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Jan 2017 DOI 10.14302/issn.2476-1710.jdt-16-1324
An important step in the validation of disorder-specific etiological models is the examination of the predictive specificity of proposed vulnerability factors. It may advance the understanding of the emergence of comorbidity and the identification of at risk-populations for mental disorders. To enhance the currently limited evidence on the specificity of Beck´s cognitive diathesis-stress model of depression, the present study investigated longitudinal effects of dysfunctional attitudes and stressful life events on the development of depressive, eating disorder and aggressive symptoms in children and adolescents. A large sample of initially asymptomatic children and adolescents completed self-report symptom measures at study entrance and again approx. 20 months later, and reported stressful life events during the study interval. Stressful life events proved to be a risk factor to all investigated symptom domains. Dysfunctional attitudes at T1 were prospectively related to depressive symptoms, aggressive behavior and weight concerns at T2. However, types of associations varied as dysfunctional attitudes showed linear associations with weight concern, but nonlinear effects on depressive and aggressive symptomatology. Findings of the current study thus suggest that dysfunctional attitudes are not uniquely related to the development of depressive symptomatology in children and adolescents, but may contribute to adverse outcomes in various symptom domains. Thus, intervention efforts based on Beck´s vulnerability - stress model of depression may turn out to be useful in reducing vulnerability to a variety of outcomes in children and adolescents.
Nov 2025 DOI 10.14302/issn.2690-4721.ijcm-25-5786
In the Republic of the Congo, tuberculosis (TB) remains a major public health concern. Although the GeneXpert MTB/RIF assay is the WHO-recommended first-line diagnostic test, smear microscopy is still used for treatment monitoring and in facilities where molecular testing is limited. Evaluating the diagnostic accuracy of smear microscopy compared to GeneXpert and MGIT culture is essential to guide diagnostic strategies and strengthen TB control in the country. A cross-sectional study was conducted among 92 presumptive pulmonary TB patients at Makelekele Hospital. Sputum samples were analyzed by smear microscopy, GeneXpert MTB/RIF, and MGIT culture. Sensitivity, specificity, positive and negative predictive value were calculated for smear microscopy and GeneXpert, using culture as the reference standard. Culture detected more Mycobacterium tuberculosis than microscopy (49% vs. 32%, P<0.001). Smear microscopy showed a sensitivity of 58% (95% CI: 43–71%) and specificity of 92% (95% CI: 80–97%). GeneXpert detected more MTB (62% vs. 49%, P<0.001) with a sensitivity of 98% (95% CI: 89–100%) and specificity of 72% (95% CI: 58–83%). GeneXpert showed superior sensitivity for TB detection, while microscopy remained specific. Expanding GeneXpert testing across the Republic of the Congo will improve TB management.
Sep 2023 DOI 10.14302/issn.2692-1537.ijcv-23-4660
SARS-CoV-2 real-time reverse-transcription PCR (rRT-PCR) is the most effective testing system available to combat COVID-19, given the absence of any specific treatment or vaccine. Moreover, numerous SARS-CoV-2 rRT-PCR kits have been approved under emergency-use-authorization (EUA) worldwide. In this article, we present a comparison of important performance features among five commercial RT-PCR assays. A total of consecutive nasopharyngeal (NPS) samples and oropharyngeal (OP) swabs were collected from 50 COVID-19 patients to analyze sensitivity and specificity. The results showed variations in sensitivity among all the RT-PCR kits examined. The Pishtaz teb assays demonstrated the highest positive percent agreement (PPA) of 95.2% (40/42), followed by Covitech (90.5% - 38/42), DaAn Gene (83.3% - 35/42), Sansure (66.66% - 28/42), and Power check of SARS- CoV-2 panel (64.3% - 27/42). Conversely, all five molecular assays demonstrated a negative percent agreement (NPA) of 100% (8/8). These findings provide a technical baseline for assessing the performance of five distinct commercial PCR assays for detecting SARS-CoV-2. They could prove practical and useful for laboratories seeking to purchase any of these assays for further clinical validation. Highlights ·Compared five COVID-19 RT-PCR kits approved and available by Iran Ministry of Health. ·Pishtaz teb's kit identified the highest number of positive clinical samples.
Feb 2023 DOI 10.14302/issn.2641-5526.jmid-23-4450
Acute appendicitis is one of the most common surgical emergencies globally, with a lifetime incidence of 8.6% in men and 6.7% in women. While acute appendicitis should be managed promptly to reduce the morbidity associated with perforated appendicitis, morbidity from negative appendicectomy is similar to morbidity from uncomplicated appendicitis. Computer tomography is widely used to aid in the diagnosis of acute appendicitis, however, is costly, often has a slow turn around time, and is associated with exposure to ionising radiation. In contrast, ultrasound is cheap, widely available, requires minimal patient preparation, and does not require exposure to ionising radiation. Ultrasonography is becoming increasingly used for adult patients in emergency settings. The literature has estimated the sensitivity of ultrasound for acute appendicitis in adult patients as between 39-96.4%. The sensitivity and specificity of ultrasound for the diagnosis of acute appendicitis is significantly increased when the appendix is visualised. In cases of a non visualised appendix, indirect ultrasound signs can improve the sensitivity to 93.9% and specificity to 85.7%. The variation in sensitivity and specificity for ultrasound in the diagnosis of acute appendicitis in adults may be due to multiple factors. Ultrasonographer experience, a retrocaecal appendix and obesity have all been described. Given the availability, cost and potential to reduce the rate of negative appendicectomy, ultrasound should be considered as the first line imaging modality for adult patients presenting with suspected AA.
