Fabrication and optimization of chitosan-g-m-PEG-NH2 copolymer for advanced glioblastoma therapy using surface engineered lentinan loaded nanovesicles for nasal delivery.

Glioblastoma multiforme (GBM) exhibits a high mortality with an incidence rate of 3-5 per 100,000 each year, which demands existence of newer approach for its treatment. The current study focuses on synthesis of novel lipidic nanovesicles (LNs) loaded with highly potent macromolecule Lentinan (LNT) and surface modified with methoxy poly (ethylene glycol; PEG) amine (m-PEG-NH2)-grafted-chitosan (CS) for intranasal delivery. The grafting procedure was optimized using Box Behnken design (BBD) to limit the use of organic solvents. The fabricated polymer showed enhanced aqueous solubility, biodegradability and mucoadhesion, resulting in higher nasal mucosa permeation (z = 53.52 µm). The presence of PEG enabled the sustained release of LNT till 48 h and assisted in achieving higher accumulation of LNT in CSF (41.7 ± 3.1 µg/mL) and a higher brain targeting potential of 96.3 ± 2.31 % (p < 0.05). In-vitro cellular studies showed the enhanced anti-GBM effect of LNT on U87 MG cells by reducing the cell viability (~2 times reduction in IC50 value) accompanied with large number of cells undergoing late apoptosis and death (p < 0.05) because of the higher cellular uptake (63.22 ± 3.01 ng/100 cells) of novel formulation. The copolymer comprising LNs were biocompatible, stable and can be used as an effective tool in the management of GBM.

A bioanalytical assay for estimation of thymoquinone in rats cerebrospinal fluid and brain tissues of nasally administrated thymoquinone loaded lipo-polymeric nanoshells and its pharmacokinetic profiling.

Thymoquinone (TH) has been one of the major phytochemical used in the treatment of cancers since long time, especially in the management of glioblastoma multiforme (GBM). The formulation of lipo-polymeric nanoshells (LPNs) and their nasal delivery are fascinating approaches for overcoming the drawbacks of low solubility and poor bioavailability of TH. Hence targeting LPNs to the brain requires a validated bioanalytical method for the assessment of TH concentration in Cerebrospinal fluid (CSF) and brain tissue homogenates (BTH). Therefore, the current work focuses on the development and validation of high-performance liquid chromatography (HPLC) method in CSF by employing nasal simulated fluid (NSF) as one of the major components of the mobile phase. The developed method was checked for linearity in the range of 0.05 to 1.6 µg/mL, having an r2 value of 0.999 with mean % recovery >95% and % RSD values below <2.0%. The developed method gave a clear separation of TH at 6.021 ± 0.17 min with an internal standard at 4.102 ± 0.09 min and a CSF spike at 2.170 ± 0.12 min. The developed method assisted in determining the in-vitro and in-vivo drug release study of LPNs, pharmacokinetic profiling, qualitative in-vivo brain uptake study, in-vitro cellular uptake, and generating stability data of formulated LPNs proposed for intranasal administration in rats.

Development of topical thymoquinone loaded polymer-lipid hybrid vesicular gel: in-vitro and ex-vivo evaluation.

Thymoquinone (TH), a naturally occurring moiety shows excellent anticancer properties and in the present study, TH loaded polymer hybrid lipid vesicles (TH PLH) were formulated, and in-vitro cytotoxicity towards breast cancer cell lines (MCF-7 and MBD-MB 231 cells) was determined. TH PLH was primed by the ethanol spraying method and were characterized for various attributes like particle size, morphology, percentage drug entrapment, elasticity, rheological, zeta potential, etc. The prepared TH PLH Vesicles showed an average particle size from 344.7 ± 3.7 nm to 351.9 ± 2.3 nm and showed very narrow distribution with polydispersity index ranging from 0.245 ± 0.36 to 0.271 ± 0.66. The surface charge on the vesicles was confirmed by zeta potential (þ -21.3 ± 1.08 mV to þ -19.98 ± 3.4 mV). The globular moulded vesicles were in the nano-size range and had high flexibility and an approximately 10-fold increase in elasticity properties. TH PLH Vesicular gel was formulated by means of Carbopol 934 and was evaluated for physical appearance, pH, rheological behaviour, and skin irritation test. Cytotoxicity study reveals paramount inhibition on cells by TH vesicle-loaded gel. These results showed the high potential of TH PLH vesicles for topical anticancer application.

