In modernized lifestyle smoking is one of the trendy, psychological, and socioeconomic scenarios of young adolescents mainly in the age of the reproductive stage. Based on a number of cigarettes smoked, age, and duration of the smoke, the study aims to search for the profound effects of smoking and its impact on semen parameters, sperm-DNA integrity, and fragmentation of sperm DNA with cotinine and apoptotic caspase-3 marker in the seminal plasma of fertile and infertile smokers. To determine oxidative damage by 8-hydroxy deoxyguanosine (8-OHdG) from isolated sperm DNA (steps: reactive oxygen species washing by nitro blue tetrazolium (NBT), sperm lysis, salt digestion, ethanol washing, and finally with high-performance liquid chromatography analysis). Level of DNA fragmentation (percentage) in native and intact DNA, the activity of caspase-3 in infertile smokers will be compared with the control group of nonsmokers. Also, the sperm viability was visualized by eosin-nigrosin and aniline blue staining. Cotinine is one of the best markers of smoking. The cotinine level (2224.24 ± 1.19 *** ng/mL), when abundant it negative correlates with morphology and rapid motility in infertile smokers than nonsmokers. Gel preprogram measured the sperm integrity and was found to be less in smokers than nonsmokers. The spermatic oxidative marker 8-OHdG was high and gave an R 2 value of 0.9104 with morphology and 0.9007 for rapid motility of infertile sperm, respectively. Infertile smoking subjects (<10 cigarettes/day) had significant changes increase in sperm fragmentation, caspase-3, and cotinine while negative impact with motility, morphology, and pH of semen compared with fertile, infertile nonsmoking subjects. K E Y W O R D S 8-hydroxy deoxyguanosine, caspase-3, nicotine, sperm-DNA fragmentation

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Augmented escape of nanostructures to the ecosystem has necessitated the comprehensive study of their impact, especially on plants. In the current study, hematite nanoparticles were prepared by employing garlic extract and checked for their cytogenetic effect on onion roots and germination characteristics of five agricultural crops (Vigna radiata, Triticum aestivum, Trigonella foenum-graecum, Cicer arietinum and Vicia faba) in the concentration range of 20-100 mg/L. Onion roots exhibited an increased mitotic index till 60 mg/L dosage, beyond which trend decreased marginally. Percentage of aberrant chromosomes reported for 100 mg/L exposure was very low (3.358 ± 0.13%) and included common defects such as clumped/sticky metaphase, ring chromosomes, laggards, spindle abnormality, chromosome bridges etc. Moreover, comet assay, DNA laddering experiment and electron micrograph study confirmed negligible damage to onion roots. Seed germination study indicated a positive response in different agronomic traits (germination index, root length, fold change in weight and vigour index) up to 60 mg/L, beyond which either negative or neutral effect was observed. However, none of the samples showed 50% inhibition in germination index; highest being 33.33% inhibition for V. faba, compared to the control. In brief, biogenic hematite nanoparticles caused insignificant phytotoxicity and were likely assimilated as iron source at lower dosage.

Highlights As a first time upto 6 months the sperm cells were stored with new medium from green formulation. TPP-TAB of E4 medium reduces ROS damage.