A Comparative Oncology Study of Iniparib Defines Its Pharmacokinetic Profile and Biological Activity in a Naturally-Occurring Canine Cancer Model

Development of iniparib as an anti-cancer agent was hindered in part by lingering questions regarding its mechanism of action, the activity of its metabolites, and their potential accumulation in tumors. Due to strong similarities in metabolism of iniparib between humans and dogs, a veterinary clinical trial in pet dogs with spontaneous cancers was designed to answer specific questions pertaining to pharmacokinetic exposures and tolerability of iniparib. Dogs were treated with iniparib alone and in combination with carboplatin chemotherapy. Iniparib doses ranged between 10–70 mg/kg intravenously (IV). Plasma, tumor and normal tissue samples were collected before and at various time points scheduled after exposure for pharmacokinetic and biologic analysis. The primary endpoints included characterization of dose-limiting toxicities (DLT) and determination of the drug exposures that could be achieved in both normal and tumor tissues. Nineteen dogs were treated. DLT included fever, anorexia, diarrhea, neutropenia, and thrombocytopenia; most effects were attributable to carboplatin based on the timing of adverse event onset. The maximum tolerated dose (MTD) of iniparib was not identified. Moderate to high variability in plasma exposure was noted for iniparib and all metabolites between animals. When quantifiable, iniparib and metabolite plasma:tumor ratios were < 0.088 and <1.7, respectively. In this study, iniparib was well tolerated as a single agent and in combination with carboplatin over a range of doses. However, clinically relevant concentrations of the parent drug and selected metabolites were not detectable in canine tumor tissues at any studied dose, thus eliminating expectations for clinical responses in dogs or humans. Negative clinical trials in humans, and the uncertainties of its mechanism of action, ultimately led to the decision to stop clinical development of the drug. Nevertheless, the questions that can be asked and answered within the comparative oncology approach are evident from this successfully executed comparative clinical trial and exemplify the value of such studies in drug development.     

Recent trends in non-viral vector-mediated gene delivery

Nucleic acids-based next generation biopharmaceuticals (i.e., pDNA, oligonucleotides, short interfering RNA) are potential pioneering materials to cope with various incurable diseases. However, several biological barriers present a challenge for efficient gene delivery. On the other hand, developments in nanotechnology now offer numerous non-viral vectors that have been fabricated and found capable of transmitting the biopharmaceuticals into the cell and even into specific subcellular compartments like mitochondria. This overview illustrates cellular barriers and current status of non-viral gene vectors, i.e., lipoplexes, liposomes, polyplexes, and nanoparticles, to relocate therapeutic DNA-based nanomedicine into the target cell. Despite the awesome impact of physical methods (i.e., ultrasound, electroporation), chemical methods have been shown to accomplish high-level and safe transgene expression. Further comprehension of barriers and the mechanism of cellular uptake will facilitate development of nucleic acids-based nanotherapy for alleviation of various disorders.     

Micropropagation of Curry Leaf Tree [Murraya koenigii (L.) Spreng.] by axillary proliferation using intact seedlings

An efficient and reproducible procedure for the large scale propagation of Murraya koenigii (L.) Spreng. (Curry Leaf Tree) is described. High-frequency direct shoot proliferation was induced in intact seedlings of M. koenigii on modified Murashige and Skoog (1962) (MS) medium supplemented with 5.0 mg/l benzyladenine. Shoot buds originated from the region adjacent to the apex of the primary shoot and the epicotyledonary node of the intact seedling. Shoots elongated following transfer to MS medium without plant growth regulators. The shoot-forming capacity of intact seedlings was influenced by explant orientation. Maximum shoot proliferation was obtained when the shoot-forming region was in direct contact with the medium surface or slightly embedded into the medium. Proliferating shoot cultures were established by repeatedly subculturing mother seedlings on fresh medium of the same composition after excising all newly formed shoots. Roots were formed on excised shoots when they were transfered to half-strength MS containing 1.0 mg/l indole-3-butyric acid. Plantlets were acclimatized and established in soil where they exhibited normal growth. Received: 25 September 1996 / Revised version received: 5 May 1997 / Accepted: 22 May 1997     

Point of Care Tuberculosis Sero-Diagnosis Kit for Wild Animals: Combination of Proteins for Improving the Diagnostic Sensitivity and Specificity

Tuberculosis is a significant problem globally for domestic animals as well as captive and free ranging wild life. Rapid point of care (POC) serology kits are well suited for the diagnosis of TB in wild animals. However, wild animals are invariably exposed to environmental non-pathogenic mycobacterium species with the development of cross reacting antibodies. In the present study, POC TB diagnosis kit was developed using a combination of pathogenic Mycobacteria specific recombinant antigens and purified protein derivatives of pathogenic and non-pathogenic Mycobacteria. To benchmark the TB antibody detection kit, particularly in respect to specificity which could not be determined in wildlife due to the lack of samples from confirmed uninfected animals, we first tested well-characterized sera from 100 M. bovis infected and 100 uninfected cattle. Then we investigated the kit’s performance using sera samples from wildlife, namely Sloth Bears (n = 74), Elephants (n = 9), Cervidae (n = 14), Felidae (n = 21), Cape buffalo (n = 2), Wild bear (n = 1) and Wild dog (n = 1).In cattle, a sensitivity of 81% and a specificity of 90% were obtained. The diagnostic sensitivity of the kit was 94% when the kit was tested using known TB positive sloth bear sera samples. 47.4% of the in-contact sloth bears turned seropositive using the rapid POC TB diagnostic kit. Seropositivity in other wild animals was 25% when the sera samples were tested using the kit. A point of care TB sero-diagnostic kit with the combination of proteins was developed and the kit was validated using the sera samples of wild animals.     

