Regulation of IMP3 by EGFR signaling and repression by ERß: implications for triple-negative breast cancer.

Insulin-like growth factor II (IGF-II) mRNA-binding protein 3 (IMP3) is emerging as a useful indicator of the progression and outcome of several cancers. IMP3 expression is associated with triple-negative breast carcinomas (TNBCs), which are aggressive tumors associated with poor outcome. In this study, we addressed the hypothesis that signaling pathways, which are characteristic of TNBCs, impact the expression of IMP3 and that IMP3 contributes to the function of TNBCs. The data obtained reveal that IMP3 expression is repressed specifically by estrogen receptor ß (ERß) and its ligand 3ßA-diol but not by ERα. Epidermal growth factor receptor (EGFR) signaling and consequent activation of the mitogen-activated protein kinase pathway induce IMP3 transcription and expression. Interestingly, we discovered that the EGFR promoter contains an imperfect estrogen response element and that ERß represses EGFR transcription. These data support a mechanism in which ERß inhibits IMP3 expression indirectly by repressing the EGFR. This mechanism relates to the biology of TNBC, which is characterized by diminished ERß and increased EGFR expression. We also demonstrate that IMP3 contributes to the migration and invasion of breast carcinoma cells. Given that IMP3 is an mRNA-binding protein, we determined that it binds several key mRNAs that could contribute to migration and invasion, including CD164 (endolyn) and MMP9. Moreover, expression of these mRNAs is repressed by ERß and enhanced by EGFR signaling, consistent with our proposed mechanism for the regulation of IMP3 expression in breast cancer cells. Our findings show that IMP3 is an effector of EGFR-mediated migration and invasion and they provide the first indication of how this important mRNA-binding protein is regulated in cancer.

Insulin sensitizing property of Indigofera mysorensis extract.

Indigofera mysorens is a shrub used for its antidiabetic activity in rural India. Here, we elucidate the antidiabetic potential of Indigofera mysorensis extract. Ethanolic extract of the whole shrub of Indigofera (EEI) at 300 mg/kg for 10 days, produced a 63% reduction in plasma glucose, 41% reduction in plasma triglyceride and 77% reduction in plasma insulin levels in insulin resistant db/db mice, which is better than insulin sensitizer, troglitazone (400 mg/kg). EEI unlike sulphonylureas failed to show any acute hypoglycemic effect in normoglycemic Swiss albino mice (SAM). Even in a chronic study (10 days) in SAM, EEI (300 mg/kg) like insulin sensitizers showed no effect on plasma glucose, but an 81% reduction in plasma insulin levels. When challenged with 3 gm/kg sucrose, SAM treated with EEI (300 mg/kg, 7 days) failed to show any effect on the absorption of sugar, whereas standard drug, acarbose (10 mg/kg) showed 52% reduction in the area under the plasma glucose curve. EEI failed to show any significant transactivation of PPARgamma, a proposed target of synthetic insulin sensitizers. Taken together, our data indicate that the antidiabetic effect of the ethanolic extract of Indigofera is due to its insulin sensitizing property and is clearly different from that of sulfonylurea or acarbose.

Antidiabetic and hypolipidemic activity of Helicteres isora in animal models.

Helicteres isora (Sterculiaceae) root juice has been used in the treatment of diabetes by several ethnic groups in different parts of India. A program was initiated to elucidate the scientific basis for the antidiabetic activity of H. isora. Ethanolic extract of H. isora root caused significant reduction in plasma glucose, triglyceride and insulin levels at 300 mg/kg dose after 9 days of administration to insulin resistant and diabetic C57BL/KsJdb/db mice. In normoglycemic and mildly hypertriglyceridemic Swiss albino mice, the extract also showed significant reduction in plasma triglyceride and insulin levels, without affecting plasma glucose level. An ethanolic extract showed activity distinctly different from glybenclamide and acarbose but similar to troglitazone in these models. In high fat fed hamster model, the extract showed significant reduction in plasma lipid levels. In order to identify the active pharmacophore, the ethanolic extract was further subjected to sequential partitioning with low, medium and high polarity solvents, which yielded a semipurified fraction having both euglycemic and lipid-lowering activity. Our study suggests that the extract of H. isora has insulin-sensitizing and hypolipidemic activity and has the potential for use in the treatment of type-2 diabetes.

