Mesoporous organosilicas with highly-content tyrosine framework as extractive phases for non-steroidal anti-inflammatory drugs in aquatic media.

BACKGROUND: Attentions regarding ordered mesoporous silica materials (OMSs), with large specific surface areas and narrow pore size distribution, which are prepared via self-assembly techniques, have been raised in sorption, separation, and sample preparation. However, in order to extend and improve their applications, a functionalization step is required. Organic units can be anchored on the inner or outer surface as well as in the silica wall framework by co-condensation-, grafting-, and periodic mesoporous organosilica (PMO) preparation approaches. Apparently, by synthesizing PMO with extensive and flexible organic bridging groups within the mesoporous wall, an efficient extractive phase can be achieved. RESULTS: We employed tyrosine amino acid to synthesize a PMO-based extractive phase. The FT-IR, 1H NMR, HR-ESI-MS, Low angle-XRD, TEM, FESEM, BET, and EDX-MAP analyses confirmed the successful synthesis of PMO within the salt-assisted templating method. A comprehensive study on sorption behavior of PMO was performed and its efficiency was evaluated against the grafting and co-condensation methods. Then, it was implemented to the pipette tip-micro solid phase extraction (PT-µ-SPE) of widely used non-steroidal anti-inflammatory drugs (NSAIDs) in water/wastewaters. Limits of detection and quantification were obtained in the range of 0.1-1.5 and 0.3-5 µg L-1, respectively. The calibration plots are linear in the 1-1000, 3-1000, 10-750, and 3-750 µg L-1, respectively. The intra-and inter-day precision at 50 and 200 µg L-1 levels are 2.9-7.1 % and 3.5-8%, while recoveries are between 84 and 111 %. SIGNIFICANCE: High-capacity tyrosine functionalized PMO with 2D hexagonal symmetry silica mesoporous structures found to be highly efficient extractive media. Despite the bulkiness and flexibility of the bridging group within the mesoporous wall, the synthesis condition was optimized in order to load more organic content in the PMO structure. The PMO performance was superior over organically modified ordered mesoporous silica materials prepared by the grafting and co-condensation methods.

Temporal trends and prediction of bovine tuberculosis: a time series analysis in the North-East of Iran.

Background: Bovine tuberculosis (BTB) is a disease with high economic relevance. Aims: This study aimed to determine a fast alert surveillance system for bTB before the outbreak in the epidemic region of Iran. Methods: This cross-sectional study was conducted using the Auto-Regressive Integrated Moving Average (ARIMA) model for monthly bTB detections (reactors). These reactor cases result from the positive Tuberculin Purified Protein Derivative (PPD) test on cattle farms for the period between April 2007 and March 2019 in Razavi Khorasan province. Autocorrelation functions (ACF) and partial autocorrelation functions (PACF) plots were used to determine model parameters. The Akaike Information Criteria (AIC) were employed to select the best-fitted model. The root mean square error (RMSE) was applied for the evaluation of the models. Then, the best-fitted model was hired to predict the cases for 12 oncoming months. The data were analysed by STATA (ver. 14) software with a significant level at P≤0.05. Results: ARIMA (3, 0, 3) 12 was introduced as a recommended fitted model according to white noise residual test (Q=22.87 and P=0.98), lower AIC (541.85), and more precise model RMSE (1.50). However, the forecast values were more than the observed values. Conclusion: The application and interpretation of ARIMA models are straightforward, and may be used as immediate tools for monitoring systems. However, we proposed an Auto-Regressive Integrated Moving Average with Exogenous Input (ARIMAX) model with some measurable exotic factors such as economic fluctuations, climate changes, and pulmonary tuberculosis to introduce a more precise and accurate model for the fast alert surveillance system.

Differential regulation of CXC ligand 1 transcription in melanoma cell lines by poly(ADP-ribose) polymerase-1.