Sep 2022 DOI 10.14302/issn.2692-1537.ijcv-22-4328
Objectives To evaluate the diagnostic accuracy of chest CT for the diagnosis of COVID-19 associated with the clinical presentation and in relation to the PCR-RT. Sensitivity, specificity, positive predictive value and negative predictive value, gender, age group and degree of lung involvement will be evaluated. Methods We evaluated 1545 patients with chest CT, delineating the age range and degree of lung involvement, and 306 patients with chest CT and PCR-RT. Results Of the 1545 examinations, 53% were men and 47% were women, there was greater involvement in the 50-59 age group. In the pulmonary study, 55.05% were COVID-19. In the degree of lung involvement 37.70% were mild, 35.76% were moderate, and 26.54% were severe. In the distribution by age, there was a greater involvement between 50-59 years with 56% between moderate (27.6%) and severe (28.0%). Between tomography and PCR-RT, the sensitivity was 68.8%, specificity 59.5%, accuracy 91.3%, with prevalence 31.9%, positive predictive value 44.3% and negative predictive value 80.3%, in females, sensitivity 55.3%, positive predictive value 37.1%, negative predictive value 75.3%, in males, sensitivity 81.6%, positive predictive value 50, 6 and negative predictive value 86.6%.The sensitivities are different between the genders with p of 0.005 and specificity of 0.938, with age effect, starting at 45 years we have a p of 0.057 that decreases to 0.006 at 80 years for sensitivity and specificity. Conclusions The sensitivity and accuracy of CT scan in relation to PCR-RT was significant. Sensitivity increases with prevalence and in the older age group and in men.
Jun 2022 DOI 10.14302/issn.2328-0182.japst-22-4193
The strategy for safe drug discovery and development has limited clinical success as compared to wasted time and resources annually. This is due to the fact that the results of multiphase preclinical trials are less likely to make an accurate early prediction on the safety of test compounds to progress into the clinic as a valuable therapeutic agent. A lot of time and resources has been wasted in the multistage processes of drug discovery and development that does not work at the end of the procedure every year. During pre-marketing stage, for instance, the number of unsuccessful clinical trials are greater than the successful one because of safety issues. A toxicity study at different stages of preclinical and clinical trials is a routine procedure to investigate the undesirable side effects of test compounds being manifested on the natural processes of living things. It deals with the effect and mechanism of toxicity of test compounds that triggers different biological responses on different organ systems. The biological responses that would be manifested as a result of interaction between the receptors and active molecules of a test compound could be desirable pharmacological effect or undesirable side effect or both responses are manifested simultaneously depending on the selectivity or specificity of the molecule of a test compound for its receptor subtype which makes safe drug discovery and development very challenging. The response efficiency of the body (the net outcome of the body’s biological reaction against the side effect) would determine the potency of a test compound to manifest undesirable pharmacologic effect. In other words, the amount of a drug required to cause a biological harm or injury depends on the magnitude of the body’s biological reaction in which the immune response plays a great pharmacological role by neutralizing and harmonizing xenobiotics with the biological molecules. The dose of a test compound at 100 mg/kg body weight, for instance, could be lethal to some of the study animals while it is still non-lethal to some other study animals depending on the response efficiency of the body. The immune system is well connected to each and every biological systems of the body which allows it to detect undesirable side effects being manifested through immunoglobulins signalling and activation mechanisms. This complex communication network helps to localize the diverse side effects of a test compound being manifested on different organ systems into the immune system which makes a toxicity study relatively simple to monitor. The cellular immune system becomes active following the molecule-receptor interaction and start producing antibodies which is also known as immunoglobulins to protect bodily harm and destruction. Under normal biological circumstances, the amount of immunoglobulins produced by the cellular immune system following exposure to a test compound is proportional to the number of harmful molecules interacted with its receptor subtype. Thus, with the reference to the changes in the immune response against the administered dose, it would be able to deal with the diverse undesirable side effects of a test compound being manifested on treated study animals using computational systemic biology.
May 2022 DOI 10.14302/issn.2766-8681.jcsr-22-4162
Alveolar-arterial oxygen pressure difference (P(Aa)O2) can reflect pulmonary ability to exchange oxygen; it shows good correlation with the oxygenation index (OI), which is important in diagnosing acute respiratory distress syndrome (ARDS). This study explored the ability of P(Aa)O2 in diagnosing ARDS in pneumonia patients. Methods We selected patients with community-acquired pneumonia and sepsis in the intensive care unit (ICU) of the People’s Hospital of Qiandongnan Miao and Dong Autonomous Prefecture; we measured P(Aa)O2 and the OI under anoxic conditions upon their admittance to the ICU. We divided the patients into ARDS and non-ARDS groups. We compared the differences in P(Aa)O2 and OI; we analyzed the correlation between P(Aa)O2 and ARDS. To assess the diagnostic ability of P(Aa)O2 for ARDS, we drew the receiver operating characteristic (ROC) curve. Result We found that P(Aa)O2 in the ARDS group was greater than in the non-ARDS group (t = 8.875, P <0.001); the OI in the ARDS group was smaller than in the non-ARDS group (t = –6.956, P <0.001). There was a positive correlation between P(Aa)O2 and ARDS (r = 0.718, P <0.001). The area under the ROC curve for P(Aa)O2 in the diagnosis of ARDS was 0.931 (0.873–0.988); the cutoff value was 214.70 mmHg, the sensitivity was 89.50%, and the specificity was 85.00%. Conclusion We conclude that P(Aa)O2 is a good reference index in diagnosing ARDS.
Aug 2021 DOI 10.14302/issn.2326-0793.jpgr-21-3917
50 years ago the Enzyme Immunoassay Enzyme-Linked Immunosorbent Assay, mostly known as ELISA was developed. This is a powerful but simple method that is very widely used in the diagnostic practice, as well as in biomedical research. During this time a number of ELISA modification were developed that significantly increased its properties, especially the senstivity, such as avidin-biotin assay, immuno-PCR, nano-ELISA and finally, the digital ELISA. This short review describes the principles of ELISA and the evolution from a conventional assay to the modern ultra-sensitive method. Most of the immunological methods have two components: antigen and antibody. The high specificity of their interaction gives a possibility to detect one of them if other one is included in the reaction as a specific partner. The simplest method for antigen detection in the presence of the antibody is immune diffusion (radial immune diffusion in that case), which practically the formation of precipitate of the “antigen-antibody” complex, when the target antigen diffuses from well into agarose containing the specific antibody. Unfortunately, this assay, as well as other traditional methods, like hemagglutination or complement fixation, have a low sensitivity and are unwieldy.
