Oligospermines For Non-Viral Sirna Delivery

In this thesis, delivery systems for siRNA delivery are introduced with special attention to non-viral vectors. Many successful vectors used in vivo were reviewed. Our work focused on the effect of different architectures of oligospermine polmers on their suitability for siRNA delivery in lung cancer cells. Different archituctures showed different polyplex structures and variable transfection efficiencies. Moreover, we presented a review on nanoimprint lithography techniques with an outlook on possible biological applications in the field of gene and drug delivery

Mastitogenic Bacteria Isolated from Clinical Mastitis Cases Associated with Teat Lesions and their Antimicrobial Sensitivity

Mastitis is the most common and most expensive disease that impacts dairy farms in Egypt. In this study, we investigated bacterial mastitis cases that did not respond to antimicrobial treatment from buffaloes and cows. Milk samples (11) were collected from diseased animals (5 buffaloes and 6 cows). The samples were tested microbiologically to isolate and identify the causative bacteria and antimicrobial susceptibility. The antimicrobial sensitivity of the isolated bacteria was tested by using minimum inhibitory concentration technique according to the clinical national laboratory standards.  The total number of bacterial isolates from the cases was 15 isolates. Staphylococcus aureus (S. aureus) was isolated alone in two cases (13.33% of the isolates), and S. aureus isolated in combination with Arcanobacterium pyogenes (A. pyogenes) in one case (6.67%), and S. aureus isolated in combination with both A. pyogenes and Streptococcus agalactiae (S. agalactiae) in two cases (13.33%), and Coagulase negative staphylococci (CNS) isolated alone in one case (6.67%), and CNS isolated in combination with Klebsiella pneumoniae in one case (6.67%), and A. pyogenes isolated alone in two cases (13.33%). In two cases were not able to isolate any bacterial pathogen. In terms of susceptibility to antibiotics, the isolated strains were sensitive to clindamycin with MICs ranging from 0.125 to <0.03125 µM. The same isolates exhibited medium sensitivity to gentamicin and ciprofloxacin with MICs ranging from 0.5 to 8 µM. Most S. aureus isolates were resistant to oxytetracycline with a MIC of 128 µM. Most of the isolated bacteria were resistant to polymyxin B with a MIC > 128 µM. With the exception of the A. pyogenes isolates, all the isolated bacteria were sensitive to enrofloxacin with a MIC <1 µM.       &nbsp

Non-manifesting AHI1 truncations indicate localized loss-of-function tolerance in a severe Mendelian disease gene

Determination of variant pathogenicity represents a major challenge in the era of high-throughput sequencing. Erroneous categorization may result if variants affect genes that are in fact dispensable. We demonstrate that this also applies to rare, apparently unambiguous truncating mutations of an established disease gene. By whole-exome sequencing (WES) in a consanguineous family with congenital non-syndromic deafness, we unexpectedly identified a homozygous nonsense variant, p.Arg1066*, in AHI1, a gene associated with Joubert syndrome (JBTS), a severe recessive ciliopathy. None of four homozygotes expressed any signs of JBTS, and one of them had normal hearing, which also ruled out p.Arg1066* as the cause of deafness. Homozygosity mapping and WES in the only other reported JBTS family with a homozygous C-terminal truncation (p.Trp1088Leufs*16) confirmed AHI1 as disease gene, but based on a more N-terminal missense mutation impairing WD40-repeat formation. Morpholinos against N-terminal zebrafish Ahi1, orthologous to where human mutations cluster, produced a ciliopathy, but targeting near human p.Arg1066 and p.Trp1088 did not. Most AHI1 mutations in JBTS patients result in truncated protein lacking WD40-repeats and the SH3 domain; disease was hitherto attributed to loss of these protein interaction modules. Our findings indicate that normal development does not require the C-terminal SH3 domain. This has far-reaching implications, considering that variants like p.Glu984* identified by preconception screening (‘Kingsmore panel') do not necessarily indicate JBTS carriership. Genomes of individuals with consanguineous background are enriched for homozygous variants that may unmask dispensable regions of disease genes and unrecognized false positives in diagnostic large-scale sequencing and preconception carrier screenin

Yield Responses to Total Water Input from Irrigation and Rainfall in Six Wheat Cultivars Under Different Climatic Zones in Egypt