Differential Induction of β-1,3-Glucanase Gene in Expression of Partial Resistance to Rust (<Emphasis Type="Italic">Uromyces fabae</Emphasis> (Pers.) de-Bary) in Pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

The present study highlights differential induction of pathogenesis related protein PR-2 (β-1,3- glucanases) in expression of rust resistance in pea using different resistant and susceptible recombinant inbred lines of pea (Pisum sativum L.). The enhanced levels of glucanase expression was noted in resistant genotypes at 24 h post inoculations that was negatively correlated (–0.54) with Area Under Disease Progress Curve (AUDPC) and positively correlated (0.67) with lignin accumulations. A significant role of structural defence mechanism in rust resistance in pea was evident from reduced colony size and lesser number of haustorium per colony in resistant lines as well as their negative correlations with lignin accumulation and AUDPC. Gene specific markers indicated constitutive nature of glucanase and peroxidase genes in test genotypes, though differential expression of the glucanase activity was observed in the resistant and susceptible genotypes. However, association of peroxidases with resistance to pea rust is yet to be established due to its non-specific role in slow rusting in pea. The result showed a significant role of β-1,3-glucanase in expression of rust resistance in pea.     

Interactive effects of UV-B and Cu on photosynthesis,uptake and metabolism of nutrients in a green alga Chlorella vulgaris under simulated ozone column

This study demonstrated a general reduction in photosynthesis (carbon fixation, O(2)-evolution and photochemical electron transport chain), the uptake of NH(4)(+), NO(3)(-), urea and PO(4)(3+), and activities of nitrate reductase, urease, acid phosphatase and ATPase following UV-B and copper exposure of Chlorella vulgaris in the absence or presence of 1 and 2 ppm concentrations of a 4-inch-thick ozone layer. Though the effect of stressors used in combination was very detrimental to the above processes, selected concentrations of ozone not only counteracted the UV-B-induced inhibition of the above processes, but also stimulated O(2)-evolution and the photochemical electron transport chain. Kinetics of nutrient uptake and enzyme activities demonstrated that UV-B causes structural change(s) in the enzymes/carriers responsible for the uptake of NH(4)(+), NO(3)(-), urea and PO(4)(3+) as well as their assimilatory enzymes. Except for nitrate reductase, copper was found to compete for the binding sites of all the above enzymes. Synergistic inhibition of photosynthetic activity, nutrient (except NH(4)(+)) uptake, and enzyme activities by UV-B+Cu seems to be due to increased Cu uptake as a consequence of altered membrane permeability brought about by the peroxidation of membrane lipids in UV-B-exposed cells.     

Experimental Absidia corymbifera infection in rabbits: Clinicopathological studies

Absidiosis was produced experimentally in rabbits by intravenous inoculation of 1.4×10⁵ spores of Absidia corymbifera. Infected rabbits exhibited a rise in body temperature, anorexia, dullness, listlessness, diarrhoea, occasional blindness, convulsions and death in some cases. Mortality occurred mainly between 6 to 9 days post infection (DPI) and overall mortality was 50 per cent during the three week observation period. No significant difference was observed in erythrocytic indices viz., Hb, PCV, TEC in control and infected rabbits. However, erythrocyte sedimentation rate was considerably increased in the infected rabbits. A state of leucocytosis was observed in the infected rabbits, which was due to increase in the relative percentage of neutrophils and decrease in lymphocytes. There was a significant increase in blood urea nitrogen concentrations of infected rabbits from 3 to 14 DPI as compared to controls, but serum creatinine values were not significantly altered at any stage of infection. The cause of death was attributed to kidney failure and uraemia in infected rabbits. The rabbit was found to be a suitable model for the study of absidiosis.     

Rapid clonal propagation of three mulberries,Morus cathayana HemsL,M. lhou Koiz. andM. serrata Roxb., through in vitro culture of apical shoot buds and nodal explants from mature trees

High-frequency bud break and multiple shoots were induced in apical shoot buds and nodal explants ofMorus cathayana, M. lhou andM. serrata on Murashige and Skoog (MS) medium containing 0.5–1.0 mg/l 6-benzylaminopurine (BAP). Addition of gibberellic acid (0.4 mg/l) along with BAP induced faster bud break both in apical shoot buds and nodal explants and also enhanced the frequency of bud break in all three species. Shoot culture initiation was greatly influenced by explant type, explant age and explanting season. The shoots were successfully rooted on half-strength MS medium containing a combination of indole-3-acetic acid, indole-3-butyric acid and indole-3-propionic acid, each at 1.0 mg/l. The plantlets were successfully acclimated and eventually established in soil.Abbreviations BAP 6-Benzylaminopurine - GA ₃ Gibberellic acid - IAA Indole-3-acetic acid - IBA Indole-3-butyric acid - IPA Indole-3-propionic acid - Kn Kinetin - MS Murashige and Skoog (1962) medium - NAA 1-Naphthalene acetic acid     

In Vitro Culture of Pimpinella anisum L. (anise)

A simple in vitro protocol has been developed for large scale multiplication of plants from various explants of Pimpinella anisum L., a medicinally important plant belonging to family Apiaceae. Browning of cultures was observed during the maintenance. Frequent subculture at an interval of about 15–17 days was essential for obtaining embryogenic callus cultures and preventing browning of cultures. High frequency of multiple shoot formation was achieved from callus cultures derived from shoot apices, root and stem explants, and also from seed-derived calli. Somatic embryogenesis was observed in callus cultures derived from seeds and shoot apices. Complete plants developed from these embryoids. Direct regeneration of plantlets from shoot apices was also observed. Roots formation occurred in all the cultures. The requirement for exogenous auxin and cytokinin for differentiation was found to be varying in different tissues.