Characterization of phytochemicals and evaluation of anti-cancer potential of Blumea eriantha DC.

The aim of the present study was to evaluate the in vitro anti-cancer and anti-oxidant potential of methanol extract of Blumea eriantha as well as its phytochemical characterization. The in vitro cytotoxic and antiproliferative activities of B. eriantha methanolic extract of leaves were evaluated using MTT assay on HeLa and B16F10 cell lines, and wound scratch and colony formation assays on B16F10 cell lines. The expressions of p53 and Bcl-2 genes were also determined by reverse transcriptase-polymerase chain reaction to establish apoptosis. Isolation and identification of chemical constituents were carried out by various chromatographic and spectroscopic analytical techniques including HPTLC and LC-MS. The methanol extract of the leaves of B. eriantha showed potent in vitro antioxidant and anticancer properties. Moreover, the extract showed significant loss of wound healing, thus suggesting that it could prevent a possible metastasis. Hence, B. eriantha could be explored as a potential anticancer plant with antimetastatic properties.

Design and evaluation of self-nanoemulsifying pellets of repaglinide.

The aim of study was to develop self-nanoemulsifying pellets (SNEP) for oral delivery of poorly water soluble drug, repaglinide (RPG). Solubility of RPG in oily phases and surfactants was determined to identify components of self-nanoemulsifying drug delivery system (SNEDDS). The surfactants and cosurfactants were screened for their ability to emulsify oily phase. Ternary phase diagrams were constructed to identify nanoemulsification area for the selected systems. SNEDDS formulations with globule size less than 100 nm were evaluated for in vivo anti-hyperglycemic activity in neonatal streptozotocin rat model. A significant reduction in glucose levels was produced by optimized SNEDDS formulation in comparison to the control group. The optimized SNEDDS formulations were pelletized via extrusion/spheronization technique using microcrystalline cellulose and lactose. SNEP were characterized by X-ray powder diffraction and scanning electron microscopy. X-ray diffraction study indicated loss of crystallinity of RPG in SNEP. The SNEP exhibited good flow properties, mechanical strength and formed nanoemulsion with globule size less than 200 nm. SNEP showed in vitro release of more than 80% RPG in 10 min which was significantly higher than RPG containing reference pellets. In conclusion, our studies illustrated that RPG, a poorly water soluble drug can be successfully formulated into SNEP which can serve as a promising system for the delivery of poorly water soluble drugs.

Morphine pharmacokinetics and pharmacodynamics in preterm and term neonates: secondary results from the NEOPAIN trial.

BACKGROUND: Relationships between plasma morphine concentrations and neonatal responses to endotracheal tube (ETT) suctioning are unknown in preterm neonates. METHODS: Ventilated preterm neonates (n=898) from 16 centres were randomly assigned to placebo (n=449) or morphine (n=449). After an i.v. loading dose (100 microg kg(-1)), morphine infusions [23-26 weeks postmenstrual age (PMA) 10 microg kg(-1) h(-1); 27-29 weeks 20 microg kg(-1) h(-1); and 30-32 weeks 30 microg kg(-1) h(-1)] were established for a maximum of 14 days. Open-label morphine (20-100 microg kg(-1)) was given for pain or agitation. Morphine assay and neonatal response to ETT suctioning was measured at 20-28 and 70-76 h after starting the drug infusion and at 10-14 h after discontinuation of the study drug. The concentration-effect response was investigated using non-linear mixed effects models. RESULTS: A total of 5119 data points (1598 measured morphine concentrations and 3521 effect measures) were available from 875 neonates for analysis. Clearance was 50% that of the mature value at 54.2 weeks PMA (CLmat(50)) and increased from 2.05 litre h(-1) 70 kg(-1) at 24 weeks PMA to 6.04 litre h(-1) 70 kg(-1) at 32 weeks PMA. The volume of distribution in preterm neonates was 190 litre 70 kg(-1) (CV 51%) and did not change with age. There was no relationship between morphine concentrations (range 0-440 microg litre(-1)) and heart rate changes associated with ETT suctioning or with the Premature Infant Pain Profile. CONCLUSIONS: A sigmoid curve describing maturation of morphine clearance is moved to the right in preterm neonates and volume of distribution is increased compared with term neonates. Morphine does not alter the neonatal response to ETT suctioning.

