Growth Of Viable Cells Research Articles

CDK4/6 inhibitors sensitize gammaherpesvirus-infected tumor cells to T-cell killing by enhancing expression of immune surface molecules

BackgroundThe two oncogenic human gammaherpesviruses, Kaposi sarcoma-associated herpesvirus (KSHV) and Epstein–Barr virus (EBV), both downregulate immune surface molecules, such as MHC-I, ICAM-1, and B7-2, enabling them to evade T-cell and natural killer cell immunity. Both also either encode for human cyclin homologues or promote cellular cyclin activity, and this has been shown to be important for proliferation and survival of gammaherpesvirus-induced tumors. CDK4/6 inhibitors, which are approved for certain breast cancers, have been shown to enhance expression of MHC-I in cell lines and murine models of breast cancer, and this was attributed to activation of interferons by endogenous retrovirus elements. However, it was not known if this would occur in gammaherpesvirus-induced tumors in which interferons are already activated.MethodsMultiple KSHV/EBV-infected cell lines were treated with CDK4/6 inhibitors. The growth of viable cells and expression of surface markers was assessed. T cell activation stimulated by the treated cells was assayed by a T-cell activation bioassay. Both viral and host gene expression was surveyed using RT-qPCR.ResultsThree CDK4/6 inhibitors, abemaciclib, palbociclib, and ribociclib, inhibited cell growth in KSHV-induced primary effusion lymphoma (PEL) and EBV positive Burkitt’s lymphoma (BL) cell lines, and KSHV-infected human umbilical vein endothelial cells (HUVECs). Moreover, CDK4/6 inhibitors increased mRNA and surface expression of MHC-I in all three and prevented downregulation of MHC-I surface expression during lytic replication in KSHV-infected cells. CDK4/6 inhibitors also variably increased mRNA and surface expression of ICAM-1 and B7-2 in the tested lines. Abemaciclib also significantly enhanced T-cell activation induced by treated PEL and BL cells. Certain gammaherpesvirus genes as well as endogenous retrovirus (ERV) 3–1 genes were enhanced by CDK4/6 inhibitors in most PEL and BL lines and this enhancement was associated with expression of gamma interferon-induced genes including MHC-I.ConclusionsThese observations provide evidence that CDK4/6 inhibitors can induce expression of surface immune markers MHC-I, B7-2, and ICAM-1 in gammaherpesvirus-infected cell lines and induce virus-specific immunity. They can thus thwart virus-induced immune evasion. These effects, along with their direct effects on KSHV- or EBV-induced tumors, provide a rational for the clinical testing of these drugs in these tumors.

Kefir Culture-Mediated Fermentation to Improve Phenolic-Linked Antioxidant, Anti-Hyperglycemic and Human Gut Health Benefits in Sprouted Food Barley

The bioprocessing strategy is an effective approach to improve bioavailability and stability of bioactive compounds for designing functional foods and ingredients. In this study, food barley was bio-transformed to improve functional bioactives by sprouting, coupled with beneficial lactic acid bacteria (LAB)-based fermentation. Dairy Kefir culture with mixed beneficial LAB strains was targeted to ferment aqueous slurries of sprouted hulless food barley flour (unpigmented, purple, and black barley) for 72 h, and modulation of phenolic-linked antioxidant and anti-hyperglycemic functionalities were evaluated using in vitro assay models. The biochemical parameters analyzed were total soluble phenolic (TSP) content, profile of phenolic compounds, total antioxidant activity, and anti-hyperglycemic property-relevant α-amylase and α-glucosidase enzyme inhibitory activities. Furthermore, human gut health benefits of relevant properties of fermented slurries of barley flour were also evaluated based on growth of Kefir culture and subsequent determination of anti-bacterial potential against pathogenic human ulcer causing bacteria Helicobacter pylori. Kefir culture-mediated fermentation of 48-h sprouted barley flours improved the TSP content and associated antioxidant and anti-hyperglycemic functionalities. Additionally, anti-bacterial potential against H. pylori and sustaining active growth of viable LAB cells above the minimum level required for probiotic activity were also observed in fermented food barley flour slurries.

