Effect of precursors feeding and media manipulation on production of novel anticancer pro-drug camptothecin from endophytic fungus

We have established methodology for the isolation and characterization of a novel endophytic fungus from the inner bark of medicinal plant Nothapodytes foetida, which produced camptothecin in Sabouraud broth (SB) under shake flask conditions. Camptothecin and its related compounds are at present obtained by extraction from intact plants, but fungal endopytes may be an alternative source of production. In present study we have observed the effect of different nutrient combinations and precursors (tryptophan, tryptamine, geraniol, citral, mevalonic acid and leucine) on the accumulation of camptothecin by endophytic fungus Entrophospora infrequens. The precursors were fed either alone or in combinations (tryptophan and geraniol, tryptophan and citral, tryptophan and mevalonic acid, tryptophan and leucine). The highest camptothecin content was observed in the range of 503 ± 25µg/100g dry cell mass in Sabouraud medium. Camptothecin content in the medium was increased by 2.5 folds by the presence of tryptophan and leucine whereas the production with trytophan was also significantly different from other treatments. Furthermore, the effect of fungal camptothecin on the morphology of human cancer cell lines was also studied. The treated cells showed reduction in size, condensation of nucleus and the protoplasmic extensions were reduced. All these characteristics are found in apoptotic cells.

Ocular Manifestations of Meningitis in Children.

Introduction: Meningitis is the most common central nervous system disease affecting children leading to focal neurological deficits and various oculovisual anomalies including blindness in children. The objective of this study was to evaluate the oculovisual anomalies occurring in Nepalese children suffering from different types of bacterial meningitis. Materials and Methods: A Prospective, study was undertaken for 18 months at B.P.Koirala Lion’s Center for ophthalmic studies, TU Teaching Hospital to study the children suffering from bacterial meningitis admitted at Kanti Children’s Hospital for ocular involvement. A through history, anterior and posterior segment ocular examination and investigations like blood, CSF and CT scan were done. Results: A total of 182 cases of bacterial meningitis were screened. Tubercular meningitis cases were 40 (21.97%) and Pyogenic were 142 (78.02%). Oculovisual anomalies were seen in 70 (38.46%) cases. The ocular abnormalities included pupillary changes (34.28%), Cranial Nerve Palsy (22.86%), Fundus changes (35.72%), Cortical Blindness (4.28%), Panophthalmitis and Proptosis (1.43%). Third nerve involvement was seen in 17.14% cases, sixth nerve in 4.29% cases, Papilledema in 11.43 % and Optic atrophy in 22.86 %. Risk factors included late presentation; hydrocephalous and increased CSF cell count and protein level. Conclusion: Oculovisual anomalies formed an important group of clinical manifestations of bacterial meningitis. Incidence of oculovisual anomalies was more frequently seen in Tubercular meningitis (55%).Children with early presentation and intervention had better prognosis. Hence, timely intervention and health education is important.

DAZL 260A>G and MTHFR 677C>T variants in sperm DNA of infertile Indian men.

DAZL (deleted in azoospermia-like) 260A>G and MTHFR (methylene tetrahydrofolate reductase) 677C>T are two important autosomal variants associated with impaired spermatogenesis. In this study, we investigated DAZL 260A>G and MTHFR 677C>T variants in sperm DNA and their frequency in oligozoospermic infertile men of Indian origin. The study on sperm DNA was performed, since it is more prone to oxidative stress-induced damage and mutation. One hundred oligozoopsermic infertile men having normal chromosomal complement with intact Y chromosome and 100 age- and ethnically-matched fertile controls were investigated for these variants in their sperm genome. Spermatozoa were separated by gradient centrifugation and DNA was isolated and analyzed for the single nucleotide polymorphisms (SNPs) by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). The results showed no significant differences in the frequency of DAZL AG (P = 0.58) and MTHFR CT (P = 0.44) between oligozoospermic infertile men and controls. However, 8% (8/100) oligozoospermic infertile men harbored both the variants and showed significantly (P<0.0001) lower sperm count (3.28 ± 1.1 vs 12.50 ± 4.09) compared to infertile men with either of the single variant. None of the fertile controls showed the presence of the both variants. In conclusion, the combined effect of both DAZL 260A>G and MTHFR 677C>T variants may have role in compromised sperm count. However, further studies are required to find the pathological role of these combined variants in male infertility.

HER-2 targeted immunonanoparticles for breast cancer chemotherapy.

The success of anti-cancer drug targeting is depends on the ability of the therapeutics to reach their desirable cellular and intracellular sites and minimizing action at the nonspecific sites. In the present study, anti-human epidermal growth factor receptor (HER-2, ErbB2) antibody anchored nanoparticles were prepared and evaluated for the assessment of targeting potential in breast cancer cell. In an attempt for comparison of carrier system for site-selective delivery, docetaxel loaded PLGA nanoparticles, PLGA-PEG nanoparticles and PLGA-PEG immunonanoparticles capable of targeting breast cancer were prepared by emulsion solvent evaporation technique. The drug-loaded nanoparticles were characterized for their size and size distribution, surface charge, drug encapsulation efficiency and in vitro drug release. Our results demonstrate that docetaxel loaded PLGA-PEG immunonanoparticles strongly enhances the site specific uptake and high cytotoxic effect at targeted sites, as compared with PLGA, PLGA-PEG nanoparticles. In conclusion polymeric immunonanoparticles could be a promising carrier for the treatment of HER2-overexpressing breast cancers.

Fine-Needle Aspiration Versus Non Aspiration Technique of Cytodiagnosis in Thyroid Lesions.

The two sampling techniques were studied in 50 patients presenting with enlargement of thyroid gland. They were sampled by fine needle aspiration (FNA) and by non-aspiration (NA - a needle without application of aspiration pressure). Cell samples were cytologically assessed and were scored (0, 1 & 2) using five objective parameters which includes background blood or clot, amount of cellular material, degree of cellular degeneration, degree of cellular trauma and retention of appropriate architecture. Non-aspiration performed better than FNAC for criteria background blood or clot and retention of appropriate architecture with statistically significant difference. Non-aspiration also scored better than FNAC for the other parameters as well as cumulatively. Smears were then cytologically assessed as diagnostically inadequate, adequate and superior. Greater number of diagnostically adequate specimens were obtained by FNAC than by NA but the number of diagnostically superior specimen were obtained significantly more frequently by the non-aspiration technique. Thus, the non-aspiration technique combined with FNAC can result in obtaining good quality cellular material in thyroid lesions.

Idiopathic lobular panniculitis (Weber Christian disease): a case report.

Weber Christian disease, an idiopathic relapsing febrile subcutaneous and visceral panniculitis is a rare disease in children. We report a case of Weber Christian disease for he first time from Kanti Children's Hospital.