291, indicating that there was no recombination between the 334 RILs and this marker. In addition, a single nucleotide polymorphism (SNP) marker developed by high-resolution melting curve analysis and a cleaved amplified polymorphic sequence (CAPS) marker with Rsa1 recognition site were used for the genetic mapping. These two markers were also mapped to the same genomic location with the PMD resistance gene. We validated three tightly linked markers to the PMD resistance gene using 38 BC(6)F(2) lines and corresponding BC(6)F(2:3) families. The three marker genotypes of the backcross lines predicted the observed PMD phenotypes of the lines with complete accuracy. We have mapped a putatively novel single dominant PMD resistance gene in PI 567301B and developed three new molecular markers closely linked to the gene. Molecular markers developed from this study may be used for high-throughput marker-assisted breeding for PMD resistance with the gene from PI 567301B.


Population PK/PD modeling of eltrombopag in healthy volunteers and patients with immune thrombocytopenic purpura and optimization of response-guided dosing.
The relationship between plasma eltrombopag concentrations and increases in platelet counts (PLTC) was characterized in healthy volunteers (HVs) and patients with immune thrombocytopenic purpura (ITP) using population pharmacokinetic/pharmacodynamic (PK/PD) models. The semiphysiological model included 3 PK, 1 precursor production, 2 maturation, and 1 blood platelet compartments and assumed a linear increase in platelet production rate with eltrombopag concentrations. Thrombopoiesis was assumed to be the same in HVs and patients, whereas platelets degraded more rapidly in patients. A mixture model was used, with nonresponders accounting for 19% of the patients. The following covariates were predictive of higher PLTC in ITP patients based on PK or PD differences in descending order of magnitude: East Asian race, age 65 years or older, baseline PLTC greater than 15 Gi/L, female, and concurrent corticosteroid. Simulations support starting eltrombopag at a dose of 50 mg once daily, except in East Asian patients, for whom 25 mg once daily is warranted. Doses can be titrated at 2-week intervals (or longer) to achieve target PLTC.


Long acting growth hormone (LAGH), an update.
In 1957, Maurice Raben at Yale was able to isolate and purify growth hormone from cadaveric pituitary glands. Pituitary growth hormone was the only way to treat children with growth hormone (GH) deficiency, until 1985 when recombinant GH became available for daily subcutaneous injection. For many years, the pediatric endocrine community longed for a long-acting recombinant GH formulation that would decrease the inconvenience of daily injections. Several mechanisms were employed to develop a GH that is rapidly absorbed into the blood stream after subcutaneous injection, but provides slow removal from the circulatory system to potentially optimize patient adherence to GH therapy. Four long-acting growth hormones are currently available in the world, or are close to regulatory approval. They are: (1) Pegylated formulations, (2) Prodrug formulations which are converted into active drug, (3) Nonvalent transient albumin binding GH compounds and (4) GH fusion proteins where a protein si fused with GH. All four formulations have undergone detailed phase 3 studies and were found to show non-inferiority in these clinical studies. All four demonstrate a safety and tolerability profile that is comparable to that of daily somatropin with an excellent adherence profile.


A multicenter study of histoplasmosis and blastomycosis after solid organ transplantation.
A review of the clinical presentation, diagnosis, treatment and outcomes of 30 solid organ transplant recipients (SOTRs) with histoplasmosis or blastomycosis from 3 Midwestern academic medical centers. The endemic fungal pathogens, Histoplasma capsulatum and Blastomyces dermatitidis, may cause severe infection in SOTRs. In this report, we describe the clinical presentation, diagnosis, treatment, and outcomes of these endemic fungal infections (EFIs) among SOTRs at 3 academic transplant centers. A retrospective review was conducted of SOTRs with histoplasmosis or blastomycosis from 3 Midwestern medical centers in the United States. Data collected included demographics, immunosuppression, clinical presentation, method of diagnosis, antifungal treatment, response to therapy, and patient and graft survival. Between 1996 and 2008, 30 transplant recipients with histoplasmosis or blastomycosis were identified, giving a cumulative incidence of infection of 0.50% (30/5989); 73% of the study patients were renal transplant recipients, and the median time to disease onset after transplantation was 10.5 months. The lungs were the most common site of infection (83%), and 60% had disseminated disease. Urine antigen testing was positive in all patients in whom it was performed (23/23). Initial antifungal therapy consisted of amphotericin B in 70%, and 87% received azoles, typically itraconazole (83%). Two patients developed relapsed infection and 7 patients had graft failure after EFI. Overall mortality was 30%, with an attributable mortality of 13%. As in several previous single-center studies, the incidence of post-transplant histoplasmosis and blastomycosis was <1%, but often resulted in disseminated infection. In this cohort, EFI was associated with a high rate of allograft loss and overall mortality.


Application of microfluidic technologies in forensic analysis.
The application of microfluidic technology in forensic medicine has steadily expanded over the last two decades due to the favorable features of low cost, rapidity, high throughput, user-friendliness, contamination-free, and minimum sample and reagent consumption. In this context, bibliometric methods were adopted to visualize the literature information contained in the Science Citation Index Expanded from 1989 to 2022, focusing on the co-occurrence analysis of forensic and microfluidic topics. A deep interpretation of the literature was conducted based on co-occurrence results, in which microfluidic technologies and their applications in forensic medicine, particularly forensic genetics, were elaborated. The purpose of this review is to provide an impartial evaluation of the utilization of microfluidic technology in forensic medicine. Additionally, the challenges and future trends of implementing microfluidic technology in forensic genetics are also addressed.


[The applications and challenges of liquid chromatography tandem mass spectrometry in maternal and child health].
Due to its ultra-high sensitivity, specificity and throughput, liquid chromatography tandem mass spectrometry (LC-MS/MS) has become an important analytical tool in clinical laboratories in quantifying various small molecules, such as vitamins, bile acids, steroids and other internal metabolites relevant to maternal diseases. As an effective means of screening and diagnosing diseases in preventive medicine, LC-MS/MS has been widely used in maternal and child health, contributing to the reduction of the incidence of maternal and child diseases and premature morbidity and mortality. At present, LC-MS/MS is an emerging and powerful platform in laboratory testing in China, facing both challenges and opportunities. In this article, the representative applications in the field of maternal and child health are summarized and discussed, along with the major hurdles of LC-MS/MS in clinical recognition and implementation.


Buried Probe in Complex Congenital Nasolacrimal Duct Obstructions: Clinical Profiles and Outcomes.
To report the clinical profiles and outcomes of buried probe variant of complex congenital nasolacrimal duct obstruction (CNLDO). Retrospective chart review of all patients endoscopically diagnosed as a buried probe variant of complex CNLDO, over a 3.5 year period from a single surgeon's (MJA) database were included in the study. A detailed lacrimal system evaluation was performed and intraoperative findings including nasal endoscopy were documented. A minimum follow up of 3 months following the final intervention was considered for analysis. Anatomical and functional success of the interventions was assessed at the final follow up. Twenty-two eyes of 21 patients with buried probes were studied. The mean age at presentations was 41.2 months. Epiphora and discharge were the commonest presenting symptoms noted in 77.2% (17/22). Associated lacrimal anomalies include punctal agenesis, incomplete punctal canalization and atonic lacrimal sac. All patients underwent irrigation and probing under nasal endoscopic guidance. Further, 3 patients underwent endoscopic dacryocystorhinostomy for persistent CNLDO. At a mean follow up of 4.9 months, the final anatomical and functional successes were noted in 90.9% and 81.8%, respectively. Buried probe is a variant of complex CNLDO, noted more commonly in older children. Endoscopic guidance is crucial for its diagnosis and satisfactory outcomes.


Can Tooth Preparation Design Affect the Fit of CAD/CAM Restorations?
The purpose of this study was to evaluate if the marginal fit of computer-aided design and computer-aided manufacturing (CAD/CAM) restorations manufactured with CAD/CAM systems can be affected by different tooth preparation designs. Twenty-six typodont (plastic) teeth were divided into two groups (n = 13) according to the occlusal curvature of the tooth preparation. These were the group 1 (control group) (flat occlusal design) and group 2 (curved occlusal design). Scanning of the preparations was performed, and crowns were milled using ceramic blocks. Blocks were cemented using epoxy glue on the pulpal floor only, and finger pressure was applied for 1 minute. On completion of the cementation step, poor fits between the restoration and abutment were measured by microphotography and the silicone replica technique using light-body silicon material on mesial, distal, buccal, and lingual surfaces. Two-way ANOVA analysis did not reveal a statistical difference between flat (83.61 ± 50.72) and curved (79.04 ± 30.97) preparation designs. Buccal, mesial, lingual, and distal sites on the curved design preparation showed less of a gap when compared with flat design. No difference was found on flat preparations among mesial, buccal, and distal sites (P < .05). The lingual aspect had no difference from the distal side but showed a statistically significant difference from mesial and buccal (P < .05). Difference in occlusal design did not significantly impact the marginal fit. Marginal fit was significantly affected by the location of the margin; lingual and distal locations exhibited greater margin gap values compared with buccal and mesial sites regardless of the preparation design.


[Perception of autonomy in the elderly who live with a family member].
In gerontological research and practice, autonomy is a phenomenon of central interest. Numerous researchers have considered autonomy as an elder's problem to be defined and resolved, and have studied the phenomenon from the psychological, sociological, or another discipline's perspective. The purpose of this phenomenological study was to describe the experience of autonomy as lived by 80 plus elders who share a household with a family member. Parse's human becoming theory oriented the nursing perspective to the study of autonomy toward the meanings of autonomy given by older persons within the rhythmicity of interpersonal relationships. The analysis of data for four women participants revealed the phenomenon essential themes as being able, still being someone, and chosen beyond changes. These themes are linked to the principle of cotranscendence from the human becoming theory: being able reveals the dynamic potential of each participant who, through their relationships, creates an original way of being still someone, and of choosing beyond life changes. Implications for nursing practice are discussed.


