Induction of apoptosis in leukemic cells by the alkaloid extract of garden cress (Lepidium sativum L.) / 中西医结合学报

OBJECTIVE@#Garden cress (Lepidium sativum L.) is an important herb in traditional medicine used to improve production of breast milk in women and semen in men. In the present research the authors evaluated its ability to destroy leukemic cancer (Jurkat E6-1) cells, using the alkaloid extract of this plant.@*METHODS@#Constituents of the alkaloid extract were analyzed by gas chromatography-mass spectrometry (GC-MS) and their cytotoxicity in leukemic cancer cells and healthy peripheral blood mononuclear cells (PBMCs) was assessed. Cell death via apoptosis was confirmed by DNA laddering, caspase-3 activity, annexin V-fluorescein isothiocyanate and mitochondrial toxicity assays. The specific course of gene activation in treated cells was determined through quantitative polymerase chain reaction (qPCR).@*RESULTS@#GC-MS analysis identified six alkaloids and proto-alkaloids, namely, benzyl isothiocyanate (1), 2-ethoxy-4H-3,1-benzoxazin-4-one (2), (4R)-2-(2-aminophenyl)-4-phenyloxazoline (3), 5-acetyl-1,2-dihydro-6-methyl-2-oxo-4-phenyl-3-pyridinecarbonitrile (4), benzo[b][1,8]-naphthyridin-5(10H)-one,2,4,7-trimethyl (5) and 1,4-diaminoanthraquinone (6), in the alkaloid extract of L. sativum. Of these, compound 1 was previously identified in the seeds of L. sativum. Exposure to the alkaloid extract caused death of Jurkat E6-1 cells, with median lethal concentration (LC) of 75.25 µg/mL. However, the alkaloid extract also showed a nontoxic and proliferative (1.6-fold) effect in healthy PBMCs. Further experiments performed with Jurkat cells at LC and sub-LC doses demonstrated DNA fragmentation, activation of caspase-3 and time-dependant phosphatidylserine translocation (apoptosis) from inner to outer cell membranes. Cell toxicity and assessment of adenosine triphosphate level, together with using qPCR to evaluate expression profile of major apoptosis genes, revealed that apoptosis may be induced by disruption in the mitochondrial outer membrane potential, through activation of extrinsic and intrinsic apoptosis pathways in Jurkat cells.@*CONCLUSION@#The ability of the alkaloid extract of L. sativum seeds to induce apoptosis indicates a potential pharmacological use in cancer chemotherapy. The separation of individual active compounds and further in-depth exploration of the molecular mechanism of apoptosis may lead to novel chemotherapeutic compounds in our future antineoplastic research.

Outcome of Bipolar I (mania) disorders, In Relation with Personality Profile.

This prospective semi structured study evaluated the relations of symptomatology and outcome of bipolar manic patients with personality vulnerability. Methods: 52 patients of bipolar (mania) disorder, out of total 430 admitted patients in psychiatry ward, from January 10, to July 9, 2005 were included in the study. The patients with organic diseases or on any drugs for last two weeks were excluded from the study. All the patients were diagnosed as per ICD-10 diagnostic criteria. Patients of bipolar mania were administered Young Mania Rating Scale (YMRS) to assess the severity of mania. The personality traits and disorders were assessed by the help of ICD-10 module of International Personality Disorder Examination (IPDE). The stress in preceding one month was evaluated by using 41 items Presumptive Stressful Life Event Scale. Initial response to lorazepam was monitored to determine outcome categories. Results: The clinical and demographic variables of the study sample were analyzed with initial response to IV lorazepam as quick responder (grade-I), moderate and poor responders (grade II, III). Sociodemographic variables like marital status (x2 = 1.62, df = 2, NS) and education status (X2 = 4.57, df = 2, NS) did not approach to statistical significance in outcome. However, the outcome of the low income group patients was significantly better ( X2 = 16.84, df = 2, p < 0.001). Out of 14 (26.92%) patients of first manic episode, only 3 patients showed good response to initial lorazepam treatment (Grade I) and 9 and 2 patients assigned outcome category II and III respectively. Patients with history of multiple episodes had shown better response (X2=11.59, df=1, p<0.001, highly significant). Presence of stressful life events was positively correlated with better response to lorazepam treatment (x2==6.73, df=1, p<0.01 significant). Anxious (avoidant) or dependent traits alone or in combination with emotionally unstable personality traits in manic patients significantly determined better episode recovery with lorazepam at one hand proneness for relapses on the other hand.

Clinico-cytopathological spectrum of hepatocellular carcinoma, its correlation with serum alpha-fetoprotein level, and hepatitis B and C viral markers.

