Liver fibrosis and MAFLD: the exploration of multi-drug combination therapy strategies.
In recent years, the prevalence of metabolic-associated fatty liver disease (MAFLD) has reached pandemic proportions as a leading cause of liver fibrosis worldwide. However, the stage of liver fibrosis is associated with an increased risk of severe liver-related and cardiovascular events and is the strongest predictor of mortality in MAFLD patients. More and more people believe that MAFLD is a multifactorial disease with multiple pathways are involved in promoting the progression of liver fibrosis. Numerous drug targets and drugs have been explored for various anti-fibrosis pathways. The treatment of single medicines is brutal to obtain satisfactory results, so the strategies of multi-drug combination therapies have attracted increasing attention. In this review, we discuss the mechanism of MAFLD-related liver fibrosis and its regression, summarize the current intervention and treatment methods for this disease, and focus on the analysis of drug combination strategies for MAFLD and its subsequent liver fibrosis in recent years to explore safer and more effective multi-drug combination therapy strategies.


Nursing in a changing NHS.
In the past few weeks, I have had the privilege of attending a series of seminars exploring the themes emerging from the Mid Staffordshire Public Inquiry to date. They are highly relevant to nurses and to the future of nursing. They have explored system and professional regulation, organizational culture, board leadership, information systems, nurse training, patient experience and commissioning.


Nucleotide sequence of rat liver tyrosine aminotransferase gene fragment.
The sequence of a 3677 nucleotide EcoRI fragment was determined that codes for part of the rat liver tyrosine aminotransferase gene. The sequence was compared with the previously determined cDNA sequence and the intron and exon boundaries were deduced.


The role of renal surgery in the era of targeted therapy: the urologist's perspective.
Renal cell carcinoma (RCC) is increasingly diagnosed at an early stage in many countries, which has likely contributed to the recent leveling of RCC mortality. Nevertheless, despite these advances in diagnosis, up to 30% of patients have metastatic RCC at the time of diagnosis, and around 20-30% of subjects undergoing surgery will suffer recurrence. Following the recent approval and introduction in clinical practice of targeted therapies, the role and timing of surgery is being debated. In particular, targeted therapy agents have shown more favorable response rates than immunotherapy in subjects with advanced disease, and this increased efficacy raises the question whether cytoreductive nephrectomy is still beneficial in this subset of patients. On the basis of the current clinical studies, renal surgery seems to play a fundamental role also in the modern targeted therapy era. The optimal timing of surgery and the role of neoadjuvant treatment in advanced cases are still unclear. Furthermore modern urologists must acquire clinical skills in administering neoadjuvant and adjuvant therapy with noncytotoxic therapies, according to a multidisciplinary model of care for the management of patients with advanced RCC.


Risk assessment and dose-effect of co-exposure to benzene, toluene, ethylbenzene, xylene, and styrene (BTEXS) on pulmonary function: A cross-sectional study.
Inhalation is the most frequent route and the lung is the primary damaged organ for human exposure to benzene, toluene, ethylbenzene, xylene, and styrene (BTEXS). However, there is limited information on the risk and dose-effect of the BTEXS mixture on pulmonary function, particularly the overall effect. We conducted a cross-sectional study in a petrochemical plant in southern China. Spirometry and cumulative exposure dose (CED) of BTEXS were used to measure lung function and exposure levels for 635 workers in 2020, respectively. Forced vital capacity (FVC) and forced expiratory volume in 1 s (FEV1) were tested and interpreted as percentages to predicted values [FVC or FEV1% predicted], and FEV1 to FVC ratio [FEV1/FVC (%)]. We found the reduction in FVC% predicted and the risk of lung ventilation dysfunction (LVD) and its two subtypes (mixed and restrictive ventilation dysfunction, MVD, and MVD) were significantly associated with BTEXS individuals. In addition, pulmonary function damage associated with BTEXS was modified by the smoking status and age. Generalized weighted quantile sum (gWQS) regressions were used to estimate the overall dose-effect on lung function damage induced by the BTEXS mixture. Our results show wqs, an index of weighted quartiles for BTEXS, was potentially associated with the reduction in FVC and FEV1% predicted with the coefficients [95% confidence intervals (CI)] between -1.136 (-2.202, -0.070) and -1.230 (-2.265, -0.195). Odds ratios (ORs) and 95% CIs for the wqs index of LVD, MVD, and RVD were 1.362 (1.129, 1.594), 1.323 (1.084, 1.562), and 1.394 (1.096, 1.692), respectively. Furthermore, xylene, benzene, and toluene in the BTEXS mixture potentially contribute to the development of lung function impairment. Our novel findings demonstrated the dose-response relationships between pulmonary function impairment and the BTEXS mixture and disclosed the potential key pollutants in the BTEXS mixture.


[Nitric Oxide Signaling Pathway and Vascular Activity].
Nitric oxide-cyclic guanosine 3',5'-monophosphate (cGMP)-cGMP dependent protein kinase signaling pathway is a key mechanism for the modulation of vascular function. The progress in this area of recent years was briefly summarized in this article, in particular, regarding the redox-dependent dimerization of the relevant signaling molecules and their physiological significance as well as the role of soluble guanylyl cyclase-derived cyclic inosine 3',5'-monophosphate as a new messenger for vasoconstriction.


Market Impact of Foot-and-Mouth Disease Control Strategies: A UK Case Study.
Foot-and-mouth disease (FMD) poses a serious threat to the agricultural sector due to its highly contagious nature. Outbreaks of FMD can lead to substantial disruptions to livestock markets due to loss of production and access to international markets. In a previously FMD-free country, the use of vaccination to augment control of an FMD outbreak is increasingly being recognized as an alternative control strategy to direct slaughtering [stamping-out (SO)]. The choice of control strategy has implications on production, trade, and hence prices of the sector. Specific choice of eradication strategies depends on their costs and benefits. Economic impact assessments are often based on benefit-cost framework, which provide detailed information on the changes in profit for a farm or budget implications for a government (1). However, this framework cannot capture price effects caused by changes in production due to culling of animals; access to international markets; and consumers' reaction. These three impacts combine to affect equilibrium within commodity markets (2). This paper provides assessment of sectoral level impacts of the eradication choices of FMD outbreaks, which are typically not available from benefit-cost framework, in the context of the UK. The FAPRI-UK model, a partial equilibrium model of the agricultural sector, is utilized to investigate market outcomes of different control strategies (namely SO and vaccinate-to-die) in the case of FMD outbreaks. The outputs from the simulations of the EXODIS epidemiological model (number of animals culled/vaccinated and duration of outbreak) are used as inputs within the economic model to capture the overall price impact of the animal destruction, export ban, and consumers' response.


Retrieval of a dropped corneal button from a vitrectomized eye.
A successful retrieval of a dropped corneal button from an aphakic vitrectomized eye during penetrating keratoplasty is reported. A 52-year-old female patient underwent penetrating keratoplasty for aphakic corneal edema in a vitrectomized eye. The donor button inadvertently dropped into the vitreous cavity and was retrieved successfully. The dropped corneal button was visualized after focusing the operating microscope on the retina. The visualization significantly improved after aspiration of the fluid from the vitrectomized eye. The donor button was picked up by McPherson's forceps and subsequently sutured to the recipient's cornea. Corneal surgeons need to be aware of this rare complication in aphakic vitrectomized eyes and its retrieval after accurate visualization.


Clinical course of factor VIII inhibitors developed after exposure to a pasteurised Dutch concentrate compared to classic inhibitors in hemophilia A.
After the introduction of a new pasteurised factor VIII concentrate (Factor VIII CPS-P) in The Netherlands in June 1990, an increase in the occurrence of inhibitors in hemophilia A patients was reported. The clinical course of this group of inhibitors (n = 12) was compared with hemophilia patients in whom an inhibitor developed before June 1990 (classic inhibitors) (n = 32). Striking differences were found between both groups not only in patient age (median 22 years versus 8 years) and number of exposure days (< 50 - > 1000 versus < 50), as described in previous reports, but also in clinical course and response to treatment. In the recent group of inhibitors antibody titers showed a rapid decline when product was changed which was not the case in the group with classic inhibitors. In the group of classic inhibitors immune tolerance therapy with low dose factor VIII succeeded in 83%. Success was to a high degree dependent on the inhibitor level. In the group of recent inhibitors immune tolerance with the same concentrate was only successful in a single patient. However, once the patients were switched to another concentrate, antibody levels dropped to less than 2 BU/ml within 8 months in all patients. It seems likely that in this group of product associated inhibitors, treatment success was due to elimination of antigen stimulation rather than induction of immune tolerance.


[Quantitative study of vascular endothelial growth factor expression in primary lung cancer].
To study the expression levels of vascular endothelial growth factor (VEGF) and its relation with intratumoral microvessel density (MD), cellular proliferation activity (PI), and lymph node metastasis of primary lung cancer. Surgical samples of 53 patients with primary lung cancer were measured. The VEGF was immunofluorescence stained with VEGF polyclonal antibody. The propidium iodide was used to stain cellular DNA. VEGF expression levels and DNA content were analyzed quantitatively using flow cytometry. Intratumoral microvessel density was detected by factor VIII polyclonal antibody using immunohistochemistry. VEGF expression levels (FI) were high in lung cancer (1.3 +/- 0.6, positive ratio 73.58%). The cellular proliferation activity (PI) in VEGF-positive tumors was significantly higher than that in VEGF-negative tumors (29.8% +/- 9.2% vs 18.7% +/- 6.5%, P < 0.01). The intratumoral microvessel density (MD) in VEGF-positive tumors was higher than that in VEGF-negative tumors (19.4 +/- 11.3 vs 12.3 +/- 8.0, P < 0.05). No association was found between the MD and PI (r = 0.169, P > 0.05). The VEGF expression levels in lung cancer tissues with lymph node metastasis (n = 33) were significantly higher than that in those without lymph node metastasis (n = 20) (1.5 +/- 0.6 vs 1.0 +/- 0.5, P < 0.01). VEGF plays an important role in the growth and metastasis of lung cancer.