The utility of targeted perioperative transthoracic echocardiography in managing an adult patient with anomalous origin of the left coronary artery-pulmonary artery for noncardiac surgery.
Congenital coronary artery anomalies as a whole are uncommon. Abnormal origin of the left coronary artery from the pulmonary artery (ALCAPA) is probably the most common congenital coronary defect. An overwhelming majority of the patients with untreated ALCAPA do not survive to adulthood. As yet, there is no consensus on the management of adults with ALCAPA. We describe a patient with breast malignancy and incidentally detected ALCAPA; primacy was given to treatment of the oncologic condition as a first step. Anesthesia management was focused on maintaining adequate collateral coronary perfusion and avoidance of excessive loading of the left ventricle. This was achieved using a simplified transthoracic echocardiography (TTE) protocol at the time of induction of anesthesia; TTE was also used to reconfirm the absence of disturbances in myocardial function at the end of surgery. We sugggest the routine use of tte in managing perioperative care in low resource settings when the underlying cardiac disease is rare and the evidence base if often insufficient.


White adipose tissue re-growth after partial lipectomy in high fat diet induced obese wistar rats.
The effects of partial removal of epididymal (EPI) and retroperitoneal (RET) adipose tissues (partial lipectomy) on the triacylglycerol deposition of high fat diet induced obese rats were analyzed, aiming to challenge the hypothesized body fat regulatory system. Male 28-day-old wistar rats received a diet enriched with peanuts, milk chocolate and sweet biscuits during the experimental period. At the 90th day of life, rats were submitted to either lipectomy (L) or sham surgery. After 7 or 30 days, RET, EPI, liver, brown adipose tissue (BAT), blood and carcass were obtained and analyzed. Seven days following surgery, liver lipogenesis rate and EPI relative weight were increased in L. After 30 days, L, RET and EPI presented increased lipogenesis, lipolysis and percentage of small area adipocytes. L rats also presented increased liver malic enzyme activity, BAT lipogenesis, and triacylglycerol and corticosterone serum levels. The partial removal of visceral fat pads affected the metabolism of high fat diet obese rats, which leads to excised tissue re-growth and possibly compensatory growth of non-excised depots at a later time.


Entacapone does not induce conformational changes in liver mitochondria or skeletal muscle in vivo.
Entacapone and tolcapone, novel catechol-O-methyl-transferase (COMT) inhibitors, have been developed for the treatment of Parkinson's disease in combination with levodopa. Three fatal cases of drug-induced hepatitis, one with hepatic necrosis and mitochondrial changes have been reported in clinical use of tolcapone. In vitro tolcapone has been shown to induce uncoupling of oxidative phosphorylation. Liver and skeletal muscle tissues from an oral rat toxicity study were used to investigate the influence of entacapone, tolcapone (300 and 500 mg/kg/day) or a known uncoupling agent, 2,4-dinitrophenol (DNP), (20 mg/kg/ day) on the cell morphology. Centrolobular hypertrophy was revealed in the histopathology of the liver in tolcapone-treated rats. Transmission electron microscopy (TEM) of the liver and skeletal muscle tissue, revealed mitochondrial swelling and reduced matrix density with deformation of cristae in the tolcapone and DNP groups. Intermyofibrillar edema was characteristic of the skeletal muscle tissue of DNP- and tolcapone-exposed animals. In the tolcapone group, also the sarcomeres were prominent. Treatment-related light microscopic or TEM findings were not observed either in entacapone-treated or control animals. The similarity of structural damages induced by both tolcapone- and DNP suggests that uncoupling of oxidative phosphorylation may contribute to the toxicity of tolcapone in the rat.


Air pollution, UV irradiation and skin carcinogenesis: what we know, where we stand and what is likely to happen in the future?
The link between air pollution, UV irradiation and skin carcinogenesis has been demonstrated within a large number of epidemiological studies. Many have shown the detrimental effect that UV irradiation can have on human health as well as the long-term damage which can result from air pollution, the European ESCAPE project being a notable example. In total, at present around 2800 different chemical substances are systematically released into the air. This paper looks at the hazardous impact of air pollution and UV and discusses: 1) what we know; 2) where we stand; and 3) what is likely to happen in the future. Thereafter, we will argue that there is still insufficient evidence of how great direct air pollution and UV irradiation are as factors in the development of skin carcinogenesis. However, future prospects of progress are bright due to a number of encouraging diagnostic and preventive projects in progress at the moment.


Characterization of the complete chloroplast genome of Lonicera tangutica (Caprifoliaceae), an ornamental and medicinal plant in China.
In the present study, the complete chloroplast genome of Lonicera tangutica is presented and characterized for the first time. The complete chloroplast genome was 156,121 bp in length, including 23,899 bp inverted repeat (IR) regions, an 89,466 bp large single-copy (LSC) region, and an 18,851 bp small single-copy (SSC) region. A total of 129 genes, including 37 tRNA genes, eight rRNA genes, and 84 protein-coding genes, were annotated, and the overall GC content of the chloroplast genome was 38.35%. Two introns in the ycf3 gene and a single intron in another gene were detected. Maximum-likelihood phylogenetic analysis indicated that L. tangutica has a very close evolutionary relationship with Lonicera praeflorens, Lonicera hispida, Lonicera fragrantissima, and Lonicera stephanocarpa. These results are valuable for studying the evolution and genetic diversity of L. tangutica.


Topology evaluation of models for difficult targets in the 14th round of the critical assessment of protein structure prediction (CASP14).
This report describes the tertiary structure prediction assessment of difficult modeling targets in the 14th round of the Critical Assessment of Structure Prediction (CASP14). We implemented an official ranking scheme that used the same scores as the previous CASP topology-based assessment, but combined these scores with one that emphasized physically realistic models. The top performing AlphaFold2 group outperformed the rest of the prediction community on all but two of the difficult targets considered in this assessment. They provided high quality models for most of the targets (86% over GDT_TS 70), including larger targets above 150 residues, and they correctly predicted the topology of almost all the rest. AlphaFold2 performance was followed by two manual Baker methods, a Feig method that refined Zhang-server models, two notable automated Zhang server methods (QUARK and Zhang-server), and a Zhang manual group. Despite the remarkable progress in protein structure prediction of difficult targets, both the prediction community and AlphaFold2, to a lesser extent, faced challenges with flexible regions and obligate oligomeric assemblies. The official ranking of top-performing methods was supported by performance generated PCA and heatmap clusters that gave insight into target difficulties and the most successful state-of-the-art structure prediction methodologies.


A periplasmic phospholipase that maintains outer membrane lipid asymmetry in Pseudomonas aeruginosa.
The outer membrane of Gram-negative bacteria is unique in both structure and function. The surface-exposed outer leaflet is composed of lipopolysaccharide, while the inner leaflet is composed of glycerophospholipids. This lipid asymmetry creates mechanical strength, lowers membrane permeability, and is necessary for virulence in many pathogens. Glycerophospholipids that mislocalize to the outer leaflet are removed by the Mla pathway, which consists of the outer membrane channel MlaA, the periplasmic lipid carrier MlaC, and the inner membrane transporter MlaBDEF. The opportunistic pathogen Pseudomonas aeruginosa has two proteins of the MlaA family: PA2800 and PA3239. Here, we show that PA2800 is part of a canonical Mla pathway, while PA3239 functions with the putative lipase PA3238. While loss of either pathway individually has little to no effect on outer membrane integrity, loss of both pathways weakens the outer membrane permeability barrier and increases production of the secondary metabolite pyocyanin. We propose that mislocalized glycerophospholipids are removed from the outer leaflet by PA3239 (renamed MlaZ), transferred to PA3238 (renamed MlaY), and degraded. This pathway streamlines recycling of glycerophospholipid degradation products by removing glycerophospholipids from the outer leaflet prior to degradation.


Enantioselective separation of the sunscreen agent 3-(4-methylbenzylidene)-camphor by electrokinetic chromatography: Quantitative analysis in cosmetic formulations.
3-(4-Methylbenzylidene)-camphor (MBC) is a chiral sunscreen agent used in cosmetic products. In this work, the enantioseparation of MBC has been performed by EKC and applied to the analysis of the MBC enantiomers in cosmetic creams. Different experimental conditions (type and concentration of the chiral selector, temperature, and sample solvent) have been optimized. Due to the neutral nature of this compound, anionic CD derivatives were investigated as chiral selectors. Carboxymethylated-beta-CD (CM-beta-CD) showed the highest chiral separation power, observing that a 15 mM concentration of this CD at a working temperature of 15 degrees C enabled to obtain the highest enantioresolution. However, under these conditions, tailing of peaks obtained for the enantiomers was observed. The addition of increasing concentrations of the neutral alpha-CD to CM-beta-CD at a 15 mM concentration in a 100 mM borate buffer at pH 9.0 improved the enantiomeric separation and decreased peak tailing. The use of DMF for the total dissolution of the cosmetic creams, and methanol:water (1:1 v/v) for appropriate dilution enabled to observe good shape and size for the peaks of the MBC enantiomers. After optimizing a method for the preconditioning of the capillary, the analytical characteristics of the chiral separation method for the analysis of MBC were investigated. Linearity, LODs and LOQs, precision (instrumental repeatability, method repeatability, intermediate precision), accuracy, and selectivity were evaluated. The method was applied to analyze MBC enantiomers contained in two commercial cosmetic creams containing racemic MBC and to study the skin absorption of this compound with time.


Artificial cultivation of true morels: current state, issues and perspectives.
Morels (Morchella, Ascomycota), which are some of the most highly prized edible and medicinal mushrooms, are of great economic and scientific value. Morel cultivation has been a research focus worldwide for more than 100 years, and the outdoor cultivation of morels has succeeded and expanded to a large scale in China in recent years. In this study, we review the progress in recent research regarding the life cycle and reproductive systems in the genus Morchella and the current state of outdoor cultivation. Sclerotia formation and conidia production are two important phases during the life cycle. The morel species cultivated commercially in America is M. rufobrunnea based on molecular phylogenetic analysis. The species currently cultivated in China are black morels, including M. importuna, M. sextalata and M. eximia. The field cultivation of morels expanded in the majority of the provinces in China with a yield of fresh morels of 0-7620 kg per ha. The key techniques include spawn production, land preparation and spawning, the addition of exogenous nutrition, fruiting management and harvesting. The application of exogenous nutrition is the most important breakthrough in the field of morel cultivation, but the mechanism remains unclear. It was estimated that the total amount of field cultivated fresh morels was ∼500 t in 2015-2016. We also discuss the potential issues remaining in the current literature and suggest directions for future studies.


