The association of TMPRSS6 gene polymorphism with iron status in Egyptian children (a pilot study).

Several studies have shown association of single nucleotide polymorphisms (SNPs) of hepcidin regulatory pathways genes with impaired iron status. The most common is in the TMPRSS6 gene. In Africa, very few studies have been reported. We aimed to investigate the correlation between the common SNPs in the transmembrane protease, serine 6 (TMPRSS6) gene and iron indicators in a sample of Egyptian children for identifying the suitable candidate for iron supplementation.Patients and methods One hundred and sixty children aged 5-13 years were included & classified into iron deficient, iron deficient anemia and normal healthy controls. All were subjected to assessment of serum iron, serum ferritin, total iron binding capacity, complete blood count, reticulocyte count, serum soluble transferrin receptor and serum hepcidin. Molecular study of TMPRSS6 genotyping polymorphisms (rs4820268, rs855791 and rs11704654) were also evaluated.Results There was an association of iron deficiency with AG of rs855791 SNP, (P = 0.01). The minor allele frequency for included children were 0.43, 0.45 & 0.17 for rs4820268, rs855791 & rs11704654 respectively. Genotype GG of rs4820268 expressed the highest hepcidin gene expression fold, the lowest serum ferroportin & iron store compared to AA and AG genotypes (p = 0.05, p = 0.05, p = 0.03 respectively). GG of rs855791 had lower serum ferritin than AA (p = 0.04), lowest iron store & highest serum hepcidin compared to AA and AG genotypes (p = 0.04, p = 0.01 respectively). Children having CC of rs11704654 had lower level of hemoglobin, serum ferritin and serum hepcidin compared with CT genotype (p = 0.01, p = 0.01, p = 0.02) respectively.Conclusion Possible contribution of SNPs (rs855791, rs4820268 and rs11704654) to low iron status.

Role of adiponectin, resistin and monocyte chemo-attractant protein-1 in overweight/obese asthma phenotype in children.

BACKGROUND: Asthma is a chronic inflammatory disorder of the airways with diverse overlapping pathologies and phenotypes contributing to a significant heterogeneity in clinical manifestations. Obesity may modify asthma risk, phenotype, and prognosis. A suggested mechanism linking obesity and asthma is through systemic inflammation. Adipokines secreted by adipose tissue were suggested to provide a link between obesity and asthma. OBJECTIVE: To have an understanding for the contribution of adiponectin, resistin and MCP-1 to development of distinct asthma phenotype in overweight/obese children through assessment of their serum level and correlation to pulmonary function tests. SUBJECTS AND METHODS: The study included 29 normal weight asthmatics, 23 overweight/obese asthmatic children and 30 controls. All cases were subjected to detailed history taking, thorough examination and pulmonary function tests. Serum adiponectin, resistin, MCP-1 and IgE were assessed to all recruited subjects. RESULTS: Adiponectin level was significantly higher in overweight/obese asthmatics (24900 ± 1600 ng/ml) compared to normal weight asthmatics (21700 ± 1700 ng/ml) and control (23000 ± 3200 ng/ml), (p < 0.001 & 0.051 respectively). Normal weight asthmatics had significantly lower adiponectin level than control, (p = 0.039). A significant low level of MCP-1 in overweight/obese asthmatics (149.5 (20-545) ng/L) compared to control (175 (28 -1123.5) ng/L), p = 0.037. No significant difference was found regarding resistin. Normal weight asthmatics had significantly lower FEV1% and FVC% compared to overweight/obese asthmatics (p = 0.036, 0.016 respectively). A significant positive correlation was found between (FEV1%, FVC) and BMI in normal weight asthmatics (P = 0.01, < 0.01 respectively) and a significant negative correlation between PEF and BMI (-0.42, p = 0.05) in obese/overweight asthmatics. Resistin/adiponectin ratio was not affected by sex, degree of asthma severity or level of asthma control in either normal weight or overweight/obese asthmatic. CONCLUSION: This work could suggest that adiponectin may play a role in overweight/obese asthma phenotype where it is possible to have a dual action (pro & anti- inflammatory). It seems that resistin had no role in asthma pathogenesis.

Ultrasound risk stratification for malignancy using the 2015 American Thyroid Association Management Guidelines for Children with Thyroid Nodules and Differentiated Thyroid Cancer.