Canonical Wnt signaling in the notochordal cell is upregulated in early intervertebral disk degeneration.
The notochordal cell (NC) of the nucleus pulposus (NP) is considered a potential NP progenitor cell, and early intervertebral disk (IVD) degeneration involves replacement of NCs by chondrocyte-like cells (CLCs). Wnt/β-catenin signaling plays a crucial role in maintaining the notochordal fate during embryogenesis, but is also involved in tissue degeneration and regeneration. The canine species, which can be subdivided into non-chondrodystrophic and chondrodystrophic breeds, is characterized by differential maintenance of the NC: in non-chondrodystrophic dogs, the NC remains the predominant cell type during the majority of life, with IVD degeneration only occurring at old age; conversely, in chondrodystrophic dogs the NC is lost early in life, with concurrent degeneration of all IVDs. This study investigated Wnt/β-catenin signaling in the healthy, NC-rich NP and early degenerated, CLC-rich NP of both breed types by immunohistochemistry of β-catenin and relative gene expression of brachyury and cytokeratin 8 (notochordal markers) and Wnt targets axin2, cyclin D1, and c-myc. Both NCs and CLCs showed nuclear and cytoplasmic β-catenin protein expression and axin2 gene expression, but β-catenin signal intensity and Wnt target gene expression were higher in the CLC-rich NP. Primary NCs in monolayer culture (normoxic conditions) showed Wnt/β-catenin signaling comparable to the in vivo situation, with increased cyclin D1 and c-myc gene expression. In conclusion, Wnt/β-catenin signaling activity in the NC within the NC-rich NP and in culture supports the role of this cell as a potential progenitor cell; increased Wnt/β-catenin signaling activity in early IVD degeneration may be a reflection of its dual role.


Relative contributions of expressive behavior and contextual information to the judgment of the emotional state of another.
This study used a technique for assessing the relative impact of facial-gestural expressions, as opposed to contextual information regarding the elicitor and situation, on the judgment of emotion. In Study 1, 28 undergraduates rated videotapes of spontaneous facial-gestural expressions and separately rated the emotionally loaded color slides that elicited those expressions. The source clarities of the expressions and slides were matched using correlation and distance measures, and 18 expressions and 9 slides were selected. In Study 2, 72 undergraduate receivers were shown systematic pairings of these expressions and slides and rated the emotional state of the expresser, who was supposedly watching that slide under public or private situational conditions. Expressions were found to be more important sources for all emotion judgments. For female receivers slides were relatively more important in the public than in the private situation.


Analysis of one-sided marker segregation patterns resulting from mammalian gene targeting.
The double-strand break repair (DSBR) model is currently accepted as the paradigm for acts of double-strand break (DSB) repair that lead to crossing over between homologous sequences. The DSBR model predicts that asymmetric heteroduplex DNA (hDNA) will form on both sides of the DSB (two-sided events; 5:3/5:3 segregation). In contrast, in yeast and mammalian cells, a considerable fraction of recombinants are one sided: they display full conversion (6:2 segregation) or half-conversion (5:3 segregation) on one side of the DSB together with normal 4:4 segregation on the other side of the DSB. Two mechanisms have been proposed to account for these observations: (i) hDNA formation is restricted to one side of the DSB or the other, and (ii) recombination is initially two sided, but hDNA repair directed by Holliday junction cuts restores normal 4:4 segregation on that side of the DSB in which the mismatch is closest to the cut junction initiating repair. In this study, we exploited a well-characterized gene-targeting assay to test the predictions that these mechanisms make with respect to the frequency of recombinants displaying 4:4 marker segregation on one side of the DSB. Unexpectedly, the results do not support the predictions of either mechanism. We propose a derivation of mechanism (ii) in which the nicks arising from Holliday junction cleavage are not equivalent with respect to directing repair of adjacent hDNA, possibly as a result of asynchronous cleavage of the DSBR intermediate.


Temporal patterns recognized by a network of coordinated time delays and coincidence detectors.
A computational model of a neuronal network is described which performs a fundamental task of general perception: recognition of temporal patterns in continuous and uncued neuronal spike trains. The presented network is able to recognize each pattern element (100 ms interval composed of sets of 10, 20, 30 and 40 ms interspike intervals combined in linear order) as it arrives. Its operation is based upon biologically plausible filtering mechanisms and population neurodynamics.


Pulse radiolytic measurement of redox potentials: the tyrosine and tryptophan radicals.
With the technique of pulse radiolysis we have measured the redox midpoint potentials of the tryptophan side chain neutral indolyl radical (1.05 +/- 0.01 V vs NHE, pH 7.0 and 25 degrees C) and the tyrosine side chain neutral phenoxy radical (0.94 +/- 0.01 vs NHE, pH 7.0 and 25 degrees C). These potentials were obtained by using a variety of inorganic reference compounds in both kinetic and equilibrium protocols. We compare these results with others already in the literature, and we also present data useful in establishing a pulse radiolysis redox reference scale over the range 0.42-1.28 V.


Blood Transfusion During Total Ankle Arthroplasty Is Associated With Increased In-Hospital Complications and Costs.
Total ankle arthroplasty (TAA) is an increasingly used, effective treatment for end-stage ankle arthritis. Although numerous studies have associated blood transfusion with complications following hip and knee arthroplasty, its effects following TAA are largely unknown. This study uses data from a large, nationally representative database to estimate the association between blood transfusion and inpatient complications and hospital costs following TAA. Using the Nationwide Inpatient Sample (NIS) database from 2004 to 2014, 25 412 patients who underwent TAA were identified, with 286 (1.1%) receiving a blood transfusion. Univariate analysis assessed patient and hospital factors associated with blood transfusion following TAA. Patients requiring blood transfusion were more likely to be female, African American, Medicare recipients, and treated in nonteaching hospitals. Average length of stay for patients following transfusion was 3.0 days longer, while average inpatient cost was increased by approximately 50%. Patients who received blood transfusion were significantly more likely to suffer from congestive heart failure, peripheral vascular disease, hypothyroidism, coagulation disorder, or anemia. Acute renal failure was significantly more common among patients receiving blood transfusion ( P < .001). Blood transfusions following TAA are infrequent and are associated with multiple medical comorbidities, increased complications, longer hospital stays, and increased overall cost. Level III: Retrospective, comparative study.


Targeting Kupffer cells in non-alcoholic fatty liver disease/non-alcoholic steatohepatitis: Why and how?
Mechanisms for non-alcoholic steatohepatitis (NASH) development are under investigation in an era of increased prevalence of obesity and metabolic syndrome. Previous findings have pointed to the role of adipose tissue, adipose tissue macrophages and their secretory products in the development of a chronic inflammatory status inducing insulin resistance and a higher risk of liver steatosis called non-alcoholic fatty liver disease. The activation of resident macrophages [Kupffer cells (KC)] and the recruitment of blood derived monocytes/macrophages into the diseased liver have now been identified as key elements for disease initiation and progression. Those cells could be activated through gut flora modifications and an altered gut barrier function but also through the internalization of toxic lipid compounds in adjacent hepatocytes or in KC themselves. Due to the role of activated KC in insulin resistance, fibrosis development and inflammation amplification, they became a target in clinical trials. A shift towards an anti-inflammatory KC phenotype through peroxisome proliferator activator-receptorδ agonists, an inhibition of macrophage recruitment through anti-C-C chemokine receptor 2 action and a specific blocking of internalization of toxic lipoxidation or glycation compounds into KC by galectin-3 receptor inhibitors are now under investigation in human NASH.


Gamification to Support Adherence to a Therapeutic Ambylopia Treatment for Children: Retrospective Study Using a Focal Ambient Visual Acuity Stimulation Game.
The gold standard treatment for amblyopia is patching the better eye. Improvement of visual acuity in the amblyopic eye is significantly impacted by the adherence to the patching therapy. It is known that the overall adherence is rather low. This retrospective study evaluated whether an updated version of attention-binding digital therapeutic games based on the principle of focal ambient visual acuity stimulation (FAVAS) would result in improved patient adherence in 4- to 16-year-old patients with amblyopia associated with anisometropia or strabismus. We analyzed electronically pseudonymized recorded data from patients treated with occlusion therapy and FAVAS therapeutic games. One group used an older version (2015) and the other group used a newer version (2020) that provided more attractive therapeutic games with tablet computer functionality. Objective adherence was calculated by comparing the number of minutes using the therapeutic games as monitored in the automatized logbook versus the prescribed number of minutes for using the games. Children in group 2015 (n=138) spent on average 2009.3 (SD 1372.1; range 36-5556) minutes using FAVAS; children in group 2020 (n=129) spent on average 2651.2 (SD 1557.1; range 38-5672) minutes using the newer version. Group 2020 spent on average 641.9 more minutes on FAVAS than group 2015 (t255.49=3.56, P<.001, d=0.45; 95% CI 0.69-0.20). Although patient adherence was very variable, compared to the 55.0% (SD 29.4%) in group 2015, it significantly improved up to 68.5% (SD 33.7%) in group 2020 (t254.38=3.48, P=.001, d=0.44; 95% CI 0.68-0.19). FAVAS 2020, with improved gamification aspect as well as tablet computer functionality, increased adherence significantly compared to the earlier version of FAVAS 2015, indicating that FAVAS 2020 could be an effective approach to support adherence to amblyopia treatment. German Clinical Trials Register (DRKS) DRKS00017633; https://drks.de/search/de/trial/DRKS00017633.


Protein sequence-structure alignment based on site-alignment probabilities.
A protein sequence-structure alignment method for database searches is examined on how effectively this method together with a simple scoring function previously developed can identify compatibilities between sequences and structures of proteins. The scoring function consists of pairwise contact energies, repulsive packing potentials of residues for overly dense arrangement and short-range potentials for secondary structures. Pairwise contact interactions in a sequence-structure alignment are evaluated in a mean field approximation on the basis of probabilities of site pairs to be aligned. Gap penalties are assumed to be proportional to the number of contacts at each residue position, and as a result gaps will be more frequently placed on protein surfaces than in cores. In addition to minimum energy alignments, we use probability alignments made by successively aligning site pairs in order by pairwise alignment probabilities. Results show that the present energy function and alignment method can detect well both folds compatible with a given sequence and, inversely, sequences compatible with a given fold. Probability alignments consisting of most reliable site pairs only can yield small root mean square deviations, and including less reliable pairs increases the deviations. Remarkably, by this method some individual sequence-structure pairs are detected having only 5-20% sequence identity.


