Factors affecting low fetal fraction in fetal screening with cell-free DNA in pregnant women: a systematic review and meta-analysis.

BACKGROUND: Cell-Free DNA (cfDNA) is a non-invasive perinatal test (NIPT) used to assess fetal anomalies. The ability to detect fetal chromosomal aneuploidies is directly related to a sample's fetal to total DNA fraction, known as the fetal fraction (FF). The minimum FF is considered 4%, and the test result below 4% is uncertain due to low fetal fraction (LFF). This study aimed to conduct a systematic review and a meta-analysis to determine the possible factors affecting LFF in cfDNA testing for fetal screening. METHODS: PubMed, Web of Science, Google Scholar, Since Direct, Scopus, CINHAL, Cochrane Library, and Persian databases, including Scientific Information Database, Irandoc, and Magiran were searched for studies investigating factors affecting LFF in cfDNA testing from 2000 until the end of 2021. Gathered data were analyzed using Comprehensive Meta-Analysis (CMA) software version 3.3.070. The quality of the included studies was assessed using the Joanna Briggs Institute Critical Appraisal of Cohort Studies tool. RESULTS: Thirteen articles related to the topic were included, and seven related articles were reviewed for meta-analysis. The other six were reviewed qualitatively. Four factors were identified that might have a potential effect on the LFF, of which only gestational age had a significant association with LFF (Pooled mean difference= -1.111, SE = 0.515, 95% CI= -2.121, -0.101, (P-value < 0.05)). Maternal age (P-value = 0.573), maternal weight (P-value = 0.113), and Body Mass Index (P-value = 0.104) had no statically significant effect. The effect size was pooled by mean difference and 95% confidence interval. CONCLUSION: Lower gestational age is significantly associated with LFF. Thus, this factor can be considered when interpreting prenatal cfDNA screening tests.

Ultrasound Evaluation of Mid-Urethral Sling Position: A Potential Predictor of Outcomes and Adverse Effects.

OBJECTIVE: The aim of this study was to evaluate mid-urethral sling (MUS) position and its association with postoperative outcomes and complications. DESIGN: This was a prospective cohort study. Ninety-two women who underwent MUS procedure with a median follow-up period of 11 months (interquartile range 5-24 months) were recruited. PARTICIPANTS/MATERIALS, SETTING, METHODS: Two-dimensional trans-labial ultrasound with an endovaginal probe was used to visualize the urethral length (UL), sling distance to the bladder neck (BN) and to the urethra longitudinal smooth muscle (LSM). RESULTS: Mean sling-LSM, UL, and sling-BN distances were 5.97 ± 2.04 mm, 28.66 ± 3.19 mm, and 18.85 ± 4.46 mm, respectively. Sling position relative to BN (proximal vs. middle vs. distal) was not associated with surgery outcomes and complications; however, mean sling-LSM in patients with exposure (4.3%) was significantly higher compared to those who did not experience exposure (8.80 ± 1.9 mm vs. 5.8 ± 2.0 mm, p = 0.048). Moreover, the mean sling-LSM distance was associated with patient satisfaction (5.87 ± 2.0 mm in satisfied patients with VAS > 6 vs. 6.29 ± 2.1 mm in unsatisfied patients, p value = 0.043). Likewise, patients who had worsened or showed de novo overactive bladder (OAB) symptoms (8.8%) had significantly higher mean sling-LSM distance compared to patients with improved OAB symptoms (6.52 ± 2.0 mm vs. 5.37 ± 1.9 mm, p = 0.007). Mean sling-LSM distance was lower in patients with recurrent urinary tract infection (UTI), voiding dysfunction, and improved stress urinary incontinence, whereas patients with dyspareunia after surgery had higher mean sling-LSM distance; however, these differences were not statistically significant. LIMITATIONS: Both trans-obturator and retropubic procedures with or without concomitant prolapse surgery were assessed. Heterogeneity of the study population and pelvic floor ultrasound being performed by a single urogynecologist were the limitations of the current study. CONCLUSIONS: Ultrasound visualization of MUS is feasible and has the potential to predict outcomes and complications. High sling-LSM distance was associated with exposure, and low sling-LSM distance with increased satisfaction rate, probability of voiding dysfunction, and recurrent UTI.

Outcomes of the anterior approach versus posterior sacrospinous ligament fixation for pelvic organ prolapse.

