ex-vivo-treatment-response-primary-tumors-andor-associated-metastases

<em>Ex Vivo</em> Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics

The molecular analysis of established cancer cell lines has been the mainstay of cancer research for the past several decades. Cell culture provides both ...

Memorial Sloan Kettering Cancer Center

We performed gene expression profiling of normal and hepatocellular carcinoma (HCC) liver tissues using a high-density microarray that contained 3,063 human cDNA. The results of a microarray hybridization experiment from eight different HCC tissues were analyzed and classified by the Cluster program. Among these differentially-expressed genes, the galectin-3, serine/threonine kinase SGK, translation factor eIF-4A, -4B, -3, fibroblast growth factor receptor, and ribosomal protein L35A were up-regulated; the mRNAs of Nip3, decorin, and the insulin-like growth factor binding protein-3 were down-regulated in HCC. The differential expression of these genes was further confirmed by an RT-PCR analysis. In addition, our data suggest that the gene expression profile of HCC varies according to the histological types.

Gene expression profile analysis in human hepatocellular carcinoma by cDNA microarray.

Relationship between disease activity indices and colonoscopic findings in ulcerative colitis.

To identify candidate genes that could be used as diagnostic and therapeutic targets for hepatocellular carcinoma (HCC), we searched for the genes that are overexpressed in HCC by combining representational difference analysis and microarray. Genes such as glypican-3 (GPC3), insulin-like growth factor 2, long-chain fatty-acid-coenzyme A ligase 4, farnesyl diphosphate synthase were frequently identified in our screening. Northern blot analysis with these four genes confirmed their overexpression in HCC. Among them we found that GPC3 transcript is upregulated in six out of seven cases of HCC. Immunoblot and immunohistochemical staining using polyclonal anti-GPC3 antibodies further confirmed that GPC3 protein is indeed increased in HCC tumor samples. We also found that GPC3 is secreted into culture media from cell lines derived from HCC. We conclude that GPC3 is a good molecular marker for HCC.

Glypican-3 is overexpressed in human hepatocellular carcinoma.

Glypican-3 (GPC3) encodes a cell-surface heparan-sulfate proteoglycan mutated in type 1 Simpson-Golabi-Behmel syndrome (SGBS1), an X-linked overgrowth syndrome. The phenotype of SGBS1 patients and of GPC3 knockout mice suggests that GPC3 plays a negative role in cell proliferation, and an apoptosis-inducing role in specific tissues. Ectopic expression of GPC3 in some cell lines has supported the idea that GPC3 inhibits cell growth. Here we report that blocking endogenous GPC3 expression with an antisense transcript promotes the growth of Hep G2 and Hep 3B hepatoma cell lines. Moreover, antisense inhibition releases Hep 3B cells from cell cycle arrest. Hence, our data further support the notion that GPC3 is an inhibitor of cell proliferation and demonstrate that it modulates cell cycle progression.

Blocking endogenous glypican-3 expression releases Hep 3B cells from G1 arrest.

Glypican-3 (GPC3) encodes a cell-surface heparan- sulfate proteoglycan and its expression is frequently silenced in ovarian cancer, mesotheliomas, and breast cancer cell lines and ectopic expression of GPC3 inhibited the growth of these cells, suggesting that GPC3 plays a negative role in cell proliferation. In contrast, up-regulation of GPC3 is often observed in hepatoma, neuroblastoma, and Wilms' tumor. Whether GPC3 plays the same growth inhibitory role in these tumors remains to be studied. Here we report that antisense-mediated knockdown of GPC3 in the HepG2 hepatoma cells significantly promotes the growth of hepatoma cells. In addition, we show that this growth promotion is independent of insulin-like growth factor 2 (IGF2) signaling. Our data suggest that GPC3 plays a growth-suppressing role in hepatoma and provide cell biological evidence inconsistent with the hypothesis that GPC3 acts as a growth suppressor by downregulating IGF2.

Growth promotion of HepG2 hepatoma cells by antisense-mediated knockdown of glypican-3 is independent of insulin-like growth factor 2 signaling.