© 2024 The Author(s). Licensee MDPI, Basel, Switzerland. This is an open access article distributed under the Creative Commons Attribution License (CC BY), https://creativecommons.org/licenses/by/4.0/In Egypt, wheat is the most consumed cereal grain, and its availability and affordability are important for social stability. Irrigation plays a vital role in wheat cultivation, despite intense competition for water resources from the River Nile across various societal sectors. To explore how grain and above-ground biomass yields respond to total seasonal water input from sowing to maturity in six bread wheat cultivars, eight field irrigation experiments were performed at four locations representative of three agro-climatic zones in two consecutive cropping seasons. A three-replicate strip-plot design was used with cultivars nested within the main plots featuring five irrigation treatments, ranging from six to two applications. Overall, irrigation treatment significantly affected nine agronomic traits. Compared with the six irrigation ap-plications treatment (T1), the two irrigation applications treatment (T5) decreased the times to heading and maturity by 6.6 (7.3%) and 8.6 (6.3%) days, respectively. Similar-ly, T5 reduced the plant height by 14.9 cm (14.3%), flag leaf area by 12.0 cm2 (27.2%), number of spikes per square metre by 77.7 (20.1%), number of kernels per spike by 13.9 (25.2%) and thousand grain weight by 10.0 g (19.6%). T5 also decreased the overall mean grain yield and above-ground biomass yield by 2834.9 (32.0%) and 7910.4 (32.86%) kg/ha, respectively. The grain yield and above-ground biomass production were consistently greater for all six cultivars at Al Mataenah and Sids than at Nubaria and Ismailia in the two cropping seasons. All six cultivars showed significantly greater responses to total seasonal water input for the grain yield and above-ground biomass at Al Mataenah and Ismailia. These results emphasise the necessity for choosing regions with favourable soil and climatic conditions to grow wheat cultivars that respond better to irrigation to enhance the large-scale production of wheat in Egypt. The grain and above-ground biomass yields were mostly linearly and positively associated with the total seasonal water input for all six cultivars at all four locations. This sug-gests that maintaining the current irrigation schedule of six irrigations is valid and should be practised to maximise productivity, particularly in areas similar to the three representative agro-climatic zones in Egypt.Peer reviewe

APPLIED STUDY OF MICROBIOLOGICAL HAZARDS IN RAW MILK SOFT WHITE CHEESE IN EGYPT

Raw milk Domiati cheese is the main soft white pickled cheese produced in Egypt. The objective of this study was to assess the microbiological hazards, physiochemical quality and safety of some raw Domiati cheese subtypes including Double Cream, Tallaga, Baramely and Istanbully. The cheese samples were randomly collected from different cheese retailers in Cairo. The results indicated that the protein content and fat content were high in Double cream cheese. The pH value was low in Baramely cheese. Total solid, ash, and EC were high in Istanbully cheese because it is high salt level. Also, results indicated high microbiological hazards. the cheese samples were highly contaminated, having microbial load exceeding the acceptable limits such as: Total bacterial count, Total coliform, Fecal E. coli and yeast & molds. Pathogenic bacteria (Staphylococcus aureus, E. coli O157:H7, Salmonella sp., Shigella sp., Listeria monocytogenes, Bacillus cereus and Campylobacter jejuni) were detected in three cheese samples.  Except Istanbully cheese sample were found free of most pathogenic bacteria.  A high microbial load of the cheese samples present a public health hazard to the consumers and emphasize the need for improving hygienic standard. Microbiological hazards must be limited to the stander limit of food safety. Raw milk soft white cheese must be manufactured according to hygienic standard

Stability Indicating Spectrophotometric Methods Determination of Nicardipine in the Presence of its Alkaline Induced Degradation Products