The continuous production of the CXC ligand 1 (CXCL1) chemokine by melanoma cells is a major effector of tumor growth. We have previously shown that the constitutive expression of this chemokine is dependent upon transcription factors nuclear factor-kappa B (NF-kappaB), stimulating protein-1 (SP1), high-mobility group-I/Y (HMGI/Y), CAAT displacement protein (CDP) and poly(ADP-ribose) polymerase-1 (PARP-1). In this study, we demonstrate for the first time the mechanism of transcriptional regulation of CXCL1 through PARP-1 in melanoma cells. In its inactive state, PARP-1 binds to the CXCL1 promoter in a sequence-specific manner and prevents binding of NF-kappaB (p65/p50) to its element. However, activation of the PARP-1 enzymatic activity enhances CXCL1 expression, owing to the loss of PARP-1 binding to the CXCL1 promoter, accompanied by enhanced binding of p65 to the promoter. The delineation of the role of NF-kappaB-interacting factors in the putative CXCL1 enhanceosome will provide key information in developing strategies to block constitutive expression of this and other chemokines in cancer and to develop targeted therapy.

Antigen stool test for assessment of Helicobacter pylori infection in patients with upper gastrointestinal bleeding.

BACKGROUND: Re-bleeding of Helicobacter pylori-associated peptic ulcer disease is reduced by H. pylori eradication. AIM: To validate a non-invasive test, the Premium Platinum HpSA stool test, in patients with upper gastrointestinal bleeding. METHODS: Stool samples of consecutive patients with relevant bleeding from gastric or duodenal ulcers or erosions were collected at initial endoscopy and during the following week. Samples were assessed using the HpSA test. H. pylori status was defined by three biopsy-based reference methods: culture, rapid urease test and histology. It was positive if culture was positive or if rapid urease test and histology were positive. RESULTS: One hundred and fourteen patients (mean age, 66 years) were included. In accordance with the definition, 56 (49%) were H. pylori positive. The sensitivity and specificity of the first stool sample were 84% and 90%, respectively. The respective values for two samples from consecutive days were 91% and 86%. In comparison with a serum immunoglobulin G antibody enzyme-linked immunoabsorbent assay, the HpSA test showed superior specificity. CONCLUSIONS: The diagnostic accuracy, in particular the sensitivity, of the HpSA stool test is reduced by upper gastrointestinal bleeding. The positive predictive value of 89%, however, justifies the initiation of eradication therapy on the basis of a positive stool test. A negative test result should be confirmed by a further diagnostic method.

Prevalence of Helicobacter pylori infection among low socio-economic workers.

Epidemiological studies have shown that the prevalence of Helicobacter pylori infection in a community and occupational health are closely related to lifestyle and socio-economic status. There is little information on H. pylori profile in industrial workers in the literature. The aim of this study was to investigate the prevalence rate of H. pylori profiles among low socio-economic workers in the United Arab Emirates (UAE). This study was undertaken by determining IgG H. pylori antibody profiles among industrial exposed and referent workers, sera. Presence of anti-H. pylori antibodies in the frozen stored sera was determined by ELISA. Also, data on dietary and lifestyle were obtained. The result was considered positive if IgG anti-H. pylori antibody titers was > 300. People with seropositive levels of IgG antibodies to H. pylori were assumed to be infected with H. pylori. Most of the industrial workers lived in less modern accommodation, were less educated, ate their vegetable products unwashed and did not have drinking water facilities, when compared to referents. H. pylori serology by IgG was positive in 167 industrial workers (78.4%) and 137 in referent workers (64.3%) respectively, (p < 0.002). The sensitivity and specificity of the IgG serology assay were 94.5%, and 97.2% respectively. There was statistically significant difference between the exposed industrial and non-exposed control groups in respect of their H. pylori profiles.

A novel immunoassay based on monoclonal antibodies for the detection of Helicobacter pylori antigens in human stool.

BACKGROUND AND AIM: With the Premier Platinum HpSA EIAtrade mark a new enzyme immunoassay was developed for diagnosis of H. pylori infection, using polyclonal antibodies against H. pylori antigens in human stool. Here we evaluated FemtoLab H. pyloritrade mark based on the use of monoclonal antibodies in comparison to established reference methods. METHODS: 53 consecutive patients (27male, 26 female, age: 17-85 years) undergoing routine upper gastrointestinal endoscopy were enrolled in this study. The H. pylori status was determined by 4 reference methods: Histology, rapid urease test (HUT), (13)C-urea breath test ((13)C-UBT) and serology. Patients were considered to be infected with H. pylori if at least 2 of the 4 reference tests were positive. Stool samples were aliquoted after reception and stored frozen (-20 degrees C) until tested. The FemtoLab H. pyloritrade mark (Connex GmbH, Germany) and the Premier Platinum HpSA EIAtrade mark (Meridian, Connecticut, Ohio, USA) were performed according to the manufacturers protocols. RESULTS: 26 of the 53 patients were H. pylori infected. 3 were false-negative by the FemtoLab H. pyloritrade mark and one false-positive result was obtained (sensitivity 88,5 %, specificity 96,3 %). The concordance between the 2 stool tests was 94,3 % (50/53 cases). CONCLUSION: The diagnostic quality of the novel FemtoLab H. pyloritrade mark Enzyme Immunoassay is comparable with the established Premier Platinum HpSA EIAtrade mark. The differences between positive and negative results obtained with the FemtoLab H. pyloritrade mark are greater in comparison to the Premier Platinum HpSA EIAtrade mark and therefore this test system allows a better distinction.