Traboulsi Syndrome in Pakistan.
Traboulsi syndrome is characterised by facial dysmorphism, abnormal spontaneous filtering blebs, ectopia lentis and multiple anterior segment abnormalities. The constellation of abnormalities separate it from syndromes related to connective tissue abnormalities which are associated with ectopia lentis. We report five females with distinctive spontaneous filtering blebs, ectopia lentis and other anterior segment abnormalities and no systemic features other than flat cheeks and beaking of nose. The cases are being managed conservatively in the Cornea and Glaucoma departments of Al-Shifa Trust Eye Hospital, Rawalpindi, Pakistan.


The role of polycyclic aromatic hydrocarbon-DNA adducts in inducing mutations in mouse skin.
Polycyclic aromatic hydrocarbons (PAH) form stable and depurinating DNA adducts in mouse skin to induce preneoplastic mutations. Some mutations transform cells, which then clonally expand to establish tumors. Strong clues about the mutagenic mechanism can be obtained if the PAH-DNA adducts can be correlated with both preneoplastic and tumor mutations. To this end, we studied mutagenesis in PAH-treated early preneoplastic skin (1 day after exposure) and in the induced papillomas in SENCAR mice. Papillomas were studied by PCR amplification of the H-ras gene and sequencing. For benzo[a]pyrene (BP), BP-7,8-dihydrodiol (BPDHD), 7,12-dimethylbenz[a]anthracene (DMBA) and dibenzo[a,l]pyrene (DB[a,l]P), the codon 13 (GGC to GTC) and codon 61 (CAA to CTA) mutations in papillomas corresponded to the relative levels of Gua and Ade-depurinating adducts, despite BP and BPDHD forming significant amounts of stable DNA adducts. Such a relationship was expected for DMBA and DB[a,l]P, as they formed primarily depurinating adducts. These results suggest that depurinating adducts play a major role in forming the tumorigenic mutations. To validate this correlation, preneoplastic skin mutations were studied by cloning H-ras PCR products and sequencing individual clones. DMBA- and DB[a,l]P-treated skin showed primarily A.T to G.C mutations, which correlated with the high ratio of the Ade/Gua-depurinating adducts. Incubation of skin DNA with T.G-DNA glycosylase eliminated most of these A.T to G.C mutations, indicating that they existed as G.T heteroduplexes, as would be expected if they were formed by errors in the repair of abasic sites generated by the depurinating adducts. BP and its metabolites induced mainly G.C to T.A mutations in preneoplastic skin. However, PCR over unrepaired anti-BPDE-N(2)dG adducts can generate similar mutations as artifacts of the study protocol, making it difficult to establish an adduct-mutation correlation for determining which BP-DNA adducts induce the early preneoplastic mutations. In conclusion, this study suggests that depurinating adducts play a major role in PAH mutagenesis.


Impact of multiple organ system dysfunction and nosocomial infections on survival of children treated with extracorporeal membrane oxygenation after heart surgery.
To evaluate whether cardiac and noncardiac variables may be used to predict survival in children treated with extracorporeal membrane oxygenation (ECMO) after cardiopulmonary bypass and to determine when to discontinue ECMO support. Retrospective review. Neonatal and pediatric intensive care units of Kosair Children's Hospital. Fifty-nine children treated with ECMO after cardiopulmonary bypass from 1987 through 1996. None. Medical, nursing, operative, and perfusion records for each patient were reviewed. The primary outcome measure was survival to hospital discharge. Cardiac and noncardiac variables were recorded at serial times. Nineteen of 59 patients (32%) survived. No cardiac variable was a clinically useful predictor of survival or marker for when to discontinue ECMO. Among the noncardiac variables, progressive multiple organ system dysfunction and development of a nosocomial infection were significantly associated with nonsurvival. No patient with a positive blood culture (n = 3) within the first 24 hrs of ECMO survived, and 21 of 24 children with a positive culture from any site during ECMO died (p = .007). Despite their higher mortality, children with positive cultures were supported with ECMO significantly longer than those with negative cultures (275+/-168 vs. 135+/-108 hrs, respectively; p = .0004). For all patients, the longest duration of ECMO that resulted in survival was 256 hrs. For children with a positive culture, the longest duration of support that resulted in survival was 200 hrs. Support with ECMO beyond 256 hrs was not associated with survival. Progressive multiple organ system dysfunction and nosocomial infections have a negative impact on survival. Serious consideration should be given to discontinuing ECMO support whenever there is a progressive increase in the number of abnormally functioning organ systems, a nosocomial infection occurs, or native cardiac function has not improved significantly by 250 hrs of ECMO support.


Broad thumbs and halluces with deafness: a patient with Keipert syndrome.
Clinical, radiological, and audiological features are described in a boy whose condition closely parallels the profile of patients previously described with Keipert syndrome. This case represents the fourth report of this rare disorder.


Differential alternative splicing expressions of telomerase reverse transcriptase in gastrointestinal cell lines.
Telomerase is a cellular RNA-dependent DNA polymerase that serves to maintain the tandem arrays of telomeric TTAGGG repeats at eukaryotic chromosome ends. One of the human telomerase components is hTERT, which has three alternative spliced sites that introduce eight isoforms of hTERT mRNA. The expression of these isoforms in gastrointestinal cell lines is unknown. We developed a PCR-based assay for detecting these splicing variants. In gastric and hepatocellular carcinoma cell lines, the gamma deletion variant and its combination variants, alpha- and gamma-, beta- and gamma-, and alpha-, beta- and gamma-deletion variants were frequently detected, while they were not detected in colorectal carcinoma cell lines. Our results provide important information of use for more detailed studies on the regulation of telomerase activity.


Arthrodesis of the first metatarsal-phalangeal joint with flexible, ridged titanium intramedullary nails alone or supplemented with static staples and immediate weight bearing: a consecutive series of 148 procedures.
Myriad forms of fixation have been proposed for arthrodesis of the first metatarsal-phalangeal joint (MTPJ). However, nonunion of the arthrodesis site remains a common complication. The authors performed a retrospective review of all patients undergoing arthrodesis of the first MTPJ with flexible titanium intramedullary nails alone or supplemented with static staples followed by immediate protected weight bearing. Subjects were included if they had surgery performed only by one of us; not undergone bilateral surgery in the same setting; surgery for pathology of the first MTPJ other than rheumatoid arthritis; weight-bearing preoperative radiographs; postoperative weight-bearing radiographs demonstrating presence or absence of union at the arthrodesis site; initiated weight bearing on the operative foot immediately postoperative in a protective shoe; and documentation of any complications. A total of 128 patients (148 feet) met inclusion criteria and were included. There were 110 (85.9%) females and 18 (14.1%) males with a mean age ± SD of 64 ± 14.1 years. Indications included (a) 93 (62.8%) severe hallux valgus deformity, (b) 37 (25%) hallux rigidus, and (c) 18 (12.2%) failed first MTPJ surgeries. Ten different configurations where included based on the requirements needed to achieve a solid construct intraoperatively. There were a total of 9 (6.1%) complications consisting of 4 (2.7%) asymptomatic nonunions, 4 (2.7%) hardware removals, and 1 (0.7%) loss of reduction requiring revision surgery. The incidence of nonunion with flexible titanium intramedullary nails alone or supplemented with static staples followed by immediate protected weight bearing for predominantly severe hallux valgus and hallux rigidus is lower than the historic mean for most other fixation techniques. However, there is still a need for methodologically sound prospective cohort studies that focus on the use of isolated arthrodesis of the first MTPJ for purely severe hallux valgus or hallux rigidus, and comparison of the technique presented here with other modern osteosynthesis techniques. Levels of Evidence : Therapeutic, Level IV.


Attachment of Acanthamoeba to first- and second-generation silicone hydrogel contact lenses.
To investigate the attachment of Acanthamoeba to first- and second-generation silicone hydrogel contact lenses, and to determine if patient wear or the presence of a bacterial biofilm coating affects attachment characteristics. Experimental study. Attachment to the silicone hydrogel lenses was compared with that to a conventional hydrogel control lens. Sixteen replicates (n = 16) were carried out for unworn, worn, and biofilm-coated lenses of each type. Unworn, worn, and Pseudomonas aeruginosa biofilm-coated first-generation (lotrafilcon A) and second-generation (galyfilcon A) silicone hydrogel and conventional hydrogel (etafilcon A) lens quarters were incubated for 90 minutes in suspensions of plate-cultured Acanthamoeba castellanii trophozoites. Trophozoites attached to one surface of each lens quarter were counted by direct light microscopy. Logarithmic transformation of data allowed the use of parametric analysis of variance for statistical analysis. Attachment of Acanthamoeba was affected significantly by lens material type (P<0.001), with higher numbers of trophozoites attaching to the first-generation lotrafilcon A silicone hydrogel lens, compared with the second-generation galyfilcon A lens and the conventional hydrogel lens. Attachments to the latter 2 lenses did not differ significantly from each other (P = 0.126). Patient wear and the presence of a bacterial biofilm had no significant effect on attachment to the lotrafilcon A lens (P = 0.426) but did significantly increase attachment to the galyfilcon A (P<0.001) and the etafilcon A (P = 0.009) lenses; attachment to the latter 2 lenses was still significantly less than that found with the first-generation silicone hydrogel (P<0.001). Acanthamoeba demonstrated a significantly greater affinity for the first-generation silicone hydrogel lens as compared with the second-generation silicone hydrogel and the conventional hydrogel. If exposed to Acanthamoeba (e.g., when showering or swimming, through noncontinuous wear and ineffective lens care regimes), first-generation silicone hydrogel lenses may promote a greater risk of Acanthamoeba infection due to the enhanced attachment characteristics of this lens material. However, prospective studies in patients are required to determine if these experimental results are clinically significant.


Rising Interest in the Development of Metal Complexes in Cancer Immunotherapy.
Metal complexes have shown great potential in cancer immunotherapy. This review briefly introduces the basic concepts and strategies of cancer immunotherapy and summarizes the recent discoveries on the immune effects of traditional platinum-based anticancer compounds. In addition, we also outline the latest research progresses on metal complexes for cancer immunotherapy focusing on platinum, ruthenium, iridium, rhenium and copper complexes. Finally, the research perspectives and unsolved problems on the applications of metallo-anticancer agents in cancer immunotherapy are purposed.