BACKGROUND: The 2015 American Thyroid Association (ATA) Management Guidelines for Children with Thyroid Nodules and Differentiated Thyroid Cancer provides selection criteria for nodules prior to ultrasound-guided fine-needle aspiration biopsy. OBJECTIVE: To evaluate the diagnostic performance of pediatric thyroid nodule risk stratification for predicting malignancy when applying the ultrasound (US) criteria recommended. MATERIALS AND METHODS: US characteristics of 39 thyroid nodules in 33 pediatric patients who underwent US fine-needle aspiration biopsy were reviewed by two radiologists. Based on the aggregated US criteria from the ATA Guidelines, each nodule was assigned a level of malignancy risk. Kappa coefficients were estimated to assess intra- and interobserver reliability. Using each patient's largest nodule observation (n = 33), univariable exact logistic regression analyses of US parameters were then conducted to estimate the odds of a malignant pathology diagnosis. A penalized Firth correction was employed in the univariable models analyzing composition, shape and level of suspicion due to quasi-complete data separation. RESULTS: Twenty-seven nodules in 21 patients (median age: 16 years; 17 female) were benign and 12 nodules in 12 patients (median age: 16.5 years; 11 female) were malignant. Intraobserver agreement was substantial to almost perfect for composition, echogenicity, shape and margins. Interobserver agreement was almost perfect for composite level of suspicion. High level of suspicion was assigned to all 12 malignant nodules versus 9/21 (43%) of the benign nodules. Level of suspicion, solid/predominantly solid composition, irregular margins and echogenic foci emerged as significant predictors of malignancy with odds ratios (OR) of 8.5 (95% confidence interval [CI]: 1.7-1,130, P = 0.001), 10.5 (95% CI: 1.1-1,417, P = 0.04), 53.2 (95% CI: 5.1-2,988, P < 0.0001) and 3.5 (95% CI: 1.1-23.2, P = 0.03), respectively. CONCLUSION: The composite, US-based risk stratification criteria from the 2015 ATA Guidelines may provide an appropriate and reproducible method for estimating risk of malignancy for pediatric thyroid nodules.

Evaluation of rezasurin microtiter assay and high resolution melting curve analysis for detection of rifampicin and isoniazid resistance of Mycobacterium tuberculosis clinical isolates.

BACKGROUND: Several methods have been proposed for rapid detection of drug resistance of Mycobacterium tuberculosis (M. tuberculosis). High resolution melting (HRM) curve analysis has been developed for accurate and simultaneous detection of resistance of M. tuberculosis isolates to rifampicin (RIF) and isoniazid (INH). Other techniques include the resazurin microtiter assay (REMA) which is one of the new colorimetric phenotypic methods. The present study evaluated the HRM curve analysis and REMA assay in comparison to the proportional method (PM) for rapid identification of multidrug resistant (MDR) M. tuberculosis isolates. METHODS: Thirty M. tuberculosis clinical isolates of known resistance phenotypes were used. An HRM curve was generated for each isolate to scan for mutations in the rpoB and katG genes to detect RIF and INH resistance, respectively. The REMA colorimetric assay was also evaluated using the same isolates. The results of both techniques were compared to the gold standard proportional method. RESULTS: The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of the REMA assay for RIF and INH susceptibility testing were 100%. The HRM curve assay results for RIF susceptibility testing were 92.3%, 100%, 100%, 94.4%, and 96.7%, respectively, and for INH they were 85%, 100%, 100%, 76.9%, and 90%, respectively. CONCLUSIONS: REMA is a rapid non-conventional and inexpensive method and may serve as a replacement for the proportion method in resource limited settings, while the PPV and NPV of the HRM curve make this assay an ideal screening method for the TB laboratory.

Heterotopic ossification post COVID-19: Report of two cases.

Heterotopic ossification is often associated with trauma and surgery, and infrequently reported with immobilization due to critical illness. We present 2 patients who developed heterotopic ossification following severe COVID-19 infection. Both patients were middle-aged females who were hospitalized for one month or greater due to COVID-19 requiring mechanical ventilation. Both developed shoulder pain and/or stiffness a few months after discharge, with imaging studies clearly illustrating development of heterotopic ossification around the shoulders. The etiology is unclear, with immobilization and hypoxia being the primary considerations. Physical examination and radiography are essential to diagnosis. Awareness of this complication and early diagnosis may help minimize functional impairment.

C5 palsy after cervical laminectomy: natural history in a 10-year series.