Plasticity in the intrinsic excitability of cortical pyramidal neurons.

During learning and development, the level of synaptic input received by cortical neurons may change dramatically. Given a limited range of possible firing rates, how do neurons maintain responsiveness to both small and large synaptic inputs? We demonstrate that in response to changes in activity, cultured cortical pyramidal neurons regulate intrinsic excitability to promote stability in firing. Depriving pyramidal neurons of activity for two days increased sensitivity to current injection by selectively regulating voltage-dependent conductances. This suggests that one mechanism by which neurons maintain sensitivity to different levels of synaptic input is by altering the function relating current to firing rate.

Cytoplasmic localization of BAG-1 in leukoplakia and carcinoma of the tongue: correlation with p53 and c-erbB2 in carcinoma.

BACKGROUND: The present study evaluated the clinical significance of BAG-1, an antiapoptotic protein, in leukoplakia and carcinoma of the tongue. METHODS: BAG-1 expression was evaluated by immunohistochemistry in paraffin-embedded tissues of leukoplakia (n=25) and carcinoma of the tongue (n=61). RESULTS: Cytoplasmic expression was predominantly seen in 80% and 70% of patients with leukoplakia and carcinoma, respectively. BAG-1 expression was found to be significantly lower in tobacco users than in non-tobacco users. BAG-1 expression in tobacco-using leukoplakia and carcinoma patients was compared by grouping the carcinoma patients according to lymph node status and disease stage. Carcinoma patients with tumor-positive lymph nodes had significantly lower BAG-1 expression than patients with negative lymph nodes and leukoplakia. Further, a trend towards an inverse correlation was observed with p53 and c-erbB2. In univariate and multivariate survival analysis, patient subgroups with 2+ or 3+ marker positivity (BAG-1 negativity, p53 and c-erbB2 positivity) had a reduced overall survival compared with patient subgroups with 1+ marker positivity or negativity. CONCLUSION: BAG-1 negativity in association with p53 and c-erbB2 positivity identified a subgroup of tongue cancer patients with an aggressive phenotype. Hence, an antiapoptotic protein, BAG-1, was found to be down-regulated in chewing-tobacco-mediated tongue carcinogenesis.

Remediation of textile azo dye acid red 114 by hairy roots of Ipomoea carnea Jacq. and assessment of degraded dye toxicity with human keratinocyte cell line.

Bioremediation has proven to be the most desirable and cost effective method to counter textile dye pollution. Hairy roots (HRs) of Ipomoea carnea J. were tested for decolourization of 25 textile azo dyes, out of which >90% decolourization was observed in 15 dyes. A diazo dye, Acid Red 114 was decolourized to >98% and hence, was chosen as the model dye. A significant increase in the activities of oxidoreductive enzymes was observed during decolourization of AR114. The phytodegradation of AR114 was confirmed by HPLC, UV-vis and FTIR spectroscopy. The possible metabolites were identified by GCMS as 4- aminobenzene sulfonic acid 2-methylaniline and 4- aminophenyl 4-ethyl benzene sulfonate and a probable pathway for the biodegradation of AR114 has been proposed. The nontoxic nature of the metabolites and toxicity of AR114 was confirmed by cytotoxicity tests on human keratinocyte cell line (HaCaT). When HaCaT cells were treated separately with 150 µg mL(-1) of AR114 and metabolites, MTT assay showed 50% and ≈100% viability respectively. Furthermore, flow cytometry data showed that, as compared to control, the cells in G2-M and death phase increased by 2.4 and 3.6 folds respectively on treatment with AR114 but remained unaltered in cells treated with metabolites.

Two-color GFP expression system for C. elegans.

We describe the use of modified versions of the Aequora victoria green fluorescent protein (GFP) to simultaneously follow the expression and distribution of two different proteins in the nematode, Caenorhabditis elegans. A cyan-colored GFP derivative, designated CFP, contains amino acid (aa) substitutions Y66W, N146I, M153T and V163A relative to the original GFP sequence and is similar to the previously reported "W7" form. A yellow-shifted GFP derivative, designated YFP, contains aa substitutions S65G, V68A, S72A and T203Y and is similar to the previously described "I0C" variant. Coding regions for CFP and YFP were constructed in the context of a high-activity C. elegans expression system. Previously characterized promoters and localization signals have been used to express CFP and YFP in C. elegans. Filter sets designed to distinguish YFP and CFP fluorescence spectra allowed visualization of the two distinct forms of GFP in neurons and in muscle cells. A series of expression vectors carrying CFP and YFP have been constructed and are being made available to the scientific community.

