Molecular and Genetic Medicine

Nail-Patella syndrome is pleiotropic autosomal dominant disorder characterized by skeletal abnormalities and frequently renal disease. In this case report genetic analysis revealed a pathogenic heterozygous missense mutation of the LMX1B gene as c.[737G>A], p.([Arg246Gln)]. This variant is predicted to cause substitution of a basic arginine residue for a polar uncharged glutamine residue at position 246. The nucleotide at position 737 and amino acid at position 246 are both highly conserved across species. Although LMX1B mutation causing hereditary focal segmental glomerulosclerosis (FSGS) without any extra-renal involvement has been described as nail patella like renal disease (NPLRD) where there is partial impairment of transcriptional activity with these mutations leading to absence of other features of nail patella syndrome (NPS). Our case report gets interesting as patient had progressed relatively quickly to end stage renal disease. Some of heterozygous gene mutation FSGS cases are expected to recur after kidney transplantation. Our case has received a successful renal transplant from brother which is working well for the last 3 years without any evidence of recurrence. Another interesting feature of our case was the strong family history of unrelated other renal genetic disorder autosomal dominant adult polycystic kidney disease (ADPKD).

Study background: Over the past years, the number of patients diagnosed with glioma has increased. Glioma is the most widespread pediatric malignancy of the brain and a high-grade tumor is associated with a dismal prognosis. Members of the matrix metalloproteases (MMP) enzyme family (especially MMP-2 and MMP-play an important role in the degradation of the extracellular matrix (ECM) a requirement for disease progression, in addition to the miRNAs play a vital role in regulating “cancer hallmarks”.
Aim: Our aim in this study was to detect the expression level of MMP2 and MMP9 in high-grade glioma and low-grade glioma to find the differential expression in two different subtypes and to find the correlation of MMP and progression of disease and to identify any relationship between miRNA and MMP expression in gliomas.
Methods and finding: We determined the expression of all the miRNAs and mRNAs for the (MMP2 and MMP9) in over 20 high grade glioma (HGG) and 20 low grade glioma (LGG) in paraffin-embedded (FFPE) using Quantitative polymerase chain reaction (qPCR).
Result: Seven miRNAs showed high expression specifically in HGG compared to LGG with significant differential expression (p-value ≤ 0.05).
Conclusion: The mRNA for the MMP-2 and MMP-9 were highly expressed in HGG than in LGG, although the difference did not reach the statistical significance. The main limitations of this study are the small sample size of the patients and further work is needed to investigate the relativeness of our work to the clinical side.

Introduction: This is a case presentation of inherited retina dystrophy type RPE65. We enhanced signs for early diagnosis in order to prevent vision loss as this condition has treatment now-a-days.
Method opted: Phenotype and genotype investigation resulted in RPE65 mutation, one sub-retinal injection was done in each eye.
Results: Vision is improved.
Discussion and conclusion: Early diagnosis is needed for LCA and retina dystrophies. This case presentation illustrated diagnosis is delayed due to the unknown early signs. Early signs are not detected by a specialist, even though for a mother is certain that vision is not normal for first month of life. There is treatment now and this may prevent loss of vision due to the degenerative nature of this condition, particularly as during first year of life treated patients manifest improved vision.

Objectives: This paper investigated the pharmacokinetic (PK) change, optimal dose and therapeutic monitoring method of vancomycin in severe burns.
Method: This is a retrospective, matched cohort study. Nine burn patients were included, and each individual was matched with a control critically ill patient. Vancomycin concentrations were analyzed using the first order pharmacokinetic principles to calculate elimination rate constant (K), half-life (t1/2), volume distribution (Vd), total body clearance (CLvancomycin) and the 24-hour area under the curve (AUC24 h). Population PK analysis and Monte Carlo simulation was used to investigate optimal dose.
Results: The mean burn area of the burn group and age was 60% and 20 years-old respectively. The loading dose and daily maintenance dose was significantly higher in burn group than that in control group. Vancomycin clearance was significantly higher (p<0.05) in burns patients when compared to control patients. CLCr (creatinine clearance) was not significantly correlated to CLvancomycin (vancomycin clearance) in both groups. Ctrough (serum trough concentration) was significantly correlated with AUC24h in the control group (r=0.98, R2=0.96, p<0.01), but not in the burn group (r=0.63, R2=0.40, P=0.07). Daily dose of 5000 mg of vancomycin could achieve 90% probability of target attainment if target was AUC24h /MIC (minimum inhibitory concentration)>400.
Conclusion: Two steady state concentrations but not Ctrough is an appropriate reference for vancomycin therapeutic monitoring. A daily dose of at least 5000 mg is suggested in severe burn patients with normal renal function (CLCr>90 mL/min).

