Proteomics analysis of colon cancer progression.

BACKGROUND: The aim of this pilot study was to identify proteins associated with advancement of colon cancer (CC). METHODS: A quantitative proteomics approach was used to determine the global changes in the proteome of primary colon cancer from patients with non-cancer normal colon (NC), non-adenomatous colon polyp (NAP), non-metastatic tumor (CC NM) and metastatic tumor (CC M) tissues, to identify up- and down-regulated proteins. Total protein was extracted from each biopsy, trypsin-digested, iTRAQ-labeled and the resulting peptides separated using strong cation exchange (SCX) and reverse-phase (RP) chromatography on-line to electrospray ionization mass spectrometry (ESI-MS). RESULTS: Database searching of the MS/MS data resulted in the identification of 2777 proteins which were clustered into groups associated with disease progression. Proteins which were changed in all disease stages including benign, and hence indicative of the earliest molecular perturbations, were strongly associated with spliceosomal activity, cell cycle division, and stromal and cytoskeleton disruption reflecting increased proliferation and expansion into the surrounding healthy tissue. Those proteins changed in cancer stages but not in benign, were linked to inflammation/immune response, loss of cell adhesion, mitochondrial function and autophagy, demonstrating early evidence of cells within the nutrient-poor solid mass either undergoing cell death or adjusting for survival. Caveolin-1, which decreased and Matrix metalloproteinase-9, which increased through the three disease stages compared to normal tissue, was selected to validate the proteomics results, but significant patient-to-patient variation obfuscated interpretation so corroborated the contradictory observations made by others. CONCLUSION: Nevertheless, the study has provided significant insights into CC stage progression for further investigation.

Lipids characterization of ultrasound and microwave processed germinated sorghum.

BACKGROUND: Cereal crops and oilseeds provide diverse pool of fatty acids with characteristic properties. Sorghum (Sorghum bicolor (L.) Moench) provides the staple food with serving as main source of energy and protein. Germination of sorghum generally increases the nutritive value of seeds and the effects of germination on lipids composition of seeds vary greatly with processing conditions. Therefore, the current study was conducted to compare the effect of emerging processing techniques such as ultrasound (US) and microwave (MW) on fatty acids composition and oil yield of sorghum seeds before and after germination. METHODS: Initially sorghum grains were soaked with 5% NaOCl (sodium hypochlorite) for surface sterilization. Afterwards, grains were soaked in excess water for 22 h at room temperature and were divided into four portions. The first portion (100 g grains) was subjected to germination without applying any microwave and ultrasonic treatment (T0). Second portion was further divided into four groups (T1, T2, T3, T4) (100 g of each group) and grains were subjected to ultrasonic treatments using two different ultrasonic intensities (US1: 40%; US2: 60%) within range of 0-100% and with two different time durations (tUS1: 5 min; tUS2: 10 min) at constant temperature. Third portion was also divided into four groups (T1, T2, T3, T4) (100 g of each group) and exposed to microwave treatments at two different power levels (MW1: 450 watt; MW2: 700 watt) within the range of 100-900 W for two different time durations (tMW1: 15 s; tMW2: 30s). Similarly, fourth portion was divided into four groups (T1, T2, T3, T4) (100 g of each group). Each group was exposed to both MW (MW1, MW2) (100-900 watt power) & US (US1, US2) (0-100% intensity) treatments at two different time levels (tUS, tMW). Then, germination was carried out and pre-treated raw and pre-treated germinated sorghum grains were analyzed for total oil yield, fatty acid composition and unsaturated fatty acids (Un-SFA)/saturated fatty acids (SFA) ratio by gas chromatography. RESULTS: The results revealed that oil yield in sorghum before and after germination ranged from 6.55 to 7.84% and 6.28 to 7.57%, respectively. All the microwave and ultrasound processed samples showed significant difference in oil yield than the raw sorghum grains. The highest tested yield was 7.84 ± 0.31% when combination of microwave power (700 W) and ultrasound intensity (60%) was applied for 30s and 10 min, respectively. The results further demonstrate that the raw sorghum contained palmitic (13.73 ± 0.10%), palmitoleic (0.43 ± 0.02%), stearic (1.07 ± 0.04%), oleic (37.15 ± 0.10%), linoleic (43.33 ± 0.21%), linolenic (1.55 ± 0.04%), arachidic acid (0.13 ± 0.01%) and eicosenoic acid (0.37 ± 0.02%), respectively. The highest fatty acid percentage for palmitic, stearic and arachidic acid was 13.75 ± 0.07%, 1.11 ± 0.09% and 0.15 ± 0.03% at 60% US intensity for 10 min (T4), respectively. Maximum amount observed was 1.60 ± 0.09% of linolenic acid while amount of eicosenoic acid decreased from 0.37 ± 0.02% to 0.31 ± 0.01% after processing. In case of applying combination of microwave and sonication treatments, the change in eicosenoic acid increased from 0.35 ± 0.02% to 0.40 ± 0.04% while there was no significant change in other fatty acids. The ungerminated sorghum oil possessed 14.93-15.05% and 82.83-83.12% of SFA and Un-SFA, respectively. After germination, percentage of saturated fatty acids increased (16.4-16.55%) while decreased for unsaturated fatty acids (80.13-80.56%) were noted. CONCLUSIONS: The results of the present study conclude that the yield of oil from sorghum grains increased by emerging processing. Fatty acid analysis of sorghum oil suggested that pre-treatment strategies will not affect the quality of the oil with respect to essential fatty acids content. Overall, the composition of saturated fatty acid in germinated grain is improved than ungerminated grains after processing.