Hyperglycemia Associated With Targeted Oncologic Treatment: Mechanisms and Management.
: Molecularly targeted cancer therapy has rapidly changed the landscape of oncologic care, often improving patients' prognosis without causing as substantial a quality-of-life decrement as cytotoxic chemotherapy does. Nevertheless, targeted agents can cause side effects that may be less familiar to medical oncologists and that require the attention and expertise of subspecialists. In this review, we focus on hyperglycemia, which can occur with use of new anticancer agents that interact with cell proliferation pathways. Key mediators of these pathways include the tyrosine kinase receptors insulin growth factor receptor 1 (IGF-1R) and epidermal growth factor receptor (EGFR), as well as intracellular signaling molecules phosphatidylinositol 3-kinase (PI3K), AKT, and mammalian target of rapamycin (mTOR). We summarize available information on hyperglycemia associated with agents that inhibit these molecules within the larger context of adverse event profiles. The highest incidence of hyperglycemia is observed with inhibition of IGF-1R or mTOR, and although the incidence is lower with PI3K, AKT, and EGFR inhibitors, hyperglycemia is still a common adverse event. Given the interrelationships between the IGF-1R and cell proliferation pathways, it is important for oncologists to understand the etiology of hyperglycemia caused by anticancer agents that target those pathways. We also discuss monitoring and management approaches for treatment-related hyperglycemia for some of these agents, with a focus on our experience during the clinical development of the EGFR inhibitor rociletinib. Treatment-related hyperglycemia is associated with several anticancer agents. Many cancer patients may also have preexisting or undiagnosed diabetes or glucose intolerance. Screening can identify patients at risk for hyperglycemia before treatment with these agents. Proper monitoring and management of symptoms, including lifestyle changes and pharmacologic intervention, may allow patients to continue benefiting from use of anticancer agents.


Examination of neighborhood disadvantage and sleep in a multi-ethnic cohort of adolescents.
Neighborhood-level socioeconomic disadvantage and lower individual-level socioeconomic status are associated with poorer sleep health in adults. However, few studies have examined the association between neighborhood-level disadvantage and sleep in adolescents, a population at high-risk for sleep disturbances. The current study is the first to examine how objective (i.e. via census tract-level data) and subjective measures of neighborhood disadvantage are associated with sleep in a racially/ ethnically and socioeconomically diverse sample of 2493 youth [Non-Hispanic White (20%), Hispanic (46%), Asian (21%), and Multiracial/ Other (13%)]. Findings indicated that greater perceived neighborhood-level social cohesion and lower neighborhood-level poverty were associated with better sleep outcomes in adolescents. However, there was some evidence that the magnitude of the associations differed according to family-level socioeconomic status and race/ ethnicity. Findings suggest that subjective and objective neighborhood characteristics may affect the sleep health of older adolescents, with certain demographic subgroups being particularly vulnerable.


Sedation in pediatric patients undergoing diagnostic procedures.
Sedation is routinely used in pediatric patients undergoing diagnostic procedures. Meperidine with promethazine and chlorpromazine; meperidine with pentobarbital; and meperidine, morphine, pentobarbital, thiopental, methohexital, chloral hydrate, and benzodiazepines as single agents have been used at different doses in patients for various procedures including computed tomography, endoscopy, electroencephalography, and bone marrow biopsies. Most available studies, however, have not compared these drugs in a controlled and blind manner, and the data have often been collected retrospectively. In addition, the degree and duration of sedation required may depend on the procedure. Thus, it is difficult to recommend the drug of choice for producing sedation. Serious cardiac and respiratory effects and excessive sedation have been associated with these drugs, even when normal doses were used. Controlled studies and specific guidelines are needed for optimal use and monitoring of these drugs in pediatric patients.


[Research progress of the drug screening technology in type 2 diabetes mellitus].
With the incidence of type 2 diabetes mellitus increasing year after year, the technology of drug screening of type 2 diabetes mellitus progress rapidly, from the level of animal screening to cellular and molecular screening model, from the traditional drug screening technology to high efficient and throughput screening. This paper will summarize the technology of drug screening in the therapy of type 2 diabetes mellitus.


Anaerobic butanol production driven by oxygen-evolving photosynthesis using the heterocyst-forming multicellular cyanobacterium Anabaena sp. PCC 7120.
Cyanobacteria are oxygen-evolving photosynthetic bacteria. Established genetic manipulation methods and recently developed gene-regulation tools have enabled the photosynthetic conversion of carbon dioxide to biofuels and valuable chemicals in cyanobacteria, especially in unicellular cyanobacteria. However, the oxygen sensitivity of enzyme(s) introduced into cyanobacteria hampers productivity in some cases. Anabaena sp. PCC 7120 is a filamentous cyanobacterium consisting of a few hundred of vegetative cells, which perform oxygenic photosynthesis. Upon nitrogen deprivation, heterocysts, which are specialized cells for nitrogen fixation, are differentiated from vegetative cells at semiregular intervals. The micro-oxic environment within heterocysts protects oxygen-labile nitrogenase from oxygen. This study aimed to repurpose the heterocyst as a host for the production of chemicals with oxygen-sensitive enzymes under photosynthetic conditions. Herein, Anabaena strains expressing enzymes of 1-butanol synthetic pathway from the anaerobe Clostridium acetobutylicum within heterocysts were created. A strain that expressed a highly oxygen-sensitive Bcd/EtfAB complex produced 1-butanol even under photosynthetic conditions. Furthermore, the 1-butanol production per heterocyst cell of a butanol-producing Anabaena strain was fivefold higher than that per cell of unicellular cyanobacterium with the same set of 1-butanol synthetic pathway genes. Thus, our study showed the usefulness of Anabaena heterocysts as a chassis for anaerobic production driven by oxygen-evolving photosynthesis.


Cyclin-dependent kinase 2 inhibitor SU9516 increases sensitivity of colorectal carcinoma cells Caco-2 but not HT29 to BH3 mimetic ABT-737.
Colorectal carcinoma (CRC) that represents one of the major causes for cancer-related death in humans is often associated with over-expression of anti-apoptotic proteins of Bcl-2 family. The aim of presented study was to determine the effect of ABT-737 inhibitor of anti-apoptotic proteins Bcl-2, Bcl-XL and Bcl-w as well as cyclin-dependent kinase 2 (CDK2) inhibitor SU9516 alone and in combination with ABT-737 on survival of colorectal cell lines HT29 and Caco-2. We have shown that both Caco-2 and HT29 cells that are relatively resistant to ABT-737 are also partially sensitive to SU9516, which increased sensitivity of Caco-2 but not HT29 cells to ABT-737. Increased sensitivity of Caco-2 cells to ABT-737 after addition of SU9516 correlated well with SU9516-induced decrease of Mcl-1 expression while we have not observed downregulation of Mcl-1 after the treatment of HT29 cells with SU9516. Instead of this, we have shown that treatment of HT29 cells with SU9516 is associated with decreased expression of tumour suppressor protein p53. Our findings provide a rationale for clinical use of Bcl-2 family inhibitors in combination with CDK2 inhibitors for treatment of Mcl-1-dependent colorectal tumours associated with expression of Bcl-2, Bcl-XL and Bcl-w proteins. In addition, we have shown potential of CDK2 inhibitors for treatment of tumours expressing R273H mutant p53.


Conessine treatment reduces dexamethasone-induced muscle atrophy by regulating MuRF1 and atrogin-1 expression.
Conessine, a steroidal alkaloid, is a potent histamine H3 antagonist with anti-malarial activity. We recently reported that conessine treatment interferes with H₂O₂-induced cell death by regulating autophagy. However, the cellular signaling pathways involved in conessine treatment are not fully understood. Here, we report that conessine reduces muscle atrophy by interfering with the expression of atrophy-related ubiquitin ligases MuRF-1 and atrogin-1. Promoter reporter assay revealed that conessine treatment inhibits FoxO3a-dependent transcription, NF-kappaB-dependent transcription and p53-dependent transcription. We also showed that conessine treatment reduced dexamethasone-induced expression of MuRF1 and atrogin-1 by the quantitative RT-PCR and Western blot. Finally, we demonstrated that conessine treatment reduced dexamethasone-induced muscle atrophy using differentiated C2C12 cells. These results collectively suggest that conessine is potentially useful in the treatment of muscle atrophy.


The prevalence of trichomoniasis and associated factors among women treated at a university hospital in southern Brazil.
Trichomoniasis is the most prevalent non-viral sexually transmitted disease (STD) in the world; however, it remains a neglected parasitic disease. This study aimed to determine the prevalence of trichomoniasis and its associated epidemiological factors among women treated at a hospital in southern Brazil. A cross-sectional study was performed to determine the prevalence of this infection in women treated at Hospital Universitário (HU) in Rio Grande, Rio Grande do Sul, Brazil, between January 2012 and January 2015. This study consisted a self-administered questionnaire regarding demographic, clinical, and behavioural data and a molecular diagnosis with polymerase chain reaction (PCR) using the TVK3/7 primer set, which was confirmed with sequence analysis. Of the 345 women surveyed, the overall prevalence of Trichomonas vaginalis (T. vaginalis) was 4.1% (14/345). The prevalence rates were 5.9% among pregnant women, 8.5% among HIV-positive women, and 10.1% among HIV-positive pregnant women. The rates for groups with other significant demographic and clinical features were as follows: 6.6% among women with white skin, 12.3% among women with an income below the minimum monthly wage, 7.4% among women with a vaginal pH greater than or equal to 4.6, and 7.9% among women with a comorbid STD. The multivariate analysis confirmed that pregnant women who were HIV-positive (p = 0.001) and had low incomes (p = 0.026) were the most likely to have this infection. A multivariate analysis confirmed that HIV-positive pregnant women with low incomes were the participants most likely to have trichomoniasis. These results are important because this Brazilian region presents a high prevalence of HIV-1 subtype C, which is associated with greater transmissibility. Additionally, low family income reveals a socioeconomic fragility that might favour the transmission of this STD.


