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Current approaches in CRISPR-Cas9 mediated gene editing for biomedical and therapeutic applications

Bhattacharjee G, Gohil N, Khambhati K, Mani I, Maurya R, Karapurkar JK, et al.

J Control Release, 2022 Feb 08.
PMID: 35149141 DOI: 10.1016/j.jconrel.2022.02.005

A single gene mutation can cause a number of human diseases that affect quality of life. Until the development of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) systems, it was challenging to correct a gene mutation to avoid disease by reverting phenotypes. The advent of CRISPR technology has changed the field of gene editing, given its simplicity and intrinsic programmability, surpassing the limitations of both zinc-finger nuclease and transcription activator-like effector nuclease and becoming the method of choice for therapeutic gene editing by overcoming the bottlenecks of conventional gene-editing techniques. Currently, there is no commercially available medicinal cure to correct a gene mutation that corrects and reverses the abnormality of a gene's function. Devising reprogramming strategies for faithful recapitulation of normal phenotypes is a crucial aspect for directing the reprogrammed cells toward clinical trials. The CRISPR-Cas9 system has been promising as a tool for correcting gene mutations in maladies including blood disorders and muscular degeneration as well as neurological, cardiovascular, renal, genetic, stem cell, and optical diseases. In this review, we highlight recent developments and utilization of the CRISPR-Cas9 system in correcting or generating gene mutations to create model organisms to develop deeper insights into diseases, rescue normal gene functionality, and curb the progression of a disease.
Fulltext Formulation Development, Characterization and Antifungal Evaluation of Chitosan NPs for Topical Delivery of Voriconazole In Vitro and Ex Vivo

Shah MKA, Azad AK, Nawaz A, Ullah S, Latif MS, Rahman H, et al.

Polymers (Basel), 2021 Dec 30;14(1).
PMID: 35012154 DOI: 10.3390/polym14010135

This study aims to develop chitosan-based voriconazole nanoparticles (NPs) using spray-drying technique. The effect of surfactants and polymers on the physicochemical properties, in vitro release, and permeation of NPs was investigated. The prepared NPs containing various surfactants and polymers (e.g., Tween 20 (T20), Tween 80 (T80), sodium lauryl sulfate (SLS), propylene glycol (PG), and Polyethylene glycol-4000 (PEG-4000)) were physiochemically evaluated for size, zeta potential, drug content, percent entrapment efficiency, in vitro release, and permeation across rats' skin. A Franz diffusion cell was used for evaluating the in vitro release and permeation profile. The voriconazole-loaded NPs were investigated for antifungal activity against Candida albicans (C. albicans). The prepared NPs were in the nano range (i.e., 160-500 nm) and positively charged. Images taken by a scanning electron microscope showed that all prepared NPs were spherical and smooth. The drug content of NPs ranged from 75% to 90%. Nanoparticle formulations exhibited a good in vitro release profile and transport voriconazole across the rat's skin in a slow control release manner. The NPs containing SLS, T80, and PG exhibited the best penetration and skin retention profile. In addition, the formulation exhibited a potential antifungal effect against C. albicans. It was concluded that the development of chitosan NPs has a great potential for the topical delivery of voriconazole against fungal infection.
The emergence of an imported variant of dengue virus serotype 2 in the Jazan region, southwestern Saudi Arabia

Dafalla O, Abdulhaq AA, Almutairi H, Noureldin E, Ghzwani J, Mashi O, et al.

Trop Dis Travel Med Vaccines, 2023 Mar 15;9(1):5.
PMID: 36922890 DOI: 10.1186/s40794-023-00188-8

BACKGROUND: Dengue virus (DENV) infection is a global economic and public health concern, particularly in tropical and subtropical countries where it is endemic. Saudi Arabia has seen an increase in DENV infections, especially in the western and southwestern regions. This study aims to investigate the genetic variants of DENV-2 that were circulating during a serious outbreak in Jazan region in 2019.
METHODS: A total of 482 serum samples collected during 2019 from Jazan region were tested with reverse transcription-polymerase chain reaction (RT-PCR) to detect and classify DENV; positive samples underwent sequencing and bioinformatics analyses.
RESULTS: Out of 294 positive samples, type-specific RT-PCR identified 58.8% as DENV-2 but could not identify 41.2%. Based on sequencing and bioinformatics analyses, the samples tested PCR positive in the first round but PCR negative in the second round were found to be imported genetic variant of DENV-2. The identified DENV-2 imported variant showed similarities to DENV-2 sequences reported in Malaysia, Singapore, Korea and China. The results revealed the imported genetic variant of DENV-2 was circulating in Jazan region that was highly prevalent and it was likely a major factor in this outbreak.
CONCLUSIONS: The emergence of imported DENV variants is a serious challenge for the dengue fever surveillance and control programmes in endemic areas. Therefore, further investigations and continuous surveillance of existing and new viral strains in the region are warranted.
Fulltext Exploring extracellular vesicles in zoonotic helminth biology: implications for diagnosis, therapeutic and delivery

Qadeer A, Wajid A, Rafey HA, Nawaz S, Khan S, Rahman SU, et al.