Bioinspired chitosan based functionalization of biomedical implant surfaces for enhanced hemocompatibility, antioxidation and anticoagulation potential: an in silico and in vitro study.

Endowing implanted biomaterials with better hemocompatibility, anticoagulation, antioxidant and antiplatelet adhesion is necessary because of their potential to trigger activation of multiple reactive mechanisms including coagulation cascade and potentially causing serious adverse clinical events like late thrombosis. Active ingredients from natural sources including Foeniculum vulgare, Angelica sinensis, and Cinnamomum verum have the ability to inhibit the coagulation cascade and thrombus formation around biomedical implants. These properties are of interest for the development of a novel drug for biomedical implants to potentially solve the current blood clotting and coagulation problems which lead to stent thrombosis. The objective of this study was to incorporate different anticoagulants from natural sources into a degradable matrix of chitosan with varying concentrations ranging from 5% to 15% and a composite containing all three drugs. The presence of anticoagulant constituents was identified using GC-MS. Subsequently, all the compositions were characterized principally by using Fourier transform infrared spectroscopy and scanning electron microscopy while the drug release profile was determined using UV-spectrometry for a 30 days immersion period. The results indicated an initial burst release which was subsequently followed by the sustained release pattern. Compared to heparin loaded chitosan, DPPH and hemolysis tests revealed better blood compatibility of natural drug loaded films. Moreover, the anticoagulation activity of natural drugs was equivalent to the heparin loaded film; however, through docking, the mechanism of inhibition of the coagulation cascade of the novel drug was found to be through blocking the extrinsic pathway. The study suggested that the proposed drug composite expresses an optimum composition which may be a practicable and appropriate candidate for biomedical implant coatings.

Antioxidant potential and oil composition of Callistemon viminalis leaves.

The present study was designed to investigate the antioxidant potential and oil composition of Callistemon viminalis leaves. GC-MS analysis of the n-hexane extract revealed the presence of 40 compounds. Leaves contained appreciable levels of total phenolic contents (0.27-0.85 GAE mg/g) and total flavonoid contents (2.25-7.96 CE mg/g). DPPH radical scavenging IC50 and % inhibition of linoleic acid peroxidation were found to be in the ranges of 28.4-56.2 µg/ml and 40.1-70.2%, respectively. The haemolytic effect of the plant leaves was found in the range of 1.79-4.95%. The antioxidant activity of extracts was also studied using sunflower oil as an oxidative substrate and found that it stabilized the oil. The correlation between the results of different antioxidant assays and oxidation parameters of oil indicated that leaves' methanolic extract, exhibiting higher TPC and TFC and scavenging power, was also more potent for enhancing the oxidative stability of sunflower oil.

Thalassaemia Patients with Polymorphism of COL1A1 Sp1 are at Greater Risk of Spine Degenerative Changes.