Rh(V) -Nitrenoid as a Key Intermediate in Rh(III) -Catalyzed Heterocyclization by C-H Activation: A Computational Perspective on the Cycloaddition of Benzamide and Diazo Compounds.
A mechanistic study of the substituent-dependent ring formations in Rh(III) -catalyzed C-H activation/cycloaddition of benzamide and diazo compounds was carried out by using DFT calculations. The results indicated that the decomposition of the diazo is facilitated upon the formation of the five-membered rhodacycle, in which the Rh(III) center is more electrophilic. The insertion of carbenoid into Rh-C(phenyl) bond occurs readily and forms a 6-membered rhodacycle, however, the following C-N bond formation is difficult both kinetically and thermodynamically by reductive elimination from the Rh(III) species. Instead, the Rh(V) -nitrenoid intermediate could be formed by migration of the pivalate from N to Rh, which undergoes the heterocyclization much more easily and complementary ring-formations could be modulated by the nature of the substituent at the α-carbon. When a vinyl is attached, the stepwise 1,3-allylic migration occurs prior to the pivalate migration and the 8-membered ring product will be formed. On the other hand, the pivalate migration becomes more favorable for the phenyl-contained intermediate because of the difficult 1,3-allylic migration accompanied by dearomatization, thus the 5-membered ring product was formed selectively.


Support for relactation among mothers of HIV-infected children: a pilot study in Soweto.
Breastfeeding is accepted as the healthiest practice for human immunodeficiency virus (HIV)-infected infants, but decisions about infant feeding are made before the child's HIV status is known. We examined the feasibility of counseling to support breastfeeding for newly diagnosed HIV-infected infants, including relactation for those who had never initiated or who had stopped breastfeeding before the infant's HIV status was known. Mothers of 30 HIV-infected infants <12 weeks of age were enrolled in Soweto, South Africa. Mothers were offered lactation counseling, including support for relactation. Mother-infant pairs were followed for 24 weeks with regular counseling. We evaluated feeding practices, attitudes, and maternal and infant outcomes, including morbidity and growth. All infants and mothers who met local eligibility criteria were started on antiretroviral therapy. Mother-infant pairs (19 of the original 30) were followed up for 24 weeks. Ten of 19 women (53%) reported some breastfeeding at enrollment, two had stopped, and seven had never breastfed. At 24 weeks post-enrollment, 11 of 19 (58%) were providing breastmilk for all milk feeds. All women produced milk and provided some breastfeeds during the initial weeks of the study, but eight reported difficulty overcoming infant latching problems and stopped all breastfeeding. Attitudes toward breastfeeding were positive at the outset but became more negative in those who did not establish or sustain breastfeeding. Three of the seven who had never breastfed before enrollment into the study were fully breastfeeding at 24 weeks post-enrollment. Support for breastfeeding and relactation is possible among mothers of newly diagnosed HIV-infected infants but requires motivation from mothers and clinicians. Lactation counseling at the time of infant diagnosis is challenging as other issues predominate at this time. Improvements in antenatal infant feeding counseling are essential.


Molecular profiling of urinary extracellular vesicles in chronic kidney disease and renal fibrosis.
Chronic kidney disease (CKD) is a long-term kidney damage caused by gradual loss of essential kidney functions. A global health issue, CKD affects up to 16% of the population worldwide. Symptoms are often not apparent in the early stages, and if left untreated, CKD can progress to end-stage kidney disease (ESKD), also known as kidney failure, when the only possible treatments are dialysis and kidney transplantation. The end point of nearly all forms of CKD is kidney fibrosis, a process of unsuccessful wound-healing of kidney tissue. Detection of kidney fibrosis, therefore, often means detection of CKD. Renal biopsy remains the best test for renal scarring, despite being intrinsically limited by its invasiveness and sampling bias. Urine is a desirable source of fibrosis biomarkers as it can be easily obtained in a non-invasive way and in large volumes. Besides, urine contains biomolecules filtered through the glomeruli, mirroring the pathological state. There is, however, a problem of highly abundant urinary proteins that can mask rare disease biomarkers. Urinary extracellular vesicles (uEVs), which originate from renal cells and carry proteins, nucleic acids, and lipids, are an attractive source of potential rare CKD biomarkers. Their cargo consists of low-abundant proteins but highly concentrated in a nanosize-volume, as well as molecules too large to be filtered from plasma. Combining molecular profiling data (protein and miRNAs) of uEVs, isolated from patients affected by various forms of CKD, this review considers the possible diagnostic and prognostic value of uEVs biomarkers and their potential application in the translation of new experimental antifibrotic therapeutics.


Decrease of H2O2 plasma membrane permeability during adaptation to H2O2 in Saccharomyces cerevisiae.
Contrary to what is widely believed, recent published results show that H2O2 does not freely diffuse across biomembranes. The fast removal of H2O2 by antioxidant enzymes is able to generate a gradient if H2O2 is produced in a different compartment from that containing the enzymes (Antunes, F., and Cadenas, E. (2000) FEBS Lett. 475, 121-126). In this work, we extended these studies and tested whether an active regulation of biomembranes permeability characteristics is part of the cell response to oxidative stress. Using Saccharomyces cerevisiae as a model, we showed that: (a) H2O2 gradients across the plasma membrane are formed upon exposure to external H2O2; (b) there is a correlation between the magnitude of the gradients and the resistance to H2O2; (c) there is not a correlation between the intracellular capacity to remove H2O2 and the resistance to H2O2; (d) the plasma membrane permeability to H2O2 decreases by a factor of two upon acquisition of resistance to this agent by pre-exposing cells either to nonlethal doses of H2O2 or to cycloheximide, an inhibitor of protein synthesis; and (e) erg3Delta and erg6Delta mutants, which have impaired ergosterol biosynthesis pathways, show higher plasma membrane permeability to H2O2 and are more sensitive to H2O2. Altogether, the regulation of the plasma membrane permeability to H2O2 emerged as a new mechanism by which cells respond and adapt to H2O2. The consequences of the results to cellular redox compartmentalization and to the origin and evolution of the eukaryotic cell are discussed.


Protein identification using mass spectrometry: a method overview.
With the introduction of soft ionization techniques such as Matrix Assisted Laser Desorption Ionization (MALDI), and Electrospray Ionization (ESI), proteins have become accessible to mass spectrometric analyses. Since then, mass spectrometry has become the method of choice for sensitive, reliable and inexpensive protein and peptide identification. With the increasing number of full genome sequences for a variety of organisms and the numerous protein databases constructed thereof, all the tools necessary for the high-throughput protein identification with mass spectrometry are in place. This chapter highlights the different mass spectrometric techniques currently applied in proteome research by giving a brief overview of methods for identification of posttranslational modifications and discussing their suitability of strategies for automated data analysis.


Hepatoprotective effect of bis(4-methylbenzoyl) diselenide against CCl(4)-induced oxidative damage in mice.
From a pharmacological point of view, organoseleniums are compounds with important and interesting antioxidant and biological activities. The aim of this study was to evaluate the hepatoprotective effect of bis(4-methylbenzoyl) diselenide (BMD) against carbon tetrachloride (CCl4 )-induced oxidative damage in mice. The animals received BMD (25 mg/kg p.o., for 3 days), and after 1 day, CCl4 (1 mg/kg body weight) was administered by intraperitoneal route. One day after the CCl4 exposure, the animals were euthanized for biochemical and histological analysis. Treatment with BMD (25 mg/kg p.o.) protected against aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyl transferase and lactate dehydrogenase activity increases induced by CCl4 plasma exposure. Treatment with BMD (25 mg/kg) protected against increases in thiobarbituric reactive species and decreasing non-protein thiols and ascorbic acid levels in liver of mice. Catalase and superoxide dismutase activity inhibition in the liver caused by CCl4 were protected by treatment with BMD (25 mg/kg). Glutathione S-transferase activity was inhibited by CCl4 and remained unaltered even after treatment with BMD. Sections of liver from CCl4 -exposed mice presented an intense infiltration of inflammatory cells and loss of the cellular architecture. BMD (25 mg/kg) attenuated CCl4 -induced hepatic histological alterations. The results demonstrated the hepatoprotective effects of BMD in the mouse liver, possibly by modulating the antioxidant status.


Climate change and vector-borne diseases: what are the implications for public health research and policy?
Vector-borne diseases continue to contribute significantly to the global burden of disease, and cause epidemics that disrupt health security and cause wider socioeconomic impacts around the world. All are sensitive in different ways to weather and climate conditions, so that the ongoing trends of increasing temperature and more variable weather threaten to undermine recent global progress against these diseases. Here, we review the current state of the global public health effort to address this challenge, and outline related initiatives by the World Health Organization (WHO) and its partners. Much of the debate to date has centred on attribution of past changes in disease rates to climate change, and the use of scenario-based models to project future changes in risk for specific diseases. While these can give useful indications, the unavoidable uncertainty in such analyses, and contingency on other socioeconomic and public health determinants in the past or future, limit their utility as decision-support tools. For operational health agencies, the most pressing need is the strengthening of current disease control efforts to bring down current disease rates and manage short-term climate risks, which will, in turn, increase resilience to long-term climate change. The WHO and partner agencies are working through a range of programmes to (i) ensure political support and financial investment in preventive and curative interventions to bring down current disease burdens; (ii) promote a comprehensive approach to climate risk management; (iii) support applied research, through definition of global and regional research agendas, and targeted research initiatives on priority diseases and population groups.


Axonal growth and target selection during development: retinal projections to the ventrobasal complex and other "nonvisual" structures in neonatal Syrian hamsters.
In newborn hamsters, there is a direct retinal projection to the ventrobasal complex, the principal thalamic somatosensory nucleus. The projection decreases precipitously between the second and third postnatal days. A few retinofugal axons remain dorsally along the lateral border of the nucleus on day 4, and none are present thereafter. In neonatal hamsters, retinofugal axons project to additional "nonvisual" nuclei including the periventricular and anterior nuclei of the hypothalamus, zona incerta, substantia nigra, inferior colliculus, pons, and mesencephalic tegmentum. Some of these connections remain in adult hamsters, although in apparently reduced density or relative volume, while others disappear. The contribution of transient connections to the normal morphological or functional development of the brain remains to be clarified. The combined results of this and other studies show that the normally transient retino-ventrobasal projection is a substrate for abnormal connections in that it can be permanently stabilized by appropriate neurosurgery on the day of birth.