INTRODUCTION AND HYPOTHESIS: This study aimed to compare anterior sacrospinous ligament fixation (SSLF) with the standard posterior SSLF concerning complications and outcomes in patients with apical compartment pelvic organ prolapse (POP). METHODS: This is an observational descriptive study using prospective data collected from two referral urogynecological centers. The study cohort represents all 135 women in our prospective study who underwent anterior approach bilateral anterior or unilateral posterior meshless SSLF from January 2018 to December 2020 using the PFDI-20 questionnaire and the POP quantification (POP-Q) system pre- and postoperatively. The objective success rate was assessed by the number of POP recurrence cases and total vaginal length (TVL) postoperatively. Patients were followed up for at least 6 months (range, 6 to 18 months). Data were analyzed using SPSS version 21. P < 0.05 was considered statistically significant. RESULTS: Sixty-seven (49.6%) patients underwent posterior SSLF, and 68 (50.4%) underwent anterior SSLF. The mean age of patients was 58.2 ± 9.7 and 64.9 ± 11.6, respectively (P < 0.001). Most patients who underwent the posterior approach had stage III apical prolapse (74.6%), while 65.5% of those who underwent anterior SSLF had stage II apical prolapse (P < 0.001). Following the treatment, no significant difference was detected between these two vaginal approaches in terms of women's satisfaction rate (P > 0.05). One case of postoperative recurrence was found in the posterior group, which ultimately led to surgical retreatment. There were no major intra- or postoperative complications in the groups. Postoperative TVL was higher in the anterior SSLF group (P < 0.001). The postoperative POPDI-6, CRADI-8, UDI-6, and PFDI-20 decreased significantly compared to preoperative status in both groups (P < 0.001). CONCLUSION: It appears that the anterior SSLF approach can be regarded as effective as the posterior approach in the management of apical POP. Therefore, the proper surgical technique can be chosen according to the surgeon's expertise and other compartment's prolapse status.

The Effects of home-based stabilization exercises focusing on the pelvic floor on postnatal stress urinary incontinence and low back pain: a randomized controlled trial.

INTRODUCTION AND HYPOTHESIS: Stress urinary incontinence (SUI) and low back pain (LBP) are common postnatal problems. We aimed to compare the effects of stabilization exercises focusing on the pelvic floor on postnatal SUI and LBP. METHODS: This two-arm, single-blind, parallel, randomized controlled trial was done on 80 women (mean age: 30.5, range: 20-45 years), with postnatal SUI and LBP. They were randomized into two equal control and intervention groups. The control group received no treatment while the intervention group received home-based stabilization exercises focusing on pelvic floor muscles (PFM) 3 days a week for 12 weeks, three sets a day; each set included three different types of exercise each week. Outcome measures were UI severity, assessed by ICIQ-UI-SF, low back pain functional disability, assessed by the Oswestry Disability Index (ODI), LBP severity, assessed by visual analog scale (VAS), and PFM strength and endurance, assessed by vaginal examination. Transverse abdominis (TrA) muscle strength was assessed by manometric biofeedback. All outcomes were measured directly before and after treatment. RESULTS: In the intervention group, PFM strength, TrA muscle strength, functional disability and pain severity were significantly improved (P < 0.05). Within-group results showed that all outcomes except pain severity (P = 0.06) had directly improved in the intervention group after treatment (P < 0.05), while in the control group only PFM strength and endurance and UI severity had improved (P < 0.05). CONCLUSIONS: Home-based stabilization exercises focusing on the pelvic floor muscles could be effective for postnatal LBP and SUI. TRIAL REGISTRATION: Iranian Registry of Clinical Trials (Code: IRCT2017050618760N4).

Prevalence and Risk Factors of Urinary/Anal Incontinence and Pelvic Organ Prolapse in Healthy Middle-Aged Iranian Women.