Front Cell Infect Microbiol, 2024;14:1424838.
PMID: 39165921 DOI: 10.3389/fcimb.2024.1424838

Extracellular vesicles (EVs) have emerged as key intercellular communication and pathogenesis mediators. Parasitic organisms' helminths, cause widespread infections with significant health impacts worldwide. Recent research has shed light on the role of EVs in the lifecycle, immune evasion, and disease progression of these parasitic organisms. These tiny membrane-bound organelles including microvesicles and exosomes, facilitate the transfer of proteins, lipids, mRNAs, and microRNAs between cells. EVs have been isolated from various bodily fluids, offering a potential diagnostic and therapeutic avenue for combating infectious agents. According to recent research, EVs from helminths hold great promise in the diagnosis of parasitic infections due to their specificity, early detection capabilities, accessibility, and the potential for staging and monitoring infections, promote intercellular communication, and are a viable therapeutic tool for the treatment of infectious agents. Exploring host-parasite interactions has identified promising new targets for diagnostic, therapy, and vaccine development against helminths. This literature review delves into EVS's origin, nature, biogenesis, and composition in these parasitic organisms. It also highlights the proteins and miRNAs involved in EV release, providing a comprehensive summary of the latest findings on the significance of EVs in the biology of helminths, promising targets for therapeutic and diagnostic biomarkers.
Fulltext A cross-sectional assessment of knowledge, attitudes and self-perceived effectiveness of complementary and alternative medicine among pharmacy and non-pharmacy university students

Ashraf M, Saeed H, Saleem Z, Rathore HA, Rasool F, Tahir E, et al.

BMC Complement Altern Med, 2019 May 03;19(1):95.
PMID: 31053114 DOI: 10.1186/s12906-019-2503-y

BACKGROUND: Traditional medicine has always been Pakistan's cultural heritage, providing health care to a large part of its population. Thus, we aimed to assess and compare the knowledge, attitude, and perception about complementary and alternative medicine (CAM) between pharmacy and non-pharmacy students, the results of which may be helpful in devising national health-education policy.
METHODS: A cross-sectional study was conducted by enrolling 937 students, pharmacy (437) and non-pharmacy (500), of Punjab University, Lahore. A self-administered questionnaire was used for data collection. Data were analyzed using SPSS. (IBM v22).
RESULTS: Data suggested that majority of students knew about the use of traditional herbs and considered massage (P: 84.4%, NP: 82%, p = 0.099), homeopathy, herbs (P: 86.5%, NP: 81%, p = 0.064], yoga [P: 357 (81.7%), NP: 84%), p = 0.42] and spiritual healing (P: 85.6%, NP: 86.2%, p = 0.55) as effective and least harmful CAM modalities. The pharmacy students had better knowledge about CAM modalities compared to non-pharmacy students. Despite utilizing non-reliable sources of CAM information and their belief that CAM is practiced by quacks, the majority of students had positive attitudes and perceptions about CAM usage. Students also believed that CAM had a positive impact on health outcomes [P: 3.19 ± 1.04, NP: 3.02 ± 1.09, p = 0.008] and acceded to include CAM in the pharmacy curriculum. However, non-pharmacy students scored higher in their beliefs that CAM usage should be discouraged due to the non-scientific basis of CAM (P: 3.04 ± 0.97, NP: 3.17 ± 1.02, p = 0.028) and a possible threat to public health (P: 3.81 ± 1.74, NP: 4.06 ± 1.56, p = 0.02). On the other hand, pharmacy students believed that patients might get benefits from CAM modalities (P: 4.31 ± 1.48, NP: 4.12 ± 1.45, p = 0.02). Majority of students perceived that spiritual healing is the most useful and safer CAM modality, while acupuncture (P: 25.4%, NP: 21.8%, p = 0.0005), hypnosis (P: 26.8%, NP: 19.6%, p = 0.001) and chiropractic (P: 18.8%, NP: 11.6%, p = 0.0005) were among the harmful ones.
CONCLUSION: In conclusion, despite poor knowledge about CAM, students demonstrated positive attitudes and beliefs regarding CAM. They exhibited better awareness about yoga, spiritual healing/prayer, herbs, and massage. Students also showed willingness to advance their knowledge about CAM and favored its inclusion in the curriculum.