Association of Serum Biomarkers with the Mortality of Trauma Victims in a Level -1 Trauma Care Centre of Eastern India.

OBJECTIVE: To determine correlation of important biochemical laboratory investigations in different trauma patients and their degree of injury severity and overall mortality association. METHODS: In this hospital based retrospective observational study, 238 trauma patients were divided into two groups. Group I with injury severity score (ISS)<16 and group II with ISS>16. Haemoglobin (Hb), international normalized ratio, serum creatinine, blood urea nitogen (BUN), serum electrolyte, serum uric acid and liver function parameters were recorded and statistically analyzed. RESULTS: Group II had statistically significant (p<0.0001) elevated levels for referral pulse rate, creatinine, BUN, liver enzymes and decreased level in Hb% and potassium level compared to Group I. Strong positive correlation only exists between BUN and severity score, moderate positive correlation exists between creatinine, aspartate transaminase, and alanine transaminase, alkaline phosphatase and severity score and negative correlation between potassium and severity score. In this study, higher odds of high BUN and creatinine and lower potassium to normal values are associated with bad outcome such as higher mortality in the population of high ISS (>16). CONCLUSION: The study establishes the absolute need of doing three laboratory parameters (serum creatinine, serum blood urea nitrogen and serum potassium) instead of doing laboratory tests battery at the time of trauma victims admission and predicting survival among injured patients in trauma population from Indian settings.

Anti-thyroid peroxidase antibody positivity during early pregnancy is associated with pregnancy complications and maternal morbidity in later life.

AIMS: We assessed the impact of detecting anti-thyroid peroxidase antibodies (anti-TPO-Ab) in the first trimester of pregnancy on pregnancy outcomes and maternal thyroid function during the postpartum period. MATERIALS AND METHODS: In a prospective study consisting 400 pregnant women (8-12 weeks pregnant) were screened for their thyroid profile and followed-up to 12 weeks postpartum. Patients with abnormal thyroid function at 12 weeks postpartum were further followed-up with repeated assessment of thyroid stimulating hormone (TSH) and serum-free thyroxine-4 levels at 8 weeks interval up to 12 months postpartum. RESULTS: 11.5% of the subjects were positive for anti-TPO-Ab who had mean TSH level of 2.31 µIU/ml, which was significantly (P- 0.0001) higher than pregnant women negative for anti-TPO-Ab (1.73 µIU/ml). Increased incidence of miscarriage was observed in anti-TPO positive mothers when compared to antibody negative mothers. Postpartum thyroid dysfunction developed in 4.7% cases at 12 weeks, among them antibody positivity was observed in 81.25% of subjects. In 18.75% mothers positive for anti-TPO-Ab, the thyroid dysfunction persisted up to 12 months postpartum. CONCLUSIONS: Thyroid antibodies detected in early pregnancy seems to be predicting pregnancy complications and later maternal thyroid disease related morbidity.

Evaluation of Magnesium as an adjuvant in Ropivacaine-induced supraclavicular brachial plexus block: A prospective, double-blinded randomized controlled study.

OBJECTIVE: Different additives have been used to prolong brachial plexus block. We evaluated the effect of adding magnesium sulfate to ropivacaine for supraclavicular brachial plexus blockade. The primary endpoints were the onset and duration of sensory and motor block and duration of analgesia. METHODS: One hundred patients (25-55 years) posted for elective forearm and hand surgeries under supraclavicular brachial plexus block were divided into two equal groups (Groups RM and RN) in a randomized, double-blind fashion. In group RM (n = 50), 30 ml 0.5% ropivacaine plus 150 mg (in 1 ml 0.9% saline) magnesium sulfate and in group RN (n = 50), 30 ml 0.5% ropivacaine plus 1 ml normal saline were administered in supraclavicular block. Sensory and motor block onset times and block durations, time to first analgesic use, total analgesic need, postoperative visual analog scale (VAS), hemodynamic variables, and side effects were recorded for each patient. FINDINGS: Though with similar demographic profile and block (sensory and motor) onset time, the sensory and motor block duration and time to first analgesic use were significantly longer and the total need for rescue analgesics was lower in group RM (P = 0.026) than group RN. Postoperative VAS values at 24 h were significantly lower in group RM (P = 0.045). Intraoperative hemodynamics was comparable among two groups and no appreciable side effect was noted throughout the study period. CONCLUSION: It can be concluded from this study that adding magnesium sulfate to supraclavicular brachial plexus block may increase the sensory and motor block duration and time to first analgesic use, and decrease total analgesic needs, with no side effects.