Impact of asymptomatic hypertension on left ventricular functions.
(M-Mode Echocardiography was used to study the L.V. Systolic functions and the determinants of L.V. functions in subjects with asymptomatic Hypertension in comparison with subjects with normotensives. The functions were assessed among (60) sixty healthy subject and sixty (60) hypertensive subjects. The L.V. Function Parameters assessed by Echocardiography at lateral and septal sides of Left ventricle at the level of mitral annulus. Techholz's formula was used to measure the stroke volume, ejection fraction, cardiac output and cardiac index etc... The resting stroke volume, cardiac output and cardiac index were normal but significantly high among hypertensive subjects compared to normotensive subjects (P<0.05). The fractional fiber shortening (F.F.S.) and other parameters of L.V. contractile state were significantly reduced among hypertensives compared to normotensives (P<0.05). The significant Impairment of F.F.S. is due to alteration in dimension of L.V. wall thickness, L.V. cavity, L.V. geometry and fibrous changes in L.V. myocardium. This carries prognostic implication and requires further documentations, investigations and researches. As the R.A.A.S. play role in development of structural changes in L.V. wall drugs A.C.E. inhibiters i.e. Captopril and A.II.R. antagonist i.e. Losartan play significant role in improvement of L.V. functional changes. Even in the presence of normal L.V. systolic function, the L.V. hypertrophy is associated with altered L.V. geometry. Normal or near normal F.F.S. is considered a hallmark of normal L.V. function when L.V. geometry is normal, but not when L.V. hypertrophy type of geometry. Normal limit of F.F.S. should be reset to newer value. The L.V. contractile state was negatively correlated to L.V. after load parameters. So the main objective of management of hypertensive subjects should be, to reduce the after load to improve the L.V. contractile state.)


The Southeastern Aerosol Research and Characterization Study, part 3: continuous measurements of fine particulate matter mass and composition.
Deployment of continuous analyzers in the Southeastern Aerosol Research and Characterization Study (SEARCH) network began in 1998 and continues today as new technologies are developed. Measurement of fine particulate matter (PM2.5) mass is performed using a dried, 30 degrees C tapered element oscillating microbalance (TEOM). TEOM measurements are complemented by observations of light scattering by nephelometry. Measurements of major constituents include: (1) SO4(2-) via reduction to SO2; (2) NH4+ and NO3- via respective catalytic oxidation and reduction to NO, (3) black carbon (BC) by optical absorption, (4) total carbon by combustion to CO2, and (5) organic carbon by difference between the latter two measurements. Several illustrative examples of continuous data from the SEARCH network are presented. A distinctive composite annual average diurnal pattern is observed for PM2.5 mass, nitrate, and BC, likely indicating the influence of traffic-related emissions, growth, and break up of the boundary layer and formation of ammonium nitrate. Examination of PM2.5 components indicates the need to better understand the continuous composition of the unmeasured "other" category, because it contributes a significant fraction to total mass during periods of high PM2.5 loading. Selected episodes are presented to illustrate applications of SEARCH data. An SO2 conversion rate of 0.2%/hr is derived from an observation of a plume from a coal-fired power plant during early spring, and the importance of local, rural sources of NH3 to the formation of ammonium nitrate in particulate matter (PM) is demonstrated.


Enantioselective Intermolecular [2 + 2] Photocycloaddition Reactions of 2(1H)-Quinolones Induced by Visible Light Irradiation.
In the presence of a chiral thioxanthone catalyst (10 mol %) the title compounds underwent a clean intermolecular [2 + 2] photocycloaddition with electron-deficient olefins at λ = 419 nm. The reactions not only proceeded with excellent regio- and diastereoselectivity but also delivered the respective cyclobutane products with significant enantiomeric excess (up to 95% ee). Key to the success of the reactions is a two-point hydrogen bonding between quinolone and catalyst enabling efficient energy transfer and high enantioface differentiation. Preliminary work indicated that solar irradiation can be used for this process and that the substrate scope can be further expanded to isoquinolones.


[A case of demodicosis on a Dutch dairy farm].
Demodex bovis is a burrowing mite that lives in the hair follicles of cattle, resulting in nodules, granulomatous inflammation, and the formation of scar tissue. Clinical and histopathological changes of a recently diagnosed case of demodicosis on a Dutch dairy farm are presented before and after treatment. Treatment results were disappointing. The role of demodicosis in Dutch dairy farming is discussed.


Biotransformation of phenol in synthetic wastewater using the functionalized magnetic nano-biocatalyst particles carrying tyrosinase.
Low conversion efficiency and long-processing time are some of the major problems associated with the use of biocatalysts in industrial processes. In this study, modified magnetic iron oxide nanoparticles bearing tyrosinase (tyrosinase-MNPs) were employed as a magnetic nano-biocatalyst to treat phenol-containing wastewater. Different factors affecting the phenol removal efficiency of the fabricated nano-biocatalyst such as catalyst dosage, pH, temperature, initial phenol concentration, and reusability were investigated. The results proved that the precise dosage of nano-biocatalyst was able to degrade phenol at the wide range of pHs and temperatures. The immobilized tyrosinase showed proper phenol degradation more than 70%, where the substrate with a high concentration of 2500 mg/L was subjected to phenol removal. The nano-biocatalyst was highly efficient and reusable, since it displayed phenol degradation yields of 100% after the third reuse cycle and about 58% after the seventh cycle. Moreover, the immobilized tyrosinase was able to degrade phenol dissolved in real water samples up to 78% after incubation for 60 min. It was also reusable at least seven cycles in the real water sample. The results proved the effectiveness and applicability of the fabricated nano-biocatalyst to treat phenol-containing wastewaters in a shorter time and higher efficiency even at high phenol concentration. The developed nano-biocatalyst can be promising for the micropollutants removal and an alternative for the catalysts used in traditional treatment processes.


Survival in patients with heart failure: focus on captopril.
Congestive heart failure is a serious health-care problem that is associated with a high mortality. Although drug therapy has been effective in reducing symptoms, until recently there has been little evidence of prolonged survival. Within the past few years, however, certain vasodilator therapies, when added to a conventional regimen, have demonstrated a positive effect on longevity. In patients with moderately severe to severe heart failure, the combination of isosorbide dinitrate and hydralazine reduced mortality by 34%. In patients with severe heart failure, enalapril reduced one-year mortality by 31%. Captopril has also been shown to be effective in improving survival. In a recent analysis, the mortality rate of patients with moderately severe heart failure treated with captopril was 4% over three months, compared with 21% among placebo-treated patients. In a smaller study, 83% of captopril-treated patients, compared with 20% of hydralazine-treated patients, with moderately severe to severe heart failure survived for at least one year. The mechanisms through which captopril could potentially improve survival include hemodynamic, functional, structural, and neurohormonal variables.


Assessing antimicrobial resistance in Campylobacter jejuni and Campylobacter coli and its association with antimicrobial use in Canadian turkey flocks.
Turkeys are important sources of antimicrobial-resistant Campylobacter. A total of 1063 isolates were obtained from 293 turkey flocks across Canada between 2016 and 2021 to evaluate their antimicrobial resistance (AMR) prevalence, patterns, distribution, and association with antimicrobial use (AMU). A high proportion of C. jejuni and C. coli isolates were resistant to tetracyclines and fluoroquinolones, despite the very low use of these drugs. C. jejuni isolates had a higher probability of being resistant to tetracyclines than C. coli isolates. The chance of C. jejuni isolates being resistant to fluoroquinolones, macrolides, and lincosamides was lower compared to C. coli. Isolates from the western region had a higher probability of being resistant to fluoroquinolones than isolates from Ontario. Isolates from Ontario had higher odds of being resistant to tetracyclines than isolates from Quebec. No associations were noted between the resistance and use of the same antimicrobial, but the use of certain antimicrobial classes may have played a role in the maintenance of resistance in Campylobacter (fluoroquinolone resistance - bacitracin and streptogramin use, tetracycline resistance - flavophospholipids and streptogramins use, macrolide resistance - flavophospholipid use). Low-level multidrug-resistant Campylobacter was observed indicating a stable AMR in turkeys. This study provided insights aiding future AMU and AMR surveillance.


Economical Burden of Burn Injuries in a Developing Country.
Burn injuries have economical impacts on patients in several ways. Understanding the charges of burn treatment is very important for patients, families, governmental authorities, and insurance companies. During the protocol of their treatment, they may be admitted several times for treatment of acute burn and then for reconstructive treatments of burn's complications. Calculating the hospital burn charges can serve as an objective criteria for authorities to plan for a sufficient budget for acute burn treatment, for additional management for chronic complications, and as a guide for planning preventive and public educational programs. The authors used data of their burn registry program. During more than 3 years, the authors had 912 patients with multiple admissions for burns. All of hospital costs during several admissions were recorded. Men were 71% and women were 29% of the patients. Burns caused by flame were the most frequent (50.1%) followed by scald (34.0%). Mean hospital stay was 14.1 days (range, 0-64 days). Patients with TBSA equal to or less than 10% were 38.8%, TBSA between 11 and 22% were 29.1%, and TBSA more than 23% were 32.1%. Those who were admitted for 30 days or less were 34.1%, those between 31 and 131 days were 32.7%, and those with more than 132 days of admission were 33.2%. Mean hospital cost for all patients during the 3 years was about $2766 (range, from $143 to $33,566; median = 1586.93; SE = 93.84). The patients were admitted for treatment of acute burns and later admitted for reconstruction of the burn sequels. Total number of admissions was up to six times (median = 2). About 66.27% of the total charges were the cost of first admission, 19.39% the cost of second admission, 7.34% the cost of third admission, 3.56% for fourth admission, 2.3% for fifth admission, and 1.15% for last or sixth admission. The authors conducted a multiple linear regression test. Male sex, TBSA, length of stay, and number of admissions were significantly related to total treatment charges. But "age" did not influence the charges. Mean total cost of several burn admissions in one patient was around $2766. TBSA, length of stay, male sex, and number of admissions were significantly related to the hospital costs.