Clinical evaluation of 3-day tebipenem pivoxil therapy in children with community-acquired pneumonia.
A 4mg/kg dose of tebipenem pivoxil (TBPM-PI) was administered twice daily for 3 days to 34 pediatric patients with pneumonia who had chest X-ray findings indicative of pneumonia and CRP values of at least 3.0 mg/dL. The clinical effects of this regimen were evaluated by retrospectively examining medical charts for the period from April 2012 to March 2015. The patients were 6 months to 8 years old, with serum CRP values ranging from 3.06 to 14.25 mg/dL. Fever resolved within 24 hours and respiratory symptoms improved within 3 to 5 days after the start of treatment in all 34 patients. Although CRP was positive in 28 of 30 patients at the end of the treatment period, none of these children showed worsening of pneumonia. Eight patients (23.5%) experienced adverse drug reactions including diarrhea. These results indicate that a 3-day course of TBPM-PI is useful for treating pediatric pneumonia.


Effects of intermittent positive-pressure ventilation on lungs of normal rabbits.
We ventilated 10 healthy young rabbits with warm moist air at 20 cm H2O peak airway pressure and compared their pulmonary histology to that of 10 controls. The histological changes were analysed using the Leitz Texture Analysis System. Four hundred high-power fields (240 micrometers X 240 micrometers) were scanned from each section and the mean proportions of air space, interstitium and oedema determined. Calculated bicarbonate decreased significantly in both groups (P < 0.05); this was accompanied by a modest decrease in pHa in the ventilated group. The PaO2 and PaCO2 were maintained within the normal range. The blood pressure decreased in both groups. Seven of 10 ventilated rabbits had interstitial widening while only 1 of 10 controls had a similar lesion (chi 2 = 5.21, P = < 0.05). The area occupied by interstitium ranged from 26 to 46% (mean 37.6%) in controls and 18 to 81% (mean 45.4%) in ventilated rabbits. There was a lack of correlation between clinical and histopathological findings. While 6 of 10 ventilated rabbits had intra-alveolar oedema and a decrease in air space to less than 50% of total area, none developed hypoxaemia. Two of these 6 rabbits also had pHa less than 7.35 by the end of the study, yet all were clinically stable. These data indicate that intermittent partial-pressure ventilation with warm moist air may result in pulmonary parenchymal damage within 6 h in healthy rabbits.


Intratympanic Gentamicin for Intractable Ménière's Disease - A Review and Analysis of Audiovestibular Impact.
Introduction Intratympanic gentamicin regulates the symptoms in most patients with incapacitating Ménière's disease. The treatment protocols have changed over the years from medical labyrinthectomy to preservation of vestibular function. Objectives This study aims to review the audiovestibular response related to the effect of the drug in controlling vertigo. Data Synthesis Articles were identified by means of a search in the PubMed database using the key words Meniere and intratympanic or transtympanic gentamicin . Total 144 articles were reviewed after excluding those that were technical reports, those based on experimental animal studies, those that focused on outcomes other than vertigo (tinnitus or aural fullness), those with delivery methods other than tympanic membrane injection, and those with bilateral cases. If there was more than one article by the same author(s) or institution, only the most recent one matching the aforementioned criteria and those that were not overlapping were included. Conclusion Titration methods or multiple injections on a daily basis can be preferred if the patients have profound or non-serviceable hearing, since these methods have significant incidence of hearing loss. Treatment protocols with a frequency of injection not shorter than once a week, or those with injections on a monthly basis as "needed" provide the same level of vertigo control with better preservation of hearing. Caloric testing is not an ideal tool to analyze the correlation between vertigo control and the effect of gentamicin as compared with gain asymmetry of the vestibulo-ocular reflex. Vestibular-evoked myogenic potentials and the head thrust test are more reliable than other vestibular tests for the follow-up of patients undergoing gentamicin treatment.


Strategies to combat poverty and their interface with health promotion.
The population impoverishment is a social reality whose overcoming is necessary so that we can think about health as a positive concept. This study proposes a reflection on the coping strategies adopted by the Conjunto Palmeira, a Brazilian community in the Northeast, and their interface with health promotion. This community's reality is an example of overcoming social exclusion for different regions of Brazil and other countries. The history of the Conjunto and the collective strategies of empowerment for coping with poverty and search for human development are initially presented. After that, we establish the relationship of those strategies with the action fields for health promotion. Finally, we consider that the mutual responsibility of the community with its health and its relationship with the environment in which they live are means of promoting transformation towards the conquest of a worthy social space.


[Status of libraries and databases for natural products at abroad].
For natural products are one of the important sources for drug discovery, libraries and databases of natural products are significant for the development and research of natural products. At present, most of compound libraries at abroad are synthetic or combinatorial synthetic molecules, resulting to access natural products difficult; for information of natural products are scattered with different standards, it is difficult to construct convenient, comprehensive and large-scale databases for natural products. This paper reviewed the status of current accessing libraries and databases for natural products at abroad and provided some important information for the development of libraries and database for natural products.


A Comparative Meta-Analysis and in silico Analysis of Differentially Expressed Genes and Proteins in Canine and Human Bladder Cancer.
Canine and human bladder cancer present similar anatomical, morphological, and molecular characteristics, and dogs can be considered a model for human bladder cancer. However, the veterinary literature lacks information regarding cross-validation analysis between human and canine large-scale data. Therefore, this research aimed to perform a meta-analysis of the canine literature on bladder cancer, identifying genes and proteins previously evaluated in these studies. In addition, we also performed a cross-validation of the canine transcriptome data and the human data from The Cancer Genome Atlas (TCGA) to identify potential markers for both species. The meta-analysis was performed using the following indexing terms: "bladder" AND "carcinoma" AND "dog" in different international databases, and 385 manuscripts were identified in our initial search. Then, several inclusion criteria were applied, and only 25 studies met these criteria. Among these studies, five presented transcriptome data, and 20 evaluated only isolated genes or proteins. Regarding the studies involving isolated protein analysis, the HER-2 protein was the most studied (3/20), followed by TAG-72 (2/20), COX-2 (2/20), survivin (2/20), and CK7 (2/20), and the remaining nine studies evaluated one isolated protein each. Regarding the cross-validation analysis of human and canine transcriptome data, we identified 35 dysregulated genes, including ERBB2, TP53, EGFR, and E2F2. Our results demonstrate that the canine literature on bladder cancer previously focused on the evaluation of isolated markers with no association with patient survival. This limitation may be related to the lack of a homogenous protocol for treating patients and the lack of follow-up during treatment. In addition, the lack of information regarding tumor muscle invasion can be considered an important limitation when comparing human and canine bladder tumors. Our in silico analysis involving canine and human transcriptome data provided several genes with the potential to be markers for both human and canine bladder tumors, and these genes should be considered for future studies on canine bladder cancer.


Effects of Objective and Perceived Social Isolation on Cardiovascular and Brain Health: A Scientific Statement From the American Heart Association.
Background Social isolation, the relative absence of or infrequency of contact with different types of social relationships, and loneliness (perceived isolation) are associated with adverse health outcomes. Objective To review observational and intervention research that examines the impact of social isolation and loneliness on cardiovascular and brain health and discuss proposed mechanisms for observed associations. Methods We conducted a systematic scoping review of available research. We searched 4 databases, PubMed, PsycInfo, Cumulative Index of Nursing and Allied Health, and Scopus. Findings Evidence is most consistent for a direct association between social isolation, loneliness, and coronary heart disease and stroke mortality. However, data on the association between social isolation and loneliness with heart failure, dementia, and cognitive impairment are sparse and less robust. Few studies have empirically tested mediating pathways between social isolation, loneliness, and cardiovascular and brain health outcomes using appropriate methods for explanatory analyses. Notably, the effect estimates are small, and there may be unmeasured confounders of the associations. Research in groups that may be at higher risk or more vulnerable to the effects of social isolation is limited. We did not find any intervention studies that sought to reduce the adverse impact of social isolation or loneliness on cardiovascular or brain health outcomes. Conclusions Social isolation and loneliness are common and appear to be independent risk factors for worse cardiovascular and brain health; however, consistency of the associations varies by outcome. There is a need to develop, implement, and test interventions to improve cardiovascular and brain health for individuals who are socially isolated or lonely.


Formation of DNA triplexes accounts for arrests of DNA synthesis at d(TC)n and d(GA)n tracts.
To study the mechanism of arrest of DNA synthesis at d(TC)n and d(GA)n sequences, single-stranded DNA molecules including d(TC)27 or d(TC)31 tracts or a d(GA)27 tract were used as templates for in vitro assays of complementary DNA synthesis performed by extension of a primer with the Klenow polymerase or the Taq polymerase (Thermus aquaticus DNA polymerase). Electrophoresis of the products revealed that arrests occurred around the middle of these tracts. The arrests in the d(TC)n sequences were eliminated when dATP or dGTP was replaced with the analogue 7-deaza dATP or 7-deaza dGTP, respectively, or when the templates were preincubated with the Escherichia coli single-strand binding protein (SSB). Preincubation of the template including a d(GA)27 tract with SSB has also eliminated the arrests at this sequence. Furthermore, arrests did not occur at d[G(7-deaza A)]27 or d[(7-deaza G)A]27 tracts when molecules including such tracts were used as templates. These results are compatible with the notion that the arrests were caused by formation of d(TC)i.d(GA)i.d(TC)i and d(GA)i.d(GA)i.d(TC)i triplexes, in which the bases in the uncopied portions of the d(TC)n tracts, or of the d(GA)27 tract, and the purine bases in the newly synthesized d(TC)i.d(GA)i duplexes were bound by hydrogen bonds. In the assays performed with the Taq polymerase, the pH dependence (in the range of 6.0-9.0) and the temperature dependence of the arrests were determined. As the pH was lowered, the arrests in the d(TC)27 tract were enhanced, in line with the expected properties of d(TC)i.d(GA)i.d(TC)i triplexes. The arrests in the d(GA)27 tract were enhanced by an increase in the pH. At pH 7.2 the arrests in the d(GA)27 tract persisted up to 80 degrees C, whereas the arrests in the d(TC)27 tract were eliminated at 50 degrees C; these results presumably reflect the relative stabilities of the two triplexes mentioned above at this physiological pH value and could be biologically significant.