Validity of a Helicobacter pylori stool antigen assay for the assessment of H. pylori status following eradication therapy.

BACKGROUND: A Helicobacter pylori stool antigen (HpSA) test has been proposed as a valid alternative to the 13C-urea breath test (13C-UBT) for the non-invasive detection of H. pylori infection in primary diagnosis. Published reports show conflicting results with regard to the test's diagnostic accuracy after eradication therapy. The aim of the present study was to assess the diagnostic value of the HpSA test and to determine the optimal discriminating cut-off value in patients following H. pylori eradication therapy. METHOD: Stool samples were collected and the 13C-UBT was performed in 113 patients 4-6 weeks after eradication therapy. A validated test protocol for the 13C-UBT was used. Stool specimens were analysed with the Premier Platinum HpSA enzyme immunoassay (EIA). A receiver operator characteristics (ROC) analysis was performed to define the optimal cut-off value on the basis of the results of the 13C-UBT. RESULTS: The results of the 13C-UBT showed that H. pylori eradication was successful in 83/113 (73%) patients. According to the manufacturer, the cut-off value for the HpSA test is 0.14 optical density, but this does not appear to be valid after eradication therapy (sensitivity 76.7%, specificity 98.8%). On the basis of ROC analysis, the optimal cut-off value after therapy was determined to be 0.11 optical density, giving a sensitivity of 93.3% and a specificity of 93.9%. CONCLUSION: The HpSA test is a valid test for the assessment of H. pylori status after eradication therapy, provided an adjusted cut-off value is applied.

The Helicobacter pylori vacA s1, m1 genotype and cagA is associated with gastric carcinoma in Germany.

BACKGROUND: The H. pylori vacuolating cytotoxin encoded by vacA and the cytotoxin-associated protein encoded by cagA are considered to be important virulence determinants that have been implicated in the development of peptic ulcers and gastric carcinoma. However, conflicting results regarding the association between these virulence factors and clinical disease have been reported from different geographic regions. AIMS: To determine the frequency of vacA genotypes, vacuolating cytotoxin activity, and cagA in H. pylori isolates obtained from patients with gastric cancer in Germany. METHODS: H. pylori isolates were obtained from 34 patients with gastric cancer and from 35 subjects with asymptomatic H. pylori gastritis. vacA genotypes and cagA were identified by PCR. Cytotoxic activity was determined by HeLa cell assays. Gastritis was assessed according to the updated Sydney System. RESULTS: The H. pylori vacA s1,m1 genotype was significantly more frequent in patients with gastric cancer (24/34, 70.6%) as compared with controls (12/35, 34. 3%) (p = 0.005). Cytotoxic activity was detected in 24 (70.6%) and 15 (42.9%) H. pylori isolates from gastric cancer patients and controls, respectively (p = 0.03). The cagA gene was identified in 30 (88.2%) and 21 (60%) H. pylori isolates in the respective groups (p = 0.01). CONCLUSIONS: Our study suggests a significant association between the H. pylori vacA s1,m1 genotype, cytotoxic activity, cagA, and gastric cancer. Detection of H. pylori possessing these virulence determinants may help to identify patients being at an increased risk to develop gastric cancer in our population.

Evaluation of an enzyme immunoassay for detecting Helicobacter pylori antigens in human stool samples.