Inhibitory activity of traditional plants against Mycobacterium smegmatis and their action on Filamenting temperature sensitive mutant Z (FtsZ)-A cell division protein.

The study was designed to assess whether plant extracts / phytochemical (D-Pinitol) synergistically combine with antituberculosis drugs and act on Mycobacterium smegmatis (M. smegmatis) as well as assess their mode of action on Mycobacterium tuberculosis (M.tb) Filamenting temperature sensitive mutant Z (FtsZ) protein. Resazurin microtitre plate assay (Checker board) was performed to analyze the activity of plant extracts against M. smegmatis. Synergistic behaviour of plant extracts / D-Pinitol with Isoniazid (INH) and Rifampicin (RIF) were determined by time-kill and checker board assays. Elongation of M. smegmatis cells due to this treatment was determined by light microscopy. The effect of Hexane methanol extract (HXM) plant extracts on cell viability was determined using PI/SYTO9 dual dye reporter Live/Dead assay. Action of HXM plant extracts / D-Pinitol on inhibition of FtsZ protein was done using Guanosine triphosphatase (GTPase) light scattering assay and quantitative Polymerase Chain Reaction (qPCR). The Hexane-methanolic plant extract of Acacia nilotica, Aegle marmelos and Glycyrrhiza glabra showed antimycobacterial activity at 1.56 ± 0.03, 1.32 ± 0.02 and 1.25 ± 0.03 mg/mL respectively and that of INH and RIF were 4.00 ± 0.06 μg/mL and 2.00 ± 0.04 μg/mL respectively. These plant extracts and major phytochemical exudate D-Pinitol was found to act synergistically with antimycobacterial drugs INH and RIF with an FIC index ~ 0.20. Time-Kill kinetics studies indicate that, these plant extracts were bacteriostatic in nature. D-Pinitol in conjunction with INH and RIF exhibited a 2 Log reduction in the growth of viable cells compared to untreated. Attempt to elucidate their mode of action through phenotypic analysis indicated that these plant extracts and D-Pinitol was found to interfere in cell division there by leading to an abnormal elongated cellular morphology. HXM extracts and D-Pinitol synergistically combined with the first line tuberculosis drugs, INH and RIF, to act on M. smegmatis. The increase in the length of M. smegmatis cells on treatment with D-Pinitol and HXM extract of the plants indicated that they hinder the cell division mechanism thereby leading to a filamentous phenotype, and finally leading to cell death. In addition, the integrity of the bacterial cell membrane is also altered causing cell death. Further gene expression analysis showed that these plant extracts and D-Pinitol hampers with function of FtsZ protein which was confirmed through in vitro inhibition of FtsZ-GTPase enzymatic activity.

Generation of HeLa spheroids in Ca-alginate-PEG microbeads using flicking technique as an improved three-dimensional cell culture system

In a conventional three-dimensional (3D) cell culture system based on microencapsulation, the calcium alginate microcapsules tend to rupture within 7 days of culture, causing unwanted cell leakage. The microencapsulation based on flicking model of calcium-alginate-polyethylene glycol (Ca-alg-PEG) was proposed to rectify this problem. An analytical model to simulate the flicking process based on the deflection of a needle cantilever was also successfully developed and used to predict the size of the microbeads. The size of the microbeads were ranged from 300 to 500 μm and it is controlled by varying the liquid flow rate from 4.8 to 366 μL/min and flicking speed from 70 to 120 rpm. Under a flicking force of 0.58 N, uniform sized and spherical shaped Ca-alginate-PEG microbeads were produced at a liquid flow rate of 40 μL/min and a flicking rate of 100 rpm. The microbeads were characterized by Field Emission Scanning Microscopy, Atomic Force Microscopy, Raman Spectroscopy and Nanoindentation, and the results indicated improved bio-physical properties of the ca-alginate microbeads after added with PEG. The cell viability test demonstrated that Ca-alginate-PEG microbeads were able to support the growth of viable HeLa cells into spheroids. Resilient calcium-alginate-polyethylene glycol (Ca-alg-PEG) microbeads were found to be able to last up to 15 days before rupturing and greatly reduced the cell leakage problem.