OBJECTIVES: Urinary incontinence (UI) and anal incontinence (AI) cause concern, social exclusion, and ultimately reduced quality of life in women. The aim of present study was to assess the prevalence and related risk factors of UI, AI, and pelvic organ prolapse (POP). METHODS: The present study recruited 340 menopausal women living in Tabriz in northwest Iran. The data collection tools included the Pelvic Floor Distress Inventory-20 and a personal and social information questionnaire. POP was diagnosed via clinical examination using the simplified pelvic organ prolapse quantification system. RESULTS: The prevalence of UI and POP was approximately 50%, and approximately 16% of participants reported AI. Based on the odds ratios, the most remarkable risk factor of urinary stress incontinence was the number of vaginal deliveries, whereas that of urinary urge incontinence was obesity. Episiotomy and age were the most major risk factors of AI and POP, respectively. CONCLUSIONS: The results of the present study showed that the prevalence of POP, UI, and AI is remarkably high among postmenopausal women, warranting the need to prioritize the assessment of POP and various incontinences in middle-aged women in the primary health care system. Furthermore, increased emphasis should be put on modifiable risk factors.

Long-term Outcome of Synthetic Mesh Use in Iranian Women with Genital Prolapse.

PURPOSE: To evaluate the long-term outcome of synthetic mesh use in the treatment of women with Pelvic organ prolapse (POP). MATERIALS AND METHODS: We evaluated the outcome of synthetic mesh implantation by vaginal surgery method in 153 women (mean age of 53.66±9.31 years) with POP grade >2 in the anterior compartment. Demographic findings, baseline symptoms as well as subjective and objective outcome were recorded during the follow-up period of 36.89±11.33 months.   Results: POP relapse occurred in 3.3% indicative of 96.7% anatomical success rate. Patients' common baseline findings were frequency (72.5%), stress and urge incontinence (59.5% and 47.7%). Subjective outcome were vaginal pain (13.7%), dyspareunia (9.2%) and tension feeling (8.5%), while objective outcomes were mesh exposure (3.9%), urge incontinence (11.1%) and vaginal infection (1.3%). Stress incontinence was completely treated following surgery. There was significant improvement in dyspareunia, vaginal pain, urge and stress incontinence (all p < 0.001) and fecal incontinence (p = 0.02). After surgery, 88.42% were satisfied of the surgery outcome. CONCLUSION: POP surgery with synthetic mesh has acceptable results, considerable improvement in symptoms and high rate of satisfaction during follow-up; however, side effects are not uncommon but tolerable.

The effect of Seidlitzia rosmarinus (eshnan) on the prevention of recurrent cystitis in women of reproductive age: A randomized, controlled, clinical trial.

The World Health Organization has recommended herbal medicine for treatment and control of recurrent cystitis. This study was conducted to determine effect of eshnan on the prevention of recurrent cystitis. The present triple-blind clinical trial was conducted on 126 women (63 women per group) with recurrent cystitis. The participants were assigned to the intervention and control groups. Three 500-mg eshnan or placebo tablets were taken orally with a glass full of water by the participants half an hour after each meal for 2 months. The study subjects were clinically and paraclinically evaluated by the end of the second, fourth, and sixth months after the intervention and every time they showed clinical symptoms of cystitis. The present findings showed that the incidence rate of cystitis and symptoms of cystitis was significantly lower in the eshnan group compared with the placebo group 2, 4, and 6 months after the intervention (p < .05). The incidence of recurrent was also significantly lower in the intervention group compared with the control group (p < .001). No side effects were observed in the intervention and control groups. The consumption of eshnan can improve the symptoms of cystitis and prevent the incidence of recurrent cystitis in women of reproductive age.

Pelvic floor rehabilitation in the treatment of women with dyspareunia: a randomized controlled clinical trial.

INTRODUCTION AND HYPOTHESIS: Dyspareunia, the symptom of painful sexual intercourse, is a common sexual dysfunction in reproductive-aged women. Because of its multifactorial etiology, a multidisciplinary approach may be required to treat it. Musculoskeletal factors play an important role; thus, rehabilitating the pelvic floor and modifying the tone of the pelvic floor muscles (PFMs) may be an effective way to treat this dysfunction. The aim of this randomized controlled clinical study was to evaluate the effects of pelvic floor rehabilitation techniques on dyspareunia. METHODS: Of 84 women, assessed for eligibility, 64 women with dyspareunia were randomized into two groups: the experimental group (n = 32) received electrotherapy, manual therapy, and PFM exercises and the control group (n = 32) had no treatment while on the waiting list. Evaluations of PFM strength and endurance, sexual function, and pain were made directly before and after 3 months of treatment and at the 3-month follow-up. RESULTS: Between-group changes showed significant improvement in the experimental group in comparison with control group. Mean difference in the PFM strength (according to the 0-5 Oxford scale) between groups was 2.01 and the mean difference of endurance was 6.26 s. Also, the mean difference in the Female Sexual Function Index score (the score ranges from 2 to 95) was 51.05, and the mean difference in the VAS score was 7.32. All of the changes were statistically significant (p < 0.05). CONCLUSIONS: According to the results, pelvic floor rehabilitation is an important part of a multidisciplinary treatment approach to dyspareunia.

