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Biomedical subjects

Muhammad Ashfaq

Publications and source records attributed to Muhammad Ashfaq.

9 recordsLinked to original sources

Transforming Plant Viruses into Vectors for Next-Generation Agriculture-A Review.

Plant viral vectors have evolved from tools for transient gene expression into a versatile platform for precise genetic intervention, offering a rapid, transgene-free alternative to conventional crop transformation. This review critically assesses their engineering for scalable field application, moving beyond foundational techniques like virus-induced gene silencing (VIGS). We highlight how advanced vector design, including deconstructed genomes and synthetic regulatory circuits enhances cargo capacity, specificity, and biosafety. The integration of viral delivery with CRISPR-Cas systems has unlocked virus-induced genome editing (VIGE), base editing, and prime editing, enabling heritable trait modification without tissue culture. However, the transition from proof-of-concept in model plants to robust field technology hinges on overcoming critical bottlenecks: expanding host range through chimeric vectors, ensuring environmental containment, and developing scalable delivery methods such as nano-formulations or adjusted agroinfiltration protocols. We evaluate these delivery routes and emerging synergies with nanobiotechnology for targeted and efficient applications. While challenges in regulation, public perception, and large-scale production persist, the strategic engineering of viral vectors for stability, specificity, and safety positions them as a transformative, next-generation biotechnological input for achieving sustainable crop improvement and protection under changing climatic conditions.

Plant Viruses↗

Pyrethroid-resistant diamondback moth expresses alternatively spliced sodium channel transcripts with and without T929I mutation.

This study revealed two distinct alternatively spliced exons, A1 and A2, encoding part of the domain IIS4-IIS5 of the para-sodium channel gene in the diamondback moth (DBM). Exons A1 and A2, respectively, revealed 79% and 91% identity at the nucleotide and amino acid levels. Both alternative exons included the T929I site, which has been associated with pyrethroid resistance in DBM. In the pyrethroid-resistant strain, susceptible (Thr) and resistant (Ile) amino acids were encoded at the T929I site in exons A1 and A2, respectively, but in the pyrethroid-susceptible strain, only Thr was encoded at the site in both exons. The transcripts containing exon A1 were expressed constitutively in all developmental stages. The transcripts containing exon A2 were also detected in all developmental stages, but the levels were significantly lower in the 3rd and 4th instar larvae. Tissue-specific data from the 4th instar larvae and adults showed that the expression of transcripts containing exon A2 was higher in heads than in bodies. These findings suggest that alternative splicing of the para-sodium channel gene might produce distinct channels with different sensitivities to pyrethroids, possibly in a tissue-specific manner.

Alternative Splicing↗

Cloning and nucleotide sequencing of three heat shock protein genes (hsp90, hsc70, and hsp19.5) from the diamondback moth, Plutella xylostella (L.) and their expression in relation to developmental stage and temperature.

Heat shock protein genes, hsp90, hsc70, and hsp19.5, were cloned and sequenced from the diamondback moth, Plutella xylostella (L.) by RT-PCR and RACE method. The cDNA sequence analysis of hsp90 and hsp19.5 revealed open reading frames (ORFs) of 2,151 and 522 bp in length, which encode proteins with calculated molecular weights of 82.4 and 19.5 kDa, respectively. Analysis of cDNA from hsc70 revealed an ORF of 1,878 bp coding a protein with a calculated molecular weight of 69.3 kDa. Furthermore, the analysis of genomic DNA from hsc70 confirmed the presence of introns while no introns were apparent in hsp90 and hsp19.5. Southern blot analysis suggested the presence of multiple copies of each gene family in the DBM genome. Detectable expression of hsp19.5 was observed at the pupal stage while expression of hsp90 and hsc70 was detected at both pupal and adult stages. At adult stage, females showed a higher expression of hsp90 and hsc70 than males. An increased expression was observed in all three genes after exposure to a high temperature in both sexes. These results suggest that in addition to a heat shock response, these HSP genes might be involved in other functions during the course of development in DBM.