Medical-grade polycaprolactone scaffolds made by melt electrospinning writing for oral bone regeneration \u2013 a pilot study in vitro

BackgroundThe spectrum of indications for the use of membranes and scaffolds in the field of oral and maxillofacial surgery includes, amongst others, guided bone regeneration (GBR). Currently available membrane systems face certain disadvantages such as difficult clinical handling, inconsistent degradation, undirected cell growth and a lack of stability that often complicate their application. Therefore, new membranes which can overcome these issues are of great interest in this field.MethodsIn this pilot study, we investigated polycaprolactone (PCL) scaffolds intended to enhance oral wound healing by means of melt electrospinning writing (MEW), which allowed for three-dimensional (3D) printing of micron scale fibers and very exact fiber placement. A singular set of box-shaped scaffolds of different sizes consisting of medical-grade PCL was examined and the scaffolds’ morphology was evaluated via scanning electron microscopy (SEM). Each prototype sample with box sizes of 225 μm, 300 μm, 375 μm, 450 μm and 500 μm was assessed for cytotoxicity and cell growth by seeding each scaffold with human osteoblast-like cell line MG63.ResultsAll scaffolds demonstrated good cytocompatibility according to cell viability, protein concentration, and cell number. SEM analysis revealed an exact fiber placement of the MEW scaffolds and the growth of viable MG63 cells on them. For the examined box-shaped scaffolds with pore sizes between 225 μm and 500 μm, a preferred box size for initial osteoblast attachment could not be found.ConclusionsThese well-defined 3D scaffolds consisting of medical-grade materials optimized for cell attachment and cell growth hold the key to a promising new approach in GBR in oral and maxillofacial surgery.

Nanoparticle-Based Imaging of Clinical Transplant Populations Encapsulated in Protective Polymer Matrices.

A recent clinical trial proves that autologous olfactory mucosal cell (OMC) transplantation improves locomotion in dogs with naturally occurring spinal injuries comparable to human lesions. However, not all dogs respond to the treatment, likely due to the transplantation procedures involving injections of cell suspensions that are associated with cell death, uneven cell distribution, and cell washout. Encapsulating cells in protective hydrogel matrices offers a tissue engineering solution to safely achieve 3D growth of viable transplant cells for implantation into injury sites, to improve regenerative outcomes. It is shown for the first time that canine OMCs (cOMCs) can be propagated with high viability in 3D collagen matrices. Further, a method to incorporate cOMCs pre-labeled with clinical-grade iron oxide nanoparticles into the constructs is described. Intraconstruct labeled cells are visualized using magnetic resonance imaging, offering substantial promise for in vivo tracking of cOMCs delivered in protective matrices.

English

This study evaluates the antimicrobial activities of a multi-species probiotic ingredient derived from the African opaque sorghum beer during its propagation in a starchy career model. The aim was to establish the optimum growth conditions that warrant the optimum antimicrobial activities in the product. The antimicrobial activities were tested against Gram-positive bacteria (Staphylococcus aureus ATCC 27844, S. aureus MR 825), Gram-negative bacteria (Escherichia coli ATCC 25922, E. coli O157:H7 ATCC 700728, Salmonella typhi R 30951401, Klebsiella pneumoniae ATCC 35657), as well as against yeast (Candida albicans MHMR), using agar disc diffusion method. Also, the growth of viable cells and physicochemical parameters during the propagation were monitored. The results showed that the pH and dry matter content of the probiotic ingredient decrease significantly (p˂0.05) during the propagation whereas the lactic acid, the titratable acidity, lactic acid bacteria (LAB), yeasts and moulds counts increase significantly (p˂0.05). From 0 to 12 h, the product failed to inhibit the growth of all indicator strains. From 24 h and onward, the probiotic career inhibited all indicator strains except for K. pneumoniae (ATCC 35657) which could not be inhibited. Clearly, our study showed that 36 h of the propagation were sufficient to generate a probiotic ingredient with optimum antimicrobial activities. Key words: Probiotic, antibiotic, opaque beer, antimicrobial activity.