Progesterone induces apoptosis in TRAIL-resistant ovarian cancer cells by circumventing c-FLIPL overexpression.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) holds great potential as an anticancer drug, since it induces selective cell death in cancer cells but not in normal ones. However, cancer cells often acquire resistance to TRAIL, which hinders its clinical efficacy. We previously demonstrated that progesterone triggers apoptosis in human ovarian cancer (OCa) cells. In the present study, we evaluated the prospect of utilizing progestins in combination with TRAIL to enhance cell death in TRAIL-sensitive (OVCA 420, OVCA 429, and OVCA 433) and -resistant (OVCA 432) OCa cell lines. TRAIL sensitivity (60-80% cell kill) bore no correlation with expression of the TRAIL receptors (DR4, DR5) or their decoys (DcR1 and DcR2), but was associated with activation of caspase-8 and -3, and downregulation of the long isoform of FLICE-like inhibitory protein (c-FLIP(L)), an anti-apoptosis mediator. Small interfering RNA-mediated knockdown of c-FLIP(L) expression restored TRAIL sensitivity in OVCA 432 cells. Induction of c-FLIP(L) overexpression increased TRAIL resistance in TRAIL-sensitive lines. Thus, persistent high level of c-FLIP(L) expression likely mediates TRAIL resistance in OCa cells. Treatment of OCa cells with progesterone enhanced TRAIL-induced cell death (>85%), but only in TRAIL-sensitive cell lines. Combined treatment with two progestins was superior to single progestin treatment, with progesterone plus medroxyprogesterone acetate (MPA) achieving over 85% cell kill in both TRAIL-sensitive and -resistant OCa cell lines. Significantly, unlike TRAIL, progestin-induced cell death did not involve c-FLIP(L) downregulation. Hence, combined progestin regimens, with or without TRAIL, may serve as an effective therapy for OCa by circumventing the anti-apoptotic action of c-FLIP(L).

Profiling estrogen-regulated gene expression changes in normal and malignant human ovarian surface epithelial cells.

Estrogens regulate normal ovarian surface epithelium (OSE) cell functions but also affect epithelial ovarian cancer (OCa) development. Little is known about how estrogens play such opposing roles. Transcriptional profiling using a cDNA microarray containing 2400 named genes identified 155 genes whose expression was altered by estradiol-17beta (E2) in three immortalized normal human ovarian surface epithelial (HOSE) cell lines and 315 genes whose expression was affected by the hormone in three established OCa (OVCA) cell lines. All but 19 of the genes in these two sets were different. Among the 19 overlapping genes, five were found to show discordant responses between HOSE and OVCA cell lines. The five genes are those that encode clone 5.1 RNA-binding protein (RNPS1), erythrocyte adducin alpha subunit (ADD1), plexin A3 (PLXNA3 or the SEX gene), nuclear protein SkiP (SKIIP), and Rap-2 (rap-2). RNPS1, ADD1, rap-2, and SKIIP were upregulated by E2 in HOSE cells but downregulated by estrogen in OVCA cells, whereas PLXNA3 showed the reverse pattern of regulation. The estrogen effects was observed within 6-18 h of treatment. In silicon analyses revealed presence of estrogen response elements in the proximal promoters of all five genes. RNPS1, ADD1, and PLXNA3 were underexpressed in OVCA cell lines compared to HOSE cell lines, while the opposite was true for rap-2 and SKIIP. Functional studies showed that RNPS1 and ADD1 exerted multiple antitumor actions in OVCA cells, while PLXNA3 only inhibited cell invasiveness. In contrast, rap-2 was found to cause significant oncogenic effects in OVCA cells, while SKIIP promotes only anchorage-independent growth. In sum, gene profiling data reveal that (1) E2 exerts different actions on HOSE cells than on OVCA cells by affecting two distinct transcriptomes with few overlapping genes and (2) among the overlapping genes, a set of putative oncogenes/tumor suppressors have been identified due to their differential responses to E2 between the two cell types. These findings may explain the paradoxical roles of estrogens in regulating normal and malignant OSE cell functions.

Estrogen-induced loss of progesterone receptor expression in normal and malignant ovarian surface epithelial cells.