OBJECTIVE: To explore the association of the COL1A1 Sp1 polymorphism with decreased bone density and spinal changes in ß-thalassaemia patients. STUDY DESIGN: Cross-sectional, comparative study. Place and Duration of the Study: University of Health Sciences, Lahore, from May 2020 to June 2021. METHODOLOGY: A total of 110 participants (55patients and 55 controls) of either gender, with ages ranging from 18-40 years were enrolled in the study. Bone density parameters including T-score, Z-score, bone-transmission time (BTT), and amplitude-dependent speed of sound (ADSOS) were assessed by ultrasound bone profiler. Lumbar spine radiographs were collected from patients and assessed for spine changes. Genotype analysis was done by HRM-PCR. Data were analysed using SPSS version 23. RESULTS: All bone density parameters were significantly lower in ß-thalassaemai patients (p<0.001). Spine degenerative changes were more obvious in patients with age <25 years (p=0.04). Loss of lumbar lordosis was seen in 74.5% of the patients. The frequency of mutant allele (ss) was 7.3% while heterozygous (Ss) frequency was found to be 33.6%. The polymorphism showed significant association with T-scores (p=0.03) and ADSOS (p=0.02) in patients. Radiographic grades were higher in osteopenic and osteoporotic patients (p=0.04). CONCLUSION: The association of polymorphism with decreased bone density in ß-thalassaemia patients revealed the potential role of genetics in bone changes and related disorders. KEY WORDS: Bone Density, COL1A1 Polymorphism, Osteoporosis, Thalassaemia.

Evolutionary perspective of drug eluting stents: from thick polymer to polymer free approach.

BACKGROUND: Introduction of Bare Metal Stents (BMS) was itself a revolutionary step in the history of the medical industry; however, Drug Eluting Stents (DES) maintained its superiority over BMS in every aspect from restenosis rate to late lumen loss. The reason behind the magnanimous position of the DES in the stent market is the degree of improvement with which it evolves. New and better stents come into the market every year, surpassing their predecessors by many folds. LITERATURE REVIEW: This review paper discusses the journey of DES with supporting clinical trials in detail. In the first generation, there were stainless-steel stents with thicker coatings. Although they had superior results compared to BMS, there was still room for improvement. Afterward came the second-generation stents, which had superior metal platforms with thinner struts and thin coatings. The drugs were also changed from Paclitaxel and Sirolimus to Zotrolimus and Everolimus. These stents performed best; however, there was an issue of permanent coating, which remained intact over the stent surface after complete drug elution and started to cause issues in longer-term studies. Hence, an improved version of DES was introduced to these permanent coatings called the third generation of drug eluting stents, which initially utilized biodegradable polymer and ultimately moved towards polymer free drug coatings. This generation has introduced a unique amalgam of technologies to achieve its polymer free coatings; however, researchers have numerous prospects of growth in this field. This review paper highlights the major coups of stent technology evolution from BMS to DES, from thick polymeric coatings to thin coatings and from durable polymers to polymer free DES. CONCLUSION: In conclusion, though the medical industry promptly accepted BMS as the best treatment option for cardiovascular diseases; however, DES has provided even better results than BMS. In DES, the first and second generation has ruled the technology for many years and are still on the shelves. Still, the issues aroused due to durable polymer shifted the attention towards biodegradable drug eluting stents, the third generation growing rapidly. But the scientific community has not restricted themselves and is investigating bioresorbable stents that completely eliminate the polymer intervention in drug eluting stent technology.

Recent progress in the design, synthesis and applications of chiral metal-organic frameworks.

Chiral Metal-Organic Frameworks (CMOFs) are unique crystalline and porous class of materials which is composed of organic linkers and metal ions. CMOFs surpass traditional organic and inorganic porous materials because of their tunable shape, size, functional diversity, and selectivity. Specific applications of CMOFs may be exploited by introducing desired functional groups. CMOFs have chiral recognition abilities, making them unique for chiral compound synthesis and separation. The CMOFs can be synthesized through different approaches. Two main approaches have been discussed, i.e., direct and indirect synthesis. Synthetic strategies play an essential role in getting desired properties in MOFs. CMOFs find potential applications in adsorption, asymmetric catalysis, luminescence, degradation, and enantioselective separation. The MOFs' porosity, stability, and reusability make them an attractive material for these applications. The plethora of applications of CMOFs have motivated chemists to synthesize novel MOFs and number of MOFs have been ever-escalating. Herein, the synthetic methods of CMOFs and their various applications have been discussed.

Diagnostic and Therapeutic Potential of Circulating-Free DNA and Cell-Free RNA in Cancer Management.