Anti-Neoplastic Activity of Litchi chinensis Leaf Extract Against Ehrlich’s Ascites Carcinoma in Mice

The anti-carcinogenic activity of Litchi chinensis leaf extract (LCLE) was investigated against Ehrlich’s ascites carcinoma (EAC) in mice. LCLE inhibited the growth and metabolic activity of viable EAC cells in both in vivo and in vitro studies. Treatment with LCLE showed characteristic features of apoptosis. Flow cytometric analysis showed appreciable number of cells in early and late apoptotic stages. LCLE arrested the EAC cell populations in the G2-M phase of cell cycle. LCLE induced apoptosis was found to be mediated through mitochondrial intrinsic pathway involving the release of cytochrome c into the cytosol and activation of caspase-9 and caspase-3.

굴(Crassostrea gigas) 패각 분말 첨가에 의한 배추김치의 식품학적 품질 변화

This study investigated the effects of adding oyster shell powder (OSP) from Crassostrea gigas on the food quality of Chinese cabbage Kimchi (CCK). We monitored the changes in microbial levels, pH, acidity and sensory evaluation during the fermentation of CCK treated with various contents of OSP. The microbial assay showed that adding OSP to CCK inhibited the growth of viable cells, total coliforms, and lactic acid bacteria, with the greatest growth inhibition against lactic acid bacteria over the fermentation period. After fermentation for 18 days, the lactic acid bacterial counts in CCK treated with OSP (0.3%, 0.5% and 1%) were at least 1 log CFU/g lower than those of control CCK. In addition, the pH and acidity of CCK treated with OSP were lower than in control CCK over the fermentation period. The overall sensory evaluation of CCK with 0.3% OSP was better than that of control CCK after fermentation for 24 days. In conclusion, OSP treatment, especially 0.3% OSP, enhances the food quality and extends the self-life of CCK, while minimizing the detrimental effects on its sensory characteristics.

The Inhibitory Effect of Ecklonia cava and Eisenia bicyclis Ethanol Extract on Histamine in Mackerel

Bo-Ram Kim , Koth-Bong-Woo-Ri Kim , Min-Ji Kim , Dong-Hyun Kim , Seul-A Jung , Bo-Kyeong Kang , Si-Woo Bark , Won-Min Pak , Hong-Min Park , Sung-Mee Lim , Young-Je Cho and Dong-Hyun Ahn . Microbiol. Biotechnol. Lett. 2014;42:93-8. https://doi.org/10.4014/kjmb.1310.10006

찹쌀 첨가에 따른 전통발효주의 품질 특성

찹쌀 첨가량(0, 10, 15, 20%)에 따른 술덧을 제조하여 발효 10일 동안의 알코올 함량, pH 및 총산, 총당 함량, 환원당 함량, 유기산 함량, 유리당 함량 등의 이화학적 특성과 미생물 변화를 알아보고 발효주의 관능적 특성을 조사하였다. 알코올 함량은 발효 1일째 9.1~9.7%에서 발효 종료일 10일째 15.8~17.1%로 증가하였으나 각 실험군 간에 유의적인 차이는 없었다. 총균, 유산균, 효모수는 알코올 생성속도와 비례하여 발효초기에는 증가하면서 발효후기에는 감소하는 경향을 나타내었다. pH는 담금 직후 5.30~5.46에서 발효1일째 3.89~3.91로 감소한 후, 다시 증가하기 시작하여 발효 종료일인 10일째 모두 4.5 수준이었다. 총산 함량은 발효 3일째 0.14~0.16%에서 발효 5일째 0.18~0.19%로 증가하였다. 이는 지속적인 유산균의 생장으로 인한 젖산의 누적과 효모에 의한 산 생성으로 발효 3~5일째 총산 함량도 증가한 것으로 추측된다. 총효모수는 2단 담금 이후 유의적으로 증가하였다가 발효 과정을 거치면서 발효 종료일(발효 10일째)에 유의적으로 점차 감소하였다. 환원당은 발효 1일째 1.1~1.2%에서 발효 2일째 0.9~1.1%로 유의적으로 감소하였다가 알코올 발효보다 전분분해에 의한 당 생성 속도가 증가하여 발효 종료일인 10일째 2.8~3.3%로 증가하였다. 총당 함량은 발효 초기 당분이 술덧 중에 생육하는 효모나 유산균 등의 발효기질로 이용하여 발효 종료일 10일째에는 감소하였고, 찹쌀 15%첨가군이 유의적으로 가장 높았다. 당화효소인 amylase에 의해 전분질의 분해로 포도당이 비교적 많이 생성되어 술덧의 주요 유리당이었으며, 유기산에서는 succinic acid 함량이 가장 높았다. 발효주의 관능평가에서 찹쌀 15%첨가군은 전반적인 기호도(4.8점)와 목넘김(4.3점)에서 유의적으로 가장 높게 평가되었다. 묘사분석에서는 찹쌀 15%첨가군이 단맛 강도가 8.6으로 유의적으로 가장 높았으며, 이는 찹쌀 15%첨가군의 높은 총당 함량과 관계가 있는 것으로 판단된다.