Mar 2021 DOI 10.14302/issn.2575-1212.jvhc-21-3767
Antibodies and antibody fragments, especially single-domain antibodies known as nanobodies, are important tools in diagnostics, research, and therapeutics. In a conventional antibody, light and heavy chains contribute to the formation of the antigen binding site. In addition to conventional antibodies, old and new world camels also have heavy-chain antibodies (hcAbs), which lack the light-chain antibodies that usually bind to the antigen, as well as single domain antibodies, the VHH domain, which are the smallest antigen-binding fragments and have high solubility, stability, and specificity. A VHH library against E. coli lipopolysaccharide (LPS) was produced using the camel immune system. E. coli strains from dead camel calves were isolated to extract the LPS and used to immunize a 2-year-old female camel. After isolating mononuclear lymphocytes for RNA extraction and amplification of the VHH gene, the PCR product was cloned into the pF1AT7 Flexi vector and transformed into JM109 E. coli competent cells by heat shock, resulting in a comprehensive VHHs library with 6.9 × 104 cfu/µg. The VHHs were expressed and screened with ELISA and PCR. Eleven colonies were positive by PCR, six of which were sequenced and submitted to Genbank compared with GenBank data to confirm the production of nanobodies with a similarity >90%.
Oct 2020 DOI 10.14302/issn.2379-7835.ijn-20-3418
Isolation methods that employ readily-available inexpensive supplies on the open market, which are reliable, as well as economical, such as nucleic acid amplification techniques (NAAT) based on microfluidic technology in low-resource research settings (LRRS) that meets the ASSURED guidelines are essential to develop a noninvasive diagnostic colon cancer screen in stool using micro(mi)RNA molecules. A combination of a microfluidic-based MiRNA stool test with a reliable rolling circle amplification/detection method applied to the quantification of miRNA molecules, result in an affordable sensitive and specific isothermal method for the noninvasive quantitative detection of miRNAs in LRRS. Scientists and engineers have become interested in miRNAs, and they have intensified their efforts to apply emerging simple detection tools to the important bioanalytical challenge of quantifying these small 18-26 nt long molecules. Some of the proposed approaches incorporate novel material, such as simple centrifuges and methods based on microfluidic technology, while others utilize the interesting biological properties of these molecules, such as forming branched RCA structures, allowing for the detection of these biomarker molecules at an attomolar "aM" concentration level, using low cost extraction and isothermal amplification methods in LRRS. We have been interested in studying colorectal cancer (CRC) because it is the 3rd most common malignancy worldwide, and stool can be obtained noninvasively from the patients. We have focused in this research on colon cancer (CC) because it is more common in the USA than rectal cancer (RC). The innovation of our approach lies in the exploratory use of an affordable, quantitative miRNA profiling in noninvasive stool samples in LRRS, whose extracted fragile total RNA is stabilized shortly after excretion from stool by commercially available kits, so it does not ever fragment, followed by quantitative standardized analytical tests that are neither labor intensive, nor require expensive instrumentation, in order to develop apanel of novel miRNA genes for the noninvasive diagnostic screening of early left and right sporadic colon cancers, more economically, and with higher sensitivity and specificity than any other colon cancer screening test currently available on the market. To show the clinical sensitivity and specificity of the proposed quantitative miRNA test using simple methodologies in LRRS,the miRNA results are to be correlated with FOBT, colonoscopy, and pathology data. Standardization establishes test’s performance criteria (sample selection, optimal sample running conditions, preservation and storage), in order to ensure that the assay will perform the same way in any laboratory, by any trained personnel, anywhere in low-resource laboratory settings worldwide.
Sep 2020 DOI 10.14302/issn.2575-1212.jvhc-20-3477
A total number of 100 samples from ten random broiler chicken carcasses (breast and thigh) were collected from an automatic poultry slaughtering plant in Ismailia city, Egypt. The mean values of Enterobacteriacae count were 5.9x104±9.7x103 cfu/g and 7.1x 104 ± 1.1x104 cfu/g for chicken breast and thigh samples respectively. The prevalence of E.coli were 12% and 9% breast and thigh samples examined, respectively. They are serologically identified as 33.35 and 22.2% O157:H7 (EHEC) , 16.6% and 11.1% O114:H21(EPEC), 16.6% and 33.3 %O127:H6 (ETEC) , 0% and 0% O126 (ETEC) and 33.3% and 0% O26 (EHEC) for breast and thigh samples, respectively. The incidence of E.coli O157:H7 was 100% in both serological and PCR methods from biochemical positive E.coli samples. Culture is specific and cheap whereas PCR is sensitive and expensive, hence, we recommend both culture and molecular methods, which improve sensitivity and specificity, to enhance detection of foodborne pathogens including E.coli.
Aug 2020 DOI 10.14302/issn.2835-513X.ijl-20-3457
PEGylation is a well-established strategy for improving the target specificity, circulation time and stability of liposomes, thereby improving their stealth properties. This brief review provides an insight on the composition of PEGylated liposomes and the characteristics that dictate the functionality of PEGylated liposomes such as surface density, molecular weight, presence of linkers and acyl groups. Physicochemical techniques used to characterize the PEG liposomes and test their stability are also discussed along with their clinical implications. This review provides the readers with a broad range of understanding of various PEGylated lipids, techniques to access their stability in liposomal formulations and state-of -the-art development of PEGylated liposomal formulations.