Quantitative target display: a method to screen yeast mutants conferring quantitative phenotypes by 'mutant DNA fingerprints'.

Whole genome sequencing of several microbes has revealed thousands of genes of unknown function. A large proportion of these genes seem to confer subtle quantitative phenotypes or phenotypes that do not have a plate screen. We report a novel method to monitor such phenotypes, where the fitness of mutants is assessed in mixed cultures under competitive growth conditions, and the abundance of any individual mutant in the pool is followed by means of its unique feature, namely the mutation itself. A mixed population of yeast mutants, obtained through transposon mutagenesis, was subjected to selection. The DNA regions (targets) flanking the transposon, until nearby restriction sites, are then quantitatively amplified by means of a ligation-mediated PCR method, using transposon-specific and adapter-specific primers. The amplified PCR products correspond to mutated regions of the genome and serve as 'mutant DNA fingerprints' that can be displayed on a sequencing gel. The relative intensity of the amplified DNA fragments before and after selection match with the relative abundance of corresponding mutants, thereby revealing the fate of the mutants during selection. Using this method we demonstrate that UBI4, YDJ1 and HSP26 are essential for stress tolerance of yeast during ethanol production. We anticipate that this method will be useful for functional analysis of genes of any microbe amenable to insertional mutagenesis.

Antitumor bicyclic hexapeptide RA-VII modulates cyclin D1 protein level.

A bicyclic hexapeptide, RA-VII or O-methyl deoxybouvardin, isolated from Rubia cordifolia, is known to inhibit protein biosynthesis in vitro and in vivo. We here demonstrate that the treatment of human colon cancer DLD-1 cells with RA-VII induces cell growth inhibition associated with a partial G1 arrest and a rapid decrease (below 2 h) in the level of cyclin D1 protein. Since cycloheximide, another protein synthesis inhibitor, neither decreased the amount of cyclin D1 in the cells nor arrested cells in G1 phase, it is unlikely that this RA-VII-induced reduction of cyclin D1 was fully dependent on its direct inhibitory effect of protein synthesis. Northern blot analysis revealed that RA-VII did not affect the level of cyclin D1 mRNA. Meanwhile, pre-treatment of cells with lactacystin, a proteasome inhibitor, abolished the RA-VII-induced decrease in cyclin D1. Moreover, RA-VII still decreased cyclin D1 protein in the presence of cycloheximide. These results indicate that the RA-VII-induced cyclin D1 decrease depends on cyclin D1 degradation via the ubiquitin-proteasome pathway and does not require additional protein synthesis. RA-VII might actively proceed the degradation process of cyclin D1 via the ubiquitin-proteasome pathway in DLD-1 cells.

Traumatic perforation of the tympanic membrane.

Traumatic perforations of the ear drum are often encountered in the emergency room and in the primary care setting. We review mechanisms of injury to the tympanic membrane and elucidate the appropriate evaluation and treatment of this injury.

Radiosensitization of advanced squamous cell carcinoma of the head and neck with cisplatin during concomitant radiation therapy.

Treatment regimens for advanced squamous cell carcinoma of the head and neck require both attention to local tumor burden as well as contingencies for control of disseminated disease. Relatively new protocols utilizing cisplatin for radiosensitization of tumor cells during concomitant radiotherapy have shown progress in providing effective tumor control. Cisplatin as a chemotherapeutic agent induces DNA changes in malignant cells that may be mutagenic or lethal. Additionally, when used concurrently with radiation therapy, cisplatin acts as a radiosensitizer, increasing damage to malignant nuclear DNA to enhance the anti-neoplastic capability of radiotherapy. The mechanisms by which this radiosensitization occurs remain controversial, although one leading theory involves cisplatin's ability to inhibit sublethal damage repair in radiated tumor cells. Recent investigations utilizing cisplatin with concurrent radiation for treatment of advanced squamous cell carcinomas of the head and neck are reviewed. The variations in protocols presented include route of administration, dosing, scheduling, timing with surgery, and combination therapy with 5-fluorouracil and radiation. Higher response rates, prolonged mean survival, increased survival rates, longer local recurrence-free survival rates, and considerable organ preservation with the use of concurrent cisplatin and radiation have been demonstrated by these studies. Further investigation of concurrent cisplatin and radiotherapy in patients with advanced disease is justified.