Enhancing Ocular Bioavailability of Ciprofloxacin Using Colloidal Lipid-Based Carrier for the Management of Post-Surgical Infection.
Conjunctivitis and endogenous bacterial endophthalmitis mostly occurred after ophthalmic surgery. Therefore, the present study aimed to maximize the ocular delivery of ciprofloxacin (CPX) using colloidal lipid-based carrier to control the post-surgical infection. In this study, CPX was formulated as ophthalmic liposomal drops. Two different phospholipids in different ratios were utilized, including phosphatidylcholine (PC) and dimyrestoyl phosphatidylcholine (DMPC). The physiochemical properties of the prepared ophthalmic liposomes were evaluated in terms of particle size, entrapment efficiency, polydispersity index, zeta potential, and cumulative CPX in-vitro release. In addition, the effect of sonication time on particle size and entrapment efficiency of CPX ophthalmic drops was also evaluated. The results revealed that most of the prepared formulations showed particle size in nanometer size range (460-1047 nm) and entrapment efficiency ranging from 36.4-44.7%. The antibacterial activity and minimum inhibitory concentration (MIC) were investigated. Ex vivo antimicrobial effect of promising formulations was carried out against the most common causes of endophthalmitis microorganisms. The pharmacokinetics of the prepared ophthalmic drops were tested in rabbit aqueous humor and compared with commercial CPX ophthalmic drops (Ciloxan®). Observed bacterial suppression was detected in rabbit's eyes conjunctivitis with an optimized formulation A3 compared with the commercial ophthalmic drops. CPX concentration in the aqueous humor was above MIC against tested bacterial strains. The in vivo data revealed that the tested CPX drops showed superiority over the commercial ones with respect to peak aqueous humor concentration, time to reach peak aqueous humor concentration, elimination rate constant, half-life, and relative bioavailability. Based on these results, it was concluded that the prepared ophthalmic formulations significantly enhanced CPX bioavailability compared with the commercial one.


A New Mutation Genotype of K218T in Myosin-5 Confers Resistance to Phenamacril in Rice Bakanae Disease in the Field.
In 2017 and 2018, a total of 294 Fusarium fujikuroi isolates were collected from bakanae-diseased rice plants in Jinhua, Shaoxing, and Jiaxing in Zhejiang Province, China. Phenamacril sensitivity of these isolates was determined by the 50% effective concentration value or minimum inhibitory concentration methods. Our results indicated that the phenamacril resistance frequency of F. fujikuroi increased from 18% in 2017 to 47% in 2018, and rice plants infected with F. fujikuroi-resistant isolates could not be protected effectively with 50 mg/liter of phenamacril. Phenamacril-resistant F. fujikuroi isolates obtained from rice fields showed stable resistance, because their fitness levels (i.e., mycelial growth, sporulation, and pathogenicity) were similar to the phenamacril-sensitive isolates. In addition to the point mutation at codon 219 in the myosin-5 gene that conferred resistance to phenamacril, our results also showed another point mutation at codon 218 (AAG→ACG) in myosin-5 that also conferred resistance to phenamacril. In this study, we found rapid development and persistence of diversified genotypes of phenamacril resistance, highlighting the importance of proper use of phenamacril in rice fields. Our results may also help researchers develop new fungicides or new control strategies using combinations of different fungicides in the control of phenamacril-resistant F. fujikuroi isolates.


[Epidemiology and clinical aspects of non-A, non-B hepatitis (author's transl)].
Among 288 patients with acute viral hepatitis investigated between 1970 and 1976 29 (10%) had non-A, non-B hepatitis. In 9 of these 29 patients posttransfusion hepatitis was proven, the remaining 20 patients had sporadic infections. The incubation period of posttransfusion hepatitis was 2--8 weeks in 7 patients and 12 and 16 weeks in the two other patients. Transaminases changed monophasically in 22 patients, bi- or multiphasic in 7. The disease took a subicteric course in a quarter of all cases. Out of the 29 patients 17 were followed up for 2 to 7 years, one or more recurrence occurred in 9 patients, usually within the first two years after onset of the disease. Five of these 9 patients had a biphasic rise of transaminases during the acute stage. The almost complete absence of autoantibodies in the course of the acute phase might be evidence of increased suppressor cell activity favouring a chronic course. Non-organ-specific autoantibodies were only rarely observed and were completely absent in patients with posttransfusion hepatitis.


High volume centers for esophagectomy: what is the number needed to achieve low postoperative mortality?
Aimed at reducing surgical deaths, several initiatives have attempted to establish volume-based referral strategies in high risk surgery. The detailed analysis of the literature of the last 10 years, comprising 13 papers on esophageal cancer, shows a clear reduction in postoperative mortality with increasing case volumes per year. Single papers have analyzed the main reasons for this phenomenon and showed that postoperative complication rates are lower in high-volume hospitals and management of complications is more successful. Further, long-term prognosis is also correlated to case-volume. In conclusion, the analysis shows that only with the experience of more than 20 esophagectomies per year can a significant reduction of the mortality, down to 4.9%, be achieved. Based on this survey, surgery of esophageal cancer is a task for high-volume hospitals because of decreased postoperative mortality and improved long-term prognosis compared with low volume hospitals.


Novel variants of NEK9 associated with neonatal arthrogryposis: Two case reports and a literature review.
Objective: Pathogenic variants in NEK9 (MIM: 609798) have been identified in patients with lethal congenital contracture syndrome 10 (OMIM: 617022) and arthrogryposis, Perthes disease, and upward gaze palsy (APUG and OMIM: 614262). The shared core phenotype is multiple joint contractures or arthrogryposis. In the present study, three novel variants of NEK9 associated with neonatal arthrogryposis were reported. Methods: The clinical data of two premature infants and their parents were collected. The genomic DNA was extracted from their peripheral blood samples and subjected to trio-whole-exome sequencing (trio-WES) and copy number variation analysis. Results: Using trio-WES, a total of three novel pathogenic variants of NEK9 were detected in the two families. Patient 1 carried compound heterozygous variations of c.717C > A (p. C239*741) and c.2824delA (p.M942Cfs*21), which were inherited from his father and mother, respectively. Patient 2 also carried compound heterozygous variations of c.61G > T (p. E21*959) and c. 2824delA (p. M942Cfs*21), which were inherited from his father and mother, respectively. These variants have not been previously reported in the ClinVar, HGMD, or gnomAD databases. Conclusion: This is the first report about NEK9-related arthrogryposis in neonatal patients. The findings from this study suggest that different types of mutations in NEK9 lead to different phenotypes. Our study expanded the clinical phenotype spectrum and gene spectrum of NEK9-associated arthrogryposis.


Blogging as a tool to promote reflection among dietetic and physical therapy students during a multidisciplinary international service-learning experience.
Service-learning is a guided, structured learning experience that engages students in service to a community for the mutual benefit of the student and community. There is a growing trend in healthcare to include international service-learning experiences in the training curriculum to promote cultural competence. A critical element of service-learning is reflection. There have been many research studies examining how best to support and facilitate the reflective thinking process in students. The purpose of this study was to assess the development of reflective thinking among graduate allied health students during an interdisciplinary international service-learning experience using a web-based collaborative blog. Twelve graduate students-six dietetic interns and six physical therapy doctoral students-traveled to Belize for 6 days to provide primary healthcare screenings and intervention to a community. Group blogging was found to be an effective tool to promote reflection in allied health student and short duration service-learning experience developed reflective thinking.


Drug-induced and genetic hypermelanism: effects on pigment cell differentiation.
Allopurinol, a drug that inhibits the enzyme xanthine dehydrogenase (XDH), is known to cause hypermelanism in the axolotl. The hypermelanistic condition that results from allopurinol treatment is similar in most respects to the phenotype that results from the action of the melanoid (m) gene in axolotls. On the basis of structural and biochemical studies, it now seems clear that genetic and drug-induced hypermelanism are the same in the following ways. 1) Both types of melanism result in the production of more than normal amounts of melanin and more melanin-containing cells (melanophores). 2) In both cases the amount of pteridine-associated yellow pigment declines during development, and this is associated directly with fine structural changes that occur within the pigment organelles (pterinosomes) of yellow pigment cells (xanthophores). 3) In both cases the hypermelanistic condition results in the suppression of reflecting pigment cell (iridophore) differentiation. 4) Both conditions have now been linked directly to depressed levels of XDH activity. Thus both genetic and drug-induced hypermelanism result in alterations in the normal differentiation of all three pigment cell types and the subsequent disruption of normal pigment pattern formation. The possible significance of these findings with regard to factors known or suspected to direct the migration and/or differentiation of neural crest-derived pigment cells is discussed.


The Interaction of Munc18-1 Helix 11 and 12 with the Central Region of the VAMP2 SNARE Motif Is Essential for SNARE Templating and Synaptic Transmission.
Sec1/Munc18 proteins play a key role in initiating the assembly of N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes, the molecular fusion machinery. Employing comparative structure modeling, site specific crosslinking by single amino acid substitutions with the photoactivatable unnatural amino acid p-Benzoyl-phenylalanine (Bpa) and reconstituted vesicle docking/fusion assays, we mapped the binding interface between Munc18-1 and the neuronal v-SNARE VAMP2 with single amino acid resolution. Our results show that helices 11 and 12 of domain 3a in Munc18-1 interact with the VAMP2 SNARE motif covering the region from layers -4 to +5. Residue Q301 in helix 11 plays a pivotal role in VAMP2 binding and template complex formation. A VAMP2 binding deficient mutant, Munc18-1 Q301D, does not stimulate lipid mixing in a reconstituted fusion assay. The neuronal SNARE-organizer Munc13-1, which also binds VAMP2, does not bypass the requirement for the Munc18-1·VAMP2 interaction. Importantly, Munc18-1 Q301D expression in Munc18-1 deficient neurons severely reduces synaptic transmission, demonstrating the physiological significance of the Munc18-1·VAMP2 interaction.


Chrysotherapy in psoriasis.
Efficacy of oral gold was assessed in 23 patients with psoriasis of various types with excellent or good response in 14 patients (60%) whereas patients belonging to the age group of 11-20 years showed beneficial response in 10 out of 12 patients (83%). None of the patients experienced any side effect. It was concluded that oral gold (6mg/ day) for 3 to 6 months can be used as an alternative therapy in psoriasis especially in younger age group.