Risk factors of overactive bladder syndrome and its relation to sexual function in menopausal women.

BACKGROUND: Overactive bladder syndrome is a common and annoying complication worldwide that could negatively affect the quality of life of afflicted individuals. We aimed to determine the prevalence and risk factors of overactive bladder syndrome and its relation to sexual function in healthy menopausal women. METHODS: This cross-sectional study was done on 340 women aged 45-60 years in Tabriz, northwest Iran, during 2015-2016. Data were collected using a demographic data questionnaire, the Overactive Bladder Syndrome Score, and the McCoy Female Sexuality Questionnaire. RESULTS: Fifty-six (16.5%), 63 (18.5%), and 10 (2.9%) of the participating women had mild, moderate, and severe overactive bladder syndrome, respectively. Predictors of overactive bladder included: night sweats, central prolapse, episiotomy, varicose disease, illiteracy or education at the primary level, systolic blood pressure >140 mmHg and lack of physical activity. We found a significant difference between the women with and without overactive bladder with respect to the total score and sub-domain scores related to sex partner (p = 0.029) and sexual interest (p = 0.049). CONCLUSIONS: The prevalence of overactive bladder was quite high in this study. Since sexual dysfunction is not an easy topic to talk about and can affect women's quality of life, physicians should consider talking about these issues besides urinary issues to all middle-aged women.

Risk factors of overactive bladder syndrome and its relation to sexual function in menopausal women.

BACKGROUND: Overactive bladder syndrome is a common and annoying complication worldwide that could negatively affect the quality of life of afflicted individuals. We aimed to determine the prevalence and risk factors of overactive bladder syndrome and its relation to sexual function in healthy menopausal women. METHODS: This cross-sectional study was done on 340 women aged 45-60 years in Tabriz, northwest Iran, during 2015-2016. Data were collected using a demographic data questionnaire, the Overactive Bladder Syndrome Score, and the McCoy Female Sexuality Questionnaire. RESULTS: Fifty-six (16.5%), 63 (18.5%), and 10 (2.9%) of the participating women had mild, moderate, and severe overactive bladder syndrome, respectively. Predictors of overactive bladder included: night sweats, central prolapse, episiotomy, varicose disease, illiteracy or education at the primary level, systolic blood pressure >140 mmHg and lack of physical activity. We found a significant difference between the women with and without overactive bladder with respect to the total score and sub-domain scores related to sex partner (p = 0.029) and sexual interest (p = 0.049). CONCLUSIONS: The prevalence of overactive bladder was quite high in this study. Since sexual dysfunction is not an easy topic to talk about and can affect women's quality of life, physicians should consider talking about these issues besides urinary issues to all middle-aged women.

The Future of Diabetes Management by Healthy Probiotic Microorganisms.

BACKGROUND: Diabetes mellitus, a condition of multifactorial origin, is related to the intestinal microbiota by numerous molecular mechanisms. Controlling the vast increase in the prevalence of diabetes needs a natural and safe solution. Probiotics, known as live microorganisms that exert health benefits to the host, have anti-diabetic property. OBJECTIVE: This review will highlight the current evidences in probiotic effectiveness and future prospects for exploring probiotic therapy in the prevention and control of diabetes. METHODS: We searched Pub Med and Science Direct by using "Probiotics" and "Diabetes" for searching the studies aiming the application of probiotics and the beneficial effects of probiotics in diabetes prevention and control. RESULTS: It has been shown that probiotics can increase insulin sensitivity and reduce autoimmune responses by modulating intestinal microbiota and decreasing the inflammatory reactions and oxidative stress. Recent evidences show that probiotics influences the host through modulating intestinal permeability and mucosal immune response, manipulating eating behaviors by appetite-regulating hormones and controlling gut endocannabinoid (eCB) system that is believed to be associated with inflammation and diabetes. Moreover, modulating the intestinal microbiota by probiotics controls host metabolism by affecting energy extraction from food and by biochemically converting molecules derived from the host or from gut microbes themselves. CONCLUSION: Experimental and clinical evidences support the hypothesis that the modulation of the gut microbiota by probiotics could be effective in prevention and management of diabetes.

