MicroRNA 21 and microRNA 10b: early diagnostic biomarkers of breast cancer in Egyptian females.

BACKGROUND: Breast cancer (BC) is one of the most prevalent cancers in developing and developed countries among women worldwide. Mammography is one of the superior methods for BC detection, but it carries up to 20% false-negative results, especially in early cases. Histological examination of tissue biopsies and fine-needle aspiration cytology are invasive techniques. Hence, minimally invasive markers are needed for the improved detection of BC. microRNAs, small, noncoding, single-stranded RNAs functioning as tumor suppressor genes or oncogenes, are attractive biomarkers for early detection. This study aimed to examine the serum levels of miR21 and miR10b in patients with BC especially in the early stages compared to healthy controls to evaluate their potential use as BC biomarkers. METHODS: This study included 90 females who were divided into two groups. Group I included 70 patients with BC and was subdivided into group Ia with 40 nonmetastatic BC patients and group Ib with 30 metastatic BC patients. Group II included 20 apparently healthy females as a control group. Serum miR21 and miR10b as biomarkers and miR16 as a housekeeping gene were evaluated using real-time polymerase chain reaction. RESULTS: The median levels of miR10b and miR21 were statistically significantly upregulated in the sera of patients with BC compared to healthy controls (P = 0.001). Receiver operating characteristic curve analyses demonstrated that serum levels of miR10b and miR21 were useful biomarkers for distinguishing between patients with BC and the control group, with an area under the curve (AUC) of 0.991 with 97.1% sensitivity and 100% specificity at a cutoff of 3.1 for miR10b and an AUC of 0.965 with 95.7% sensitivity and 85% specificity at a cutoff of 1.7 for miR21. Regarding the early stages of BC, the median levels of the fold change of serum miR21 and miR10b were statistically significantly higher in patients with BC (stages I and IIa) than in the control group (P < 0.001). CONCLUSIONS: Both miR21 and miR10b have valuable diagnostic roles in detecting the early stages of BC.

Genetic study of pediatric hypertrophic cardiomyopathy in Egypt.

Paediatric cardiomyopathy is a progressive and often lethal disorder and the most common cause of heart failure in children. Despite their severe outcomes, their genetic etiology is still poorly characterised. The current study aimed at uncovering the genetic background of idiopathic primary hypertrophic cardiomyopathy in a cohort of Egyptian children using targeted next-generation sequencing. The study included 24 patients (15 males and 9 females) presented to the cardiomyopathy clinic of Cairo University Children's Hospital with a median age of 2.75 (0.5-14) years. Consanguinity was positive in 62.5% of patients. A family history of hypertrophic cardiomyopathy was present in 20.8% of patients. Ten rare variants were detected in eight patients; two pathogenic variants (8.3%) in MBPC3 and MYH7, and eight variants of uncertain significance in MYBPC3, TTN, VCL, MYL2, CSRP3, and RBM20.Here, we report on the first national study in Egypt that analysed sarcomeric and non-sarcomeric variants in a cohort of idiopathic paediatric hypertrophic cardiomyopathy patients using next-generation sequencing. The current pilot study suggests that paediatric hypertrophic cardiomyopathy in Egypt might have a particular genetic background, especially with the high burden of consanguinity. Including the genetic testing in the routine diagnostic service is important for a better understanding of the pathophysiology of the disease, proper patient management, and at-risk detection. Genome-wide tests (whole exome/genome sequencing) might be better than the targeted sequencing approach to test primary hypertrophic cardiomyopathy patients in addition to its ability for the identification of novel genetic causes.

Increased Free Circulating DNA Integrity Index as a Serum Biomarker in Patients with Colorectal Carcinoma.