Amino Acid Sequence↗

Effectiveness of two insect growth regulators against Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) and Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) and their impact on population densities of arthropod predators in cotton in Pakistan.

Field efficacies of two insect growth regulators (IGRs) at two recommended application rates, buprofezin at 370 and 555 g AI ha(-1) and lufenuron at 37 and 49 g AI ha(-1), were determined against the sweet potato whitefly, Bemisia tabaci (Gennadius), and the cotton bollworm, Helicoverpa armigera (Hübner), in experimental plots of cotton at the Directorate of Cotton Research, Faisalabad, Pakistan. Adverse effects of the IGRs on populations of associated arthropod predators, namely geocorids, chrysopids, coccinellids, formicids and arachnids, were also assessed. Both IGRs significantly reduced populations of B. tabaci at each application rate 24, 48 and 72 h after treatment, and higher doses were more effective than lower doses. Buprofezin was not effective against H. armigera at any tested dose for any time of treatment in any spray. Lufenuron applied at 37 and 49 g AI ha(-1) effectively suppressed H. armigera populations, resulting in significant reductions in crop damage. At lower doses, both IGRs appeared safe to predator populations, which did not differ significantly in IGR-treated versus untreated control plots. Population densities of formicids and coccinellids were significantly lower at high concentrations of both IGRs in treatment plots, possibly as a result of reduced prey availability. The potential role of buprofezin and lufenuron for control of B. tabaci and H. armigera in a spray programme and the likelihood of direct toxic effects of IGRs on predatory fauna of cotton are discussed.

Animals↗

2-D PAGE analysis of pesticide-induced stress proteins of E. coli.

Logarithmically growing batch cultures of Escherichia coli were exposed to sublethal concentrations of pyrethroid and carbamate pesticides of four different technical grades. This induced 17-20 stress proteins, as observed using two-dimensional polyacrylamide gel electrophoresis. An E. coli culture growing exponentially in Luria Bertani medium (cell density approximately 2.3x10(9) cells/ml) was exposed to predetermined sublethal doses of individual pesticides. The cells were harvested after 30 minutes of induction and the stress response was developed in fresh LB medium for three hours under the same growth conditions. Cell pellets were obtained and stored in sonication buffer. Two-dimensional polyacrylamide gel electrophoresis was performed to resolve the proteins. Visualization of the protein spots by rapid silver staining showed 17-20 stress proteins which were absent in the standard protein profile of E. coli. On average 29% of these stress proteins were unique to the pollutant, while the remaining stress proteins overlapped with those of other pesticides. The iso-electric points (PIs) and molecular weights of the proteins were determined by comparing with protein markers with known PIs and molecular weights. Furthermore, upon comparing the pesticide-induced proteins within the same class and between the two different classes (pyrethroid and carbamate), it was apparent that the general nature of the stress remained the same throughout, which indirectly proved that the gene or set of genes responsible for stress expression are also the same, irrespective of the chemical nature of the substituents of the pesticides.

Electrophoresis, Gel, Two-Dimensional↗

Effects of maternal smoking on placental morphology.