Objective: Derivative, ratio spectra derivative and ratio difference spectrophotometric methods were developed and validated for simultaneous determination of Nicardipine (NIC) in the presence of its alkaline induced degradation products. Methods: In these methods the overlapped spectra of NIC and its alkaline induced degradation products were well resolved by measuring the amplitudes of first derivative (D1) spectra and the second derivative (D2) at 382.3 and 239 nm, respectively. NIC was determined by ratio spectra derivative by measuring the amplitude at 244 nm. The ratio difference spectrophotometric method was developed in which the difference between amplitudes at 237.5 nm and 260 nm of the ratio spectra were recorded. The linearity range for the applied methods was 2-18 μg/ml.Results: All the developed methods were validated according to ICH Guidelines, NIC was determined with acceptable accuracy and precision.Conclusion: These methods were suitable as stability indicating methods for the determination of NIC in the presence of its alkaline induced degradation products either in bulk powder or in a pharmaceutical formulation. Statistical analysis of the results with those obtained by applying a reported method has been carried out revealing high accuracy and good precision.Keywords: Nicardipine, Spectrophotometry, Pharmaceutical preparations, Stability indicating, derivative, ratio derivative and ratio differenc

Enhanced therapeutic efficacy of SERCA2a gene-modified adipose-derived mesenchymal stem cell exosomes in doxorubicin-induced cardiomyopathy in male albino rats

Background: Worldwide, cancer is still the primary cause of death, and one of the most widely used anthracyclines for treating cancer is doxorubicin (DOX). But a major worry is DOX-induced cardiomyopathy, which is primarily resulted from an excess of reactive oxygen species. Heart sarcoplasmic reticulum calcium ion ATPase2a (SERCA2a) controls the amount of calcium ions stored in the sarcoplasmic reticulum (SR). This study aims to evaluate and compare the efficacy of SERCA2a gene modified adipose mesenchymal stem cell-derived exosomes (AMSCs-dE) to nontransfected AMSCs-dE, in DOX induced cardiomyopathy in adult male albino rat. Materials and methods: Thirty one adult male albino rats were randomly divided into control group and DOX group that further subdivided into three DOX, AMSCs-dE and SERCA2a AMSCs-dE subgroups. AMSCs-dE were administered intravenously (IV). The levels of serum creatine kinase MB (CK-MB) were assessed after DOX injection and before sacrifice. Cardiac muscle samples were taken for histological analysis using Masson’s trichrome and hematoxylin and eosin stains two months after the experiment. Proliferating cell nuclear antigen (PCNA) and connexin 43 were stained using immunohistochemistry. The expression of TNF and SERCA2a genes and proteins was measured by real-time polymerase chain reaction (PCR) and Western blot (Wb) analysis, respectively. Fluorescent microscopy demonstrated non-transfected and transfected exosomes labeled with PKH26 and GFP, respectively, in culture and cardiac muscle. Results: DOX induced myocarditis progressing to degenerative and fibrotic changes in cardiac muscle that regressed in response to AMSCs-dE therapy. However, SERCA2a gene modified AMSCs-dE treatment reversed the mentioned parameters to nearly its normal level. Conclusions: These findings suggest that SERCA2a gene modification enhances the therapeutic efficacy of AMSCs-dE in treating DOX-induced cardiomyopathy

Identification of Differentially Expressed Genes in Human Colorectal Cancer Using RNASeq Data Validated on the Molecular Level with Real-Time PCR

Colorectal cancer (CRC) is a prevalent cancer with high morbidity and mortality rates worldwide. Late diagnosis is a significant contributor to low survival rates in a minority of cases. The study aimed to perform a robust pipeline using integrated bioinformatics tools that will enable us to identify potential diagnostic and prognostic biomarkers for early detection of CRC by exploring differentially expressed genes (DEGs). In addition to, testing the capability of replacing chemotherapy with plant extract in CRC treatment by validating it using real-time PCR. RNA-seq data from cancerous and adjacent normal tissues were pre-processed and analyzed using various tools such as FastQC, Kallisto, DESeq@ R package, g:Profiler, GNEMANIA-CytoScape and CytoHubba, resulting in the identification of 1641 DEGs enriched in various signaling routes. MMP7, TCF21, and VEGFD were found to be promising diagnostic biomarkers for CRC. An in vitro experiment was conducted to examine the potential anticancer properties of 5-fluorouracile, Withania somnifera extract, and their combination. The extract was found to exhibit a positive trend in gene expression and potential therapeutic value by targeting the three genes; however, further trials are required to regulate the methylation promoter. Molecular docking tests supported the findings by revealing a stable ligand-receptor complex. In conclusion, the study’s analysis workflow is precise and robust in identifying DEGs in CRC that may serve as biomarkers for diagnosis and treatment. Additionally, the identified DEGs can be used in future research with larger sample sizes to analyze CRC survival