Jun 2020 DOI 10.14302/issn.2379-7835.ijn-19-3123
Isolation methods that employ readily-available inexpensive supplies on the open market, which are reliable, as well as economical, such as nucleic acid amplification techniques (NAAT) based on microfluidic technology in low-resource research settings (LRRS) that meets the ASSURED guidelines are essential to develop a noninvasive diagnostic colon cancer screen in stool using micro(mi)RNA molecules. A combination of a microfluidic-based MiRNA stool test with a reliable rolling circle amplification/detection method applied to the quantification of miRNA molecules, result in an affordable sensitive and specific isothermal method for the noninvasive quantitative detection of miRNAs in LRRS. Scientists and engineers have become interested in miRNAs, and they have intensified their efforts to apply emerging simple detection tools to the important bioanalytical challenge of quantifying these small 18-26 nt long molecules. Some of the proposed approaches incorporate novel material, such as simple centrifuges and methods based on microfluidic technology, while others utilize the interesting biological properties of these molecules, such as forming branched RCA structures, allowing for the detection of these biomarker molecules at an attomolar "aM" concentration level, using low cost extraction and isothermal amplification methods in LRRS. We have been interested in studying colorectal cancer (CRC) because it is the 3rd most common malignancy worldwide, and stool can be obtained noninvasively from the patients. We have focused in this research on colon cancer (CC) because it is more common in the USA than rectal cancer (RC). The innovation of our approach lies in the exploratory use of an affordable, quantitative miRNA profiling in noninvasive stool samples in LRRS, whose extracted fragile total RNA is stabilized shortly after excretion from stool by commercially available kits, so it does not ever fragment, followed by quantitative standardized analytical tests that are neither labor intensive, nor require expensive instrumentation, in order to develop apanel of novel miRNA genes for the noninvasive diagnostic screening of early left and right sporadic colon cancers, more economically, and with higher sensitivity and specificity than any other colon cancer screening test currently available on the market. To show the clinical sensitivity and specificity of the proposed quantitative miRNA test using simple methodologies in LRRS,the miRNA results are to be correlated with FOBT, colonoscopy, and pathology data. Standardization establishes test’s performance criteria (sample selection, optimal sample running conditions, preservation and storage), in order to ensure that the assay will perform the same way in any laboratory, by any trained personnel, anywhere in low-resource laboratory settings worldwide.
Mar 2020 DOI 10.14302/issn.2639-3166.jar-20-3260
The management of the inoculation of a plant’s roots, by means of biofertilizers (BF) containing arbuscular mycorrhizal (AM) fungi, is aimed at inducing modifications of the quality of the seeds. It is here shown that a seed-soil treatment can be elicited in the fingerprints of a symbiotic treatment using Near Infra Red (NIR)-SCiO NIR-SCiO spectra collections of single kernels: overall, a sensitivity of 73% and a specificity of 73% have been achieved, thus suggesting that it may be possible to assign the symbiotic origin of corn from just twenty kernels, provided that the dataset is adequately representative of the cultivar and AM. A global correlation study has shown a positive general trend (R2 0.45) of quality vs. quantity, in the sense that an increase in yield corresponded to an increase in the spectral differences between the symbiotic spectra and the control ones, but the inverse was also true, as a result of the parasitic behaviour of the BF treatments. The efficacy of the symbiosis can be back predicted from the NIR spectra; in fact, around 90% of the positive yield outcome results were discriminated from the negative ones. A reduction in the foliar pH (R2 0.37) and an increase in the foliar protein (R2 0.43) were observed as immediate phenotypic signs of a productive symbiosis. The commercial raw composition of the kernels appeared to only be affected slightly by the BF treatments; thus, till now uncharted secondary compounds of the maize kernels are involved, as supported by animal performances.
Jan 2020 DOI 10.14302/issn.2372-6601.jhor-20-3186
Introduction The anti-HCV RIBA test verifies the presence of anti-HCV serum antibodies detected by the Elisa test. In Côte d'Ivoire, screening for hepatitis C is done exclusively by enzyme immunoassays. In order to reduce the number of HCV positive blood donor exclusions on ELISA, we conducted this study which aimed to demonstrate the value of the RIBA test in confirming diagnosis of viral hepatitis C to blood donors. Methods Our study, which took place from 02 to 23 February 2008 in the laboratory of Abidjan NBTC, focused on 200 sera of blood donors anti-HCV positive (Elisa test) selected according to the ratio. The DECISCAN HCV PLUS confirmation test of BIORAD was used. Results Among the 200 HCV samples positive by EIA, 49% (98/200) were confirmed positive. RIBA gave an indeterminate result in 40% of cases (80/200); and negative in 11% of cases (22/200) corresponding to false ELISA devices. In RIBA 96 samples had a low ELISA ratio of which 21% (20/96) were RIBA negative, and 79% (76/96) were indeterminate. RIBA positive samples (98/200) had a high ratio in 82% of cases (80/98). The presence of NS3 (C33) and NS4 (C100) was noted in 100% of cases (98/98, C2 in 37% (36/98) of cases and C1 in 18% of cases (18/98). RIBA indeterminate noted the presence of NS3 in 98% of cases (78/80) and NS4 in 30% of cases (24/80). Proteins C1, C2 and NS4 are essential for the diagnosis of confirmation of viral hepatitis C by RIBA. Conclusion These results attest to the lower specificity of enzyme immunoassays (ELISAs); hence the benefit of using RIBA confirmatory tests. A significant number of donors are excluded from blood donation in Côte d'Ivoire on the basis of false positive results obtained by the ELISA technique.
Dec 2019 DOI 10.14302/issn.2372-6601.jhor-19-3094
Prostate specific antigen (PSA) does not provide the high reliability and precision that is required for an accurate screening for prostate cancer (PCa). The aim of our study was to search for a simple, rapid, direct, preferably non-invasive, and highly accurate biomarker and procedure for the screening for PCa. For this purpose the levels of bromine (Br) and zinc (Zn) were prospectively evaluated in expressed prostatic fluid (EPF). Also Zn/Br concentration ratio was calculated for EPF samples, obtained from 38 apparently healthy males and from 33, 51, and 24 patients with chronic prostatitis (CP), benign prostatic hyperplasia (BPH) and PCa, respectively. Measurements were performed using an application of energy dispersive X-ray fluorescent (EDXRF) microanalysis developed by us. It was found that in the EPF of cancerous prostates the levels of Zn and Zn/Br were significantly lower in comparison with those in the EPF of normal, inflamed, and hyperplastic prostates. It was shown that “Sensitivity”, “Specificity” and “Accuracy” of PCa identification using the Zn and Zn/Br levels in the EPF samples were all significantly higher than those resulting from of PSA tests in blood serum. It was concluded that the Zn and Zn/Br levels in EPF, obtained by EDXRF, is a fast, reliable, and non-invasive diagnostic tool that can be successfully used by local, non-urologist physicians at the point-of-care to provide a highly effective PCa screening and as an additional confirmatory test before a prostate gland biopsy.