Effects of probiotics on the recurrence of bacterial vaginosis: a review.

OBJECTIVE: Bacterial vaginosis (BV) is a common cause of genital discomfort in women in reproductive ages, which causes many complications. Bacterial vaginosis is usually treated by metronidazole and clindamycin. However, this protocol does not prevent its recurrence, which is a main complaint of the patients. The number of lactobacilli in the vagina of women with BV is significantly lower than that in healthy women. Hence, efforts have been made to normalize vaginal flora by oral or vaginal administration of lactobacilli. The objective of the present study was to review clinical evidences available regarding the efficacy of probiotics in the prevention and treatment of BV. MATERIALS AND METHODS: Published randomized controlled trials were searched in PubMed, Science Direct, and the Cochrane Database between 1990 and 2011. Search terms included bacterial vaginosis, urinary tract infection, lactobacillus, and probiotics. RESULTS: Orally consumed probiotics are believed to ascend to the vaginal tract after they are excreted from the rectum; vaginal administration allows for direct replacement of the probiotics for unhealthy vaginal microbiota and occupation of specific adhesion sites at the epithelial surface of the urinary tract, which consequently results in maintenance of a low pH and production of antimicrobial substances like acids and hydrogen peroxide. Receiving Lactobacillus acidophilus, Lactobacillus rhamnosus GR-1, and Lactobacillus fermentum RC-14 at a dose of at least 10 CFU/day for 2 months has been shown to present the patients with better results. CONCLUSIONS: Although the results of different studies are controversial, most studies have been in favor of the probiotics in the prevention or treatment of BV, and no adverse effects have been reported. Therefore, it may be helpful to recommend daily consumption of probiotic products to improve public health among women.

Modification and Evaluation of Avidity IgG Testing for Differentiating of Toxoplasma gondii Infection in Early Stage of Pregnancy.

OBJECTIVE: Toxoplasma gondii infection, an intracellular parasite, is often asymptomatic or is caused by different clinical diseases without being detected. Evaluation of IgG, IgA, and IgM in order to diagnose the pending Toxoplasmosis may confront some problems. Several researches has showned that Toxo IgG avidity can be useful in the recent active Toxoplasmosis. In current study, modification and importance of improved Toxoplasma Avidity IgG testing has been evlauated for differentiating Toxoplasma gondii infection in early stage of pregnancy. MATERIALS AND METHODS: This experimental study included 300 pregnant women with risk of Toxoplasmosis in their initial months of pregnancy. We randomly divided 300 serum samples into A group (n=60) with high avidity and B group (n=40) with borderline avidity. The samples with Toxo IgG levels were classified to four subgroups. IgG avidity was evaluated by enzyme-linked immunosorbent assay (ELISA) method. RESULTS: The mean absorbance of 100 samples in two groups was calculated, and then, two dilutaion curves with plotted absorbance against dilution were drawn for each serum sample. The results of this study showed that in groups with different concentrations of toxo IgG, appropriate dilution of serum is suitable for testing of Avidity. Our findings revealed the subgroups of 1, 2, 3, and 4 with serum dilutios of 1/3 , 1/6, 1/9, and 1/18 respectively, had real and good avidity. CONCLUSION: : One of the issues affectig the results of avidity is high concenteration of Toxo IgG in serum sample. As shown in this study, the best points of dilution for well avidity in both high and borderline avidities are marked with arrows in figures 1-8. This study confirmed that improved methods of measuring Toxo Avidity IgG are very important.

Comparison of neonatal and maternal outcomes associated with head-pushing and head-pulling methods for impacted fetal head extraction during cesarean delivery.