Elevated growth of Saccharomyces cerevisiae ATH1 null mutants on glucose is an artifact of nonmatching auxotrophies of mutant and reference strains.

Yeast strains disrupted for ATH1, which encodes vacuolar acid trehalase, have been reported to grow to higher cell densities than reference strains. We showed that the increase in cell density is due to the URA3 gene introduced as a part of the disruption and concluded that the misinterpretation is a result of not using a control strain with matching auxotrophic markers.

Personality traits and performance of military parachutist trainees.

Fifty-two Indian military parachutist trainees were studied throughout the training period to investigate effects of stress in parachutist training on performance efficiency. Stress attributable to fear, anxiety, and apprehension about the inherent risk of paratrooping was studied along with physiological and biochemical changes. An attempt is made to correlate personality traits with performance efficiency. In cognitive and psychomotor tests, maximum deterioration was seen before the tower jump and aircraft jump. The deterioration was correlated with the level of anxiety and urinary catecholamine levels. Inter-correlations between anxiety, catecholamine, and blood pressure were positive, and the level of anxiety was seen to be affecting the quality of performance adversely.

Role of dehydration in heat stress-induced variations in mental performance.

Variation in mental performance under different levels of heat stress-induced dehydration was recorded in 11 subjects heat acclimatized to the tropicals. Dehydration was induced by a combination of water restriction and exercise in heat. The psychological functions--arithmetic ability, short-term memory, and visuomotor tracking--were assessed in a thermoneutral room after the subjects recovered fully from the effects of exercise in heat, as reflected by their oral temperature and heart rate. The results indicated significant deterioration in mental functions at 2% or more body dehydration levels.

Differential effects of hot-humid and hot-dry environments on mental functions.

Twenty five subjects acclimatised to heat artificially were exposed to "basic effective temperatures" (BET) of 25.0 degrees, 29.6 degrees, 32.2 degrees, 33.3 degrees and 35.0 degrees C BET under conditions of both humid and dry heat. The object of the investigation was to ascertain the nature of effects of varying degrees of heat stress on mental alertness, associative learning, reasoning ability and dual-performance efficiency. A further aim was to determine the temperature levels at which impairment of psychological functions was severe enough to be of practical concern. The duration of each exposure was four hours, during which subjects performed physical exercise followed by rest every 30 min. It was found that all the psychological functions tested were adversely affected under extreme heat, and that a significant drop in various psychological functions was seen at effective temperatures of 32.2 degrees C and 33.3 degrees C in hot-humid and hot-dry conditions respectively. It is concluded that at the same effective temperatures the magnitude of the overall effect on psychological functions under humid conditions is relatively greater than that under dry conditions.

Ethnic variations in psychological performance under altitude stress.

Sea-level performance of 120 subjects equally drawn from Gorkha, Madrasi, and Rajput groups was compared with that at an altitude of 4,000 m after 10 months' stay. Alterations in concentration, eye-hand coordination, anxiety, depression, and social interaction were recorded to ascertain ethnic differences in withstanding the altitude stress. Results showed a uniform fall in the scores for eye-hand coordination and social interaction at altitude for the three ethnic groups. However, the effects on concentration, anxiety, and depression were differential for these groups indicating a better toleration of altitude stress by Gorkhas. Climatic similarity between the region of natural habitat of Gorkhas and altitude environment has been suggested to be responsible for their better performance over the other two groups.