Background: Duchenne Muscular Dystrophy (DMD) is a progressive neuromuscular disorder characterized by a relentless clinical course with diagnosis usually established around three to four years of age. DMD is caused by mutations in the dystrophin gene, where deletions and duplications of one or more exons represent the bulk of related genetic aberrations.
Aims and Methods: Our aim in the current study is to analyze the frequency and the distribution pattern of deletions/duplications associated with dystrophin gene exons and assess the mean diagnostic age of DMD in a small Lebanese group of dystrophic patients suspected with DMD/BMD based on observed clinical features.
Results and Discussion: Among 52 samples analyzed, we identified 33 cases (63%) with deletions and two cases (4%) with duplications. Deletions were of variable sizes, ranging from 1 to 47 exons and occurred mostly (78%) in two deletion hotspots (HS), HS1 (18%) and HS2 (60%), covering exons 6-19 and 45-52 respectively. Single exon deletions were even further restricted (90%) to the deletion hotspots, mainly to HS2 (80%). The average age of DMD molecular diagnosis in our subject study was 7 years of age.
Conclusion: Molecular analyses were consistent with those obtained in previous studies, with however an average age of DMD diagnosis significantly later than what is usually reported. Our study illustrates the need to implement early molecular diagnosis in order to institute optimal care, including available targeted treatments, for our patients.

Apparently balanced reciprocal translocations are a common type of chromosome rearrangements with an estimate incidence range from about 1 in 500 to 1 in 625 human newborns. Rearrangements were found both in clinically unaffected individuals and patients with phenotypic abnormalities. Most are inherited, but approximately one in five are de novo events and introduce a risk of abnormal phenotype in 6.1% of prenatal genetic counseling.

Alterations in the MET proto-oncogene, such as amplification or mutations causing exon 14 skipping, promote tumor growth, cellular transformation and invasion and are associated with poor prognosis in many types of cancers. There are several c-Met-targeted agents currently in clinical trials, highlighting the importance on developing accurate, efficient and sensitive technologies for the analysis. This short communication will provide an update on three of the most useful technologies for the detection of c-Met exon 14 skipping abnormalities, namely New Generation Sequencing (NGS), nCounter and qPCR.

Cytokines are a class of pro-inflammatory immune responses in the peripheral and central nervous system. Elevated cytokine levels contribute to appetite and weight dysregulation, anxiety, depression and other psychiatric conditions, and may underlie eating disorder (ED). Recently, two meta-analyses of cytokine levels in people with EDs – particularly anorexia nervosa (AN) – confirm elevated levels of cytokines within the 5-LOX inflammatory pathway, namely interleukin 1 (IL-1), interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF-α). IL-1, IL-6 and TNF-α are leukotrienes that stimulate the prolonged response of nuclear factor kappa beta (NF-κβ) – the major
inflammatory gateway molecule – which influences brain development and function within the hypothalamicpituitary- adrenal (HPA) axis, hippocampus and prefrontal cortex. The structure and function of these brain areas are shown to be aberrant in neuroimaging studies of EDs; thus, neuroinflammatory processes are significant biomarkers for weight and cognitive disturbances in EDs, particularly AN. Against this background, this brief article summarises the current knowledge of IL-1, IL-6 and TNF-α in EDs. Thereafter, the significance of inhibiting the NF-κβ 5-LOX inflammatory pathway with a low-risk, Cochrane-reviewed, anti-inflammatory known as Boswellia serrata is considered. Brief discussion of the clinical role for Boswellia serrata in weight recovery and reduction of comorbid mental disorder in ED is provided to stimulate further research into natural anti-inflammatory treatment interventions.

The advancements in understanding of biological mechanisms in RCCs have led to the development and application of targeted therapies. The FDA approved targeted therapies are mainly directed against angiogenesis, mTOR pathways, and immune checkpoint inhibition. VEGF overexpression due to VHL gene mutation or alteration in majority of RCCs makes it one of the most suitable targets for development of therapy. Inhibitors and antibodies against VEGF or VEGFR have shown promising results in metastatic RCCs. mTOR pathway inhibition has also proven to be an efficacious strategy as well as antibodies targeting the immune checkpoints allowing T-Cells to attack tumour cells. However, there is need for improvement as certain proportion of patients may either not respond or develop resistance against targeted therapies.

