Molekulare und genetische Medizin

Epitope sequences are unique combination of amino acids sequence positioned on exposed domains of proteins. Molecular imprinting is a promising technique for creating molecular receptors with recognition and binding sites that are chemically and sterically complementary in shape, size and functionality to the predetermined target molecules in synthetic polymer. This approach creates template-shaped cavities in polymer matrices with memory of template molecules to be used in molecular recognition. Imprinting whole protein denatures the tertiary and quaternary structures of protein in the polymer matrix and complexity and flexibility of its structure cannot be sustained in the polymer matrix. Epitope approach offers a way out of such snags. The epitope-imprinted film revealed high selectivity over the target protein and allow tolerance for even a single amino acid mismatch between the epitope and target protein. MIP sensors are ideal candidates for replacing biosensors as well as natural receptors in many sensing applications such as ELISA. In spite of advantages and burgeoning research in the field of MIPs, imprinting fraternity has not yet achieved commercial success. Substitution of antibodies used in diagnostic tools with synthetic analogues will cut down cost as well as time period for sample analysis. MIP sensing layers have proven to be highly economical and they have shown almost parallel feat as bio-sensing elements (antibody/antigen/enzyme) incorporated in ELISA. Rapid and accurate determination of disease biomarker proteins is vital for clinical diagnosis and medical abnormalities. Hence MIP-sensors of certain proteins will be useful in early diagnosis of diseased state.

Neurons, by virtue of their complex and continuously changing signaling roles in brain, must be able to regulate access to energy in order to maintain their ability to communicate meaningful frequency-encoded information. This is accomplished by release of neurotransmitters to astrocytes that in turn signal the vascular system to increase cerebral blood flow (CBF). This process has been termed “neurovascular coupling” (NVC). It has also been observed that NVC is bimodal in that there are two separate mechanisms for control of CBF. One type is rapid [phasic] in response to changes in glutamatergic synaptic activity and release of glutamate (Glu), K+ and nitric oxide (NO). Uptake of Glu and K+ by astrocytes induces Ca2+ waves activating regional astrocyte syncytium have to liberate prostaglandins which in turn dilate capillaries by relaxing surrounding pericytes. The NO dilates arterioles by relaxing surrounding smooth muscle cells. These agents acting in concert sharply increase CBF within 1-3 seconds. The other type is slow [tonic] reflecting ongoing neuronal metabolic activity of all neuron types independent of changes in synaptic activity or astrocyte Ca2+ waves and eliciting modest oscillations in CBF in 10’s of seconds. In this review, we describe two neuronal signaling mechanisms that match the criteria for phasic and for tonic regulation of CBF. The difference is being the nature and source of the “Glu” released and of their targeted astrocyte receptors. Dependence on synaptic activity limits phasic responses to gray matter, but tonic responses can regulate CBF in both gray matter and white matter and may be the primary regulator of CBF in white matter.

Purpose: Report an infant patient of propionic acidemia with two mutations in the PCCB gene identified by genetic diagnosis.

Method: The patient received gas chromatograph-mass spectrometry and liquid chromatography-tandem mass spectrometry examination, electroencephalogram (EEG), MRI and genetic tests. He was diagnosed as propionic academia.

Results: The boy was admitted in hospital at 8 months of age because of dyspnea, depression, seizures. The EEG was abnormal. MRI showed abnormal signal in bilateral basal ganglia. The gas chromatograph-mass spectrometry and liquid chromatography-tandem mass spectrometry showed glycine, 3-hydroxypropionate, tiglyglycine, methylcitric acid, propionyl carnitine increased. The genetic tests demonstrated that the patient carried the mutations c.337C>T and c.1127 G>T in the PCCB gene. The mutations were inherited from his parents individually.

Conclusion: The patient carried two compound heterozygous mutations in PCCB gene which resulted in propionic academia. The metabolomics screen and brain MRI also played significant roles in the diagnosis of propionic acidemia.

Development of sequencing techniques allowed us to determine genomic sequences in many organisms. Such a determined genome consists of not only protein-coding genes but also noncoding RNA (ncRNA) genes. We should analyze evolutional histories of such genes to estimate evolutional directions in future. Meanwhile, recent studies showed that some ncRNA genes are located in intragenic regions in protein-coding genes, which are called host genes. We considered that such information can help us to discuss gene evolutions. In this study, we constructed a database to analyze evolutions of protein-coding and noncoding genes based on gene locations in genomic sequences. We found that 547 out of 2,691 human host genes are orthologous to 546 out of 1,633 mouse host genes. Such orthologous host genes are involved in similar biological functions but some non-orthologous host genes have different functions. For example, non-orthologous host genes in human are annotated as neuron-related terms but such genes in mouse are not. Meanwhile, similarity searches for intronic microRNA (miRNA) genes between human and mouse showed that 85 out of the orthologous host genes have retained miRNA genes in the intronic regions. 64 out of such genes have retained intronic miRNA genes among human, mouse and rat. These results suggest that some orthologous genes have retained ncRNA genes in the intronic regions in the evolutionary process.