PCR-Based Evaluation of Tuberculous Endometritis in Infertile Women of North India.

BACKGROUND: Tuberculous endometritis is often a diagnostic dilemma for gynecologists in the evaluation of infertility. This study evaluated and compared different diagnostic methods in tuberculous endometritis. METHODS: 100 infertile women were investigated for tuberculous endometritis. The endometrial samples obtained by curettage were evaluated by Ziehl-Neelsen (ZN) staining of the smears for acid-fast bacilli (AFB), the samples were also cultured on Lowenstein-Jensen medium, and histopathological examination and nested PCR targeting 'hupB' gene (Rv 2986c in M. tuberculosis and Mb3010c in M. bovis) which can differentiate between M. tuberculosis and M. bovis were performed. Antibodies against 38-kDa and 16-kDa mycobacterial antigens were detected in serum using ELISA. Statistical analysis was done by online GraphPad Prism software, version 5.0. McNemar's test was applied and Kappa agreement coefficient was calculated for agreement between various methods. A p-value < 0.05 was considered significant. RESULTS: Among the 100 evaluated endometrial samples, one was AFB smear positive, none was positive by culture, four were positive by histopathology and 13 were positive by PCR. Of the 13 PCR-positive cases, 38.4% were positive for M. tuberculosis, 23.07% for M. bovis, and 38.4% showed co-infection with both species. 40% of the patients had raised IgG against M. tuberculosis 38-kDa antigen. McNemar's test was applied to PCR and the conventional methods of TB diagnosis (AFB, Culture and histopathology) and the p-value was < 0.001 (highly significant) for PCR. Detection by PCR showed a fair agreement with detection by Mantoux test and ELISA. CONCLUSION: In paucibacillary endometrial tuberculosis, the positive detection rate was found to be significantly higher for PCR compared to other methods. The 'in-house' nested PCR assay targeting the hupB gene and used in this study, can serve as a rapid diagnostic aid for tubercular endometritis. It can also differentiate between members of the Mycobacterium tuberculosis complex, namely M. tuberculosis and M. bovis.

Influence of menopause on biochemical markers of endothelial dysfunction-A case-control pilot study in North Indian population.

OBJECTIVE: Menopause, an estrogen deficient state, is known to increase the cardiovascular risk. Lipid changes accompanying menopause account for only few cases of coronary artery disease (CAD). Endothelium-dependent nitric oxide-mediated vasodilatory mechanisms are also known to play a role in development of coronary artery disease, but studies in menopausal women are very few. This study was hence undertaken to see if nitric oxide (NO)-cyclic guanidine monophosphate (c-GMP) pathway is influenced by menopause. DESIGN: This study was a hospital-based case-control study involving 100 women in age group 40-55 years. Of these, 50 women were postmenopausal and 50 were premenopausal. Women with known risk factors for CAD were excluded. Fasting blood samples from these women were collected and analyzed for estradiol levels, lipid profile, apolipoprotein B, plasma nitric oxide, c-GMP and platelet nitric oxide using standard kits and reagents. Statistical analysis was done on SPSS and two-tailed p-value <0.05 was considered significant. RESULT: Postmenopausal women had significantly lower estradiol, plasma NO, and c-GMP levels as compared to premenopausal women (p<0.05). Cholesterol, low-density lipoprotein (LDL) cholesterol and apolipoprotein B (apo-B) levels were higher and HDL levels were lower in postmenopausal as compared to premenopausal women (p<0.05). Plasma NO showed a significant positive correlation with estradiol, HDL levels and negative correlation with apo-B levels. CONCLUSION: Menopause tends to downregulate NO-c-GMP pathway resulting in endothelial dysfunction. The mechanism may be directly through estrogen receptors or indirectly through potentiation of dyslipidemia.