Isolation and characterization of cell-associated nucleases related to streptococcal extracellular deoxyribonuclease D.
A group of at least four distinct nucleases designated DcI through DcIV were isolated from cellular extracts of group A streptococcal strain S43 and shown to be antigenically similar to streptococcal extracellular deoxyribonuclease (DNase) D. These cellular endonucleases degraded single- and double-stranded deoxyribonucleic acid (DNA) as well as ribonucleic acid (RNA) to acid-soluble oligonucleotides. The products of digestion of DNA bore 5'-terminal phosphates, and in partial digests pdX-pdG linkages were most susceptible and pdA-pdX linkages were most resistant to nuclease action. The enzymes had pH optima of 8.0 to 8.5, were inhibited by NaCl, were unaffected by sulfhydryl modifying reagents, and absolutely required a divalent cation. Nucleases DcIII and DcIV were apparently hydrophobic in nature since they required the presence of detergents for migration on nondenaturing polyacrylamide gels. All four nucleases were electrophoretically distinct on such gels, from each other, and from DNase D. Molecular weights of DcI and DcII were similar to that of DNase D, suggesting that the mobility differences of these enzymes at least are reflections of differing net charges. It is suggested that the cellular nucleases represent a group of processing intermediates in the maturation and excretion of DNase D.


Some aspects of the teaching and learning of nursing.
The argument on which this paper is based is that the activities of teaching and learning nursing draw on a number of disciplines and fields of knowledge. To support this argument, elements in the process of teaching and learning in nursing are set out and each one is analysed and discussed. The elements so analysed are later put together to give a synthesis of the intended outcomes of the process of nursing education. The paper is supported with an extensive bibliography.


Effects of overexpression of huntingtin proteins on mitochondrial integrity.
Huntington's disease (HD) is caused by an expansion of a CAG trinucleotide sequence that encodes a polyglutamine tract in the huntingtin (Htt) protein. Expansion of the polyglutamine tract above 35 repeats causes disease, with the age of onset inversely related to the degree of expansion above this number. Growing evidence suggests that mitochondrial function is compromised during HD pathogenesis, but how this occurs is not understood. We examined mitochondrial properties of HeLa cells that expressed green fluorescent protein (GFP)- or FLAG-tagged N-terminal portions of the Htt protein containing either, 17, 28, 74 or 138 polyglutamine repeats. Immunofluorescence staining of cells using antibodies against Tom20, a mitochondrion localized protein, revealed that cells expressing Htt proteins with 74 or 138 polyglutamine repeats were more sensitized to oxidative stress-induced mitochondria fragmentation and had reduced ATP levels compared with cells expressing Htt proteins with 17 or 28 polyglutamine repeats. By measuring changes in fluorescence of a photoactivated GFP protein targeted to mitochondria, we found that cells expressing red fluorescent protein (RFP)-tagged Htt protein containing 74 polyglutamine repeats had mitochondria that displayed reduced movement and fusion than cells expressing RFP-Htt protein with 28 polyglutamine repeats. Overexpression of Drp-1(K38A), a dominant-negative mitochondria-fission mutant, or Mfn2, a protein that promotes mitochondria fusion, suppressed polyglutamine-induced mitochondria fragmentation, the reduction of ATP levels and cell death. In a Caenorhabditis elegans model of HD, we found that reduction of Drp-1 expression by RNA interference rescued the motility defect associated with the expression of Htt proteins with polyglutamine repeats. These results suggest that the increase in cytotoxicity induced by Htt proteins containing expanded polyglutamine tracts is likely mediated, at least in part, by an alteration in normal mitochondrial dynamics, which results in increased mitochondrial fragmentation. Furthermore, our results suggest that it might be possible to reverse polyglutamine-induced cytotoxicity by preventing mitochondrial fragmentation.


Japanese summer-type hypersensitivity pneumonitis: studies using Cryptococcus antigen.
Sixty-six patients, diagnosed as Japanese summer-type hypersensitivity pneumonitis at Osaka Prefectural Habikino hospital between 1973 and 1980, were studied. The diagnosis was based on the clinical features and summer-seasonal nature of the disease. The presence of an aetiological agent within patients' home environment was suggested by the recurrence of acute symptoms of high fever, cough and dyspnoea 5-8 hr after coming home from hospital, and by spontaneous improvement on leaving home. Immunological studies revealed the presence of anti-Cryptococcus antibody in sixty-four of sixty-five patients' sera, by indirect immunofluorescence against Cryptococcus neoformans. Precipitating antibody against culture supernatant protein-antigen of Cr. neoformans was detected in more than 80% of sera obtained from patients during the active stage of the disease. The positive result on inhalation provocation-challenge, using culture supernatant protein-antigen, suggested that Cr. neoformans or antigenically related Cryptococcus species may cause Japanese summer-type hypersensitivity pneumonitis.


Development and Evaluation of a Rapid Neutralizing Antibody Assay for COVID-19 Vaccination.
SARS-CoV-2 neutralizing antibodies have excellent application prospects in the prevention and treatment of COVID-19. This study established a competitive colloidal gold immunochromatography assay (GICA) to detect neutralizing antibodies against the receptor-binding domain (RBD) of SARS-CoV-2 in postvaccination serum. The sensitivity, stability, and specificity of GICA were evaluated using neutralizing antibody solution reference material and positive serum. The consistency and correlation between GICA, pseudovirus neutralization (PN) assay, and ELISA were compared. Consistency analysis of serum neutralizing antibody and specific IgG antibody titers was conducted, and changes in neutralizing antibodies and specific IgG antibodies in serum after inoculation with the homologous booster inactivated vaccine and recombinant vaccine were noted. The sensitivity of the reagent was 20.66 IU/L, and the specificity was 100%. There was a strong consistency and correlation between GICA and PN (κ = 0.886, n = 165; r = 0.918, P < 0.001). The correlation coefficient of serum anti-RBD antibody and specific IgG antibody titers was 0.5253 (P < 0.001). The specific IgG antibody titers in serum after (W4) inoculation with homologous booster inactivated vaccine were 10.80 (S/CO).The anti-RBD antibody titers were 28.33. The anti-RBD omicron variant (B.1.1.529) antibody titers were 11.67. After inoculation with the recombinant vaccine, the specific IgG antibody titers in the serum of W4 were 10.68. The serum anti-RBD antibody titers of W4 were 103.30. The serum anti-RBD omicron variant (B.1.1.529) antibody titers of W4 were 56.67. Therefore, vaccination of the third dose of the homologous booster inactivated vaccine and recombinant vaccine can enhance the level of neutralizing antibodies against the omicron variant (B.1.1.529). This study demonstrates that a GICA kit for neutralizing antibodies against the RBD of SARS-CoV-2 can be used for COVID-19 vaccine evaluation. Changes in titers enable long-term monitoring of a population's immunity and guide interventions when their immunity declines.


Susceptibilities of bacterial and fungal urinary tract isolates to desferrioxamine.
Desferrioxamine, a safe and effective iron chelator, was evaluated for its antibacterial and antifungal activity. The susceptibilities of 124 urinary tract isolates and 28 clinical isolates of Neisseria gonorrhoeae to desferrioxamine concentrations that are readily achievable in urine were determined. Of all isolates, 27% were inhibited by a concentration of less than or equal to 12.5 mM. Proteus mirabilis and N. gonorrhoeae isolates were particularly susceptible to the chelator. Desferrioxamine appears to have limited potential as an antibacterial agent.


Ca(2+) sensor S100beta-modulated sites of membrane guanylate cyclase in the photoreceptor-bipolar synapse.
This study documents the identity of a calcium- regulated membrane guanylate cyclase transduction system in the photoreceptor-bipolar synaptic region. The guanylate cyclase is the previously characterized ROS-GC1 from the rod outer segments and its modulator is S100beta. S100beta senses increments in free Ca(2+) and stimulates the cyclase. Specificity of photoreceptor guanylate cyclase activation by S100beta is validated by the identification of two S100beta-regulatory sites. A combination of peptide competition, surface plasmon resonance binding and deletion mutation studies has been used to show that these sites are specific for S100beta and not for another regulator of ROS-GC1, guanylate cyclase-activating protein 1. One site comprises amino acids (aa) Gly962-Asn981, the other, aa Ile1030-Gln1041. The former represents the binding site. The latter binds S100beta only marginally, yet it is critical for control of maximal cyclase activity. The findings provide evidence for a new cyclic GMP transduction system in synaptic layers and thereby extend existing concepts of how a membrane-bound guanylate cyclase is regulated by small Ca(2+)-sensor proteins.


[Grey scale and power Doppler ultrasonographic assessment of bone erosion and disease activity in early rheumatoid arthritis].
To evaluate the sensitivity and predictive value of grey scale and power Doppler ultrasound assessment of bone erosionin disease activity in patients with early rheumatoid arthritis (RA). Fifty-six patients with early RA underwent blinded sequential clinical, laboratory and ultrasound assessments, and at the same time 20 of these patients underwent X-ray and enhanced MRI. For each patient, 28-joint disease activity score (DAS28), erythrocyte sedimentation rate (ESR), C reactive protein (CRP) and health assessment questionnaire (HAQ) were recorded. The presence of bone erosion and synovitis was investigated in 28 joints by gray-scale and power Doppler ultrasonography. The ultrasound joint count and index for active synovitis with power Doppler signal were calculated. The number of bone erosions detected by ultrasonography was 5.7 times that of X-ray, while both MRI and ultrasonography were consistent (91.5%). The number of synovitis detected by ultrasonography was 1.6 times as much as by physical examination, and consistent MRI (95.7%). PDUS parameters demonstrated a highly significant correlation with DAS28, ESR and CRP, while a negative correlation with HAQ. Grey scale and power Doppler ultrasonography is a sensitive and reliable method to assess bone erosion and inflammatory activity in early RA. PDUS findings may have a predictive value in disease activity.