Objective: To present a prospective case study on the increase of telomere length, improvement in nocturnal polyuria, neck and mid-back pain, autonomic nervous system adaptability, and health-related quality of life following correction of the sagittal cervico-thoracic spinal alignment and posture using chiropractic biophysics® (CBP®) technique.

Clinical features: A 35-year-old white female elementary school teacher presented with chronic neck and midback pain for 5 years following a motor vehicle collision as well as nocturnal polyuria. Examination and radiography revealed forward head posture and loss of cervical lordosis consistent with vertebral subluxation. Patient telomere length was derived from nucleated white blood cells obtained from a blood test. Quality of life measures were determined by the Short-Form 36 health survey and heart rate variability was measured.

Intervention and outcome: The patient received CBP® spinal care including Mirror Image® corrective spinal exercises, adjustments, and traction. Full spine and drop table adjustments were administered. After 36 visits, she reported improvement in her nocturnal polyuria, neck and mid-back pain, and quality of life. Cervical x-rays showed correction of cervical lordosis and forward head posture. A blood test showed significant improvement in patient telomere length and heart rate variability improved from a health risk to within normal limits.

Conclusion: Our case suggests that correction of cervical lordosis and forward head postures by CBP® Mirror Image® methods improved the sagittal spinal alignment and posture and was temporally associated with lengthened telomeres, improved nocturnal polyuria, neck and mid-back pain, quality of life, and autonomic nervous system adaptability.

Purpose: Based on aesthetics and long-term stability, call-assisted autologous lipotransfer is a promising method for post-oncological breast reconstruction. Thereby, autologous fat grafts enriched with autologous adiposederived stem cells are transferred to the breast. However, as adipose-derived stem cells (ASC) have the ability to secrete exosomes and growth factors, they could communicate with residual breast cancer cells.

Methods: We performed a systematic review of the published literature to evaluate whether adipose-derived stem cells- based treatments for breast cancer are associated with tumor growth of rest breast cancer cells and therefore that it could exist an oncological risk. An electronic database search was performed.

Results: As described, all current studies can prove that the gene expression is influenced by the direct cell contact between adipose-derived stem cells and breast cancer cells.

Conclusion: These observations suggest that information is substituted via exosomes which can migrate intercellularly and transfer genetic material between both cell types.

RAS-RAF-MEK-MAPK pathway comprises a group of kinases which regulates the activities of effector proteins in growth, proliferation and apoptosis. The extracellular signals from growth factors, cytokines and other stimuli transmitted by surface receptors and upstream signaling molecules are integrated by this cascade of kinases whose activity is regulated by the interaction of oncoproteins and tumor suppressors. The anomaly of the signaling pathway would lead to occurrence of malignancies. The RAS-RAF-MEK-MAPK pathway is therefore targeted by anticancer therapeutic agents. The present paper discussed the interaction of individual component with tumor suppressors and the impact of their inhibitor on the efficacy of anticancer therapy, and improvement on small molecule inhibitor of RAS-RAF-MEK-MAPK pathway with modified targeting has been proposed.

Cyclic vomiting syndrome (CVS) is an episodic disorder that may be associated with migraine. It characterized by recurrent, stereotypic episodes of vomiting and nausea separated by intervals of comparative wellness. We report the first pediatric case of 22q 11 deletion syndrome (22q11DS) and CVS.

Background: Vitamin D receptor (VDR) gene polymorphisms are possibly involved in the development of type 2 diabetes mellitus (T2DM). However, the data to date have been inconclusive. Previous studies have suggested an influence of vitamin D receptor alleles on glucose metabolism and on susceptibility to type 2 diabetes mellitus in different ethnic populations through the action of vitamin D endocrine system which related with calcification and lipolysis, insulin secretion, and may be associated with many complicated disease including diabetes. To investigate the relationship between a single nucleotide polymorphism (SNP) of VDR gene and T2DM more studies had been done. However, different results have been found in different spots of the world. Therefore, more studies are needed to understand the variation in these results. This is the first study that shows the implication of the SNP of VDR gene in T2DM in Iraqi patients.

Objective: To assess the correlation between serum 25(OH)D3 levels, and vitamin D receptor (VDR) polymorphisms (Fok1, BsmI, TaqI and ApaI), and glycemic control in obese T2DM Iraqi population, this study was performed.