Ciprofloxacin is a widely used fluoroquinolone drug with broad spectrum antibacterial activities. Clinical experience has shown incidences of adverse effects related to skin, hepatic, central nervous system, gastrointestinal and phototoxicity. India is a tropical country and sunlight is abundant throughout the day. In this scenario exposure to ambient levels of ultraviolet radiation (UV-R) in sunlight may lead to harmful effects in ciprofloxacin users. Phototoxicity assessment of ciprofloxacin was studied by two mouse fibroblast cell lines L-929 and NIH-3T3. Generation of reactive oxygen species (ROS) like singlet oxygen (1O2), superoxide anion radical (O2*-) and hydroxyl radical (*OH) was studied under the exposure of ambient intensities of UV-A (1.14, 1.6 and 2.2 mW cm(-2)), UV-B (0.6, 0.9 and 1.2 mW cm(-2)) and sunlight (60 min). The drug was generating 1O2, O2*- and *OH in a concentration and dose-dependent manner. Sodium azide (NaN3) and 1,4-diazabicyclo 2-2-2-octane (DABCO) inhibited the generation of 1O2. Superoxide dismutase (SOD) inhibited 90-95% O2*- generation. The drug (5-40 microg mL(-1)) was responsible for linoleic acid peroxidation. Quenching study of linoleic acid peroxidation with SOD (25 and 50 U mL(-1)) confirms the involvement of ROS in drug-induced lipid peroxidation. The generation of *OH radical was further confirmed by using specific quenchers of *OH such as mannitol (0.5 M) and sodium benzoate (0.5 M). 2'-deoxyguanosine (2'-dGuO) assay and linoleic acid peroxidation showed that ROS were mainly responsible for ciprofloxacin-sensitized photo-degradation of guanine base. L-929 cell line showed 29%, 34% and 54% reduced cell viability at higher drug concentration (300 microg mL(-1)) under UV-A, UV-B and sunlight, respectively. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay in NIH-3T3 cell line at higher drug concentration (300 microg mL(-1)) showed a decrease in cell viability by 54%, 56% and 59% under UV-A, UV-B and sunlight, respectively. Results of neutral red uptake assay (NRU) in L-929 cell line were in accordance with MTT assay. The NIH-3T3 cell line showed a higher photosensitizing potential than L-929. The phototoxicity end point shows a time- and concentration-dependent statistically significant (P<0.001) damage. Ciprofloxacin produced ROS by Type I and Type II photodynamic reactions, interacted with nucleic acid moiety and inhibited cell viability. Further, UV-induced photo-peroxidation of linoleic acid accorded the involvement of ROS in the manifestation of drug phototoxicity. Appearance of ciprofloxacin-induced phototoxicity at the ambient level of sunlight is a real risk for the people of India and for those of other tropical countries. We suggest that sunlight exposure should be avoided (especially peak hours) during ciprofloxacin treatment.

Photosensitizing potential of ciprofloxacin at ambient level of UV radiation.

The cosmetics are nontoxic or less toxic in perse but photoactivation may then sensitize and could produce additional phototoxicity. Phototoxicity assessment of ten different lipsticks and eight facial creams was conducted. Results revealed that six lipsticks and five facial creams generated reactive oxygen species (ROS), produced haemolysis and caused lipid peroxidation in human erythrocytes (in vitro) under sunlight exposure. Seven creams and one lipstick were alkaline while one cream and two lipsticks were acidic. The test lipsticks and creams showed absorption in UV/visible range. The study demonstrated synergistic action of cosmetic products and sunlight. Therefore, sunlight exposure should be avoided after the use of photosensitive cosmetics.

Assessment of the phototoxic potential of cosmetic products.

Early onset psoriasis and late onset psoriasis are known to have different clinical patterns in Caucasian population. However, there is paucity of data among Asian patients.

Presentation of early onset psoriasis in comparison with late onset psoriasis: a clinical study from Pakistan.

To study mental and physical health of pregnant women and compare the differences between those residing in urban and rural settings.

Health related quality of life in pregnant women: a comparison between urban and rural populations.