Complications associated with digoxin therapy in low-birth weight infants.

Eighteen infants, each weighing less than 1,500 gm, were treated with low dose digoxin therapy for patent ductus arteriosus and signs of circulatory congestion. Nine of the 18 developed one or more signs of clinical deterioration felt to be related to digoxin therapy: eight infants experienced frequent episodes of bradycardia, six had cardiac arrhythmias, and six experienced feeding difficulties. All signs disappeared when digoxin therapy was discontinued. Digoxin, even in relatively low dosages, can have deleterious complications in seriously ill low-birth-weight infants. Alternatives to digoxin in this patient population should be considered before institution of digoxin therapy.

Classroom performance and social factors of children with birth weights of 1,250 grams or less: follow-up at 5 to 8 years of age.

Of 43 long-term survivors with birth weights of 1,250 g or less, 33 were compared with peers and school-aged siblings for educational levels and needs. Of the 33 children in school, three (9.1%) were in classes for children with major handicaps, whereas 30 (90.9%) were found to be comparable to their classmates by teachers and/or test scores, but 14 (47%) were receiving remedial instruction to perform at grade level. Of 13 children with school-aged siblings, three required more hours of assistance by specialized teaching staff than their siblings. The group without the need for specialized teaching staff had older mothers and tended to reside in higher socioeconomic households. Overall, our children with birth weights of 1,250 g or less (51.5%) required more special education efforts than the general school population (24.1%), thereby enabling most to compare favorably with their peers.

Active transport of nitrofurantoin into human milk.

STUDY OBJECTIVE: To determine the extent to which nitrofurantoin is transferred into human milk. DESIGN: Prospective, single-dose pharmacokinetic study. SETTING: University-affiliated clinical research center. PATIENTS: Four healthy lactating women 8-26 weeks postpartum. INTERVENTION: All subjects received a single, oral, 100-mg dose of nitrofurantoin macrocrystals with food. Serial serum and milk samples were obtained and analyzed by high-performance liquid chromatography. MEASUREMENTS AND MAIN RESULTS: Milk pH, milk fat partitioning, and protein binding in serum and milk were determined. Predicted milk:serum ratio (M:S) was compared with the observed M:S. Nitrofurantoin M:S predicted was 0.28+/-0.05, whereas M:S observed was 6.21+/-2.71. Average milk concentration was 1.3 mg/L, and estimated suckling infant dosage was 0.2 mg/kg/day or 6% of maternal dose (mg/kg). CONCLUSIONS: Nitrofurantoin is actively transported into human milk, achieving concentrations in milk greatly exceeding those in serum. Concern is warranted for suckling infants younger than 1 month old, or for infants with a high frequency of glucose-6-phosphate dehydrogenase deficiency or sensitivity to nitrofurantoin.

Housestaff experience, workload, and test ordering in a neonatal intensive care unit.

BACKGROUND: Little is known of how the inexperience or the clinical workload of housestaff affects the decisions they make in the neonatal intensive care unit (NICU). The authors hypothesized that less experienced interns would order more tests per infant than more experienced residents, especially under conditions of heavy workload. METHOD: The NICU at the University of Kentucky A. B. Chandler Medical Center has either an intern or a resident on call by himself or herself at night, a natural setting to compare test ordering by interns and residents. The authors counted the numbers of X rays, analyses of arterial blood gases (ABGs), and electrolyte determinations ordered by the house officers on call for 321 infants from July through November 1993. Data for nine interns and seven residents were compared using multivariate linear regression. RESULTS: When workload became heavier (about five NICU patients), the interns increased their ordering of ABGs per infant to a significantly greater degree than did the residents (p = .01), with the difference being even greater on weekends when the housestaff were under less supervision (p = .004). CONCLUSION: As workload becomes greater in the NICU, interns order more ABGs per infant than do more experienced residents, especially when the interns are less supervised.