Dec 2019 DOI 10.14302/issn.2690-4829.jen-19-3105
Trichoderma reeseiβ-glucosidase (Bgl1) is one of four enzymes demonstrated to act synergistically to degrade cellulose both in vitro and in vivo. Our work attempted to better understand the substrate specificity and potential biotechnological applications of Bgl1. T. reesei Bgl1H cleaves over 80% of the β-(1-4) and β-(1-3) linkages in β-glucan and 14% of the β-(1-4) linkages in amorphous cellulose, significantly more than any tested bacterial β-glucosidase. Bgl1H cleaves 50% of the β-(1-4) linkages in xyloglucan when supplemented with cellulase and α-xyloside. Approximately 20% conversion to glucose was obtained from insoluble β-(1,3)-linked curdlan using only Bgl1H; addition of a curdlanase resulted in conversion of approximately 70% of the curdlan to glucose. Bgl1H also produces xylose from xylooligosaccharides and debranched xylans. For both glucans and xylans, the relative rates of hydrolysis increase with increasing polysaccharide chain lengths. Bgl1H is able to partially degrade β-glucan in a variety of grain components; addition of endo-acting enzymes improved the enzyme’s performance on these grain components. The ability of this enzyme to produce monosaccharides from undigestible polysaccharides suggest it may have potential in improving utilization of carbohydrates in animal feed, fermentations, and other biotechnological applications.
Nov 2019 DOI 10.14302/issn.2372-6601.jhor-19-3084
Background Identifying biomarkers for early detection of hepatocellular carcinoma (HCC) remains quite challenging. In this study we aimed to estimate the number of TIE2-expressing monocytes (TEMs) cells, which display pro-tumoral activities and are defined as CD14+, TIE2+, and angiopoietin-2; and its potential use as a possible diagnostic marker in HCC patients complicating HCV induced cirrhosis. Methods Current study was conducted on 112 patients. They were divided into two groups: Group I (78 patients) with HCC complicating HCV induced cirrhosis; and group II chronic hepatitis C patients (34 patients). Both groups were compared to (age and sex-matched) healthy persons as group III (38 persons). Result The number of the circulating TEMs: CD14+ and TIE2+ monocytes were significantly higher in the peripheral blood of HCC patients than HCV LC patients and healthy controls, sensitivity and specificity for HCC diagnosis were respectively: CD14 (89.7%, 83.3%), TIE 2 (76.9%, 83.3%), and Ang-2 (76.9%, 66.7%). Moreover, analysis of the P-value and the odd’s ratio showed that CD14 has the highest predictive value for HCC. Conclusion Our results suggest that TEMs and Ang-2can be used as diagnostic markers for HCC, especially among the high-risk group of patients.
Apr 2019 DOI 10.14302/issn.2470-0436.jos-19-2693
Given limited knowledge regarding validity of the Titmus vision screener, we sought to compare visual acuity measurements obtained from the Titmus with that from the Snellen chart and assess the validity properties of the Titmus as a screening instrument to detect vision impairment. Visual acuity was measured in 150 participants recruited from an academic ophthalmology practice, using the Snellen chart as well as the Titmus vision screener. Visual acuities from the Titmus and Snellen were compared and validity of the Titmus vision screener was assessed by computing sensitivity and specificity. Using Snellen visual acuity as the reference standard, the sensitivity of the Titmus vision screener to detect vision impairment, defined as visual acuity worse than 20/40, was 92% (95% CI (72.5, 98.6)) and the specificity was 64% (95% CI (57.9, 70.1)). Comparisons of the precise visual acuity level revealed poor agreement between the two methods (weighted Kappa: 0.15, 95% CI (0.08, 0.21)). Visual acuities obtained from the Titmus were, on average, two lines worse than Snellen visual acuities. ((logMAR Snellen – logMAR Titmus) = - 0.19 ± 0.29, 95% confidence interval (CI) (-0.23, -0.16)). Titmus vision screener is a sensitive tool to detect visual impairment. However high false positive results and poor agreement with Snellen limits its widespread use in clinical applications.