The advent of low-cost sequencing and genotyping technologies has made DNA data widely accessible to individuals. Ancestry DNA tests (from companies like 23andMe, Ancestry DNA, Family Tree DNA etc.) continue to be one of the most popular genetic tests in this space. Over 15 million genetic tests have been sold so far by ancestry genetic testing companies. These companies also empower the end consumer by sharing their raw DNA data with them. While predicting ancestry and haplogroups (Mt and Y) is just one component of doing the test, the more actionable applications of having raw DNA data lie in the area of health, wellness, nutrition, fitness, precision medicine and more.

Dorcas gazelles are critically endangered mammals on the Arabian World. Past conservation efforts have been plagued by confusion about the phylogenetic relationship among various ‘phenotypically discernable’ populations, and even the question of species boundaries was far from being certain. This lack of knowledge has had a direct impact on conservation measures. Here, we provide a phylogenetic framework, based on the analysis of mtDNA sequences. We applied molecular methods to document for the first-time patterns of genetic diversity and population structure of Gazella dorcas in Bahry and East of the Nile (Sudan) using 421 bp fragment of mitochondrial DNA in seventeen populations.

Background: Wolfram syndrome is a neurodegenerative disorder characterized by the acronym DIDMOAD (Diabetes Insipidus (DI), Diabetes Mellitus (DM), Optic Atrophy (OA) and Deafness). Homozygous/compound heterozygous mutations in WFS1 gene causes autosomal recessive form of Wolfram syndrome (AR-WS) whereas heterozygous mutations are associated with autosomal dominant-low-frequency non-syndromic hearing loss (ADLFNSHL). Clinical symptoms and degree of severity is reported to be heterogeneous in WS patients.
Aim: To characterize clinical features and molecular gene mutations in patients with WS in India and compare them with data from other countries.
Patients and Methodology: Eleven patients from 6 families were enrolled. In nine patients from 4 families with phenotypic features of diabetes mellitus, optical atrophy and hearing loss WFS1 gene was sequenced. Two patients of the other 2 families presented with hearing loss only and were analysed for targeted deafness genes panel by next generation sequencing.
Results: Nine patients from 4 families had biallelic mutations in WFS1 gene. Two patients harboured heterozygous mutation in WFS1 gene. Seven different mutations WFS1 were identified, of which 5 mutations were novel. All the identified mutations were present in exon 8 of WFS1 gene.
Conclusion: Pathogenic variations in WFS1 gene can cause both AR-WS and AD-LFNSHL. We recommend a protocol in which patients with WS should be first sequenced for the hotspot exon 8. If no mutation is identified, then the full gene should be sequenced. Further, for patients with hearing loss with/without diabetes and/or optical atrophy, WS should be considered as one of the differential diagnosis.

Mutations in the WDR45 gene have recently been identified as causative for the only X-linked beta-propeller protein-associated neurodegeneration (BPAN), a subtype of neurodegeneration with brain iron accumulation (NBIA) with phenotypically and genetically heterogeneous condition. The clinical features include early-onset global developmental delay, progressive accumulation of iron in the basal ganglia, resulting in physical and neurological deterioration. We herein reported a novel mutation (c.1040-1041del) in exon 12 of WDR45 gene in a 3-year-old Chinese girl, exhibiting developmental delay, seizures, by whole-exome sequencing. Sanger sequencing confirmed the heterozygous mutation was absent in both her parents, and thus it was concluded as a de novo frameshift mutation.

Healthcare data continues to explode in size and complexity and collection and harmonization of disparate clinical, molecular, patient research and biospecimen data is key to effective research, but presents challenges in terms of access, organization, security and compliance. When designing an experimental clinical study, it is important to plan how genetic and molecular data will be collected and recorded in the course of the study. With the recent advent of internet of things (IoT) the direct recording of data could be widened to mobile devices such as wearable sensors which could capture biomedical information directly from patients’ body, and to other type of remote devices internet connected such as digital scales for body fat and weight monitoring. Despite the diffusion of the electronic data capture, paper collection data sheets are still widely employed. Electronic data capture (EDC) employs different technologies and methods for data collection. For these reasons EDC (electronic data capture) and eCRF (case report form) are becoming more widespread. Their production is complex and needs to comply with strict regulations both in Europe and in the United States, which comprise system validation, security policy and data backup.

Endometriosis is a common estrogen-dependent inflammatory disease and a major contributor to pelvic pain and subfertility in women of reproductive age. Its etiology and pathogenesis are not well known yet, but different studies had highlighted its multifactorial nature, influenced by both genetic and environmental components. This minireview analyzes the prevalence of the disease and the concordance for the presence and stage of endometriosis in monozygotic (MZ) and dizygotic (DZ) twins. A higher concordance for the presence of the disease in MZ twins rather than DZ twins was observed, with a statistically significant difference (p-value < 0.05). Only one study had evaluated the concordance for endometriosis stage within MZ twins, observing a high rate of concordance. Genetic component seems to play a pivotal role in the development of endometriosis and its severity.