BACKGROUND CONTEXT: C5 palsy is a well-known complication following cervical laminectomy, however the cause of this complication remains elusive, with many studies providing conflicting reports on prognosis and the impact of specific risk factors. PURPOSE: To describe the natural history of and risk factors for C5 palsy after first time cervical laminectomy involving C4 and/or C5, in a large series with a high rate of postoperative palsy. STUDY DESIGN/SETTING: This is a retrospective case series. PATIENT SAMPLE: Patients undergoing first time cervical laminectomy for degenerative spine pathologies at a single institution between January 2008 and July 2018. Adult patients were included if a complete laminectomy was performed at C4 or C5 for degenerative pathology and pre- and postoperative upright lateral x-rays were performed. OUTCOME MEASURES: The primary outcome measure was postoperative C5 palsy, defined as a decrease in strength of at least one point in deltoid and/or biceps within 30 days of operation. The secondary outcome measure was recovery of function. METHODS: A retrospective database of patients who underwent posterior cervical spine surgery was created and further focused by utilizing specific Common Procedural Technology (CPT) codes associated with our desired patient population. Patients were excluded from our study if they had inadequate pre- and postoperative imaging, as well as patients with a history of prior cervical spine surgery, concurrent anterior surgery, intradural pathology, spinal tumor, or spinal trauma. Patient history, surgical specifics, and neurologic function were recorded. RESULTS: A total of 190 patients were treated by 13 surgeons. 53 (27.9%) developed C5 palsy postoperatively. Of patients with C5 palsy, 40 (75.5%) recovered to baseline strength, 46 (86.6%) had at least grade 4 strength at last follow up, and 4 (7.5%) had strength worse than baseline and motor grade less than 4. Median time to recovery was 2.0 (IQR: 0.18 to 8.24) months. Age, gender, preoperative motor score, number of levels decompressed, smoking history, and comorbidities were not associated with a significant increase in the odds of C5 palsy. Risk of C5 palsy increased by 35% for every additional level fused below C4. CONCLUSION: The risk of C5 palsy is increased with instrumentation caudal to C5 in operations addressing degenerative cervical pathology. This should be taken into consideration during operative planning. Overall prognosis of C5 palsy is good; however, incidence of this condition may be greater than previously reported.

Low Dose Iron Therapy in Children with Iron Deficiency: DNA Damage and Oxidant Stress Markers.

Conflicting data are available regarding oral iron therapy in iron deficiency (ID), iron deficiency anemia (IDA) and its relation to DNA damage, oxidative stress and antioxidant markers. Our aim was assessment of DNA damage, oxidative stress and anti-oxidant markers in children with ID and IDA before and after low dose iron therapy. The study was conducted in two stages, first stage was assessment of DNA damage using comet assay, malondialdehyde (MDA) and anti-oxidant enzymes levels (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) & total antioxidant capacity (TAC) in thirty-nine children with IDA, forty-five children with ID without anemia and sixty healthy controls. Second stage was assessment of previous markers together with hematological response following oral therapy with 10 mg/day ferric ammonium citrate for 8 weeks. Before treatment, there was no significant difference between the three groups regarding MDA, GPx, SOD, CAT and TAC. A significant increase was detected in the DNA damage in the 2 groups compared to control (p < 0.005). Following iron therapy, hematological parameters was improved together with a significant increase in GPx (P = 0.04), SOD (p = 0.002), TAC (P = 0.001) and non-significant reduction in DNA damage in IDA group. There was a significant increase in SOD (p = 0.001) & TAC (p = 0.001) and significant decrease in DNA damage (p = 0.001) in ID group. Low dose iron therapy could be sufficient to improve antioxidant status and DNA damage together with correction of hematologic indices.

A practical radiographic visual estimation technique for the prediction of developmental narrowing of cervical spinal canal.