Feb 2019 DOI 10.14302/issn.2471-7061.jcrc-18-2526
There is currently no validated micro(mi)RNA diagnostic stool test to screen for colon cancer (CC) on the market because of the complexity of fecal density, vulnerability of stool to daily changes, and the presence of three sources of miRNAs in stool (cell-free from fecal homogenates, exsosomal miRNAs from fecal exosomes, and fecal colonocytes). To address these complexities, we have first carried out a microarray miRNA experiment, using Affymetrix GeneChip miRNA 2.0 Arrays, on immunocaptured and enriched stool colonocytes of 15 subjects (three healthy controls and twelve colon cancer patients [three TNM stage 0-1 (e.g., polyps◻ ³ 1 cm, villous or tubvillous, or with high grade dysplasia), three stage 2, three stage 3, and three stage 4 in triplicates to select a smaller panel of 14 preferentially expressed mature miRNAs associated with colon cancer (12 Up-Regulated, miR-19a, miR-20a, miR-21, miR-31, miR-34a, miR-96, miR-106a, miR-133a, miR-135b, miR-206, miR-224 and miR-302; and 2 Down-Regulated, miR-143 and miR-145). In a subsequent validation study carried out on total small RNA extracted by immunocapture, followed by RT that employed TaqMan® miRNA Reverse Transcription (RT) Kit and a Custom TaqMan RT Primer Pool, absolute quantification of miRNAs, in copies/µl, was measured using a chip-based Absolute QuantStudio 3D Digital PCR analysis. To ensure that we have chosen human and not bacterial small total RNA, we have carried out coextraction protocols with E. coli K1 strain RS18, compare Agilent electrophoretic patterns, and also sequenced random samples throughout this research using mRNA/miRNA sequencing. Our initial quantitative dPCR miRNA data presented herein showe that the quantitative changes in the expression of a few mature miRNA genes in stool, which are associated with right and left colon cancer, would provide for a more convenient, sensitive and specific diagnostic screening markers thatare more useful than those test markers currently available on the market, such as the low-sensitivity (<15%) fecal occult blood test (FOBT); result in better compliance; and is more economical than the invasive and expensive colonoscopy exam in colon cancer, which can be cured if that cancer is detected at the early TNM stages, and that becomes incurable and deadly if not diagnosed before metastasis. Initial test performance characteristics of the miRNA approach showed that the test has a high numerical predictive value in colon cancer. Moreover, underpinning of the miRNA markers as a function of total RNA showed that the test can numerically differentiate between control subjects and colon cancer patients, particularly at the early stages of that curable cancer. We propose to extend our initial research results to a larger prospective and randomized five-years nested case-control study, to validate the expression of the above 14 miRNAs, in stool of 180 individuals in an epidemiologically designed study, using (30 controls and 150 colon cancer patients (thirty precancerous polyps (stage 0-1), forty five stage 2, and seventy-five colon cancer stages 3 or 4). chosen randomly by an epidemiological method from 900 control and CC subjects to allow for an adequate time to collect the required 900 stool samples, as well as allowing for statistically valid analysis, standardized test conditions, and to provide a mean for determining the true sensitivity and specificity of a miRNA-screening approach in noninvasive human stool. Power-analysis has indicated that a total of 180 individuals, which will take us 5 years to enroll in testing, is an appropriate number of subjects to standardize and validate our proposed miRNA screening test. We may find out at the end of the proposed validation study in stool that fewer miRNAs, or even one miRNA, may suffice to serve as an efficient and a quantitative marker for the non-invasive diagnostic screening of colon cancer in human stool. The above approach when combined with bioinformatics analysis, to correlate miRNA seed data with our previously published messenger (m)RNA target data in stool, allows for a thorough mechanistic understanding of how miRNA genes regulate mRNA expression, and would offer a better comprehensive diagnostic screening test for the non-invasive early detection stage (0-1) of colon cancer. In order to show the clinical sensitivity and specificity of the proposed miRNA test, the absolute miRNA PCR values, in copies/µl, will be correlated with FOBT, colonoscopy, and pathology data. Standardization will establish test’s performance characteristics (sample selection, optimal sample running conditions, preservation and storage) to ensure that the assay will perform the same way in any laboratory, by any trained personnel, anywhere in the World. Ultimately, a smaller number of selected validated miRNAs (<10) showing increased and reduced expression could suffice to give quantitative miRNAs colon cancer expression values, useful for the early diagnostic screening of that curable cancer.
Dec 2018 DOI 10.14302/issn.2326-0793.jpgr-18-2422
This review summarizes tumor development in p53‑deficient mouse models, discussing tissue specificity, cooperating pathways, and insights into human cancer biology.
Aug 2018 DOI 10.14302/issn.2690-4721.ijcm-18-2217
Background Overuse of beta-lactam antibiotics has lead to selection for extended-spectrum β-lactamase (ESBL) producing Enterobacteriaceae, a major cause of antibiotic resistant urinary tract infections (UTIs). Standard detection methods are time-consuming, with disputed accuracy. This study describes a novel real-time PCR method to detect CTX-M, SHV, OXA and TEM. Methods 179 Enterobacteriaceae isolates from UTIs were collected from the Leicester Royal Infirmary, UK. A multiplex Plexor®-based real-time PCR assay detected ESBLs using their specific amplicon melting temperature, during each cycle, removing the need for a melt-curve analysis. Validation was achieved by end-point PCR and disk diffusion. Results The method was able to produce rapid and accurate results, achieving a sensitivity and specificity of 94.9% and 72% respectively, and the assay can differentiate between the different ESBL genes, with ease. Conclusions With further investigation, a Plexor®-based assay could form the basis of a high-throughput kit that health services could use to detect ESBLs or other antibiotic resistance genes.
Aug 2017 DOI 10.14302/issn.3070-2313.jeh-17-1552
Introduction. Obesity often coexists with insulin resistance, which is related to cardiometabolic risk. However, some obese individuals exhibit comparable insulin sensitivity (IS) to that of normal-weight subjects, a state associated with a reduced cardiometabolic risk. We aimed to determine the efficacy of a panel of surrogate markers of insulin sensitivity (IS) for the identification of insulin sensitive obese (ISO) vs. insulin resistant obese (IRO) with similar total fat mass (FM) and body mass index (BMI). Methods. This is a cross-sectional analysis among 144 overweight and obese post-menopausal women. IS was determined by the hyperinsulinemic-euglycemic clamp (HEC) and by surrogate indices such as Matsuda index, the simple index assessing insulin sensitivity using oral glucose tolerance test (SIisOGTT), Abdul-Ghani liver IS index, HOMA-IR and Abdul-Ghani muscle IR index. Results. When using upper and lower quartiles values or the median as cut-off for IS determined by the reference HEC to define ISO vs. IRO, Matsuda index, SIisOGTT and Abdul-Ghani indices classification identified ISO vs. IRO individuals with similar FM and BMI. With HOMA-IR, the two groups were similar for FM and had borderline significant difference in BMI. Using, receiver operating characteristic curves, Matsuda index AUC was similar to that of SIisOGTT and both indices AUCs were significantly higher than Abdul-Ghani indices AUCs. The best cut-off value for the Matsuda index was 2.5 (83.1% specificity, 54.2% sensitivity) and 0.25 for SIisOGTT (64.8% specificity, 70.8% sensitivity). Conclusion. Whole body IS indices, Matsuda and SIisOGTT indices seem to be reliable indices for the identification of ISO vs. IRO individuals.