Over time, molecular biology and genomics techniques have been developed to speed up the early diagnosis and clinical management of cancer. These therapies are often most effective when administered to the subset of malignancies harboring the target identified by molecular testing. Important advances in applying molecular testing involve circulating-free DNA (cfDNA)- and cell-free RNA (cfRNA)-based liquid biopsies for the diagnosis, prognosis, prediction, and treatment of cancer. Both cfDNA and cfRNA are sensitive and specific biomarkers for cancer detection, which have been clinically proven through multiple randomized and prospective trials. These help in cancer management based on the noninvasive evaluation of size, quantity, and point mutations, as well as copy number alterations at the tumor site. Moreover, personalized detection of ctDNA helps in adjuvant therapeutics and predicts the chances of recurrence of cancer and resistance to cancer therapy. Despite the controversial diagnostic values of cfDNA and cfRNA, many clinical trials have been completed, and the Food and Drug Administration has approved many multigene assays to detect genetic alterations in the cfDNA of cancer patients. In this review, we underpin the recent advances in the physiological roles of cfDNA and cfRNA, as well as their roles in cancer detection by highlighting recent clinical trials and their roles as prognostic and predictive markers in cancer management.

Body Mass Index and Other Risk Factors Effects on Colon Cancer Prognosis in Pakistan.

Introduction: Asian developing countries share the burden of colorectal cancer (CRC) with rising mortality rates. This prospective study aims to apprehend the clinical relevance of age, gender, lifestyle choices (dietary habits and addiction) and body mass index (BMI) to the occurrence and progression of colon cancer (CC). Methods: A cohort of non-cancer (NC) and CC patients of South-Central Asian origin registered for screening colonoscopy or surgery at Shaukat Khanum Memorial Cancer Hospital and Research Centre (SKMCH and RC), Lahore, Pakistan, from 2015 to 2020 was identified. BMI (Kg/m2) was classified according to the World Health Organization criteria as underweight (<18.5 Kg/m2), normal weight (18.5-24.9 Kg/m2) and overweight (≥25 Kg/m2). Results: Among 236 participants, 99 (41.9%) belonged to the NC group, and 137 (58.1 %) participants had CC Overall, participants included 74 women and 162 men aged 20-85 years (mean ± SD; 49.9 ± 14.9). Notably, 46.0% of cancer patients had a family history of cancer. There was a direct relationship between CC with abnormal BMI (underweight and overweight), positive smoking history and positive family history of cancer. Conclusion: Being underweight or overweight is a potential risk factor for CC patients. The overall survival in patients with CC is clinically associated with lifestyle choices before CC diagnosis. A balanced diet, walking and other forms of exercise should be strongly recommended to the community and those undergoing screening colonoscopy.

Investigating the structural properties and in vitro digestion of rice flours.

The physicochemical properties, swelling power, solubility, and digestibility of flour from four rice varieties (black, brown, white, and waxy rice flour) were analyzed. The results showed that the black and brown rice had high-amylose percentage (21.8% and 20.5%), a relatively low percentage of starch content (68.1% and 79.1%), and lower swelling power (6.6% and 7.6%) and solubility (13.5% and 15.7%), respectively. Waxy rice flour attributed to lower gelatinization temperatures and higher enthalpy values. Meanwhile, the brown, black, and white rice showed higher gelatinization temperature and lower enthalpy value. The black and brown rice flour exhibited lower pasting and viscosity values as compared to waxy rice flour. The results showed that all rice flour had an A-type X-ray diffraction pattern, and after cooking all rice flour showed V-type polymorphs except waxy rice flour. Brown and black rice flour after cooking have lower digestion rate than white rice and waxy rice flour, probably due to its lower expansion and solubility rates, and higher gelatinization temperature.

Honey and cancer: A mechanistic review.

BACKGROUND: Globally, cancer ranks among the most common causes of death. Multiple experimental and clinical studies have investigated anticancer effects of honey with promising results. This study focused on potential background mechanisms of this effect. METHODS: The current literature was reviewed for potential anticancer pathways which are suggested for honey and its ingredients. RESULTS: Flavonoids (kaempferol, catechin, and quercetin) and phenolic acids (caffeic acid and gallic acid) are the most important ingredients of honey with known anti-cancer activity. The main suggested mechanisms for anti-cancer activity of honey and its ingredients are antioxidant, apoptotic, tumor necrosis factor inhibiting, antiproliferative, immunomodulatory, anti-inflammatory and estrogenic effects. CONCLUSION: This review collates the current scientific understanding on the mechanism of anti-cancer activity of honey.

