scholarly journals Screening and identification of specific markers for bladder transitional cell carcinoma from urine urothelial cells with suppressive subtractive hybridization and cDNA microarray

2013 ◽  
Vol 5 (6) ◽  
pp. 129
Author(s):  
Baihong Guo ◽  
Tuanjie Che ◽  
Baoguang Shi ◽  
Lijun Guo ◽  
Yongsheng Yin ◽  
...  

Objective: The objective of this study was to screen and identifydifferentially expressed genes in invasive bladder transitional cellcarcinoma (BTCC).Methods: Voided urine samples were collected from consecutivepatients with BTCC and patients under surveillance for bladdercancer recurrence; voided urine samples from patients with nonmalignantdiseases served as control. We identified the differentiallyexpressed genes by comparing urine samples of bladdercarcinoma to that of the control group with suppressive subtractivehybridization (SSH) and cDNA microarray. The differentiallyexpressed genes were verified by quantitative real-time polymerasechain reaction (QPCR).Results: From the 762 white colonies, a total of 449 positive cloneswere obtained in which 112 were found to be upregulated in BTCC.Sequencing and homology analysis were performed for these 112clonies. The detection rates of some known genes (including IGF-1, human telomerase reverse transcriptase [hTERT], bladder cancerspecific nuclear matrix protein 4 [BLCA-4] and homeobox A13[HOXA13]) for BTCC at the Ta, T1 and >T1 stages were 48%, 90%and 100%, respectively, with a specificity of 85%. The test specificitywas 80% for the 30 control patients with urinary tract infections. Thecombination of BLCA-4 and HOXA13 could distinguish betweenlow- and high-grade tumours, with specificity and sensitivity of 80%.Conclusion: We successfully constructed a reliable SSH library ofBTCC and found that combination detection insulin-like growthfactor 1 (IGF-1), hTERT, BLCA-4 and HOXA13 genes could helpto evaluate BTCC at different stages.

2021 ◽  
Author(s):  
Lalainasoa Odile RIVOARILALA ◽  
Victor JEANNODA ◽  
Tania CRUCITTI ◽  
Jean Marc COLLARD

Abstract Background: Timely and accurate identification of uropathogens and determination of their antimicrobial susceptibility is paramount to the management of urinary tract infections (UTIs). The main objective of this study was to develop an assay using LAMP (Loop mediated isothermal amplification) technology for simple, rapid and sensitive detection of the most common bacteria responsible for UTIs, as well as for the detection of the most prevalent genes (encoding cefotaximases from CTX-M group 1) responsible for resistance to 3rd generation of cephalosporins. Method: We designed primers targeting Proteus mirabilis, while those targeting Escherichia coli, Klebsiella pneumoniae and Enterococcus faecalis and the CTX-M group 1 resistance gene were benchmarked from previous studies. The amplification reaction was carried out in a warm water bath for 60 min at 63±0.5 °C. The amplicons were revealed by staining with Sybr Green I. Specificity and sensitivity were determined using reference DNA extracts spiked in sterile urine samples. The analytical performance of the assays was evaluated directly on pellets of urine samples from patients suspected of UTI and compared with culture.Results: We found a high specificity (100%) for LAMP assays targeting the selected bacteria (P. mirabilis, E. coli, K. pneumoniae, E. faecalis) and the CTX-M group 1 when using DNA extracts spiked in urine samples. The sensitivities of the assays were around 1.5 103 Colony Forming Units (CFU) /mL corresponding to the cut-off value used to define bacteriuria or UTIs in patients with symptoms. Out of 161 urine samples tested, using culture as gold standard, we found a sensitivity of the LAMP techniques ranging from 96 to 100 % and specificity from 95 to 100 %.Conclusion: We showed that the LAMP assays were simple and fast. The tests showed high sensitivity and specificity using a simple procedure for DNA extraction. In addition, the assays could be performed without the need of an expensive device such as a thermal cycler. These LAMP assays could be useful as an alternative or a complementary tool to culture reducing the time to diagnosis and guiding for more effective treatment of UTIs but also as a powerful diagnostic tool in resource-limited countries where culture is not available in primary health care structures.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Lalainasoa Odile Rivoarilala ◽  
Jeannoda Victor ◽  
Tania Crucitti ◽  
Jean Marc Collard