Prognostic significance of DNA aneuploidy and p21 ras oncoprotein expression in colorectal cancer and their role in the determination of treatment modalities.

The purpose of the present study was to investigate the prognostic significance of DNA ploidy, S-phase fraction and p21 ras oncoprotein expression in patients with colorectal cancer and to correlate these factors with the clinical behavior of the tumors and their response to therapy. Of 79 patients with colorectal cancer 57% (45/79) had early stage disease. Forty-one percent (32/79) had aneuploid tumors while 30% (24/79) of the tumors had a high (>10%) S-phase fraction. p21ras oncoprotein expression was detected in 38% (30/79) of tumors. Patients with aneuploid tumors had a worse prognosis than patients with diploid tumors (p=0.0002). Similarly, patients with high S-phase fraction tumors had a shorter survival than those with low S-phase fraction tumors (p=0.005). No such difference was found between p21 raspositive and p21 ras-negative tumor subgroups. In early stage colorectal cancer, aneuploidy was closely correlated with disease outcome (p=0.029). Early stage patients with diploid tumors who received radiotherapy and chemotherapy had a better prognosis than patients with aneuploid tumors. In conclusion, DNA ploidy is a significant and independent prognostic factor in colorectal cancer. Aneuploidy and genetic alteration of the p21 ras oncoprotein are important in determining the biological aggressiveness of colorectal cancer. Furthermore, DNA ploidy may identify those subgroups of patients with early stage disease who may benefit from more aggressive treatment.

Prognostic significance of tumor angiogenesis in advanced breast carcinoma: an Indian experience.

Angiogenesis is essential for tumor growth and metastasis. In the present study we investigated the prognostic significance of microvessels (MV) density using immunohisto-localization of factor VIII antigen in 51 breast cancer patients. We counted microvessels per 200x field in the most active areas of neovascularization by staining factor VIII related antigen and graded MV density and correlated with stage, LN involvement and histologic grade. Patients who subsequently developed metastases had significantly high MV counts than patients without metastatic disease (p < 0.001). Patients who subsequently died of the disease had significantly high mean microvessels counts than patients who remained alive at the end of 5 years (p < 0.001). As density of factor VIII antigen staining increased the survival decreased (p < 0.001). All the patients having > 25 MV per 200x field had tumor recurrence faster as compared with patients having < 25 MV (p < 0.02). Thus, the MV count correlates with the prediction for metastasis and poor survival. Such an indicator would be useful in selection of a subgroup of patients with breast cancer who are at high risk for having occult metastasis at presentation and subsequently would benefit from aggressive therapy.

Nonpathologic ventriculomegaly in the premature intrauterine growth retarded infant.

OBJECTIVE: To determine any neurosonographic differences between premature intrauterine growth retarded (IUGR) neonates and premature appropriate for gestational age (AGA) infants. STUDY DESIGN: We retrospectively reviewed the head sonograms of 36 premature IUGR infants and 32 premature AGA matched controls. RESULTS: Seven of the 36 (19%) IUGR infant head sonograms revealed mild ventriculomegaly with no evidence of hemorrhage or ischemia. Only one of the 32 control scans (3%) had similar findings. Mild ventriculomegaly in the IUGR infant versus the AGA control neonate was statistically significant (p value of 0.05). Follow-up head sonograms (4 to 12 weeks after the initial head sonogram) revealed resolution of the mild ventriculomegaly in five of the seven IUGR infants. Clinically, all of the IUGR infants with mild ventriculomegaly were asymptomatic during their stay in the nursery and in clinical follow-up ranging from 3 to 5 years. CONCLUSION: Mild ventriculomegaly on the head sonograms of asymptomatic premature IUGR infants (with no associated hemorrhage or ischemic change) should be recognized as a transient, nonpathologic finding.

Purpura fulminans in three cases of early-onset neonatal group B streptococcal meningitis.

The diagnosis of purpura fulminans was associated with three cases of early-onset group B beta-hemolytic streptococcal (GBS) disease. All three infants had confirmed bacterial disease, extensive purpuric lesions involving the extremities, and laboratory evidence of a consumptive coagulopathy. All three children survived but had markedly compromised neurologic outcomes. Purpura fulminans has not been previously reported with early-onset GBS disease.