The nanoparticulate Quillaja saponin KGI exerts anti-proliferative effects by down-regulation of cell cycle molecules in U937 and HL-60 human leukemia cells.

Cancer cells are characterized by uncontrolled replication involving loss of control of cyclin dependent kinases (CDKs) and cyclins, and by abolished differentiation. In this study we introduce KGI, which is a nanoparticle with a Quillaja saponin as an active molecule. By the use of RNA array analysis and confirmation at the protein level, we show that KGI affects myeloid leukemia cells (in particular, the U937 monoblast cancer cell) by the following mechanisms: (A) ceasing cell replication via proteasome degradation, (B) down-regulation of key molecules at check points between G1/S and G2/M phases, (C) reduction of thymidine kinase activity, followed by (D) exit to differentiation and production of interleukin-8 (IL-8), eventually leading to apoptosis. Leukemia cell lines (U937 and HL-60 cells) were exposed to KGI for 8 h, after which the drug was removed. The cancer cells did not revert to replication over the following 10 days. Thus our findings suggest that the nanoparticle KGI inhibits proliferation and promotes differentiation in leukemic cells by interfering with the cell cycle process.

Phenotype-based drug screening in primary ovarian carcinoma cultures identifies intracellular iron depletion as a promising strategy for cancer treatment.

Primary cultures of patient tumor cells (PCPTC) have been used for prediction of diagnosis-specific activity and individual patient response to anticancer drugs, but have not been utilized as a model for identification of novel drugs in high throughput screening. In the present study, ovarian carcinoma cells from three patients were tested in response to a library of 3000 chemically diverse compounds. Eight hits were retrieved after counter screening using normal epithelial cells, and one of the two structurally related hit compounds was selected for further preclinical evaluation. This compound, designated VLX 50, demonstrated a broad spectrum of activity when tested in a panel of PCPTCs representing different forms of leukemia and solid tumors and displayed a high tumor to normal cell activity. VLX 50 induced delayed cell death with some features of classical apoptosis. Significant in vivo activity was confirmed on primary cultures of human ovarian carcinoma cells in mice using the hollow fiber model. Mechanistic exploration was performed using gene expression analysis of drug exposed tumor cells to generate a drug-specific signature. This query signature was analyzed using the Gene Set Enrichment Analysis and the Connectivity Map database. Strong connections to hypoxia inducible factor 1 and iron chelators were retrieved. The mechanistic hypothesis of intracellular iron depletion leading to hypoxia signaling was confirmed by a series of experiments. The results indicate the feasibility of using PCPTC for cancer drug screening and that intracellular iron depletion could be a potentially important strategy for cancer therapy.

The cytotoxic agents NSC-95397, brefeldin A, bortezomib and sanguinarine induce apoptosis in neuroendocrine tumors in vitro.

UNLABELLED: The aim of this study was to investigate the apoptosis resulting from NSC 95397, brefeldin A, bortezomib and sanguinarine in neuroendocrine tumor cell lines. MATERIALS AND METHODS: A multiparametric high-content screening assay for measurement of apoptosis was used. The human pancreatic carcinoid cell line, BON-1, human typical bronchial carcinoid cell line NCI-H727 and the human atypical bronchial carcinoid cell line NCI-H720 were tested. After incubation with cytotoxic drugs, the DNA-binding dye Hoechst 33342, fluorescein-tagged probes that covalently bind active caspase-3 and chloromethyl-X-rosamine to detect mitochondrial membrane potential were added. Image acquisition and quantitative measurement of fluorescence was performed using automated image capture and analysis instrument ArrayScan. In addition, nuclear morphology was examined on microscopic slides stained with May-Grunewald-Giemsa. RESULTS: A time- and dose-dependent activation of caspase-3 and increase in nuclear fragmentation and condensation were observed for the drugs using a multiparametric apoptosis assay. These results were confirmed with nuclear morphological examination on microscopic slides. CONCLUSION: NSC 95397, brefeldin A, bortezomib and sanguinarine induced caspase-3 activation with modest changes in nuclear morphology.

Identification of AKN-032, a novel 2-aminopyrazine tyrosine kinase inhibitor, with significant preclinical activity in acute myeloid leukemia.

