Fruit and vegetable diets rich in phenolic compounds reduce the risk of various cancers and offer multiple other health benefits due to their bioactivity and powerful antioxidant properties. However, the human health benefits of most phenolic compounds are restricted due to their limited aqueous solubility, low absorption, restricted passive cellular efflux, and poor gastrointestinal stability. Nanotechnology has been used to deliver various therapeutic drugs to specific targets overcoming many of the limitations of direct treatments. This study was designed to develop poly(lactic-co-glycolic acid) (PLGA) nanoencapsulated phenolic-rich extracts from Callistemon citrinus and berberine and to evaluate their effectiveness against extremely invasive MDA-MB 231, moderately invasive MCF-10A, and minimally invasive MCF-7 breast cancers. We have achieved about 80% encapsulation of phenolics from C. citrinus. Most encapsulated nanoparticles were polygonal with particles sizes of 200 to 250 nm. Release of phenolics from encapsulation during storage was biphasic during the first week and then levelled off thereafter. Nanoencapsulated phenolics from C. citrinus extract, berberine, and combination of both enhanced their bioactivity against the three breast cancer cell lines by nearly 2-fold. Growth inhibition of cells was a linear curve relative to phenolic concentration, with a maximum inhibition of nearly 100% at 0.1 mg/ml compared to control.

Although DNA barcoding is an efficient tool for species identification, however, its efficiency is uncertain for samples having degraded DNA and incomplete isolation/amplification of COI gene fragment (>500 bp). DNA mini-barcoding is a solution to this problem because small DNA fragment of COI genes is used for species identification. Twelve highly processed, chemically treated and finished animal skin (coats, tanned skins) and fur (mufflers) samples, received from the Sindh Wildlife Department, Pakistan, were subjected to DNA mini-barcoding. Eight mufflers belonged to Vulpes vulpes, one coat to Ursus thibetanus, one tanned skin to Lutra sumatrana, and one muffler to Vulpes sp. Origin of only one coat sample remained unidentified, success rate of 92% indicative of the fact that the mini barcoding technique can be used as a substitute of conventional barcoding where full length barcode (∼650 bp Folmer region) cannot be generated.

BackgroundThe Hindu Kush and Karakoram mountain ranges in Pakistan’s northern areas are a natural habitat of the snow leopard (Panthera uncia syn. Uncia uncia) but the ecological studies on this animal are scarce since it is human shy by nature and lives in difficult mountainous tracts. The pilot study is conducted to exploit the genetic diversity and population structure of the snow leopard in this selected natural habitat of the member of the wildcat family in Pakistan.MethodAbout 50 putative scat samples of snow leopard from five localities of Gilgit-Baltistan (Pakistan) along with a control sample of zoo maintained male snow leopard were collected for comparison. Significant quality and quantity of genomic DNA was extracted from scat samples using combined Zhang–phenol–chloroform method and successful amplification of cytochrome c oxidase I gene (190 bp) using mini-barcode primers, seven simple sequence repeats (SSR) markers and Y-linked AMELY gene (200 bp) was done.ResultsCytochrome c oxidase I gene sequencing suggested that 33/50 (66%) scat samples were of snow leopard. AMELY primer suggested that out of 33 amplified samples, 21 (63.63%) scats were from male and 12 (36.36%) from female leopards. Through successful amplification of DNA of 25 out of 33 (75.75%) scat samples using SSR markers, a total of 68 alleles on seven SSR loci were identified, showing low heterozygosity, while high gene flow between population.DiscussionThe low gene flow rate among the population results in low genetic diversity causing decreased diversification. This affects the adaptability to climatic changes, thus ultimately resulting in decreased population size of the species.

Improved feeding during pre-calving phase affects the Body Condition Score (BCS) that subsequently dictate the postcalving productive and reproductive performance. The objective of this study was to investigate the effect of three levels of concentrate supplementation [0.5, 1.0 and 1.5% of body weight (BW)] during pre-calving period on post-calving reproductive and lactation performance in Nili Ravi buffalo heifers. Twenty-six pregnant Nili-Ravi buffalo heifers at 150 ± 20 [Mean ± standard deviation (SD)] days in pregnancy with (BW=516± 57 KG, BCS (1-5) =3.98± 0.18) were selected and divided into three treatment groups; Low (n=8), Medium (n=9) and High (n=9) and allocated three dietary treatments. The treatments were basal diets composed of green fodder [dry matter (DM) = 2.0% of body weight (BW)] with three levels of Concentrates (CP=20.9 %, ME= 2.62 Mcal/kg) i.e. 0.5 (Low), 1.0 (Medium), and 1.5% (High) of the BW. The animals received same feed (forage DM at the rate of 2% of BW and concentrate at 0.5 kg per liter of milk produced on as fed basis) after calving. Increasing concentrate supplies from 0.5 to 1.5% of BW increased pre-calving dry matter intake (DMI), however, it didn't affect BW and BCS at calving. Increased concentrate supply at 1.5% of BW resulted in decrease in post calving conception rate to 33% as compared to 75 and 77.8% at 0.5 and 1.0% of BW concentrate supply. The treatment High decreased lactation length (15%) and yield (17%) as compared to Low and Medium treatment. Pre-calving supplementation of concentrate beyond 0.5% of BW showed no improvement of postcalving productive and reproductive performance. In the given dietary situation, the Nili Ravi heifers didn't require more than 0.5% of BW concentrate along with 2.0% of BW green fodder DM during post 150 days of pregnancy.

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