Stem cell derived cardiomyocytes generated either from human embryonic stem cells (hESC-CMs) or human induced pluripotent stem cells (hiPSC-CMs) hold great promise for the investigation of early developmental processes in human cardiomyogenesis and future cell replacement strategies. We have analyzed electrophysiological properties of hESC-CMs (HES2) and hiPSC-CMs, derived from reprogrammed adult foreskin fibroblasts that have previously been found to be highly similar in terms of gene expression. In contrast to the similarity found in the expression profile we found substantial differences in action potentials (APs) and sodium currents at late stage (day 60) of in vitro differentiation with higher sodium currents in hiPSC-CMs. Sensitivity to lidocain was considerably reduced in hESC-CMs as compared to hiPSC-CMs, and the effect could not be explained by differences in beating frequency. In contrast, sensitivity to tetrodotoxin (TTX) was higher in hESC-CMs suggesting different contributions of TTX-sensitive and TTX-resistant sodium channels to AP generation. These data point to physiological differences that are not necessarily detected by genomics. We conclude that novel pharmacological screening-assays using hiPSC-CMs need to be applied with some caution.

Human pluripotent stem cell-derived cardiomyocytes: response to TTX and lidocain reveals strong cell to cell variability.

Sand fly surveillance and control on Camp Ramadi, Iraq, as part of a leishmaniasis control program.

Tick-borne diseases pose significant risks to U.S. military personnel who conduct operations, both domestic and abroad. To determine the feasibility of protecting personnel from tick vectors during short-term field deployments, acaricides cypermethrin (Demon WP, Syngenta, Greensboro, NC) and lambda-cyhalothrin (Surrender Pestabs, CSI, Pasadena, TX) were applied to plots within two separate field sites on Camp Blanding Joint Training Center in Starke, FL, from May to June 2011. We analyzed their effectiveness in reducing tick counts for 6 wk after application. In total, 8,193 ticks were identified and counted, of which > 99% were a mix of nymphs and adult-stage Amblyomma americanum (L.). Our results indicate that both cypermethrin and lambda-cyhalothrin were effective in significantly reducing tick numbers and preventing entry into treated plots for 6 wk after application. Thus, these two acaracides can be used to effectively suppress tick populations and provide residual protection in small geographic areas of recreation or public health significance.

Article	Year
Gene expression profile analysis in human hepatocellular carcinoma by cDNA microarray. Molecules and cells\| PubMed ID: 12521301	2002
Relationship between disease activity indices and colonoscopic findings in ulcerative colitis. Journal of the College of Physicians and Surgeons--Pakistan : JCPSP\| PubMed ID: 12685962	2003
Glypican-3 is overexpressed in human hepatocellular carcinoma. Cancer science\| PubMed ID: 12824919	2003
Blocking endogenous glypican-3 expression releases Hep 3B cells from G1 arrest. Molecules and cells\| PubMed ID: 12872992	2003
Growth promotion of HepG2 hepatoma cells by antisense-mediated knockdown of glypican-3 is independent of insulin-like growth factor 2 signaling. Experimental & molecular medicine\| PubMed ID: 14508064	2003
Photosensitizing potential of ciprofloxacin at ambient level of UV radiation. Photochemistry and photobiology\| PubMed ID: 17880519	2007
Assessment of the phototoxic potential of cosmetic products. Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association\| PubMed ID: 18282649	2008
Presentation of early onset psoriasis in comparison with late onset psoriasis: a clinical study from Pakistan. Indian journal of dermatology, venereology and leprology\| PubMed ID: 19172029	2009
Health related quality of life in pregnant women: a comparison between urban and rural populations. JPMA. The Journal of the Pakistan Medical Association\| PubMed ID: 21465959	2011
Human pluripotent stem cell-derived cardiomyocytes: response to TTX and lidocain reveals strong cell to cell variability. PloS one\| PubMed ID: 23029342	2012
Sand fly surveillance and control on Camp Ramadi, Iraq, as part of a leishmaniasis control program. Journal of vector ecology : journal of the Society for Vector Ecology\| PubMed ID: 24581374	2013
Suppression of Amblyomma americanum (Ixodida: Ixodidae) for short-term field operations utilizing cypermethrin and lambda-cyhalothrin. Journal of medical entomology\| PubMed ID: 24897866	2014

Mohammad Farooq

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