New species of Mallocybe (Agaricales, Inocybaceae) from Pakistan, based on morphological and molecular evidence.

Within the family Inocybaceae, many species of Mallocybe have been reported, but there are only a few reports of this genus from Pakistan. In this study, six collections of Mallocybe were studied by morphological and phylogenetic methods. Phylogenetic analyses, based on sequence data from two different loci (ITS and LSU) using Maximum Likelihood and Maximum Parsimony methods, have been performed to infer species relationships within Mallocybe. Results indicated that these six collections encompass two new species of Mallocybe i.e. M.pakistanica and M.pinicola, from Pakistan. Their detailed morphological descriptions and illustrations are also provided. In addition, comparison with morphologically closely-related taxa is also discussed. Previously, only two species of this genus have been recorded from Pakistan and, with this addition, the total number of reported taxa of Mallocybe has been raised to four from Pakistan. A key to the described taxa of Mallocybe from Pakistan is also provided.

Pollen morphological assessment of specific selected species of district Sheikhupura, Punjab, Pakistan under LM and SEM.

Around 30 pollen samples, 16 families and 25 genera that were gathered, identified and preserved from 6 different localities of District Sheikhupura. Light and scanning electron microscopy was done for all collected flora. Out of the total samples, 43% specie was identified as herbs, 23% as shrubs, and 33% were identified as trees. The flora was both wild and cultivated. Wild plants were 33%, cultivated 40% and 26% of them were both wild and cultivated. Qualitative and Quantitative survey of pollens revealed polar shape of pollens were 37% spheroidal, 20% prolate spheroidal, 17% oblate spheroidal, remaining were rectangular and pecolpate. Sexine and nexine were available in 100% of samples. Thirty-three percentage of pollen grains were having spines on them. The presence of spines on pollen grains represents the evolutionary relationship amongst the plants. Current study was conducted to record and analyze palynomorphic diversity in district Sheikhupura, Punjab, Pakistan.

Identification of a Novel Homozygous Missense (c.443A>T:p.N148I) Mutation in BBS2 in a Kashmiri Family with Bardet-Biedl Syndrome.

BACKGROUND: Bardet-Biedl syndrome (BBS) is a rare autosomal recessive inherited disorder with distinctive clinical feature such as obesity, degeneration of retina, polydactyly, and renal abnormalities. The study was aimed at finding out the disease-causing variant/s in patients exhibiting clinical features of BBS. METHODS: The identification of disease-causing variant was done by using whole exome sequencing on Illumina HiSeq 4000 platform involving the SeqCap EZ Exome v3 kit (Roche NimbleGen). The identified variant was further validated by Sanger sequencing. RESULTS: WES revealed a novel homozygous missense mutation (NM_031885: c.443A>T:p.N148I) in exon 3 of the BBS2 gene. Sanger sequencing confirmed this variant as homozygous in both affected subjects and heterozygous in obligate parents, demonstrating autosomal recessive inheritance pattern. To the best of our knowledge, this variant was not present in literature and all publically available databases. The candidate variant is predicted to be pathogenic by a set of in-silico softwares. CONCLUSION: Clinical and genetic spectrum of BBS and BBS-like disorders is not completely defined in the Pakistani as well as in Kashmiri population. Therefore, more comprehensive genetic studies are required to gain insights into genotype-phenotype associations to facilitate carrier screening and genetic counseling of families with such disorders.

Light microscopy and scanning electron microscopy of bacterial colonies isolated from ras malai samples from different markets of Lahore, Pakistan.