Persistent pulmonary hypertension of the newborn infant. Comparison of conventional versus extracorporeal membrane oxygenation in neonates fulfilling Bartlett's criteria.

Persistent pulmonary hypertension of the newborn (PPHN) is a challenge for the neonatologist and a common indication for treatment with extracorporeal membrane oxygenation (ECMO) when medical management fails. We observed 132 neonates born between January 1985 and December 1988 with the diagnosis of persistent pulmonary hypertension of the newborn: 73 (55%) met the Bartlett criteria for treatment with ECMO with 80% predicted mortality; 21 (29%) deteriorated despite conventional medical treatment, were thought to be dying, and were sent for ECMO. Among the 52 patients who were medically treated 40 (77%) survived, a marked difference compared with a predicted 20% survival. All ECMO-treated neonates survived. Although conventionally treated infants showed a trend toward less dependence on supplemental oxygen at > 28 days of life, this study failed to detect a significant difference between those two groups. We conclude that mortality was lower for ECMO-treated infants than for those who were medically treated (0 of 21 vs 12 of 52, p < 0.05); mortality for infants with persistent pulmonary hypertension of the newborn who met Bartlett's criteria and were medically treated was lower than published data; and there was no significant difference in oxygen dependence at > 28 days between the survivors who received ECMO and those who received medical therapy.

Comparison of two neonatal intravenous amino acid formulations in preterm infants: a multicenter study.

Two neonatal intravenous amino acid solutions (Aminosyn-PF and Troph-Amine) were compared in 44 preterm infants. The rate of weight gain, nitrogen balance, and changes in plasma aminograms were determined over 7 days to ascertain whether different outcomes could be identified for the two solutions. At study entry, the infants received a minimum infusion of 2 g amino acid/kg/d with 50 or more nonprotein kcal/kg/d. Group mean amino acid intake over the study period was approximately 2.6 g/kg/d for both groups; nonprotein caloric intake approximated 90 kcal/kg/d. Results showed no significant differences between solutions for the rate of weight gain, nitrogen balance, and nitrogen retention, which approximated intrauterine rates. The rate of weight gain averaged nearly 15 g/kg/d for both solutions. Differences between day 0 and day 7 plasma aminograms showed significant changes between solutions for histidine, lysine, methionine, phenylalanine, threonine, and glutamic acid. However, day 7 plasma aminograms for both solutions compared favorably with those from enterally fed preterm infants reported in the literature. Failure to identify significant differences for the rate of weight gain, nitrogen balance, or nitrogen retention between the two groups suggests that differences in plasma aminograms resulting from use of one solution or the other had no short-term clinical consequences in the premature infants studied.

Methods of assessment and findings regarding pulmonary function in infants less than 1000 grams.

Information regarding lung mechanics and pulmonary function may be used to influence the selection of tiny infant ventilator settings and thus minimize the risk for lung injury during mechanical ventilation. Data regarding compliance and resistance are presented in comparison to various ventilator settings.

Ischemic extremities due to compartment syndromes in a septic neonate.

The clinical presentation and diagnostic evaluation of a septic neonate with sclerema neonatorum and impending tissue loss secondary to compartment syndromes is presented. Vascular disease leading to tissue loss is uncommon in children. To our knowledge, this is the only reported case of a compartment syndrome leading to tissue loss in a neonate.

Occult spinal dysraphism in the infant.

The progressive neurologic dysfunction caused by occult spinal dysraphism can be prevented with early clinical recognition, radiographic diagnosis, and neurosurgical treatment. However, detection of occult spinal dysraphism in the infant is difficult because neurologic symptoms often are not apparent until the child becomes ambulatory. Occult spinal dysraphism, however, can be suspected in the asymptomatic neonate when cutaneous stigmata, such as hemangiomas, hairy patches, deep and/or eccentric dimples, or subcutaneous masses are seen over the lumbosacral spine. Because of the serious, often irreversible, sequelae of a delayed diagnosis, spinal sonography of high-risk infants with midline, lumbosacral, cutaneous stigmata should be considered as an effective, noninvasive screening method.