BACKGROUND: Cell-free DNA circulating in blood is a candidate biomarker for malignant tumors. Unlike uniformly truncated DNA released from apoptotic non diseased cells, DNA released from necrotic cancer cells varies in size. OBJECTIVES: To measure the DNA integrity index in serum and the absolute DNA concentration to assess their clinical utility as potential serum biomarkers for colorectal carcinoma (CRC) compared to CEA and CA19-9. MATERIALS AND METHODS: Fifty patients with CRC, 10 with benign colonic polyps and 20 healthy sex and age matched volunteers, were investigated by real time PCR of ALU repeats (ALU q-PCR) using two sets of primers (115 and 247 bp) amplifying different lengths of DNA fragments. The DNA integrity index was calculated as the ratio of q-PCR results of ALU 247/ ALU 115bp. RESULTS: Serum DNA integrity was statistically significantly higher in CRC patients compared to the benign and control groups (p<0.001). ROC curves for differentiating CRC patients from normal controls and benign groups had areas under curves of 0.90 and 0.85 respectively. CONCLUSIONS: The DNA integrity index is superior to the absolute DNA concentration as a potential serum biomarker for screening and diagnosis of CRC. It may also serve as an indicator for monitoring the progression of CRC patients. Combining CEA and CA19-9 with either of the genetic markers studied is better than either of them alone.

Amino acid and acylcarnitine profiles in premature neonates: a pilot study.

OBJECTIVE: To investigate the longitudinal changes in amino acid (AA) and acylcarnitine (AC) profiles of preterm neonates over the first 2 wk of life, and to detect any significant deviation from full term values that requires change of cut-off values used for detection of metabolic disorders in preterm neonates. METHODS: This observational analytical longitudinal study was conducted on 131 premature neonates (gestational age ranged from 27 to 36 wk) and 143 healthy full-term neonates. Dried blood spots were taken on the 5th and 14th postnatal day from the premature neonates and on day 5 from full term neonates for neonatal screening. Samples were analyzed for AA and AC using tandem mass spectrometer. RESULTS: Most AA significantly decreased on day 14 compared to day 5 among preterm neonates (p < 0.05). The combined values of total carnitine (TC), total acylcarnitine (tAC) and short-chain acylcarnitines on day 5 among preterm neonates were statistically significantly higher compared to the day 14 sample (p 0.0001), whereas no statistically significant difference was found regarding the values of medium-, long-chain acylcarnitines, tAC/FC, and FC/TC (p > 0.05). The levels of AA of preterm neonates were statistically significantly higher than that of the controls (p < 0.05). The values of TC, tAC, short-, medium- and long-chain acylcarnitines, were significantly higher than those of the controls (p < 0.05). The reference ranges for preterm neonates were determined using the 1st and 99.9th percentiles. CONCLUSIONS: AA and AC showed an age-related distribution of their concentrations. This underlines the importance of using appropriate reference values when working with a prematurely born population.

Seminal plasma anti-Müllerian hormone level correlates with semen parameters but does not predict success of testicular sperm extraction (TESE).

AIM: To assess seminal plasma anti-Müllerian hormone (AMH) level relationships in fertile and infertile males. METHODS: Eighty-four male cases were studied and divided into four groups: fertile normozoospermia (n = 16), oligoasthenoteratozoospermia (n = 15), obstructive azoospermia (OA) (n = 13) and non-obstructive azoospermia (NOA) (n = 40). Conventional semen analysis was done for all cases. Testicular biopsy was done with histopathology and fresh tissue examination for testicular sperm extraction (TESE) in NOA cases. NOA group was subdivided according to TESE results into unsuccessful TESE (n = 19) and successful TESE (n = 21). Seminal plasma AMH was estimated by enzyme linked immunosorbent assay (ELISA) and serum follicular stimulating hormone (FSH) was estimated in NOA cases only by radioimmunoassay (RIA). RESULTS: Mean seminal AMH was significantly higher in fertile group than in oligoasthenoteratozoospermia with significance (41.5 +/- 10.9 pmol/L vs. 30.5 +/- 10.3 pmol/L, P < 0.05). Seminal AMH was not detected in any OA patients. Seminal AMH was correlated positively with testicular volume (r = 0.329, P = 0.005), sperm count (r = 0.483, P = 0.007), sperm motility percent (r = 0.419, P = 0.021) and negatively with sperm abnormal forms percent (r = -0.413, P = 0.023). Nonsignificant correlation was evident with age (r = -0.155, P = 0.414) and plasma FSH (r = -0.014, P = 0.943). In NOA cases, seminal AMH was detectable in 23/40 cases, 14 of them were successful TESE (57.5%) and was undetectable in 17/40 cases, 10 of them were unsuccessful TESE (58.2%). CONCLUSION: Seminal plasma AMH is an absolute testicular marker being absent in all OA cases. However, seminal AMH has a poor predictability for successful testicular sperm retrieval in NOA cases.