BACKGROUND: Maternal smoking is one of leading causes of premature labour and low birth weight babies. Nicotine and carbon monoxide both induce degenerative changes as well as premature aging of placenta. Degenerative changes induce increased amount of collagen in chorionic villi and increased thickness of subtrophoblastic basement membrane. Premature aging is indicated by increased number of syncytial buds and higher percentage of apoptosis in smoker's placentae. Premature aging and degenerative changes may reduce the functional component of placenta and lead to abnormal outcome of pregnancy. This study was designed to determine the effects of maternal smoking on placental morphology. METHODS: Total 40 full term placentae, 20 from normal and 20 from smoker mothers were studied histologically. Full thickness pieces of each placenta from standard area were taken for paraffin embedment. Four micron thick sections were cut on rotary microtome and stained with haematoxylin and Eosin, Malloryis trichrome and hexamine silver for syncytial buds, Apoptotic cells, chorionic villous collagen and Subtrophoblastic basement membrane. RESULTS: This study demonstrates that there is extensive aging and degenerative changes in smoker's placentae. The aging process is shown by increased syncytial buds per unit area and high percentage of apoptosis. Degenerative changes are indicated by increased amount of collagen in chorionic villi and increased thickness of subtrophoblastic basement membrane. CONCLUSION: Extensive premature aging and degenerative changes in smoker's placentae decrease the functional component of an organ, reducing its nutritive and excretory functions. This may be the cause of low birth weight babies in smokers. Extensive loss of trophoblasts by apoptosis and syncytial buds may lead to hormonal imbalance and premature labour in smokers.

Adult↗

Effects of ciprofloxacin on secondary ossification centers in juvenile Wistar albino rats.

BACKGROUND: Administration of quinolone therapy is controversial during juvenile age as stated by earlier workers. The fluroquinolones are currently not indicated for young children, because of the arthropathy and adverse effect on growing cartilage shown by studies. However the effects of ciprofloxacin on secondary ossification centers has remained undocumented. This study is therefore aimed to determine the risk of Ciprofloxacin administration on neonatal skeletal differentiation by a prospective and comparative animal study model using Wistar albino rats. METHODS: Ciprofloxacin was administered to newly born Wistar albino rat pups at a dose of 20 mg/kg body weight intraperitoneally twice daily from day-1 to day-14 after birth. These animals were killed by deep ether anaesthesia and fixed in 80% alcohol. They were then bulk stained with Alizarin red and Alcian blue. Finally they were cleared in 4% KOH and stored in glycerin. The fore and hind limbs were disarticulated from the axial skeleton and observed under stereomicroscope for evidence of skeletal differentiation in the form of presence of secondary ossification centers in long hones (left humerus and left femur). The time of appearance of these centers were noted and compared statistically with those in control animals. RESULTS: The study revealed that the skeletal differentiation in long bones was delayed by 2.4 +/- 0.2 days at both proximal and distal ends in humerus and 2.4 +/- 0.2 days at proximal end and 2.2 +/- 0.2 days at distal end of femur in experimental animals as compared with controls. CONCLUSION: The ciprofloxacin administration during post-natally presents a risk to skeletal differentiation and therefore to its growth up to the age of six weeks is albino rate pups.

Animals↗

Effect of gestational diabetes and maternal hypertension on gross morphology of placenta.

BACKGROUND: Gestational diabetes is much more common than pre-existing diabetes i.e. it complicates 2% to 5% of pregnancies. When metabolic control is good, perinatal mortality should be no higher than in general population. However, macrosomia continuous to be a problem in higher than average proportions of such cases. Macrosomia also involves placenta within the chronic hypertensive disease, the most common diagnosis is essential vascular hypertension. METHODS: Total 60 full term placenta, 20 from normal and 20 each from gestational diabetics and chronic hypertensive mothers were studied grossly. Shape, attachment of umbilical cord, weight, diameter and central thickness of all placentas were noted. RESULTS: The study demonstrates that there is change of shape i.e. two lobes in one placenta from diabetic group. All other placentae were singly lobed and discoidal shape with central attachment of umbilical cord to the foetal surface of placenta. Weight central thickness and diameter were significantly greater in diabetic group as compared to normal and hypertensive group. Hypertensive group shows non significant decrease in weight of placentae while there was no change in central thickness and diameter of placenta in hypertensive than the normal group. CONCLUSIONS: On the basis of results of present study, it is concluded that diabetic's placentae showed increase in weight, central thickness and diameter. One out of 20 placentae in diabetic group also showed change of shape and attachment of umbilical cord to one love. Hypertensive's placentae showed no significant change in weight, shape central thickness and attachment of umbilical cord when compared with normal group.

Adult↗