Aberrant signal transduction by mutant or overexpressed protein kinases has emerged as a promising target for treatment of acute myeloid leukemia (AML). We here present a novel low molecular weight kinase inhibitor, AKN-032, targeting the FMS-like tyrosine kinase 3 (FLT3) and discovered in a new type of screening funnel combining the target therapy approach with sequential cellular screens. AKN-032 was identified among 150 selected hits from three different high throughput kinase screens. Further characterization showed inhibitory activity on FLT3 enzyme with an IC(50) of 70 nM. Western blot analysis revealed reduced autophosphorylation of the FLT3-receptor in AML cell line MV4-11 cells after exposure to AKN-032. Flow cytometry disclosed cytotoxic activity against MV4-11, but not against non-malignant 3T3-L1 fibroblast cells. Using a fluorometric microculture cytotoxicity assay, AKN-032 was tested against 15 cell lines and displayed a potent cytotoxic activity in AML cell lines MV4-11 (IC(50)=0.4 µM) and Kasumi-1 (IC(50)=2.3 µM). AKN-032 was also highly cytotoxic in tumor cells from AML patients in vitro. Furthermore, AKN-032 demonstrated significant antileukemic effect in a relatively resistant in vivo hollow fiber mouse model. No major toxicity was observed in the animals. In conclusion, AKN-032 is a promising new kinase inhibitor with significant in vivo and in vitro activity in AML. Results from the hollow fiber mouse assay suggest a favorable toxicity profile. Future studies will focus on pharmacokinetic properties, toxicity as well as further clarifying the mechanisms of action of AKN-032 in AML.

Combination analyses of anti-cancer drugs on human neuroendocrine tumor cell lines.

PURPOSE: There is a large need for better pharmacological treatment of neuroendocrine tumors. The aim of this study was to investigate and quantify the cytotoxic potentiating effects resulting from a combination of five substances, NSC 95397, emetine, CGP-74514A hydrochloride, Brefeldin A and sanguinarine chloride, chosen from a previous screening of 1,280 pharmacologically active agents on neuroendocrine tumor cells, with standard cytotoxic agents currently used in the treatment of neuroendocrine tumors. METHOD: The human pancreatic carcinoid cell line BON-1, human typical bronchial carcinoid cell line NCI-H727 and the human atypical bronchial carcinoid cell line NCI-H720 were used. Combinations between doxorubicin, etoposide, oxaliplatin, docetaxel, and each one of the five agents were studied and simultaneous exposures were explored using the median-effect method. RESULTS: Most of the combinations of NSC-95397 and emetine with doxorubicin, etoposide, docetaxel, and oxaliplatin showed synergism, and their remaining combinations were additive. Almost all of the CGP-74514A hydrochloride interactions were additive, while brefeldin A and sanguinarine displayed less synergy but more additive and antagonistic interactions in combination with the standard drugs. CONCLUSION: The synergistic and additive interactions make NSC-95397, emetine, and CGP-74514A hydrochloride potential candidates for incorporation into combination chemotherapy regimens and these drugs might be the suitable candidates for further clinical studies in patients with bronchial carcinoids and pancreatic endocrine tumors.

Cytotoxic effect in vivo and in vitro of CHS 828 on human myeloma cell lines.

CHS 828 is a pyridyl cyanoguanidine with promising antitumor activity both in vitro and in vivo, and has previously been found especially active against tumor cells obtained from patients with B cell chronic lymphocytic leukemia. In the present study the cytotoxic effect in vitro of CHS 828 was investigated on a panel of 10 human myeloma cell lines using the fluorometric microculture cytotoxicity assay. CHS 828 induced a concentration-dependent, but variable decrease in tumor cell survival in the cell line panel with inhibitory concentrations 50% (IC50) in the range 0.01-0.3 microM. These concentrations are below those achievable in vivo. There was no detectable dependence on P-glycoprotein-mediated or GSH-associated drug resistance and the drug showed low to moderate cross-resistance with standard drugs, including melphalan, vincristine and doxorubicin. Furthermore, sensitivity to CHS 828 showed no apparent relationship to growth factor dependence, tumor progression or phenotypic variability. CHS 828 was also tested in vivo using a hollow fiber model in rats with three of the cell lines. The results indicate a high cytotoxic activity of CHS 828. Overall, the results show a high cytotoxic activity of CHS 828 in the myeloma models, which might warrant its further development against myeloma.