Aug 2017 DOI 10.14302/issn.2575-7881.jdrr-17-1701
Functional nucleic acids are a kind of nucleic acid sequences with special functions, which can specifically bind with the target substances or catalyze the reactions. Many target, including mycotoxins, small RNA, heavy metal ions and DNA segment, can bind to particular selected oligonucleotides, and then realized the detection. The uses of functional nucleic acids to detect the genetically modified organism (GMO) have been pursued using different approaches. Meanwhile, the flanking sequence, which was the most specific target in the GMO detection, was also usually separated with the help of functional nucleic acid. During the detection, the functional nucleic acid provided superior sensitivity, specificity and success rate compared with the traditional methods. In this report, we described different functional nucleic acids used in the GMO detection, they were classified based on their structures, and some of them were developed in our lab. The principle, structural composition, advantage, and the comparisons of the functional nucleic acids were reported. Considering most of the functional nucleic acids are fluorescently-labeled, in order to reduce the cost, more and more functional nucleic acids without labeling are under research.
Aug 2017 DOI 10.14302/issn.2471-7061.jcrc-17-1624
Background Despite the existence of the statutory early cancer detection program in Germany and the removal of financial barriers, which is frequently reported in the literature to be the main obstacle in screening, uptake of colorectal cancer screening remains quite low. The campaign for colorectal cancer screening in German companies reported in this article started in 2010. It was initiated because of the low compliance with opportunistic public colorectal cancer screening efforts. Its goal is to improve participation by offering an organized screening program using a simple test (FIT). Methods An offer for company employees is publicized through posters, company newsletters and the intranet. The difference between the positivity rates of those who returned the kits within 20 days and later than 20 days was assessed using the Z-test. The average time between a positive result and colonoscopy was estimated using the Poincaré plot method. The positive predictive values were calculated for carcinomas, advanced adenomas or any lesions. The sensitivity and specificity of immoCare-C published by Vogel et al. and Hundt et al. were used to derive the confidence intervals for the positive likelihood ratio (for carcinoma and any kind of adenoma). Results A total of 312,147 kits were returned and analyzed (return rate 70.2%). 5.6% gave a positive result. The PPV for cancer aged between 55 and 74 was 4.6% for men and women (95% CI: 2.38%-6.76% and 1.28%-7.99%, respectively), but 22% for men (95% CI: 17.93%-26.65%) and 8% for women (95%CI: 3.63%-12.26%) for advanced adenomas. The PPV for any lesion was higher for those with familial risk (49.3%) and 42.6% for those without familial risk (95% CI: 40.2%-45.0%), but with overlapping confidence intervals. Conclusions The reported sample is not representative. Although, offering CRC screening in companies may be an effective way of increasing uptake in the target population. Differences in the test performance between men and women need further evaluation.
Jan 2017 DOI 10.14302/issn.2379-8572.joa-16-1399
Aim: The relation between inflammation and cancer has been known since the 19th century. However, investigations on the pathogenesis and pathophysiology of this relation have begun recently. It was demonstrated that increased neutrophil/lymphocyte ratio is a poor prognostic factor in some malignancies. The present study aimed to determine whether preoperative neutrophil/lymphocyte ratio has a prognostic value in larynx cancer. Method: Preoperative blood analyses of 139 patients, who underwent subtotal or total laryngectomy for larynx cancer between 2003 and 2013 at Marmara University School of Medicine, Department of ENT, were retrospectively evaluated. Neutrophil/lymphocyte ratio (NLR) was calculated dividing absolute neutrophil count by absolute lymphocyte count. Optimal cut-off value for NLR was determined by receiver operating characteristics (ROC) curve analysis. Statistical analyses were done using IBM SPSS statistic 22.0 (IBM SPSS, Turkey) and Med Calc 12.3.0 package programs. Results: The sensitivity of NLR in predicting advanced-stage (Stage 3 and 4) squamous-cell carcinoma of the larynx (LSSC), T4 LSSC and lymph node metastasis at different cut-off values were 66.2%, 83.9% and 73.8%, respectively and the specificity was 76.7%, 66.2% and 65.2%, respectively. Staging according to T classification revealed that NLR significantly increases with tumor stage (p<0.001). Statistically significant relation was determined between lymph node metastasis of tumor and neutrophil/lymphocyte ratio (p=0.003). Comparing overall survival (OS) and disease-free survival (DFS) between the cases with NLR <3.02 and the cases with NLR >3.02, it was demonstrated that OS and DFS are significantly lower in the cases with NLR<3.02 (p: 0.001 vs. p<0.05 for OS and p: 0.013 vs. p<0.05 for DFS) Conclusion: NLR increases with the stage of disease in LSSC. NLR is a simple, cheap, repeatable and valuable parameter that can be obtained from routine analyses, gives information about poor prognosis and survival, and is able to predict T4 LSSC, advanced-stage LSSC (stage 3-4) and lymph node metastasis.
Oct 2016 DOI 10.14302/issn.2574-450X.jom-16-1003
Introduction: The Roux-en-Y Gastric Bypass (RYGB) has been one of the most popular surgeries in the USA for years. While many models have been made to investigate the factors that affect weight loss, these factors are still highly debated. Objective: To create a model that predict performance of RYGB patients. Methods: 110 out of 344 patients who received a RYGB at a single institution between Jan 2010 and April 2014 were included in this study. Data was collected retrospectively. Patients were included if they had greater than 1 year follow up with at least three follow up points and could be modeled with r2>0.95. All patients were one year beyond surgery, while 40 were completely lost to follow up, 104 at 1 month, 138 at 3 months, 188 at 6 months, and 225 at one year. 9 patients were not included because they did not meet the criteria of the study. Patients were divided into quartiles based on percentage excess weight loss (%EWL) at one year. Multivariate analysis was performed to determine the significant factors that influence patients being in the first quartile of weight loss (17-60% %EWL). Results: Only males with a Body Mass Index (BMI) above 44 and females with a BMI above 64 were found to be predictive of patients being in the first quartile. Our model has Positive and Negative predictor values of 66% and 80% respectively with sensitivity and specificity of 29% and 95% respectively. Conclusions: An model to predict %EWL was created, only gender and pre-operative BMI were found to be significant factors. In general females have better outcomes with higher BMI’s than do males. This information should be discussed with patients when deciding a procedure. However, more studies are needed for validation of these results.