BACKGROUND AND AIM: So far, the detection of Helicobacter pylori (Hp) infection by stool analysis appeared to be almost impossible. With the Premier Platinum HpSA EIA a new enzyme immunoassay was developed for diagnosis of Hp infection, using polyclonal antibodies against Hp antigens in human stool. We evaluated this new test in its diagnostic accuracy in comparison to established reference methods. METHODS: From 54 consecutive patients (29 male, 25 female, age: 19 to 85 years) undergoing routine upper gastrointestinal endoscopy antral and corpus biopsies were taken for histology and Helicobacter urease test (HUT). Endoscopy, 13C-urea breath test (13C-UBT), serology, and stool probes sampling were performed within two days. Stool samples were aliquoted after reception and stored frozen (-20 degrees C) until tested. The Premier Platinum HpSA test (Meridian, Connecticut, Ohio, USA) was performed according to the manufactures protocol. Patients were considered to be infected with Hp if two of the four reference tests were positive. RESULTS: 28 of the 54 patients were Hp-infected. Only one of these was found to be false-negative by the HpSA EIA. Two false-positive results were obtained in the noninfected group (sensitivity 96.4%, specificity 92.3%). CONCLUSION: In this group of patients investigated, the novel HpSA Enzyme Immunoassay (EIA) proved to be highly accurate for diagnosis of Hp infection. Collection and testing of stool are noninvasive and easy to perform, therefore this test will become an important tool for diagnosing Hp infection in clinical practice.

The global conformation of an active hammerhead RNA during the process of self-cleavage.

The RNA "hammerhead" domain is a small element of secondary structure found in the genomes of certain plant pathogens. It possesses a core of conserved sequence at the conjunction of three helix stems, and is capable of undergoing self-cleavage in the presence of divalent cations. Both crystallographic and solution studies suggest that the domain is highly structured, with the three stems assuming a Y-shaped global conformation; however, such studies have employed either RNA analogues that were catalytically inactive, or conditions of temperature and pH for which rates of self-cleavage are slow. Thus, it was unknown whether such species represented the principal conformers during the cleavage process itself. In order to address this issue, a series of time-resolved, transient electric birefringence measurements was conducted in an effort to define the global conformation of an RNA hammerhead in real time throughout the process of self-cleavage. The current study demonstrates that the angular relationship between the two helices that flank the cleavage center is essentially unchanged between the pre-cleavage and post-cleavage forms. Moreover, despite the fact that at least one kinetic intermediate is formed during the self-cleavage reaction, there is no evidence for the existence of a significant population of intermediates with altered global conformation during cleavage. Thus, any conformational isomerism that may occur is likely to be relatively localized to the active center. Finally, it was observed that sequence elements lying outside of the conserved region, at the base of stem I, influence interhelix geometry. The current results are consistent with a structural model in which the active center possesses similar conformations pre-cleavage and post-cleavage. Such a model would help to explain the significant rate of reversal of the cleavage reaction (self-ligation).

Hammering away at RNA global structure.

A major goal of the study of RNA tertiary structure is an understanding of the rules relating sequence and global conformation. This goal has been furthered during the past year for two important structural elements: yeast tRNAPhe and the self-cleaving hammerhead RNA. In both cases, a combination of solution and crystallographic studies has yielded strongly concordant views of their global conformations.

Global conformation of a self-cleaving hammerhead RNA.

Phylogenetic and biochemical studies of RNA have provided a wealth of information regarding secondary structure; however, knowledge of tertiary structure has been more difficult to obtain. In the current work, electrophoretic and hydrodynamic measurements have yielded the global conformations of a self-cleaving "hammerhead" RNA, both prior to and following self-cleavage. The pre- and post-cleavage structures appear to have nearly identical conformations in which the three helices assume a roughly coplanar arrangement with well-defined interhelix angles. Following self-cleavage, one of the three helix stems (stem I) undergoes a slight (< 10 degrees) realignment, although the relative positions of the other two stems remain unchanged. Extension of stem III from (nominally) three base pairs to nine base pairs, known to dramatically enhance the rate of self-cleavage, results in a substantial (approximately 70 degrees) realignment of stems I and II. The current study represents an approach for studying other nonhelical elements in RNA.

Fibrillarin-like proteins occur in the domain Archaea.

Fibrillarin is found in the nucleolus of Eucarya and associated with small nucleolar RNAs. It is involved in the processing of precursor rRNA. Two genes, encoding fibrillarin-like proteins from Methanococcus voltae and Methanococcus vannielii, have been isolated. The genes were named flpA (fibrillarin-like protein).

Interhelix geometry of stems I and II of a self-cleaving hammerhead RNA.