Analysis of whole genome sequencing data using a human reference genome such as GRCh38 may miss certain classes of population-specific variations. The advancement of long-read sequencing techniques has enabled efficient and effective construction of ethnically relevant reference genomes across populations. In this mini-review, we discuss recent endeavours to build ethnicity-specific reference genomes and summarize findings from these studies.

Bloom syndrome is a rare autosomal recessive disorder that attributes a chromosomal instability leading to a high risk of cancer at an early age. We report a case of Bloom syndrome in a 30-year-old man with colonic cancer due to polyposis degeneration. The diagnosis of BS was suspected at the age of five years behind growth retardation, facial sun-sensitive telangiectatic erythema and repeated respiratory infections. The patient underwent a total colectomy. The patient recovered without complication and is currently being followed with endoscopic surveillance of the rectal stump. We present a case report about CRC diagnosed in a Bloom syndrome patient at an early age to suggest the necessity of CRC screening for these patients. Then, we recommend the necessity of doing at least annual examinations and colonoscopy as well as a rapid and thorough investigation of any new symptoms.

In April 2015, Northern corn leaf blight (NCLB) symptoms were observed in corn plants (Zea mays L.) in cold highland areas at Titi Gantong, Lembah Bertam farms, Malaysia. The typical symptom of NCLB disease showed abundant lesions on the leaves of susceptible cultivars (5-20 cm long), greyish to tan in colour and elliptical in shape. Exserohilum turcicum (Pass.) was isolated from diseased leaf and cultured on PDA media for further morphological investigation. The colonies of E. turcicum on PDA are dark grey in colour and grow faster. Conidial shapes were straight to slightly curved, the hilum protrudes as a truncate at the base of the conidia. Conidial colour was pale to olivaceous brown, while the conidial size ranged from 56 to 89 μm in length, 9 to 13 μm in width, and the number of septa ranged from 6-10. Conidia were 4-9 distosptate, with a thicker wall at the base of each conidium. Based on description reported by Sivanesan (1987) the isolate was identified as E. turcicum. To verify the morphological findings, PCR amplification was conducted using universal primers, TUBUF2 forward (5' CGGTAACAACTGGGCCAAGG-3') and TUBUR1 reverse (5'-CCTGGTACTGCTGGTACTCAG-3') and yielded a 1000-bp product. Blast result showed that the sequence was having 97% similarity to published sequence of Setosphaeria turcica, teleomorph of E. turcicum (Accession No. XM_008032318). To our belief and knowledge, this is the first report of E. turcicum as the causative agent of NCLB disease of corn in Malaysia. There is need for future studies to focus on assessing management options for this disease to avoid corn yield losses in Malaysia.

Over 3.7 billion people worldwide under the age of 50 are estimated to have some form of Herpes simplex virus (HSV) infection in their lifetime. Despite mostly presenting as asymptomatic, HSV infection has been associated with worsening symptoms and biological conditions in various diseases, some of which are neurological, such as Alzheimer’s disease (AD). This mini-review thus aims to showcase the varying researches carried out in the past decades which sought the connection between the infection and the neurodegenerative disease. Various members of the Herpes viridae family have been associated with worsening AD pathology and symptoms, namely HSV-1, HSV-2, human herpes virus 6 (HHV-6), and HHV-7. The virus affects many aspects of the nervous system to promote neurodegeneration, including fragmentation of amyloid precursor proteins (APP), inducing production of excess amyloid beta (Aβ) in response to the viral infection, and formation of tau neurofibrillary tangles (NFT), all of which point to further degeneration of the nervous system structures and cognitive decline in AD patients. HSV has been significantly reported to induce concurrent alterations in the context of AD. Although more in-depth analysis still needs to be done to consider other factors such as age, viral load, and the degree of neurodegeneration, HSV infection remains an interesting drug target through the means of antivirals in ameliorating a certain degree of neurodegeneration, cognitive decline, and memory loss in AD.

The trend of personalized medicine will require more genetic studies to be done. Nurses are perfectly suited to conduct these types of studies. Designing these types of studies requires using a theoretical framework. Differential Susceptibility to Environmental Influence is a framework that describes a step-wise approach to designing these types of studies. Test of the independence of the susceptibility factor and the predictor, test of the association between the susceptibility factor and the outcome, comparison of the regression plot, test of the specificity of the model by replacing susceptibility factors and outcomes are the steps required to evaluate differential susceptibility.