Sep 2016 DOI 10.14302/issn.2381-862X.jwrh-15-771
Objective The tocodynamometer (TOCO) has poor sensitivity and specificity in monitoring uterine contractions, especially in obese patients. The intrauterine pressure catheter (IUPC) can be used to monitor adequacy of contractions, but only after amniotomy. Transabdominal uterine electromyography (EMG) and the TOCO were compared to the gold standard IUPC for monitoring uterine contractions. Methods Forty term pregnant women in labor with ruptured membranes were consented for this study. The root mean square (RMS) plot from EMG signals was compared to IUPC and TOCO recordings for 20 to 40 minutes. A comparison between the total contraction number, frequency, and the difference in contraction peak time was made using Student-t test or ANOVA (P<0.05 was significant). Results There was no significant difference in the contraction number and frequency when comparing the RMS, TOCO, and IUPC. The paper tracings had a greater standard deviation (8.57) than the digitally saved data (3.93). The mean peak time difference between TOCO and IUPC was 0.74 seconds (P=0.78; SD 5.2). For RMS vs. IUPC peaks, the mean peak difference between was 0.13 seconds (P=0.95; SD 3.93). Conclusions Uterine electrical activity measured with transabdominal uterine EMG may be used to monitor labor in patients as an alternative to the TOCO and the IUPC.
May 2016 DOI 10.14302/issn.2470-5020.jnrt-15-908
Objective: We aimed to investigate homocysteine levels and carotid intima-media thickness (CIMT) in Parkinson’s Disease (PD), to determine relationship of these parameters and as well as to determine whether CIMT in patients with PD was associated to age, disease duration, age of disease onset, stage, the Unified Parkinson Disease rating scale (UPDRS), the drugs used in therapy. Methods: The study population consisted of 55 PD patients (37 male) and 25 healty subjects. The severity of neurological impairment was assessed with UPDRS and the Hoehn-Yahr scale. CIMT and homocysteine levels were measured. Anti-parkinsonian treatments were recorded and the total daily dose of levodopa was calculated for each patient. Results: Homocysteine levels were significantly higher in the patient group compared to the control group (p=0.002). A positive correlation was found between CIMT and homocysteine (r=0.29 p=0.03), but no a relationship between CIMT and UPDRS scores, disease duration, age of disease onset, and stage. Mean levodopa dosage did not predict CIMT 0.6 mm (AUC: 0.546, 95%CI 0.372-0.720, p=0.59). Homocysteine 14 µmol/l predicted CIMT 0.6 mm with 64% sensitivity and 69% specificity (AUC: 0.654, 95%CI 0.488-0.819, p=0.07). Dıscussion: This study revealed that homocysteine levels in levodopa + dopa decarboxylase enzyme inhibitor (DDEI) group were increased which was correlated with a mild increasement of CMIT. This might indicate to the importance of clinical and radiological follow up of PD patients who are under treatment of levodopa + DDEI. Conclusion: Our Findings May Suggest The Role Of CIMT As A Meaningful Clinical Marker For Follow-Up Of Patients With PD
Mar 2016 DOI 10.14302/issn.2329-9487.jhc-13-335
Objective: To find out the diagnostic accuracy of admission ECG against coronary angiography (CAG) to predict the ARA in patients with USAP /NSTEMI. Background: In USAP even though multiple active plaques are documented, one critical lesion would be responsible for the index episode of angina. Contrary to STEMI there is no standard methodology to identify the Angina related artery (ARA) in USAP.Therefore we plan to determine whether admission ECG could predict the ARA in patients with USAP/NSTEMI. Patients and Methods: 250 eligible patients with USAP/NSTEMI undergoing coronary angiography were enrolled in the study. After locating the ARA , the patient’s admission ECG was compared with CAG finding to study whether it has any predictive value for identifying ARA. Results: Sensitivity of the admission ECG for LAD as ARA was 73.6%, Specificity was 93.5%, PPV was 90%, NPV was 81.8%, +LR 10.4, Posterior probability 0.89, -LR 0.29, Posttest odds 0.22. Similarly for RCA and LCx ,Sensitivity was 63.1%, Specificity was 93.7%, PPV was 90.5%, NPV was 72.7%, +LR 9, Posterior Probability 0.89, -LR 0.35, Posttest odds 0.33. For the LM and TVD, Sensitivity was 66.6%, Specificity was 98.5%, PPV was 91.4%, and NPV was 92.5%, +LR 33, Posterior Probability 0.89, -LR 0.33, Posttest odds 0.08. Conclusion: With high +LR, Admission ECG is moderately sensitive and highly specific for detecting culprit artery.
Mar 2016 DOI 10.14302/issn.2379-8572.joa-15-891
Background: Despite its expense, labor and intrusiveness, polysomnography (PSG) is the gold standard for diagnosing obstructive sleep apnea (OSA). Recently, commercially available electronic activity monitors, such as Fitbit, have become widely accepted and can provide an estimate of sleep patterns for screening children with possible OSA. A previous study demonstrated Fitbit to be valid compared to PSG in adults. To date, these devices have not been extensively utilized for research in children with sleep disordered breathing (SDB). Objective: To evaluate the validity of the Fitbit activity monitor compared to PSG in children and adolescents with SDB. Methods: Data was collected from 14 children, ages 3 through 11, who were scheduled for a PSG during the study period. Fitbit was worn concurrently during the night of the PSG. Analyses were performed by comparing total sleep time, number of awakenings, sleep efficiency and wake after sleep onset (WASO) Fitbit parameters with the corresponding parameters measured by PSG using Spearman’s rho. Fitbit movement epochs were also compared to PSG epochs showing movement behavior. Results: Pilot data suggest that Fitbit demonstrates a high sensitivity for sleep, a low specificity for wake and a trend suggesting good association of movement measurements. Conclusion: Although Fitbit is not as accurate as PSG for determining wakefulness, it may be a useful screening device to assess gross sleep quality in children. Further studies are indicated to validate these findings.