Quantification and reduction of organic sulfur compound formation in a commercial wood pulping process.
This study experimentally quantified the formation of organic sulfur compounds in a commercial SuperBatch kraft pulping process using a laboratory pilot-scale digester. The results indicate that wood chips not only can adsorb HS- but also methyl mercaptan (MM) in the black liquor used for pretreatment during the two pretreatment stages. The absorption rate of MM is much faster than that of HS-. In the third stage of SuperBatch pulping, the rate of formation of MM and dimethyl sulfide (DMS) is very similar to that found in conventional batch kraft pulping processes. The study examined the applicability of the phase transition cooking (PTC) concept for reducing organic sulfur compounds in pulping, previously developed in laboratory batch pulping using a bomb-type digester, in a SuperBatch process. It was confirmed that there is a phase transition point (PTP) corresponding to PTC in SuperBatch pulping beyond which further delignification significantly increases the formation of volatile organic sulfur compounds. The results indicate that a 40% reduction of TRS formation can be achieved by using PTC.


Aedes aegypti and Aedes albopictus Habitat Preferences in South Texas, USA.
The South Texas region has a historical record of occasional dengue outbreaks. The recent introduction of chikungunya virus to the Caribbean suggests that this disease may be a concern as well. Six different cities and three field habitat types (residential, tire shops, and cemeteries) were examined for evidence of habitat and longitudinal preference of two vector species, Aedes aegypti and Aedes albopictus. A. aegypti was more prevalent in tire shop sites, while A. albopictus was more prevalent in cemetery sites. In residential sites, the relative abundance of the two species varied with longitude, with A. albopictus being more abundant near the coast, and A. aegypti being more abundant inland. There was also a temporal variation, with A. aegypti declining in frequency over time in residential sites. These results have implications for control strategies and disease risk and suggest a greater need for increased surveillance and research in the region.


Estimated time spent on preventive services by primary care physicians.
Delivery of preventive health services in primary care is lacking. One of the main barriers is lack of time. We estimated the amount of time primary care physicians spend on important preventive health services. We analyzed a large dataset of primary care (family and internal medicine) visits using the National Ambulatory Medical Care Survey (2001-4); analyses were conducted 2007-8. Multiple linear regression was used to estimate the amount of time spent delivering each preventive service, controlling for demographic covariates. Preventive visits were longer than chronic care visits (M = 22.4, SD = 11.8, M = 18.9, SD = 9.2, respectively). New patients required more time from physicians. Services on which physicians spent relatively more time were prostate specific antigen (PSA), cholesterol, Papanicolaou (Pap) smear, mammography, exercise counseling, and blood pressure. Physicians spent less time than recommended on two "A" rated ("good evidence") services, tobacco cessation and Pap smear (in preventive visits), and one "B" rated ("at least fair evidence") service, nutrition counseling. Physicians spent substantial time on two services that have an "I" rating ("inconclusive evidence of effectiveness"), PSA and exercise counseling. Even with limited time, physicians address many of the "A" rated services adequately. However, they may be spending less time than recommended for important services, especially smoking cessation, Pap smear, and nutrition counseling. Future research is needed to understand how physicians decide how to allocate their time to address preventive health.


2,2-Dialkylnaphthalen-1-ones as new potassium channel activators.
A new series of 2,2-dialkylnaphthalen-1-one potassium channel activators has been prepared, and their in vitro relaxant activities in isolated rat portal vein and guinea pig tracheal spirals as well as their oral antihypertensive effect in spontaneously hypertensive rats have been evaluated. The group of 1,2-dihydro-4-(1,2-dihydro-2-oxo-1-pyridyl)-2,2-dimethylnaphthalen -1- ones with an electron-withdrawing substituent at the 6-position contain the most active compounds and 1,2-dihydro-4-(1,2-dihydro-2-oxo-1-pyridyl)-2,2-dimethyl-1-oxonaphtha lene-6- carbonitrile, 17f (UR-8225), has been selected for further pharmacological development.


The Role of Recollection and Familiarity in Nondemented Parkinson's Patients.
The aim of the current study was to examine if recollection and familiarity decline in nondemented Parkinson's patients. To do so we compared a sample of older people with Parkinson's disease (n = 32) to a control sample of healthy older people (n = 32) on an associative recognition task in which we manipulated the repetition of the pairs during the study phase (half of the pairs were presented once and half twice) to obtain corrected estimates of recollection, familiarity, and false recognition based on the logic of the process-dissociation procedure. The results clearly show that recollection is impaired but familiarity is preserved in nondemented Parkinson's patients. The results show that memory for pairs in Parkinson's patients relies largely on the familiarity of each item and not on a precise recollection of associative information, supporting the idea that recollection-based monitoring processes are impaired in these patients.


Enteropathogenic Escherichia coli infection leads to appearance of aberrant tight junctions strands in the lateral membrane of intestinal epithelial cells.
Infection of intestinal epithelial cells with enteropathogenic Escherichia coli (EPEC) disrupts tight junction (TJ) architecture and barrier function. The aim of this study was to determine the impact of EPEC on TJ protein interactions and localization. Human intestinal epithelial cells (T84) were infected for 1, 3 or 6 h with EPEC. To probe the TJ protein-protein interactions, co-immunoprecipitations were performed. The associations between ZO-1, occludin and claudin-1 progressively decreased after infection. Corresponding morphological changes were analysed by immunofluorescence confocal microscopy. Tight junction proteins progressively lost their apically restricted localization. Freeze-fracture electron microscopy revealed the appearance of aberrant strands throughout the lateral membrane that contained claudin-1 and occludin as determined by immunogold labelling. These structural alterations were accompanied by a loss of barrier function. Mutation of the gene encoding EspF, important in the disruption of TJs by EPEC, prevented the disruption of TJs. Tight junction structure normalized following eradication of EPEC with gentamicin and overnight recovery. This is the first demonstration that a microbial pathogen can cause aberrant TJ strands in the lateral membrane of host cells. We speculate that the disruption of integral and cytoplasmic TJ protein interactions following EPEC infection allows TJ strands to form or diffuse into the lateral plasma membrane.


[Preparation of giant RNA from bovine thyroid gland by agarose gel filtration].
Giant RNA's were prepared by a rapid gel-filtration technique. DNAase, pronase and thyroid homogenate were successively placed on the column. Proteins and DNA were degraded during filtration. Void volume contained pure giant RNA's. Giant RNAs' yield was 7 times higher with this method than with phenol method.


Human mesenchymal stromal cells and their derivative, SB623 cells, rescue neural cells via trophic support following in vitro ischemia.
Cell transplantation is a promising treatment strategy for many neurological disorders, including stroke, which can target multiple therapeutic mechanisms in a sustained fashion. We investigated the ability of human mesenchymal stromal cells (MSCs) and MSC-derived SB623 cells to rescue neural cells via trophic support following an in vitro stroke model. Following oxygen glucose deprivation, cortical neurons or hippocampal slices were cocultured with either MSCs or SB623 cells separated by a semiporous membrane (prohibits cell-cell contact) or with MSC- or SB623 cell-conditioned medium. MSCs, SB623 cells, MSC-conditioned media, and SB623 cell-conditioned media all significantly reduced neural cell damage/death compared to untreated conditions, and the rescue effect of the conditioned media was dose dependent. We identified 11 neurotrophic factors secreted by MSCs and/or SB623 cells. This study emphasizes the importance of trophic support provided by marrow-derived cells, which likely contributes to the efficacy of cell therapy for brain injury.


An electrochemical gate based on a stimuli-responsive membrane associated with an electrode surface.
The electrochemical gate based on a chemical signal-responsive membrane was assembled on a Au electrode surface. The polyelectrolyte gel membrane was capable to bind cholesterol because of the hydrogen bonding between cholesterol and the polymer backbone resulting in the gel swelling. The membrane channels were reversibly closed and opened upon addition and washing out cholesterol, respectively. Thus, the electrochemical process of a soluble redox probe, [Fe(CN)(6)](3-/4-), at the membrane-modified electrode was reversibly switched "on-off" by the cyclic addition and washing out cholesterol. The electrochemical reaction was also tuned by the variation of the concentration of the added cholesterol that controlled the extent of the channels closing. The switchable and tuneable operation of the chemically controlled electrochemical gate was characterized by Faradaic impedance spectroscopy and atomic force microscopy, indicating that the extent of the pores opening and closing is controlled by the concentration of the membrane-associated cholesterol. The chemical-responsive electrochemical gate was suggested to be a part of future biochemical/electrochemical systems with logic operations.


Comprehensive review on phytotechnology: Heavy metals removal by diverse aquatic plants species from wastewater.
Environmental pollution specifically water pollution is alarming both in the developed and developing countries. Heavy metal contamination of water resources is a critical issue which adversely affects humans, plants and animals. Phytoremediation is a cost-effective remediation technology which able to treat heavy metal polluted sites. This environmental friendly method has been successfully implemented in constructed wetland (CWs) which is able to restore the aquatic biosystem naturally. Nowadays, many aquatic plant species are being investigated to determine their potential and effectiveness for phytoremediation application, especially high growth rate plants i.e. macrophytes. Based on the findings, phytofiltration (rhizofiltration) is the sole method which defined as heavy metals removal from water by aquatic plants. Due to specific morphology and higher growth rate, free-floating plants were more efficient to uptake heavy metals in comparison with submerged and emergent plants. In this review, the potential of wide range of aquatic plant species with main focus on four well known species (hyper-accumulators): Pistia stratiotes, Eicchornia spp., Lemna spp. and Salvinia spp. was investigated. Moreover, we discussed about the history, methods and future prospects in phytoremediation of heavy metals by aquatic plants comprehensively.