Abstract Background Timely and accurate identification of uropathogens and determination of their antimicrobial susceptibility is paramount to the management of urinary tract infections (UTIs). The main objective of this study was to develop an assay using LAMP (Loop mediated isothermal amplification) technology for simple, rapid and sensitive detection of the most common bacteria responsible for UTIs, as well as for the detection of the most prevalent genes (encoding cefotaximases from CTX-M group 1) responsible for resistance to 3rd generation of cephalosporins. Method We designed primers targeting Proteus mirabilis, while those targeting Escherichia coli, Klebsiella pneumoniae and Enterococcus faecalis and the CTX-M group 1 resistance gene were benchmarked from previous studies. The amplification reaction was carried out in a warm water bath for 60 min at 63 ± 0.5 °C. The amplicons were revealed by staining with Sybr Green I. Specificity and sensitivity were determined using reference DNA extracts spiked in sterile urine samples. The analytical performance of the assays was evaluated directly on pellets of urine samples from patients suspected of UTI and compared with culture. Results We found a high specificity (100%) for LAMP assays targeting the selected bacteria (P. mirabilis, E. coli, K. pneumoniae, E. faecalis) and the CTX-M group 1 when using DNA extracts spiked in urine samples. The sensitivities of the assays were around 1.5 103 Colony Forming Units (CFU) /mL corresponding to the cut-off value used to define bacteriuria or UTIs in patients with symptoms. Out of 161 urine samples tested, using culture as gold standard, we found a sensitivity of the LAMP techniques ranging from 96 to 100% and specificity from 95 to 100%. Conclusion We showed that the LAMP assays were simple and fast. The tests showed high sensitivity and specificity using a simple procedure for DNA extraction. In addition, the assays could be performed without the need of an expensive device such as a thermal cycler. These LAMP assays could be useful as an alternative or a complementary tool to culture reducing the time to diagnosis and guiding for more effective treatment of UTIs but also as a powerful diagnostic tool in resource-limited countries where culture is not available in primary health care structures.


GYNECOLOGY ◽  
2018 ◽  
Vol 20 (6) ◽  
pp. 67-72
Author(s):  
D Yu Pushkar ◽  
M Yu Gvozdev

Material and methods. The study included patients (n=164) in peri- and postmenopause, having a laboratory-confirmed picture of the genitourinary menopausal syndrome - atrophic vulvovaginitis and suffering from recurrent lower urinary tract infections (UTI) - cystitis and urethritis. All patients underwent standard antibacterial therapy (ABT) UTI in accordance with the results of bacteriological urine culture according to Russian clinical guidelines on urology. Against the background of ABT, 164 patients received additional therapy with Trioginal® (the study group - SG) containing estriol, micronized progesterone and Lactobacillus casei rhamnosus Doderleini 35 (LCR 35) lactobacillus strain. Trioginal® was administered intravaginally in two stages: for 20 days, 2 capsules per day, then for 10 days, 1 capsule per day. In the comparison group - CG (n=67) with ABT, Ovipol Klio® (estriol monopreparation) was additionally intravaginally used in two stages: for 14 days, 1 suppository per day, then for 2 weeks, 1 suppository 2 times a week. The control group consisted of 30 patients receiving only standard ABT UTI, local hormone therapy for vulvovaginal atrophy (VVA) was not performed. The observation period was 12 months after the end of the course of therapy. At all stages of treatment, a subjective and objective assessment of the severity of urination disorders and urogenital atrophy (UDI-6 questionnaire), their impact on the quality of life of patients (questionnaire IIQ-7), indicators of bacteriuria, bacterial urine culture and vaginal biocenosis, frequency of undesirable phenomena were carried out. According to the initial characteristics of the group of patients were comparable. Results. At all stages of the study, in the group of patients who received therapy with the use of the Trioginal® drug, there was a significant improvement in the clinical picture of urination disorders compared to the CG and the control (main) group - MG (in SG - 18% at the end of therapy versus 100% before the start of therapy; CG - 26 % versus 100% respectively; Ledger - 58% versus 100% respectively; p


2018 ◽  
pp. 83-89
Author(s):  
Trung Thuan Phan ◽  
Dinh Binh Tran ◽  
Thanh Hue Dinh ◽  
Phong Son Dinh ◽  
Kieu Oanh Truong ◽  
...  