The current study investigates the total bacterial contamination in various packed and unpacked ras malai samples of 14 different localities of Lahore, Pakistan. The bacterial colonies such as Bacillus sp. and Gamella sp. were isolated from ras malai samples and grown on agar-broth media under sterile environmental conditions. Serial dilution technique was used to compose the replicates to get a viable count of bacteria in the samples. Results indicated that in case of packed ras malai samples, maximum bacterial count was observed in Sample 1 (422 × 10-2 to 402 × 10-6 ) and minimum bacterial count was in Sample 4 (21 × 10-2 to 9.3 × 10-6 ). For unpacked ras malai samples, maximum bacterial count was in Sample 3 (200.3 × 10-2 to 181.3 × 10-6 ) and minimum bacterial count was observed in Sample 1 (110 × 10-2 to 90.4 × 10-6 ). It was concluded that the marketed samples contain more bacterial count as compared to the standard sterilization values. Such products could possibly become the cause of many health problems in children.

Russula swatica: A new species of Russula based on molecular, light microscopy, and scanning electron microscopy analyses from Swat Valley of Khyber Pakhtunkhwa province of Pakistan.

Himalayan range of Pakistan is one of the diversity rich hotspots of the world. Many areas are yet to be explored here to discover new species of organisms including fungi. During present research, Swat District of Pakistan was explored for fungal diversity. One new species of mushroom Russula swatica (Russulales) is described from Himalayan range of Pakistan and analyzed by light microscopy and scanning electron microscopy as well as by molecular markers. A comprehensive description, photographs, and comparisons with morphologically similar and phylogenetically related species are provided. Conclusions of its phylogenetic relationships within the genus are provided based on the sequence of the nuclear internal transcribed spacer region.

Assessment of mold and yeast in some bakery products of Lahore, Pakistan based on LM and SEM.

The present investigation was designed to throw light on the microbial status of bakery products available in bakeries and supermarkets of Lahore. Different bakery samples such as biscuits, pizza, patties were collected from different localities such as Anar Kali, Chauburji, Faisal Town, Iqbal Town, Model Town, Muslim Town were investigated for mold and yeast using serial dilution technique inoculated over malt extract agar and potato dextrose agar under sterilized conditions. Isolated fungi were namely Alternaria alternata, Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, Aspergillus terreus, Curvularia americana, Fusarium solani, Penicillium digitatum, Saccharomyces cerevisiae, and Geotrichum candidum. Results depicted maximum fungal viable count in biscuits, collected from Model Town while minimum count was in the samples of Chauburji. In the case of pizza, the maximum fungal viable count was found in the sample of Muslim Town. In the case of patties, the maximum fungal viable count was found in the sample of Muslim Town while minimum count was found in the sample of Iqbal Town. Prevalence of microorganisms may be due to the use of contaminated raw material, use of polluted waters, human handling, and the use of contaminated containers. Contaminated food intake can lead to measurable signs of liver injury, inflammation, etc. Preventive measures like washing and drying of hands before preparing food, cleaning of food preparation areas, and the use of clean equipment can avoid microbes which cause hazards to human health. RESEARCH HIGHLIGHTS: Bakery products of Lahore, Pakistan is investigated for the first time on the basis of light microscopy (LM) and scanning electron microscopy (SEM) and found very significant to check the quality of bakery products or not.

A ChIP-cloning approach linking SIRT1 to transcriptional modificationof DNA targets.

The mammalian protein deacetylase SIRT1 (sirtuin1) is widely recognized for its link to calorie restriction and longevity. SIRT1 not only modulates the function of protein targets such as p53 or NFkappaB, but it also affects gene transcription by causing hypoacetylation of associated nucleosomal histones. However, the identification of SIRT1-specific DNA targets that confer chromosomal stability and cell longevity have remained elusive. Here, we report the usefulness of a ChIP-cloning approach for the identification of an endogenous DNA target intimately linked with SIRT1 activity. Using the aforementioned technique, we identified a gene encoding the neuro-oncological ventral antigen2 (nova2) as a SIRT1 target. Nova2 regulates the alternative splicing of scn1a, which encodes the alpha-subunit of a neuronal sodium channel targeted by antiepileptic drugs. This finding demonstrates that ChIP-cloning is an innovative approach for the identification of SIRT1-specific DNA targets.