Investigation of the Effect of Exendin-4 on Oleic Acid-Induced Steatosis in HepG2 Cells Using Fourier Transform Infrared Spectroscopy.
Non-alcoholic fatty liver disease (NAFLD) is a common liver lesion that is untreatable with medications. Glucagon-like peptide-1 receptor (GLP-1R) agonists have recently emerged as a potential NAFLD pharmacotherapy. However, the molecular mechanisms underlying these drugs' beneficial effects are not fully understood. Using Fourier transform infrared (FTIR) spectroscopy, we sought to investigate the biochemical changes in a steatosis cell model treated or not with the GLP-1R agonist Exendin-4 (Ex-4). HepG2 cells were made steatotic with 400 µM of oleic acid and then treated with 200 nM Ex-4 in order to reduce lipid accumulation. We quantified steatosis using the Oil Red O staining method. We investigated the biochemical alterations induced by steatosis and Ex-4 treatment using Fourier transform infrared (FTIR) spectroscopy and chemometric analyses. Analysis of the Oil Red O staining showed that Ex-4 significantly reduces steatosis. This reduction was confirmed by FTIR analysis, as the phospholipid band (C=O) at 1740 cm-1 in Ex-4 treated cells is significantly decreased compared to steatotic cells. The principal component analysis score plots for both the lipid and protein regions showed that the untreated and Ex-4-treated samples, while still separated, are clustered close to each other, far from the steatotic cells. The biochemical and structural changes induced by OA-induced lipotoxicity are at least partially reversed upon Ex-4 treatment. FTIR and chemometric analyses revealed that Ex-4 significantly reduces OA-induced lipid accumulation, and Ex-4 also restored the lipid and protein biochemical alterations caused by lipotoxicity-induced oxidative stress. In combination with chemometric analyses, FTIR spectroscopy may offer new approaches for investigating the mechanisms underpinning NAFLD.


Tumor Microenvironment of Hepatocellular Carcinoma: Challenges and Opportunities for New Treatment Options.
The prevalence of liver cancer is constantly rising, with increasing incidence and mortality in Europe and the USA in recent decades. Among the different subtypes of liver cancers, hepatocellular carcinoma (HCC) is the most commonly diagnosed liver cancer. Besides advances in diagnosis and promising results of pre-clinical studies, HCC remains a highly lethal disease. In many cases, HCC is an effect of chronic liver inflammation, which leads to the formation of a complex tumor microenvironment (TME) composed of immune and stromal cells. The TME of HCC patients is a challenge for therapies, as it is involved in metastasis and the development of resistance. However, given that the TME is an intricate system of immune and stromal cells interacting with cancer cells, new immune-based therapies are being developed to target the TME of HCC. Therefore, understanding the complexity of the TME in HCC will provide new possibilities to design novel and more effective immunotherapeutics and combinatorial therapies to overcome resistance to treatment. In this review, we describe the role of inflammation during the development and progression of HCC by focusing on TME. We also describe the most recent therapeutic advances for HCC and possible combinatorial treatment options.


The impact of the protein interactome on the syntenic structure of mammalian genomes.
Conserved synteny denotes evolutionary preserved gene order across species. It is not well understood to which degree functional relationships between genes are preserved in syntenic blocks. Here we investigate whether protein-coding genes conserved in mammalian syntenic blocks encode gene products that serve the common functional purpose of interacting at protein level, i.e. connectivity. High connectivity among protein-protein interactions (PPIs) was only moderately associated with conserved synteny on a genome-wide scale. However, we observed a smaller subset of 3.6% of all syntenic blocks with high-confidence PPIs that had significantly higher connectivity than expected by random. Additionally, syntenic blocks with high-confidence PPIs contained significantly more chromatin loops than the remaining blocks, indicating functional preservation among these syntenic blocks. Conserved synteny is typically defined by sequence similarity. In this study, we also examined whether a functional relationship, here PPI connectivity, can identify syntenic blocks independently of orthology. While orthology-based syntenic blocks with high-confident PPIs and the connectivity-based syntenic blocks largely overlapped, the connectivity-based approach identified additional syntenic blocks that were not found by conventional sequence-based methods alone. Additionally, the connectivity-based approach enabled identification of potential orthologous genes between species. Our analyses demonstrate that subsets of syntenic blocks are associated with highly connected proteins, and that PPI connectivity can be used to detect conserved synteny even if sequence conservation drifts beyond what orthology algorithms normally can identify.


Melanin-concentrating hormone, melanocortin receptors and regulation of luteinizing hormone release.
Melanin-concentrating hormone (MCH) is a neuropeptide, identified by its ability to either mimic or antagonize the melanin-dispersing action of alpha-melanocyte stimulating hormone (alphaMSH) on skin melanophores. MCH and alphaMSH also have antagonistic actions in the brain affecting feeding behaviour, aggression, anxiety, arousal and reproductive function through the release of luteinizing hormone (LH). It is not clear, however, how they exert their opposite effects in the central nervous system (CNS). One possibility is that they act via a common receptor. In this study we have examined the effect of a number of MC receptor antagonists, with relative selectivity for the MC3, 4 and 5 subtypes, on the actions of MCH on LH release. We confirmed that bilateral administration of MCH (100 and 200 ng/side) into the medial preoptic area of oestrogen-primed (oestradiol benzoate 5 microgram) ovariectomized anaesthetized rats, stimulated the release of LH. This effect was blocked by the concomitant administration into the medial preoptic area of the MC4/5 antagonist ([D-Arg8]ACTH(4-10) and the MC3/5 antagonist ([Ala6]ACTH(4-10)-both at 500 ng/side-but not by the MC3/4 antagonist, SHU9119 (200 ng/side). Furthermore, the MC3 agonist [Nle3]-gamma2 MSH failed to affect LH release. These results indicate that the MC3 and MC4 receptors are not involved in mediating the action of MCH but are consistent with an action via the MC5 subtype. Preputial glands, which express MC5 receptors, were also stimulated by MCH which is in keeping with this idea. In HEK293 cells transfected with the MC5 receptor MCH increased the production of IP3. However, it was much less potent than alphaMSH and unlike alphaMSH, had no effect on the production of cAMP. MCH (10-10 to 10-5 M) also failed to displace I125NDP-MSH from cells transfected with MC5 receptors indicating that it was not acting as a competitive antagonist and its binding site was distinct from that of alphaMSH. Thus while MCH may function as an agonist at the MC5 receptor, its stimulation of LH release is more likely to be mediated via a specific MCH receptor that has common properties with the MC5 receptor.


Cu-catalyzed cyanomethylation of imines and α,β-alkenes with acetonitrile and its derivatives.
We describe copper-catalyzed cyanomethylation of imines and α,β-alkenes with a methylnitrile source and provide an efficient route to synthesize arylacrylonitriles and β,γ-unsaturated nitriles. This method tolerates aliphatic and aromatic alkenes substituted with a variety of functional groups such as F, Cl, Br, Me, OMe, tert-Bu, NO2, NH2 and CO2H with good to excellent yields (69-98%). These systems consist of inexpensive, simple copper catalyst and acetonitrile with its derivatives (α-bromo/α-iodo-acetonitrile) and are highly applicable in the industrial production of acrylonitriles.


Multiplex fluorescence in situ hybridization in identifying chromosome involvement of complex karyotypes in de novo myelodysplastic syndromes and acute myeloid leukemia.
Complex chromosomal aberrations (CCA) can be detected in a substantial proportion of myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML), which are associated with very poor prognosis. Conventional cytogenetics (CC) cannot accurately define the specific alterations in CCA. Multiplex fluorescence in situ hybridization (M-FISH) allows the comprehensive identification of CCA. In this study, M-FISH was used in 16 patients with de novo MDS and 22 with AML with CCA detected by R-banding CC, and revealed 206 aberrations involved all 24 chromosomes, including 73 numerical chromosomal abnormalities and 133 structural abnormalities. The chromosomes most often involved were, by decreasing incidence, 5, 17, 8, 11, 7 and 21 in 57.9%, 55.3%, 44.7%, 36.8%, 34.2% and 34.2% of the cases, respectively. There were 98 unbalanced translocations, which were the most frequently observed aberrations in our study. Derivative chromosome 5 and 8 were implicated most often. The other derivatives were der(11), der(12), der(7), der(14), der(15) and der(17). Fourteen balanced translocations were detected in our series, and the most frequent reciprocal translocations was t(8;21). Fifty-five monosomies, 15 partial deletions, and 18 trisomies were found in all patients. The most frequently observed were -5/5q-, -17/17q-, -7, -18, -21, -19, and trisomy of chromosome 8 and 6. There were some abnormalities that have not been previously described, including two complex t(8;21) and seven unbalanced translocations. M-FISH could refine CCA, find or correct the missed or misidentified aberrations by CC analysis. Our findings confirmed that M-FISH was a powerful molecular cytogenetic tool to characterize complex karyotypes in MDS and AML.


Presentation weight indices for patients with cleft and non-affected children: A two-centre study.
Children with cleft lip and/or palate (CL/P) face many challenges in the society. They tend to suffer malnutrition and therefore poor weight gain due to lack of standard care, especially from their parents and society at large. The poor weight gain leads to delayed surgical repair of the cleft abnormality leading to prolonged nutritional and psychological challenges for all concerned. The aim of the study was to determine if children without cleft abnormalities presenting for routine immunisation and children with cleft abnormalities are both likely to be less than the 75th percentile of expected weight at the time of presentation, and if there is any difference in presenting weight amongst children with different cleft types. This was a retrospective study. Data of children that attended the immunisation clinic of the Department of Primary Health of the National Orthopaedic Hospital, Enugu, and routine surgical clinic of the National Orthopaedic Hospital, Enugu, Nigeria, and the Good Shepherd Specialist Hospital, Enugu, Nigeria, between January 2010 and December 2014 with a diagnosis of CL/P, were obtained from the medical records. The data were analysed with SPSS and the confidence interval was 95%. A total of 923 medical records were reviewed for the study, out of which 363 were for children with cleft abnormalities. There was no significant difference in the weight of the children without cleft and presenting for routine immunisation compared with their expected 75th percentile, or children with cleft abnormality compared with their expected 75th percentile (either in their first or in their second visits). With the children that presented with CL/P, there was a significant difference noted amongst children with both CL/P deformity with the expected 75th percentile weight, irrespective of whether the combination was unilateral or bilateral, with P = 0.041. This difference was noted in their first visit. Children with combined CL/P are at risk of presenting with poor weight. Nutritional rehabilitation considered early with children with cleft and particularly those children with combined CL/P.