Fine-needle aspirationbiopsy (FNAB) is now widely accepted as a diagnostic modality for the treatment of hepatocellular carcinoma (HCC). The most common diagnostic problem in HCC is distinguishing it from a metastatic carcinoma. The literature from India on HCC is scanty. Hence, we studied the cytomorphological features of HCC and metastatic carcinoma. The study included 37 cases of space-occupying lesions (SOLs) of the liver as demonstrated by ultrasound or computed tomography (CT) scan. Cytomorphological features of these SOLs were analyzed in all subsequent to FNAB. Hepatitis B surface antigen (HBsAg), anti-hepatitis C virus antibody (anti-HCV) and alpha-fetoprotein (AFP) were determined in all the cases by enzyme-linked immunosorbent assay (ELISA). The cytopathological diagnosis was HCC in 22 and metastatic carcinoma of the liver in 15. The individual cytomorphological features and which helped to make a definite diagnosis of HCC were: a high nuclear cytoplasmic ratio (81.8%), predominantly trabecular pattern (63.6%) and atypical naked nuclei (100%). Other features were prominent multiple nucleoli (63.3%), hyperchromasia (100%) and moderate anisonucleosis (59%). AFP was elevated in 81.8% of the cases with a mean of 634.8+812.7 ng/ml. HBsAg by ELISA was found to be positive in 72.7% of cases while only 1 case (4.5%) was positive for anti-HCV. In 1 case (4.5%), there was dual infection due to hepatitis B virus (HBV) and HCV. No viral cause was found in 18.3% of cases.

Failure to isolate verotoxin producing Escherichia coli from patients of haemolytic uremic syndrome.

Haemolytic Uremic Syndrome (HUS) been defined as the simultaneous occurrence of acute renal failure in children with haemolytic anemia and thrombocytopenia. This clinical condition that has been recognized is an important cause of acute renal failure in children.

Multidrug resistant Salmonella typhi in Delhi.

In 1990, we isolated 158 strains of Salmonella typhi from blood cultures of patients suffering from typhoid fever. Seventy nine (50%) of these isolates were found to be simultaneously resistant to chloramphenicol, ampicillin and cotrimoxazole. These strains were also resistant to streptomycin and tetracycline, but sensitive to gentamicin, amikacin and cephalexin. The minimum inhibitory concentrations of chloramphenicol and trimethoprim for a representative number of these strains were found to be greater than 1024 micrograms/ml and greater than 128 micrograms/ml respectively. Majority of the multidrug resistant (MDR) strains tested against cefotaxime (23/23), ciprofloxacin (38/38) and amoxycillin plus clavulanic acid (23/24) were sensitive to these drugs.

Gardnerella vaginalis: isolation and identification.

A total of 26 isolates of Gardnerella vaginalis were obtained from 248 patients attending the out patient department of Obstetrics and Gynaecology at A.I.I.M.S hospital, over a period of one year. The vaginal discharge of these patients was cultured on five different media in order to evaluate the best medium for the growth of G. vaginalis. Media containing human blood were found to be the most suitable, Human-Blood-Bilayer medium with Tween-80 being the best for the growth of the organism. A total of 12 biochemical tests were performed for the identification of each isolate. A short and rapid scheme for identification of the organism in a routine laboratory has been evolved.

SAFA test as an aid to the diagnosis of ocular tuberculosis.

A prospective double blind study was carried out to evaluate the role of soluble antigen fluorescent antibody (SAFA) test to detect ocular Tuberculosis. The study material comprised 39 patients with suspected ocular tuberculosis suffering from interstitial keratitis, sclero keratitis, granulomatous uveitis, phlyctenular keratoconjunctivitis, Eales disease and central serous retinopathy. The cases of proven ocular tuberculosis showed up as 70 percent strong reactors and 30 percent weak reactors to SAFA while none had a negative response to SAFA. Of these cases skin hypersensitivity reaction was positive only in 40 percent of the cases. The control group revealed a strong SAFA reaction in only 4 percent of cases with a weak reaction in 44 percent of cases. It thus appears that SAFA test can provide a useful addition to the routine tests in diagnosing tuberculosis.

Cockroaches as vectors of pathogenic bacteria.

One hundred and thirty two cockroaches of species Blattella germanica--96 from hospital ward (test group) and 36 from residential areas (control group) were caught during Nov. 1985 to Nov. 1986. A variety of pathogenic and non-pathogenic bacteria were isolated from test and control group of insects. Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus faecalis, and Micrococci were isolated only from the test group of cockroaches. A high percentage (98.95 per cent) of test cockroaches were found to be carriers of various microorganisms as compared to the control group (80.55 per cent), the difference being statistically significant (p less than 0.001). Quantitative analysis in this study revealed that higher number of microorganisms are carried by test group of insects in the hospital environment. This, thereby suggests that these insects can play an important role in the etiology of hospital acquired infections.

Susceptibility of clinical isolates to cephalexin, cefazolin and cefotaxime.

Three hundred and seventeen recent clinical isolates were tested for in vitro susceptibility to the three cephalosporins available in India--cephalexin, cefazolin and cefotaxime by the Kirby--Bauer disc diffusion method. Cefazolin was the most effective cephalosporin against Gram positive cocci (71.8% sensitive) followed by cefotaxime (62.7%) and cephalexin (52.7%). Cefotaxime was very effective against commonly isolated Gram negative bacilli with only 10 (8.8%) isolates being resistant to it while 44 (39%) and 65 (57.5%) were resistant to cefazolin and cephalexin, respectively. All isolates of Pseudomonas aeruginosa were resistant to cephalexin and cefazolin and only 29 (32.6%) were sensitive to cefotaxime.