In order to investigate the geometry of a self-cleaving hammerhead domain, an RNA heteroduplex has been constructed in which two of the three helix stems of the domain have each been elongated to 76 duplex base pairs (bp), resulting in an RNA molecule of ca. 160 bp. The heteroduplex molecule is capable of undergoing self-cleavage at neutral pH, upon addition of either Mg2+ or Mn2+, but does not dissociate following cleavage. Using a combination of electrophoretic and hydrodynamic methods, as employed earlier to define the geometry of a four-way DNA branch [Copper & Hagerman (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 7336-7340], we have determined that the elongated hammerhead stems are nearly collinear prior to self-cleavage. Following self-cleavage, and in the absence of Mg2+, the angle between the two stems becomes much more acute and/or flexible; however, in the presence of Mg2+, the cleaved structure appears to retain the geometry of the precleaved form (at least with respect to the interstem angle). It is also observed that the self-cleavage reaction is promoted by Mn2+ to a much greater extent than by Mg2+, consistent with earlier observations. Finally, although the elongated helices appear to be nearly collinear in the uncleaved molecule, the electrophoretic mobility of that species is dramatically reduced with respect to linear control RNAs, indicating that caution should be exercised in the quantitative assignment of branch angles solely from gel retardation experiments.

Selenium is involved in the negative regulation of the expression of selenium-free [NiFe] hydrogenases in Methanococcus voltae.

Competitive polymerase chain reactions (PCR) were used to analyze quantitatively the transcription patterns of the four different gene groups encoding [NiFe] hydrogenases in Methanococcus voltae. In cells growing in the presence of selenium, transcripts of only two of the hydrogenase transcription units could be detected in quantities above background levels. The missing transcripts encode the selenium-free F420-non-reducing and F420-reducing hydrogenases. In cells grown without selenium these transcripts are detectable, indicating the involvement of selenium in their transcriptional regulation.

Girls with fragile X syndrome: physical and neurocognitive status and outcome.

The fragile X syndrome, a common X-linked form of mental retardation and autism, affects females as well as males. Previous work has shown that approximately 35% of heterozygotes (women who carry the fragile X gene) demonstrate cognitive impairment. Thirty-two girls, 18 years or younger, who demonstrate the fragile X chromosome were evaluated and compared with 19 sisters who do not demonstrate the fragile X chromosome. Evaluations included a physical examination, behavioral assessment, and intelligence testing. Significant differences (in intellectual, behavioral, and physical features) were seen between the two groups. Twenty-five percent of fragile X-positive girls had an IQ in the mentally retarded range (IQ less than 70) and 28% had an IQ in the borderline range (70 to 84). Prominent ears, shyness, and poor eye contact were significant findings in fragile X-positive girls compared with fragile X-negative girls. Thirty-one percent of the fragile X-positive girls had significant attentional difficulties and most of these girls were successfully treated with stimulant medication. The majority of fragile X-positive girls in this study demonstrated significant behavioral and developmental problems which required identification and appropriate treatment. Pediatricians and health care providers should be aware of the frequency and manner with which fragile X affects females in order to initiate cytogenetic studies and treatment when indicated.

Heterozygous fragile X female: historical, physical, cognitive, and cytogenetic features.

Historical, physical, cognitive, and cytogenetic data were documented in 105 heterozygous fragile X [fra(X)] females and 90 controls in a prospective fashion. For comparisons, we divided heterozygotes and controls into those with cognitive impairment (IQ less than 85) and normal IQ (IQ greater than or equal to 85). The only finding that was significantly more frequent in impaired heterozygotes compared with impaired controls chi 2 analysis was shyness. Features that were more frequent in normal IQ heterozygotes compared with normal controls were voluntary thumb dislocation and hyperextensible metacarpal-phalangeal (MP) joints. Comparisons among heterozygotes demonstrated more math problems, hand biting, strabismus, high-arched palate, hyperextensible finger joints, and flat feet in impaired heterozygotes than in normal heterozygotes. Premature menopause was present in 8 of 61 normal heterozygotes and in none of the impaired heterozygotes. A multiple regression analysis demonstrated a significant inverse correlation between the percent fragility and IQ for the heterozygotes as a group. However, no correlation existed between IQ and fragility when the percent fragility was 2% or greater. However, a higher percentage of fragility was positively correlated with the total number of physical findings present.