Ferromagnetism and spinodal decomposition in dilute magnetic nitride semiconductors.
We propose a materials design for dilute magnetic semiconductors (DMS) based on first-principles calculations by using the Korringa-Kohn-Rostoker coherent potential approximation method. We develop an accurate method for calculations of the Curie temperature (T(C)) of DMS and show that the mean field approximation completely fails to predict T(C) for DMS, in particular for wide gap nitride DMS where the exchange interaction is short ranged. The T(C) calculated for homogeneous DMS by using the present method agrees very well with available experimental values. For more realistic material design, we simulate spinodal nanodecomposition by applying the Monte Carlo method to the Ising model with ab initio chemical pair interactions between magnetic impurities in DMS. It is found that by controlling the dimensionality of the decomposition, various characteristic phases occur in DMS. It is suggested that superparamagnetic blocking phenomena should be important for understanding the magnetism of wide gap DMS.


Investigating temporal properties of covert shifts of visual attention using the attentional walk task.
The attentional walk task was used to investigate the temporal properties of covert shifts of attention. Observers shifted attention within arrays of identical items in response to a series of auditory commands and reported the color of the final disk. The density of the arrays and the timing of the shift commands varied. Performance decreased as density increased, and the minimal amount of time needed to shift attention depended on the density of the display, varying from 350 to 750 msec. In addition, the observers were able to maintain attentional focus for at least 3,500 msec without a decline in performance, regardless of density. Thus, although the ability to maintain attention at a given position was found to be independent of the precision with which that location was defined, more precise attentional shifts required more time to execute.


[Usefulness of FDG-PET for the diagnosis of copper deficiency at an early stage: a case report].
For the patients under long-term tube feeding, copper deficiency is known to be a cause of neutropenia and/or anemia. It is recognized as reversible myelodysplasia, since the condition improves by giving a copper supplementation. Myelodysplasia caused by copper deficiency is difficult to be diagnosed because it is not so common, and often it takes a long time to reach correct diagnosis. We reported usefulness of FDG-PET for the diagnosis of myelodysplasia caused by copper deficiency in early stage. The case was 46 y.o. male patient in vegetative state for 2 years after traumatic brain injury. Laboratory examination revealed slight leukopenia. PET/CT demonstrated high and diffuse FDG accumulation mainly in the vertebral bone marrow. Based on the lower levels of serum copper and ceruloprasmin, the patient was diagnosed as copper deficiency. After treatment of copper supplementation, FDG accumulation of the bone marrow disappeared, and the serum copper level has normalized. From the FDG-PET findings, even in the early stage of copper deficiency, high glucose metabolism of bone marrow was shown.


Age-Related Differences in Injury Patterns and Outcome From Falls From Tree Stands.
From 2009 to 2019 there were 77 youth and 1046 adults injured in tree stand-related emergency department visits. There was no statistically significant different in sex or gender. Injury types were similar among groups and were categorized by body part. Despite the above-mentioned insignificant differences between the groups, we did identify several statistically significant findings. Adults that were not Caucasian or African-American were more likely to sustain injuries related to tree stands when compared to their youth counterparts (P-value 0.029). Adult patients were also more likely to sustain trauma to their torso (P-value 0.017). Lastly, adult patients were found to be more likely to require hospitalization in comparison to the youth population (P-value 0.018). Improved education and safety guidelines in the use of tree stands, particularly in individuals in ages 18 and up, would likely lessen the discrepancies between age groups identified in this study.


Further Studies on the 3-Ketosteroid 9α-Hydroxylase of Rhodococcus ruber Chol-4, a Rieske Oxygenase of the Steroid Degradation Pathway.
The biochemistry and genetics of the bacterial steroid catabolism have been extensively studied during the last years and their findings have been essential to the development of biotechnological applications. For instance, metabolic engineering of the steroid-eater strains has allowed to obtain intermediaries of industrial value. However, there are still some drawbacks that must be overcome, such as the redundancy of the steroid catabolism genes in the genome and a better knowledge of its genetic regulation. KshABs and KstDs are key enzymes involved in the aerobic breakage of the steroid nucleus. Rhodococcus ruber Chol-4 contains three kshAs genes, a single kshB gene and three kstDs genes within its genome. In the present work, the growth of R. ruber ΔkshA strains was evaluated on different steroids substrates; the promoter regions of these genes were analyzed; and their expression was followed by qRT-PCR in both wild type and ksh mutants. Additionally, the transcription level of the kstDs genes was studied in the ksh mutants. The results show that KshA2B and KshA1B are involved in AD metabolism, while KshA3B and KshA1B contribute to the cholesterol metabolism in R. ruber. In the kshA single mutants, expression of the remaining kshA and kstD genes is re-organized to survive on the steroid substrate. These data give insight into the fine regulation of steroid genes when several isoforms are present.


An example of problems in dose reconstruction from doses formed by electromagnetic irradiation by different energy sources.
Dose reconstruction for citizens of Belarus affected by the Chernobyl accident showed an unexpectedly wide range of doses. Using the EPR tooth enamel dosimetry method, it has been demonstrated that when the tooth enamel dose was formed due to x-rays with effective energy of 34 keV and the additional irradiation of enamel samples was performed by gamma radiation with mean energy of 1,250 keV, it led to a considerable increase in the reconstructed absorbed dose as compared with the applied. In the case when the dose was formed due to gamma radiation and the additional irradiation was performed by x-rays, it led to a considerable decrease in the reconstructed dose as compared with the applied. When the dose formation and the additional irradiation were carried out from external sources of electromagnetic radiation of equal energy, the reconstructed dose value was close to that of the applied. The obtained data show that for adequate reconstruction of individual absorbed doses by the EPR tooth enamel spectra, it is necessary to take into account the contribution from diagnostic x-ray examination of the teeth, jaw, and skull of some individuals who were exposed to a combined effect of the external gamma radiation and x-rays.


Evaluating outcomes of care and targeting quality improvement using Medicare health outcomes survey data.
The Medicare Health Outcomes Survey (HOS) provides a rich source of outcomes data on the Medicare Advantage (MA) program for the US Department of Health and Human Services, managed care organizations participating in Medicare, quality improvement organizations, and health services researchers working to improve quality of care for Medicare enrollees. Since 1998, the Centers for Medicare and Medicaid Services has collected longitudinal functional status information to assess the performance of Medicare managed care organizations. This introduction reviews the goals of the HOS program, how the HOS supports health care reform, and outlines recent HOS studies exploring data applications for monitoring outcomes and implementing quality improvement activities.


Evaluation of bupivacaine-clonidine combination for unilateral spinal anesthesia in lower limb below-knee orthopedic surgery.
The purposes of this study were to evaluate the onset, quality and duration of sensory and motor blockade between hyperbaric bupivacaine and clonidine combination with bupivacaine alone when administered intrathecally for unilateral spinal anesthesia in below-knee orthopedic surgery, efficacy of clonidine for post-operative analgesia and side-effects of clonidine, if any. Sixty ASA I and ASA II patients scheduled for elective surgery with time duration up to 90 min were studied. Patients were randomised in two equal groups by the lottery method. Group A (control group) was given Inj. bupivacaine (hyperbaric) 0.5% - 12.5 mg (2.5 ml) + 0.5 ml of normal saline intrathecally. Group B (clonidine group) was given Inj. bupivacaine (hyperbaric) 0.5% - 12.5 mg (2.5 ml) + 50 mcg clonidine in 0.5 ml volume intrathecally. The mean peak sensory block was earlier in Group B (4.7±1.23 min) as compared with Group A (6.27±1.51 min). The mean peak motor block was earlier in Group B (6.17±1.20 min) as compared with Group A (8.63±1.71 min). The two-segment regression of sensory block was longer in Group B (106.23±9.17 min) as compared with Group A (104.43±17.75 min), which is clinically significant. Requirement of rescue analgesia was considerably prolonged in Group B (450.33±95.10 min) as compared with Group A (220±36.36 min), which was also clinically highly significant. Intrathecal clonidine potentiates bupivacaine induced spinal sensory block and, motor block and reduces the analgesic requirement in the early post-operative period in unilateral spinal anesthesia for lower limb below knee surgery.


A study on the sequence of phototoxic effects of rose bengal using retinal pigment epithelial cells in vitro.
This paper describes a study on the sequence of the phototoxic effects of rose bengal (RB), a fluorescein derivative used as a vital stain in the diagnosis of certain external ocular diseases. Bovine melanotic RPE cells were grown in culture. These cells were labeled with [51Cr] and exposed to visible light in the presence of various concentrations of RB; the leakage of [51Cr] from the cells was used as a measure of cell lysis. Exposure to light of the cells with 0.3-10 microM RB induced approximately 13 to 43% cell lysis. The lysis progressively increased when the exposure time was varied from 10 to 30 min. A relatively short period of irradiation in the presence of RB was sufficient to produce sublytic cellular injury which could subsequently lead to complete cell lysis even in the absence of the photochemical treatment. The dark reaction was time-dependent, and reached a maximum for a given irradiation period. Our results thus show that there are two different processes that could eventually lead to the cell lysis: (a) a phototoxic effect which caused a sublytic damage and (b) a dark reaction that followed.


Noise and hearing loss: a review.
Noise-induced hearing loss is a major cause of deafness and hearing impairment in the United States. Though genetics and advanced age are major risk factors, temporary and permanent hearing impairments are becoming more common among young adults and children especially with the increased exposure to portable music players. Though treatment options are limited for most people with noise-related hearing loss, several modifiable health behaviors that should begin in childhood might prevent or delay the onset of hearing impairment. The purpose of this article is to review modifiable and non-modifiable risk factors, comorbidity, and the role of health education in the prevention of noise-induced hearing loss. Review of current literature in the etiology, prevention, and treatment of noise-induced hearing loss as well as the role of health education. Non-modifiable risk factors related to noise-related hearing loss include increasing age, genetics, male gender, and race. Modifiable risk factors are voluntary exposure to loud noise, nonuse of hearing protection, smoking, lack of exercise, poor diet, tooth loss, and the presence of diabetes and cardiovascular disease. As hearing impairment among children and teenagers rises due to mostly voluntary exposure to loud noise, there are many implications for health education. Health educators need to address barriers to the use of hearing protection, deliberate exposure to loud music, and other modifiable risk factors, which cause and exacerbate hearing loss among those exposed to loud noise.