Selective growth of malignant cells by in vitro incubation on Teflon

The selective growth of malignant cells on non-adhesive Teflon substrates is reported. Neoplastically transformed cell lines of human and murine origin proliferated on Teflon whereas similarly seeded normal non-tumorigenic cells were unable to undergo cell division and subsequent growth. This substrate should be especially useful in establishing primary cultures from human tumor material.     

Identification of Mycobacterium tuberculosis clinical isolates with altered phagocytosis by human macrophages due to a truncated lipoarabinomannan

Phenotypically distinct clinical isolates of Mycobacterium tuberculosis are capable of altering the balance that exists between the pathogen and human host and ultimately the outcome of infection. This study has identified two M. tuberculosis strains (i.e. HN885 and HN1554) among a bank of clinical isolates with a striking defect in phagocytosis by primary human macrophages when compared with strain Erdman, a commonly used laboratory strain for studies of pathogenesis. Mass spectrometry in conjunction with NMR studies unequivocally confirmed that both HN885 and HN1554 contain truncated and more branched forms of mannose-capped lipoarabinomannan (ManLAM) with a marked reduction of their linear arabinan (corresponding mainly to the inner Araf-alpha(1-->5)-Araf unit) and mannan (with fewer 6-Manp residues and more substitutions in the linear Manp-alpha(1-->6)-Manp unit) domains. The truncation in the ManLAM molecules produced by strains HN885 and HN1554 led to a significant reduction in their surface availability. In addition, there was a marked reduction of higher order phosphatidyl-myo-inositol mannosides and the presence of dimycocerosates, triglycerides, and phenolic glycolipid in their cell envelope. Less exposed ManLAM and reduced higher order phosphatidyl-myo-inositol mannosides in strains HN885 and HN1554 resulted in their low association with the macrophage mannose receptor. Despite reduced phagocytosis, ingested bacilli replicated at a fast rate following serum opsonization. Our results provide evidence that the clinical spectrum of tuberculosis may be dictated not only by the host but also by the amounts and ratios of surface exposed mycobacterial adherence factors defined by strain genotype.     

Macrophage colony-stimulating factor expression in retrovirally transduced cells is dependent upon both the adherence status of the target cells and its 5' flanking untranslated region

Numerous cell types retrovirally transduced with macrophage colony-stimulating factor (M-CSF) using LXSN-based vectors showed a variable expression of the transgene. Expression of M-CSF correlated with the cells' adherent status. Transduced adherent cells produced the M-CSF, whereas the non-adherent cells synthesized little M-CSF. Studies showed that the 5'-UTR of the M-CSF gene regulated transgenic M-CSF gene expression. Ligation of this 5'-UTR to the enhanced green fluorescent protein gene (EGFP) caused the expression of EGFP to show the same dichotomy as previously seen with the M-CSF. Transgenic M-CSF was expressed within non-adherent cells when the 5'-UTR was removed from the LXSN vector. Quantitative real-time polymerase chain reaction analysis confirmed that lesser production of M-CSF mRNA occurred within the non-adherent cells than in the adherent cells. This difference was eliminated when the 5'-UTR was removed from the retroviral vector. Our work suggests that this 5'-UTR of the M-CSF gene could be an important way to get transgenic expression within adherent cells, but not in non-adherent cells.     

Non-ventral lateral neuron-based, non-PDF-mediated clocks control circadian egg-laying rhythm in Drosophila melanogaster

The authors report the results of their study aimed at investigating the consequence of targeted ablation of ventral lateral neurons (LN(v)s--neurons regulating eclosion and locomotor activity rhythms) and genetic disruption of pigment-dispersing factor (PDF--an important output of circadian clocks) on the egg-laying rhythm of Drosophila melanogaster. The results clearly suggest that genetic ablation of LN(v)s and loss of function mutation of PDF abolish eclosion and locomotor activity rhythms, whereas the egg-laying rhythm continues unabated. Furthermore, the results also demonstrate that the period of egg-laying rhythm remains unchanged under different ambient temperatures and nutrition levels, suggesting that the egg-laying rhythm of D. melanogaster is temperature and nutrition compensated. Based on these results, the authors conclude that the egg-laying rhythm in D. melanogaster is regulated by non-LN(v)-based, non-PDF-mediated circadian clocks.     

Hybridization of Gossypium species through in ovulo embryo culture

An interspecific hybrid of the sexually incompatible species G. hirsutum cv. Laxmi and G. arboreum cv. Jyoti was obtained through in ovulo embryo culture. Eightto twelve-day-old ovules were excised and cultured on Beasley and Ting's medium supplemented with Indol-3 acetic acid (5×10^-6 to 7×10^-6 M), Kinetin (5×10^-6 to 5×10^-8 M), Gibberellic acid (5×10^-7 to 5×10^-9M), Ammonium chloride (5 to 15mM) and Casein hydrolysate (50 to 200mg/l) added individually and in various combinations along with sucrose. No single medium was adequate to ensure complete development of the fertilized ovules to plantlets, thus necessitating a sequential five step transfer to different media. Cytological studies confirmed the hybrid nature of the plants.Abbreviation IAA Indol-3 acetic acid - Kn Kinetin - GA₃ Gibberellic acid - CH Casein hydrolysate - NAA -Naphthalene-acetic acid - BT Beasley and Ting's basal medium - MS Murashige and Skoog's basal medium - W White's basal medium NCL Communication number 3823.     

Nested ANOVA for Repeated Surveys with Autocorrelated Errors

Diet surveys in the science of human nutrition involve longitudinal observations on the same individuals repeated at distinct time points. Consequently a common feature of such surveys is inherent correlation between successive observations. In this paper, an autoregressive model of order 1 is assumed for the successive observations on the same individual in a survey. Further, observations on the same individual in two different surveys are assumed to be correlated. The technique of analysis of variance is modified suitably for this set up and applied to two nutritional surveys carried on army recruits.     

Optimization of Dimethoate Degradation by Brevundimonas sp. MCM B-427 Using Factorial Design: Studies on Interactive Effects of Environmental Factors

Microbial cultures were isolated from soil exposed to dimethoate, an organophosphorus insecticide. The isolate that showed maximum degradation of dimethoate was identified as Brevundimonassp. MCM B-427. The curve relating biomass accumulation and degradation revealed that aeration appeared to enhance growth rather than degradation. For optimization of environmental factors, a 2⁴ factorial experimental design was used, wherein four factors namely pH, temperature, inoculum density and aeration condition were varied simultaneously. The interaction between the factors was analyzed using MINITAB package. Degradation of dimethoate was affected by a large number of factors interacting in a complex way. Six two-way and two three-way interactions of various environmental factors were found to be statistically significant. In all the two-way interactions, the effect of one factor was more pronounced when the other was at its optimum. The three-way interactions revealed that optimization of pH, inoculum size and aeration was more critical at 40 °C than at 30 °C.The results imply the importance of studying interactions of parameters rather than optimizing individual parameters one by one. These results demonstrate complexity of the interaction and thereby imply the need for better experimental and statistical models.