OBJECTIVE: To compare the morbidity and mortality of 2 current techniques during cesarean delivery of an impacted fetal head. METHODS: In a comparative setting, 59 pregnant women with obstructed labor due to impacted fetal head were recruited. The patients were categorized into 2 groups according to method of extraction: the "push" group (n=30) and the "pull" group (n=29). Uterus relaxants were used before cesarean in all cases and the incision was higher and wider than routine. Maternal and neonatal morbidities were compared between the groups. RESULTS: Maternal complications in the push and pull groups were extension of the uterine incision (15 [50.0%] vs 5 [17.2%]); T or J incision (3 [10.0%] vs 4 [13.8%]); blood transfusion (3 [10.0%] vs 1 [3.4%]); wound infection (4 [13.3%] vs 1 [3.4%]); fever (16 [53.3%] vs 3 [10.3%]); and urinary tract infection (10 [33.3%] vs 0 [0.0%]). Incidences of extension of the uterine incision, fever, and urinary tract infection were significantly higher in the push group (P=0.008). CONCLUSION: Owing to a lower rate of abnormal incision and postpartum fever/infection with the pull method, this technique is preferable to the push method.

Transvaginal ultrasonography compared with Bishop score for predicting cesarean section after induction of labor.

BACKGROUND: Reproductive health researchers are interested in finding better methods for predicting an unwanted type of delivery after induction of labor. The aim of this study was to compare the value of transvaginal ultrasonography findings and the Bishop score in predicting cesarean section after induction of labor. METHODS: Two hundred women with singleton pregnancies undergoing induction of labor at 37-42 weeks were enrolled in this prospective study. Transvaginal investigation was done for all participants prior to induction. To compare the predictive value of the methods, receiver-operating characteristic (ROC) curves were plotted and equality of the area under curve (AUC) was tested. RESULTS: The mean age of the participants was 29.9 years, mean gestational age was 39.6 weeks, and mean gravid was 1.5. The AUC calculated for Bishop score was 0.39 (95% confidence interval [CI] 0.3-0.48). The AUC for cervical length measured by ultrasonography was 0.69 (95% CI 0.6-0.77). The AUC for the posterior cervical angle measured by ultrasonography was 0.38 (95% CI 0.29-0.47). Testing equality of the ROC curves for these three methods showed the ROC for cervical length to be statistically different from both Bishop score and posterior cervical angle (P < 0.001). However, the difference in ROC area compared between Bishop score and posterior cervical angle was not statistically significant. CONCLUSION: Based on our findings and available information in the literature, it seems that cervical length measured by transvaginal ultrasonography has the potential to replace the traditional Bishop score, provided that such a facility is available when needed.

Effects of vitamin E supplementation on some pregnancy health indices: a randomized clinical trial.

OBJECTIVE: Available evidence about the role of supplementary vitamin E in normal pregnancy is inadequate. This study assessed the potential benefit of vitamin E supplementation on some pregnancy health indices. METHODS: A 1:2 weighted simple randomization technique was used to allocate 104 eligible pregnant women to receive vitamin E and 168 women (control) not to receive the drug. Treated women received capsules containing 400 IU vitamin E from week 14 of gestation to the end of the pregnancy. RESULTS: Background variables including maternal age were found to be similarly distributed between the study groups. The rate of maternal and perinatal outcomes including Apgar score and birth weight did not differ significantly between groups. Preeclampsia occurred in 1% of treated women vs 1.78% of control women. CONCLUSION: Giving supplemental vitamin E from the second trimester of pregnancy did not appear to affect the risk of pregnancy outcomes and occurrence of preeclampsia.

The influence of maternal ketonuria on fetal well-being tests in postterm pregnancy.

BACKGROUND: Although it is well-known that postterm pregnancies are associated with the risk of perinatal morbidity and mortality, a comprehensive study on its management is lacking. The aim of present study was to determine whether ketonuria is associated with abnormal fetal test results in pregnancies >40 weeks of gestation. METHODS: In this analytical cross sectional study, a total of 360 pregnant women with gestational age of >40 weeks were evaluated in two hospitals during 2003 - 2004. For each woman, urinary ketones, glucose, pH, proteins, and specific gravity were measured by total screen LSG tapes (Rapignost); in addition, biophysical profile test was recorded and the amniotic fluid index was assessed by ultrasonography. RESULTS: Ketonuria was found in 34 women (9.44%). Statistical analyses showed that maternal ketonuria was associated with oligohydramnios, abnormal fetal tests, and a significant increase in fetal heart rate decelerations. CONCLUSION: Patients with clinically-detectable ketonuria have higher risk for abnormal fetal tests in comparison with those without the disease.