While estrogens are suspected risk factors for epithelial ovarian cancer (OCa), progesterone (P4) has been shown to exert protective effects. The biological actions of P4 in target cells are mediated by progesterone receptors (PRs) that exist principally as A- and B-isoforms. We observed overexpression of PR-A and PR-B protein in two lines of OCa cells when compared to two lines of nontumorigenic, normal human ovarian surface epithelial (HOSE) cells. Treatment of HOSE or OCa cells with estrone or 17beta-estradiol at 10(-8) M for a period of 72 h induced significant loss of PR-A and PR-B mRNA and protein expression, with the regulation primarily controlled at the transcriptional level. In contrast, breast cancer cells (line MCF-7) exposed to estrogens upregulated PR-A and PR-B expression. Of significance, both the inhibitory and stimulatory actions of estrogens were blocked by the specific ER-antagonist ICI 182,780 (ICI, 10(-5) M), confirming estrogen specificity. Co-treatment of estrogen-exposed HOSE, OCa, and MCF-7 cell lines with inhibitors of type 1- and type 2-17beta hydroxysteroid dehydrogenase did not affect the previously observed changes in PR expression, suggesting that the action of each estrogen is direct and not mediated via conversion to its metabolic counterpart. Green fluorescence protein (GFP)-PR-A and GFP-PR-B were localized in the cytoplasmic compartment of untreated HOSE cells and translocated to the nucleus after P4 treatment, while both chimera PRs resided in the nuclei of OCa cells in a ligand-independent manner. In OCa cell cultures, P4 (10(-6) M), but not RU486 (10(-5) M), induced apoptosis that was blocked by co-treatment with the antiprogestin but enhanced by co-treatment with ICI. In sharp contrast, P4 induced proliferation, while ICI and RU486 caused cell death in MCF-7 cells. In conclusion, this study is first to demonstrate estrogens as negative regulators of PR expression in HOSE/OCa cells and to provide a mechanistic basis upon which to explain the antagonism of estrogens on the anti-OCa action of progestins. It also raises the possibility of using progestin and ICI as a combinational therapy for OCa treatment.

Identification of ATF-3, caveolin-1, DLC-1, and NM23-H2 as putative antitumorigenic, progesterone-regulated genes for ovarian cancer cells by gene profiling.

Although progesterone (P4) has been implicated to offer protection against ovarian cancer (OCa), little is known of its mechanism of action. The goal of this study was to identify P4-regulated genes that have anti-OCa action. Three immortalized nontumorigenic human ovarian surface epithelial (HOSE) cell lines and three OCa (OVCA) cell lines were subjected to 5 days of P4 treatment. Transcriptional profiling with a cDNA microarray containing approximately 2400 known genes was used to identify genes (1) whose expression was consistently downregulated in OVCA cell lines compared to HOSE cell lines, and (2) whose expression was restored in OCa cell lines by P4 treatment. From the candidates selected, activating transcription factor-3 (ATF-3), caveolin-1, deleted in liver cancer-1 (DLC-1), and nonmetastatic clone 23 (NM23-H2) were chosen for post hoc functional studies based on their previously reported action as tumor suppressors or apoptosis inducers. Semiquantitative RT-PCR analyses confirmed loss of or reduced transcription of these genes in OVCA cells when compared to HOSE cells and their upregulation following P4 treatment. Hormonal specificity was demonstrated by blockade experiments with a progestin antagonist RU 38486. Ectopic expression of caveolin-1, DLC-1, and NM23-H2 caused growth inhibition in OVCA cell cultures, but not in HOSE cell cultures, while forced expression of ATF-3 suppressed growth in both. Overexpression of AFT-3 also enhanced caspase-3 activity in both HOSE and OVCA cells, whereas ectopic expression of caveolin-1 and DLC-1 only activated this enzyme in OCa cells. In contrast, NM23-H2 overexpression was ineffective in activating caspase-3. Overexpression of any of the four genes in OCa cells reduced soft-agar colony formation and cell invasiveness. Taken together, we have identified four new P4-regulated, antitumor genes for OCa. However, their modes of action differ significantly; ATF-3 primarily functions as an apoptosis inducer, NM23-H2 as a suppressor of cell motility, and caveolin-1 and DLC-1 exhibiting features of classical tumor suppressors. To the best of our knowledge, except for NM23-H2, this is the first report linking P4 to the regulation of these tumor suppressor/proapoptotic genes, which could serve as future therapeutic targets.

Purification and characterization of 3-phosphoglycerate kinase from Ehrlich ascites carcinoma cells.

3-Phosphoglycerate kinase (3-PGK) has been purified to apparent homogeneity from Ehrlich ascites carcinoma (EAC) cells by (NH4)2SO4 precipitation, gel filtration and ion-exchange chromatography. The enzyme has been partially characterized and compared with the characteristics of this enzyme of other normal and malignant cells. The EAC cell 3-PGK is composed of a single subunit of 47 kDa. It has a broad pH optimum (pH 6.0-7.5) for its enzymatic activity. The apparent Km values of 3-phosphoglycerate (3-PGA) and ATP for 3-PGK have been found out to be 0.25 mM and 0.1 mM respectively. Similar to 3-PGK of other cells, the EAC enzyme requires either Mg2+ or Mn2+ for full activity; the optimum concentrations of Mg2+ and Mn2+ are 0.8 mM and 0.5 mM respectively. When ATP and 3-PGA act as substrates, ADP, the reaction product of 3-PGK-catalyzed reaction has been found to inhibit this enzyme. Kinetic studies were made on the inhibition of ADP in presence of the substrates ATP and 3-PGA. Attempts to hybridize 3-PGK and glyceraldehyde-3-phosphate dehydrogenase of EAC cells by NAD or glutaraldehyde were unsuccessful.