Laryngeal manifestations of gastroesophageal reflux disease in children.
Although the association between gastroesophageal reflux disease (GERD) and laryngeal disorders in adults is well established there is still a lack of information concerning the true extent of the laryngeal complications of GERD in children. The aim of this study was to determine the laryngeal status of children with diagnosed GERD. We sought to identify the initial appearance of their larynges and then to determine the clinical response to antireflux therapy. GERD was recognized in 90/100 children examined. Using 24-h pH monitoring we found that most of the patients experienced episodes of gastroesophageal reflux during the daytime when they were in an upright position. The hallmark of GERD affecting the larynx in our group was posterior laryngitis, which is characterized by erythema of the mucous membrane overlying the arytenoid cartilages and the posterior mucosal wall of the glottis. The findings regarding the effectiveness of therapy were that, in children with severe laryngeal alterations, voice quality improved significantly after 12 weeks of antireflux treatment (p < 0.001) and laryngeal status was significantly better after 6 weeks of treatment (p < 0.001). This study provides evidence that gastroesophageal reflux in children is the underlying cause of inflammatory and morphological lesions, and that antireflux treatment is effective in reducing or eliminating these lesions.


Negative affective spillover from daily events predicts early response to cognitive therapy for depression.
This study evaluated the predictive role of depressed outpatients' (N = 62) affective reactivity to daily stressors in their rates of improvement in cognitive therapy (CT). For 1 week before treatment, patients completed nightly electronic diaries that assessed daily stressors and negative affect (NA). The authors used multilevel modeling to compute each patient's within-day relationship between daily stressors and daily NA (within-day reactivity), as well as the relationship between daily stressors and next-day NA (next-day reactivity; affective spillover). In growth model analyses, the authors evaluated the predictive role of patients' NA reactivity in their early (Sessions 1-4) and late (Sessions 5-12) response to CT. Within-day NA reactivity did not predict early or late response to CT. However, next-day reactivity predicted early response to CT, such that patients who had greater NA spillover in response to negative events had a slower rate of symptom change during the first 4 sessions. Affective spillover did not influence later response to CT. The findings suggest that depressed patients who have difficulty bouncing back the next day from their NA reactions to a relative increase in daily negative events will respond less quickly to the early sessions of CT.


Newly Diagnosed Glomerulonephritis During COVID-19 Infection Undergoing Immunosuppression Therapy, a Case Report.
During the COVID-19 pandemic, we had a 25 years old male case without any underlying disease or history of autoimmune disease in COVID-19 Clinic, Isfahan, Iran. He presented with arthralgia and weakness so we started COVID-19 therapeutic regimen. In his hospitalization, creatinine increases and abnormalities in random urine sediment was seen. Methylprednisolone and cyclophosphamide were prescribed due to suspected glomerulonephritis. After renal biopsy the diagnose was confirmed as crescentic proliferative glomerulonephritis. The patient also, underwent plasmapheresis and intravenous immunoglobulin injection. He was discharged healthy without development of new pulmonary symptoms despite immunosuppressive treatment.


European brown hare syndrome in northern Italy: results of a virological and serological survey.
Between August 1988 and August 1991, 456 carcasses of captive or sylvatic hares from several areas of northern Italy, and 931 sera taken from adult hares in farms, in hunting and natural reserves and on importation were examined using virological (sandwich enzyme-linked immunosorbent assay [ELISA] and immuno-electron microscopy) and serological (competition ELISA) tests. The epidemiological data presented relate to the incidence of European brown hare syndrome (EBHS) in various provinces of northern Italy, the mortality caused by EBHS and the seasonal frequency of this disease. The endemic character of EBHS in Italy is proved by the large number of samples testing positive for EBHS virus (EBHSV) (47.6%) and by the results of the seroepidemiological survey, in which approximately 95% of samples tested positive for specific anti-EBHSV antibodies, showing varying titres according to the different environmental conditions.


New targeted approaches in chronic myeloid leukemia.
The treatment of chronic myeloid leukemia has changed dramatically in the last few years. Stem-cell transplantation and the use of interferon alfa had already offered the possibility of complete and durable cytogenetic responses, improving the survival over that expected with conventional chemotherapy. The introduction of imatinib mesylate has started the era of molecular therapy with remarkable results including complete cytogenetic responses in up to 90% of patients and major molecular responses in most. However, some patients, particularly those treated in the advanced stages, may develop resistance to imatinib. Thus there has been interest in developing new agents that would not only help patients for whom imatinib is ineffective or intolerable, but that could also be combined with the intention of eliminating all evidence of disease. Several approaches are being pursued. These include new and more potent tyrosine kinase inhibitors that may not be affected by the most common mutations seen in the clinic. Some of these agents also inhibit Src-related kinases that may play a role in the development of resistance to imatinib. Other agents are directed at downstream or alternative pathways in leukemic cells, exploring potential synergy with imatinib. Another approach is to pursue an immune modulation that might eliminate small amounts of residual disease. Many of these agents are already showing promising results in the clinic. This manuscript reviews some of these agents, particularly those for which clinical data are already available.


Assessment of tachyphylaxis following prolonged therapy of asthma with inhaled albuterol aerosol.
Controversy exists concerning possible tachyphylaxis of the acute bronchodilating effect of albuterol, especially with regard to the duration of its acute bronchodilating action. We evaluated 140 patients with bronchial asthma in a prospective double-blind controlled study of possible tachyphylaxis to albuterol aerosol as compared to isoproterenol aerosol. We demonstrated statistically significant tachyphylaxis with regard to duration of acute bronchodilating effect. We believe that this tachyphylaxis is not clinically significant because there was no tachyphylaxis with regard to peak bronchodilating effect and because the duration of bronchodilating effect remains significantly greater, both quantitatively and statistically, when compared to isoproterenol aerosol. Moreover, it appeared that most of the tachyphylaxis was present at four weeks of therapy. There was a small increment of tachyphylaxis after eight weeks of therapy, but no further increase in tachyphylaxis was demonstrated after 13 weeks of inhaled albuterol therapy. We therefore feel that clinically significant tachyphylaxis to inhaled albuterol aerosol must be quite unusual and that chronic therapy with inhaled albuterol aerosol is probably both safe and efficacious for bronchospastic disorders.


Diagnosis and clinical implications of pancreatobiliary reflux.
The sphincter of Oddi is located at the distal end of the pancreatic and bile ducts and regulates the outflow of bile and pancreatic juice. A common channel can be so long that the junction of the pancreatic and bile ducts is located outside of the duodenal wall, as occurs in pancreaticobiliary maljunction (PBM); in such cases, sphincter action does not functionally affect the junction. As the hydropressure within the pancreatic duct is usually greater than in the bile duct, pancreatic juice frequently refluxes into the biliary duct (pancreatobiliary reflux) in PBM, resulting in carcinogenetic conditions in the biliary tract. Pancreatobiliary reflux can be diagnosed from elevated amylase level in the bile, secretin-stimulated dynamic magnetic resonance cholangiopancreatography, and pancreatography via the minor duodenal papilla. Recently, it has become obvious that pancreatobiliary reflux can occur in individuals without PBM. Pancreatobiliary reflux might be related to biliary carcinogenesis even in some individuals without PBM. Since few systemic studies exist with respect to clinical relevance and implications of the pancreatobiliary reflux in individuals with normal pancreaticobiliary junction, further prospective clinical studies including appropriate management should be performed.


The importance of two metabolic syndrome diagnostic criteria and body fat distribution in predicting clinical severity and prognosis of acute myocardial infarction.
The interrelation between metabolic syndrome (MetS) (the revised National Cholesterol Education Program-Adult Treatment Panel III (NCEP-ATP III) and International Diabetes Federation (IDF)) and obesity indices in predicting clinical severity and prognosis of acute ST-elevation myocardial infarction (STEMI) is insufficiently known. This prospective study included 250 acute STEMI patients treated with primary percutaneous coronary intervention. The patients with/without MetS were analyzed by baseline (medical history, demography and obesity indices: overall - body mass index (BMI) vs.central - body adiposity index (BAI), conicity index (Cindex), visceral adiposity index (VAI), waist circumference (WC), waist-to-hip (WHR) and waist-to-height ratio (WHtR)), severity (clinical presentation, laboratory, echocardiography, coronary angiography and in-hospital complications) and prognostic parameters (major adverse cardiovascular events and sick leave duration during 12-month follow-up). There were 136 (54.4%) and 147 (58.8%) patients with MetS (NCEP-ATP III) and MetS (IDF), respectively. MetS (NCEP-ATP III) increased the risk of > 1 significantly stenosed coronary artery (CA), very high BAI increased the risk of dyspnea, Cindex > 1.25/1.18 increased the risk of total in-hospital complications, increased VAI increased the risk of coronary segment 3 significant stenosis, WHR ≥ 0.90/0.85 increased the risk of proximal/middle coronary segments (especially of segment 1) significant stenosis, WHtR ≥ 63/58 increased the risk of heart failure, and the number of significantly stenosed CAs increased the risk of total MACE (p < 0.05). MetS (NCEP-ATP III) and several central obesity indices are superior to BMI in predicting acute STEMI severity (clinical presentation, in-hospital complications, severity of coronary disease), while WC and MetS (IDF) have no influence on it. They all have no influence on prognosis.


Brotizolam and chronic insomnia: a multi-centre study.
A double-blind, crossover study was carried out on the acceptability of three doses of brotizolam (0.125, 0.25 and 0.5 mg) in chronic insomniacs aged between 21 and 75 years (33 men: 42 women). Patients reported a shorter time to fall asleep and less nocturnal awakenings. Improvement in sleep was evident during the first week of the study when each patient received 0.25 mg. There were no dose-related side-effects, and on withdrawal from the medication there was no evidence of disturbed sleep which would have suggested a rebound effect.