Objectives: To find a model that is effective intervention to reduce the incidence of lower genital tract infections in Khmer women. Subjects and Methods: The intervention study is carried which compared the test group with the control group of 400 Khmer women, aged 15 to 49 in Can Tho city. Results: The common knowledge of the prevention of lower genital tract infections in the intervention group increased from 30.9% in the control group to 54.5% in the intervention group. The general attitude toward the use of lower genital tract infections increased from 22.3% in the control group to 78.0% in the intervention group. This is a very significant improvement. The prevalence of subclinical infection of the intervention group (26.0%) was significantly lower than that of the control group (39.3%), the difference was statistically significant. Conclusion: Intervention effectiveness is good in both knowledge, attitudes and practices, especially the effectiveness of interventions in practice in Khmer women. Key words: Effective intervention, lower genital tract infections, Khmer women, Can Tho city


Cancers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 3167
Author(s):  
Flavia Buonaurio ◽  
Maria Luisa Astolfi ◽  
Daniela Pigini ◽  
Giovanna Tranfo ◽  
Silvia Canepari ◽  
...  

Urinary concentrations of 16 different exposure biomarkers to metals were determined at the beginning and at the end of a working shift on a group of workers in the metal carpentry industry. Five different oxidative stress biomarkers were also measured, such as the oxidation products of RNA and DNA metabolized and excreted in the urine. The results of workers exposed to metals were compared to those of a control group. The metal concentrations found in these workers were well below the occupational exposure limit values and exceeded the mean concentrations of the same metals in the urine of the control group by a factor of four at maximum. Barium (Ba), mercury (Hg), lead (Pb) and strontium (Sr) were correlated with the RNA oxidative stress biomarker, 8-oxo-7, 8-dihydroguanosine (8-oxoGuo), which was found able to discriminate exposed workers from controls with a high level of specificity and sensitivity. The power of this early diagnostic technique was assessed by means of the ROC curve. Ba, rubidium (Rb), Sr, tellurium (Te), and vanadium (V) were correlated with the level of the protein oxidation biomarker 3-Nitrotyrosine (3-NO2Tyr), and Ba, beryllium (Be), copper (Cu), and Rb with 5-methylcytidine (5-MeCyt), an epigenetic marker of RNA damage. These effect biomarkers can help in identifying those workers that can be defined as “occupationally exposed” even at low exposure levels, and they can provide information about the impact that such doses have on their health.


2021 ◽  
Vol 49 (5) ◽  
pp. 030006052110161
Author(s):  
Jing Li ◽  
Qiao-Ping Li ◽  
Bi-Hong Yang

Objective The study aim was to analyse the effect of participatory continuous nursing using the WeChat platform on the complications, family function and compliance of patients with spinal cord injuries. Methods This was a randomized controlled trial. Seventy-eight patients with stable disease treated by internal fixation were enrolled in the study from August 2017 to August 2019 and assigned equally to an observation group and a control group. The control group received regular care from the time of discharge. The observation group used the WeChat platform to participate in continuous care. Results Six months after discharge, the continuous nursing group had a significantly lower incidence of pressure ulcers, urinary tract infections, joint contractures and muscle atrophy than the control group. The continuous nursing group showed a significant improvement in family function level and compliance behaviour at 3 and 6 months after discharge. Conclusion A participation-based continuous nursing intervention using the WeChat platform can reduce the incidence of pressure ulcers, urinary tract infections, joint contracture and muscle atrophy; improve patient family function; and promote healthy compliance behaviour.


2021 ◽  
pp. 875647932098324
Author(s):  
Elif Özyazici Özkan ◽  
Mehmet Burak Ozkan ◽  
İshak Abdurrahman İsik

Objective: The objective of this study was to determine the elasticity of sternocleidomastoid muscle (SCM) in patients with congenital muscular torticollis (CMT). Methods: In all, 41 patients and 22 controls were included in the study, and the elasticity of the patients’ SCM was measured. Echogenicity, thickness, and strain values of the SCM were also obtained. Results: The thickness and strain values of the SCM were higher in the patient group than in the control group ( P = .02 and P = .15). For median values, there was no difference in echogenicity and strain. In the strain elastography evaluation of the receiver operating curve (ROC) for muscle echogenicity in the isoechoic muscle group, the specificity and sensitivity were determined to be 100% and 22%, respectively, for the area under the curve (AOC) value of 0.558 (95% confidence interval [CI], 0.424–0.6686), and the cutoff value was <1.4. In the hyperechoic muscle group, the ROC for AUC values was found to be 0.542 (95% CI, 0.411–0.6686), and the cutoff value was >1.4 with 100% sensitivity and 20.75 specificity. Conclusions: The strain elastography technique can be used in the diagnosis of CMT.