OBJECTIVES: To determine if visual spinal canal percentage of facet overlap could be used as a rapid visual pre-screening test to detect developmental bony spinal canal narrowing on lateral cervical radiographs. METHODS: Lateral cervical spine radiographs with radiology report documenting developmental narrowing of the spine canal were retrospectively identified. Cohort was supplemented with additional images without such documentation. Utilizing a radiology work station, one reader (Orthopaedics) calculated the Torg ratio at C3-C6. Two readers (Radiology, Primary Care Sports Medicine) visually estimated percentage (≥80, ≥90, 100%) of spinal canal overlaid by facet articulations C3-C6 for all cases independently. Statistical analysis included accuracy tests and kappa test for comparing the gold standard and inter-, intra-observer reliability, respectively. RESULTS: 96 lateral images were included; 25 had abnormal radiology reports. High specificity (range, 91-93%) and true negative predictive values (range, 88-93%) achieved consistent statistical significance for relationship between facet/canal overlap and Torg ratio for both combined and individual data points. Sensitivity values (range, 59-75%) showed a trend toward association, with statistical significance for only the C3 level at 80% overlap. Interobserver agreement was substantial. Intra-observer reliability was moderate to perfect. CONCLUSION: Visual estimation of percentage of facet overlap of the spinal canal is a valid tool to identify individuals with potential developmental narrowing of the spinal canal as a screening prompt to select which patients may need formal calculation utilizing accepted metrics. Advances in knowledge Visual estimation of percentage of facet overlap from lateral radiographs can predict developmental narrowing of cervical spinal canal.

Assessment of serum level of corticotropin-releasing factor in primary nocturnal enuresis.

INTRODUCTION: Primary nocturnal enuresis is one of the sleep related phenomena characterized by disruption in the relationship between arousal and urination. Corticotropin-releasing factor (CRF) is a neurohormone released from the paraventricular nucleus of the hypothalamus into the median eminence to elicit release of adrenocorticotrophin from the anterior pituitary. It may act to modulate autonomic function and behavior in concert with the endocrine effects. Conflicting animal studies about the role of CRF in micturition, either facilitating or inhibiting, have been raised. It was suggested to be a novel target for treatment of urinary disorders based on the finding that manipulation of CRF in the pontine micturition circuit could affect urodynamic function. AIM: The aim was to throw light on the possible role of CRF in primary monosymptomatic nocturnal enuresis by assessing its serum level. SUBJECTS AND METHODS: Twenty-nine children aged 8-14 years complaining of primary monosymptomatic nocturnal enuresis and 16 age- and sex-matched healthy children with good toilet control day and night were recruited to the study. History taking, clinical examination, and assessment of serum CRF levels in the morning and evening (9 a.m. and 9 p.m.) were carried out for all patients and controls. RESULTS AND DISCUSSION: A positive family history of enuresis was detected in 82.8% of enuretic patients. Serum levels of CRF (both morning and evening) were significantly lower in patients than in controls. Several animal studies suggested that CRF in descending projections from Barrington's nucleus to the lumbosacral parasympathetic neurons is inhibitory to micturition, which supports our results and the assumption that reduction of the evening serum CRF level could have a role in the occurrence of primary monosymptomatic nocturnal enuresis. No significant difference was found between morning and evening CRF serum levels in either cases or controls, which negates our assumption of having a rhythmic pattern of release (figure). No correlations with age were found. According to their history, all our enuretic patients were deep sleepers. Deep sleep and difficult arousal were found to have a major role in primary monosymptomatic nocturnal enuresis. It was proposed that CRF function may allow arousal to occur before micturition to facilitate preparative behaviors. A lower CRF level may explain deep-sleep pattern in children with enuresis. CONCLUSION: CRF was deficient in our enuretic children, which may draw attention to the possible pathophysiological implications in primary nocturnal enuresis (either at the level of loss of inhibitory effect on micturition or lack of arousal in response to bladder distension). Further proof studies are recommended.

Anti-diuretic hormone and genetic study in primary nocturnal enuresis.

OBJECTIVE: To investigate whether primary nocturnal enuresis (PNE) is related to a disturbance in anti-diuretic hormone (ADH) secretion pattern at night which may be genetically inherited. SUBJECTS AND METHODS: This study included 121 children aged 6-18 years with PNE and 45 matched healthy children as controls. Enuretic children were subjected to genetic evaluation and cytogenetic assessment (n = 99). Assay of ADH levels (9-11 am & 9-11 pm) was performed for cases (n = 99) and controls. RESULTS: There was a positive family history in 82.4% (autosomal dominant in 35.4% and autosomal recessive in 64.6%). ADH morning and evening values were reversed in cases vs controls with significant difference in morning level. Reversal of circadian rhythm was present in 71.7% of cases and normal rhythm in 28.3% of them. Morning and evening ADH levels revealed significant difference between patients with reversed rhythm and those with normal one, and with controls. Mode of inheritance had no influence on hormonal level. Chromosomal abnormality was detected in 3 cases with reversed ADH rhythm, involving chromosome 22 in two of them. CONCLUSION: PNE could be attributed in part to reversed ADH circadian rhythm which may be linked to chromosome 22.