Post-Severe-COVID-19 Cardiopulmonary Rehabilitation: A Comprehensive Study on Patient Features and Recovery Dynamics in Correlation with Workout Intensity.
The aftermath of severe COVID-19 frequently involves considerable cardiopulmonary damage, necessitating rehabilitation. This study aimed to evaluate the impact of COVID-19 on cardiopulmonary health and assess the effectiveness of various rehabilitative interventions. Conducted between September 2021 and September 2022, this prospective study included patients who had been diagnosed with severe COVID-19 and admitted at the "Victor Babes" Infectious Diseases and Pulmonology Hospital, Timisoara, Romania. The patients were stratified into low- and high-intensity rehabilitation groups. The rehabilitation protocols were individually tailored, and the patient recovery was closely monitored over a 3-month period. Our cohort comprised 84 patients, with a mean age of 56.3 years for the low-intensity group (n = 42) and 53.1 years for the high-intensity group (n = 42). Both groups showed significant improvements in the lung injury area, need for oxygen supplementation, ejection fraction, systolic pulmonary artery pressure, and forced vital capacity. Additionally, considerable enhancements were observed in maximal voluntary ventilation, FEV1, FEV1/FVC ratio, peak expiratory flow, and forced expiratory flow at 25-75%. The work intensity also demonstrated substantial improvements from the initial testing to the 3-month mark in both groups. This study provides evidence that personalized, targeted rehabilitation strategies can improve long-term cardiopulmonary health in patients recovering from severe COVID-19, proving both low-intensity and high-intensity training as sufficient to improve heart and lung function if performed correctly and over a relatively short duration of 3 months. The study findings underscore the importance of implementing comprehensive cardiopulmonary rehabilitation protocols in the care of post-COVID-19 patients and highlight the value of stratified rehabilitation intensity based on individual patient dynamics and recovery features.


From developmental disorder to heritable cancer: it's all in the BMP/TGF-beta family.
Transforming growth factor-beta (TGF-beta) regulates many cellular processes through complex signal-transduction pathways that have crucial roles in normal development. Disruption of these pathways can lead to a range of diseases, including cancer. Mutations in the genes that encode members of the TGF-beta pathway are involved in vascular diseases as well as gastrointestinal neoplasia. More recently, they have been implicated in Cowden syndrome, which is normally associated with mutations in the phosphatase and tensin homologue gene PTEN. Molecular studies of TGF-beta signalling are now showing why mutations in genes that encode components of this pathway result in inherited cancer and developmental diseases.


Analysis of the virulence-associated RevSR two-component signal transduction system of Clostridium perfringens.
Clostridium perfringens is a Gram-positive, anaerobic, spore-forming bacterium that causes human gas gangrene (clostridial myonecrosis) and food poisoning. Early studies showed that virulence was regulated by the VirSR two-component signal transduction system. However, our identification of the RevR orphan response regulator indicated that more than one system was involved in controlling virulence. To further characterize this virulence-associated regulator, gel mobility shift experiments, coupled with DNase I footprinting, were used to identify the RevR DNA binding sequence. Bioinformatics analysis suggested that an orphan sensor histidine kinase, CPE1757 (renamed RevS), was the cognate sensor of RevR. Interaction between RevS and RevR was demonstrated by use of a bacterial two-hybrid system and validated by protein-protein interaction studies using biolayer interferometry. To assess the involvement of RevS in virulence regulation, the revS gene was inactivated by Targetron insertion. When isogenic wild-type, revS and complemented revS strains were tested in a mouse myonecrosis model, the revS mutant was found to be attenuated in virulence, which was similar to the attenuation observed previously with the revR mutant. However, transcriptional analysis of selected RevR-regulated genes in the revS mutant revealed a different pattern of expression to a revR mutant, suggesting that the RevSR system is more complex than originally thought. Taken together, the results have led to the identification and characterization of the two essential parts of a new regulatory network that is involved in the regulation of virulence in C. perfringens.


Antibody response to Hepatozoon canis in experimentally infected dogs.
Canine hepatozoonosis is a disease caused by the tick-borne protozoan Hepatozoon canis. Five puppies were inoculated by ingestion of Rhipicephalus sanguineus ticks experimentally infected with H. canis, and all became infected with H. canis: gametocytes were detected in blood smears from four dogs and schizonts were observed in the spleen and bone marrow of the fifth. Antibodies reactive with H. canis gametocytes were detected by the indirect fluorescent antibody test (IFA), with IgM detected initially in all dogs 16 to 39 days post infection (PI) and IgG 22 to 43 days PI. The presence of gametocytes was first observed within peripheral blood neutrophils in Giemsa-stained blood smears between days 28 and 43 PI. Gametocyte-reactive antibodies were detected before the appearance of blood gametocytes in three of the four parasitemic dogs and also in a dog with no observed parasitemia. The detection of serum antibodies prior to the detection of blood gametocytes, or without apparent parasitemia, suggests that antibodies reactive with gametocytes may be formed against earlier forms of the parasite developing in the parenchymal tissues. Sera of dogs experimentally infected with Babesia canis, Babesia gibsoni and Ehrlichia canis exhibited no reactivity when tested with H. canis antigen. Additionally, sera positive for H. canis were not reactive with antigens of Toxoplasma gondii, Neospora caninum, Leishmania donovani and E. canis. In conclusion, incoculation of dogs with ticks infected with H. canis results in production of antibodies reactive with peripheral blood gametocytes. Detection of IgG titres would be beneficial for the diagnosis of progressive infections with undetectable parasitemia, for seroprevalence studies, and as an adjunct to IgM titres in early infections.


Organ of hearing and balance in peri- and postmenopausal women. Effects of hormone replacement therapy on hearing and balance in peri- and post-menopausal women: The current state of knowledge.
Sensory-neural hearing loss in people over 50 - senile deafness - is currently an increasing problem due to the growing proportion of the elderly people in the population. We can find a number of worldwide publications on hearing loss and dizziness in peri-menopausal women, but the data is inconsistent. Reports describing the influence of sex hormones on the ear and its aging are similarly controversial. This review attempts to summarize the current state of knowledge regarding hearing and balance in women during menopause, based on recent studies in this field. It describes the possible causes of hearing and balance impairment in post-menopausal women other than the mere physiological aging of the ear. The review concludes that impairment of hearing and balance is significantly related to osteoporosis, which results from estrogen deficiency during menopause. It also presents the results of studies on the impact of hormone replacement therapy (HRT) and of particular hormonal components of HRT on hearing and balance. The European population is aging, so accurate knowledge about the effects of HRT on hearing and balance is crucial in the context of improving the quality of life of elderly women through appropriate qualification for hormone substitution therapy. Further detailed and extensive study is necessary to verify the impact of sex hormones on hearing in post-menopausal women, taking into account the type of HRT used, serum hormone levels and the presence or absence of osteoporosis.


Effects of nicotine on zebrafish: A comparative response between a newly established gill cell line and whole gills.
A novel cell line, Danio rerio gill (DrG), derived from the gill tissue of zebrafish, was established and characterized. The cells were able to grow at a wide range of temperatures from 25°C to 32°C in Leibovitz's L-15 medium. The DrG cell line consists of epithelial-like cells with a diameter of 18-22μm. The cell line was characterized by mitochondrial 12S rRNA gene. Acute toxicity tests were conducted on D. rerio by exposing them to nicotine for 96h under static conditions. In vitro cytotoxicity of nicotine was assessed in DrG cell line using multiple endpoints such as 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), Neutral Red assay, Alamar Blue assay and Coomassie Blue protein assay. Linear correlations between each in vitro cytotoxicity assay and the in vivo mortality data were highly significant. Nicotine induced intracellular reactive oxygen species generation in DrG cell line in a concentration dependent manner. DrG cell line and zebrafish exposed to nicotine significantly increased the elevation of lipid peroxidation (LPO) while depletion of reduced glutathione (GSH), manganese superoxide dismutase (MnSOD), catalase (CAT), glutathione S-transferase (GST) and glutathione peroxidise(GPx1a) was observed. In nicotine treated fish and cells a negative correlation between reduced glutathione and LPO was observed. In addition, the production of ROS and the resulting oxidative stress resulted in increased expression of apoptosis related genes p53 and cas3.Collectively, our result suggests that nicotine has the potential to induce reactive oxygen species (ROS) production, oxidative stress and apoptosis in DrG cell line and zebrafish.


Clinical risk factors for in-hospital mortality in older adults with HIV infection: findings from a South African hospital administrative dataset.
The proportion of older South African adults (aged ≥50 years old) with HIV infection requiring hospitalization is likely to increase in the near future. Clinical risk factors for in-hospital mortality (IHM) in these patients are not well described. We aimed to identify clinical risk factors associated with IHM and their overall contribution towards IHM in older South African adults with HIV infection. Clinical data for 690 older adults with HIV infection was obtained from the hospital administrative database at the Hlabisa Hospital in KwaZulu-Natal, South Africa. Logistic regression was used to determine independent clinical risk factors for IHM. Population-attributable fractions (PAFs) were calculated for all independent clinical risk factors identified. Male gender (p=0.005), CD4 count <350 cells/mm3 (p=0.035), unknown CD4 count (p=0.048), tuberculosis (p=0.033) and renal failure (p=0.013) were independently associated with IHM. Male gender contributed the most to IHM (PAF=0.22), followed by unknown CD4 count (PAF=0.14), tuberculosis (PAF=0.12), renal failure (PAF=0.06) and CD4 count <350 cells/mm3 (PAF=0.01). Although further research is required to confirm our findings, there is potential for these clinical risk factors identified in our study to be used to stratify patient risk and reduce IHM in older adults with HIV infection.


Immunoreactivity of recombinant human glandular kallikrein using monoclonal antibodies raised against prostate-specific antigen.
The gene encoding human glandular kallikrein (KLK2) was expressed in Escherichia coli, and the corresponding protein (hK2) was produced by fermentation. The hK2 was characterized by Western blotting and epitope map using monoclonal antibodies (MAbs) specific for another protease, prostate-specific antigen (PSA) with high structural identity (80%). MAbs that recognized three different epitopes were bound to hK2, representing 7 out of 23 MAbs tested. One epitope was localized to the sequence region around amino acid position 78, which is believed to be glycosylated in hK2. The affinities of MAbs recognizing hK2 were similar to those for PSA, suggesting that common epitopes seem to contain very conserved structures. The results may help in designing specific diagnostic assays for the assessment of prostate cancer.