2021 ◽  
Vol 9 (7) ◽  
pp. 1446
Author(s):  
Ivo Hoefnagels ◽  
Josephine van de Maat ◽  
Jeroen J.A. van Kampen ◽  
Annemarie van Rossum ◽  
Charlie Obihara ◽  
...  

Lower respiratory tract infections (LRTIs) in children are common and, although often mild, a major cause of mortality and hospitalization. Recently, the respiratory microbiome has been associated with both susceptibility and severity of LRTI. In this current study, we combined respiratory microbiome, viral, and clinical data to find associations with the severity of LRTI. Nasopharyngeal aspirates of children aged one month to five years included in the STRAP study (Study to Reduce Antibiotic prescription in childhood Pneumonia), who presented at the emergency department (ED) with fever and cough or dyspnea, were sequenced with nanopore 16S-rRNA gene sequencing and subsequently analyzed with hierarchical clustering to identify respiratory microbiome profiles. Samples were also tested using a panel of 15 respiratory viruses and Mycoplasma pneumoniae, which were analyzed in two groups, according to their reported virulence. The primary outcome was hospitalization, as measure of disease severity. Nasopharyngeal samples were isolated from a total of 167 children. After quality filtering, microbiome results were available for 54 children and virology panels for 158 children. Six distinct genus-dominant microbiome profiles were identified, with Haemophilus-, Moraxella-, and Streptococcus-dominant profiles being the most prevalent. However, these profiles were not found to be significantly associated with hospitalization. At least one virus was detected in 139 (88%) children, of whom 32.4% had co-infections with multiple viruses. Viral co-infections were common for adenovirus, bocavirus, and enterovirus, and uncommon for human metapneumovirus (hMPV) and influenza A virus. The detection of enteroviruses was negatively associated with hospitalization. Virulence groups were not significantly associated with hospitalization. Our data underlines high detection rates and co-infection of viruses in children with respiratory symptoms and confirms the predominant presence of Haemophilus-, Streptococcus-, and Moraxella-dominant profiles in a symptomatic pediatric population at the ED. However, we could not assess significant associations between microbiome profiles and disease severity measures.


2021 ◽  
Vol 25 (1) ◽  
pp. 43-46
Author(s):  
Taofeeq Garuba ◽  
Nency Katrodiya ◽  
Nikita Patel ◽  
Swetal Patel ◽  
Dhanji. P. Rajani ◽  
...  

Urinary tract infections (UTI) are one of the most common form of bacterial infections but the treatment becomes cumbersome as the etiological bacteria are developing resistance against antibiotics. This present study evaluated the efficacy of antimicrobial   activity of Urena lobata against uropathogens. Six urine samples from UTI patients were collected from Pathological Laboratory, G.B. Vaghani Multispecialty Hospital, Surat. Bacteria were isolated from these samples using Nutrient agar, Mac Conkey agar plate, Blood agar, Mannitol salt agar, Eosin Methylene Blue agar and King’s agar. The bacterial isolates were identified using cultural  characteristics, microscopic features and biochemical characteristics. Leaf extract of Urena lobata was prepared using Soxhlet Extraction Method whereby methanol and distilled water were the extractants used. Herbal extract disc was prepared at  concentrations of 50,75, and100 mg/ml and tested against all the isolates. DMSO and antibiotics (Nitrofurantion, Amikacin, Levofloxacin, Norofloxacin, Ofloxacin and Cephalosporins) were used as negative and positive controls respectively.Staphylococcus aureus, Escherichia coli, Bacillus cereus, Streptococcus pneumoniae, Klebsiella spp. and Brevibacillus panacihumi were isolated from the urine samples. All concentrations of aqueous and methanolic extracts of U. lobata leaf displayed highest zone of inhibition against B. cereus. No inhibitory effect was observed against the growth of Klebsiella except at the highest concentrations. Further study is encouraged on the in-vivo study of efficacy of U. lobata on etiological agent of UTI.


Sign in / Sign up

Export Citation Format

Share Document