Materials and methods: 200 clinically diagnosed T2DM patients, distributed into three subgroups according to therapeutic pattern and 75 healthy controls from the Iraqi population were recruited in this study. The association between the VDR gene SNPs and the T2DM was determined using Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, and genotype and allele frequencies were calculated between the T2DM and control groups.

Results: No significant differences between mean age and the body mass index between both case and control groups were observed. According to current glycemic control consensus, results demonstrated only 20.5 percent of the T2DM patients met this target, which meant that 79.5 percent of the cases were suffering from poor glycemic control 25(OH)D3 levels were significantly lower in the T2DM patients than in the control group, being 17.49 ± 1.12 ng/ml and 31.26 ± 1.25 ng/ml, in the patient and control groups, respectively (p<0.001, Student’s t-test). 25(OH) D3 levels were found to be inversely associated with HbA1c levels in the T2DM patients (p<0.001, r2=0.058). In the group of T2DM patients, 160 of 200 (80 percent) as opposed to 2 of 75 (2.6 percent) in the non-diabetic l group had lower, 25(OH)D3 levels ≤ 20 ng/ml (chi-squared test, p<0.001), while in the group of type 2 patients, 15 of 200 (7.5 percent) as opposed to 5 of 75 (6.6 percent) in the control group, had vitamin D insufficiency, 25(OH)D3 levels > 20 < 30 ng/ml (chi-squared test, p<0.0001). The gene polymorphism analysis for T2DM showed that genotype and alleles frequencies for the VDR genes were in agreement with Hardy–Weinberg equilibrium in all cases.

Conclusion: VDR gene polymorphism analysis revealed that neither genotypes nor alleles of VDR BsmI, ApaI showed a significant variation between T2DM patients and controls. In contrast‚ the FF genotype of VDR FokI and TT genotype of VDR TaqI showed a significant (P<0.0001) increase in T2DM patients in comparison to controls. FF and TT homozygotes had significantly higher baseline fasting glucose and HOMI-IR levels than f allele carriers. In addition, data found significantly elevated interlukin IL-6, TNF-α, IL -1β and decreased osteocalcin in association with the Taq1 polymorphism.

Most molecular pathology laboratories perform mutational analyses to diagnose somatic cancer mutations and evaluate therapeutic options on formalin fixed paraffin embedded (FFPE) samples as daily practice using various methods. Recent studies show that targeted next-generation sequencing (NGS) is a promising diagnostic method with many benefits including simultaneous detection of multiple mutations in various genes in a single test. High quality DNA is essential for an efficient and successful NGS performance. However, low tumor tissue percentage and low DNA quality are the main limitations to use FFPE DNA for NGS assay. We reviewed and discussed the required tissue amount for the NGS assay, the effect of formalin fixation on DNA integrity, and the method for reduction of formalin-induced sequencing artifacts. We also review DNA extraction methods, DNA quality control methods and quality control workflow for nucleic acids and libraries. This review provides an overview for molecular pathology laboratories or researcher considering NGS to detect somatic cancer mutations using small FFPE samples.

This is a report on a 44-year-old woman who was referred to our hospital because of hypertension and a left adrenal mass detected by ultrasonic examination. Computer tomography scan revealed a 17 × 12 mm mass in the left adrenal region. The patient underwent surgery for suspicion of left adrenal pheochromocytoma. Histological examination revealed that the resected mass was splenic tissue. This indicated that surgeons should consider the possibility of accessory spleen when adrenal mass is suspected on imaging examination.

Background: Congenital heart disease has a great impact on a child’s morbidity and mortality as well as on the health systems cost. They represent the main cause of death among children with congenital malformations.

Method: The study was prospective, cross-sectional involving consecutive subjects from two centers which were; a tertiary hospital, a private hospital and a major cardiology center. Children with congenital malformations had transthoracic echocardiography done by a cardiologist. Data was analyzed using Microsoft excel software supplemented with Statistical Package for Social Sciences (SPSS) version 20. Means of continuous variables were compared using the Student t test and proportions using Chi-square test. Level of significance set at p<0.05.

Results: A total of 366 children with obvious congenital malformation were recruited in the study. The age range of the patients was between 1 day to 12 years with a mean age of about 6 + 508.03. Majority of the patients were neonates as shown in Table 1. Male to female ratio was 1.1:1. Up to 26.5% of the subjects had a structurally normal heart. The most common congenital heart defect observed observed was atrial septal defect (14.8%) followed by isolated ventricular septal defect (14.1%). Atrioventricular septal defect 13.7%, Patent ductus arteriosus 7.1%, tetralogy of Fallot 4.5%. Some of the patients had more than one cardiac defects.

Conclusion: The prevalence of congenital heart defects among children with congenital malformation is very high. Routine screening for cardiac defects in any patients with congenital malformation is advocated to improve quality of life, reduce morbidity and mortality in these subjects.