UVB-irradiated apoptotic cells induce accelerated growth of co-implanted viable tumor cells in immune competent mice

The presence of a solid tumor is the result of a complex balance between rejection, tolerance and regeneration in which the interactions of tumor cells with cells of the host immune system contribute strongly to the final outcome. Here we report on a model where lethally UVB-irradiated cells cause accelerated growth of viable tumor cells in vitro and in allogeneic immune competent mice. UVB-irradiated tumor cells alone did not form tumors and failed to induce tolerance for a second challenge with the same allogeneic tumor. Our data show an important role for dying cells in promoting accelerated tumor cell growth of a small number of viable tumor cells in a large inoculum of UVB-irradiated tumor cells. This occurs when viable and dying/dead tumor cells are in close proximity, suggesting that mobile factors contribute to growth promotion. The anti-inflammatory and growth promoting properties of apoptotic cells are based on several independent effects. UVB-irradiated apoptotic cells directly release a growth promoting activity and clearance by macrophages of apoptotic cells is accompanied by the secretion of IL10, TGFß, and PGE2. Growth promotion is even observed with dying heterologous cells implying a conserved mechanism. Future experiments should focus on the effects of dying tumor cells generated in vivo on the outgrowth of surviving tumor cells which is prone to have implications for cancer therapy.

PMP 모델을 활용한 시판 Salad의 Short-term Temperature Abuse 시 미생물학적 유통기한 예측에의 적용성 검토

The aim of this study was to investigate the growth of aerobic bacteria in fresh-cut salad during short-term temperature abuse (temperature for 1, 2, and 3 h) for 72 h and to develop predictive models for the growth of total viable cells (TVC) based on Predictive food microbiology (PFM). The tool that was used, Pathogen Modeling program (PMP 7.0), predicts the growth of Aeromonas hydrophila (broth Culture, aerobic) at pH 5.6, NaCl 2.5%, and sodium nitrite 150 ppm for 72 h. Linear models through linear regression analysis; DMFit program were created based on the results obtained at 5, 10, 20, and for 72 h ( >0.9). Secondary models for the growth rate and lag time, as a function of storage temperature, were developed using the polynomial model. The initial contamination level of fresh-cut salad was 5.6 log CFU/mL of TVC during 72 h storage, and the growth rate of TVC was shown to be 0.020~1.083 CFU/mL/h ( >0.9). Also, the growth tendency of TVC was similar to that of PMP (grow rate: 0.017~0.235 CFU/mL/h; $r^2

Use of starter cultures of Bacillus and Staphylococcus in the controlled fermentation of Lanhouin, a traditional fish-based condiment from West Africa

Strains of Bacillus subtilis, Bacillus licheniformis and Staphylococcus xylosus previously isolated during the spontaneous fermentation of Lanhouin were tested singly and in combination for their ability to ferment pasteurized fish flesh to produce a new type ofLanhouin, a traditional condiment product. The growth of viable cells during the fermentation was monitored according to standard microbiological methods. Changes in pH, moisture and total volatile nitrogen (TVN) were determined according to Ababouch (1995) and AOAC (1995). The results revealed that both B. subtilis and B. licheniformis used as single or mixed cultures showed similar growth potential with a bacterial load of 8.0 Log (cfu/g) and pH value ranging between 6.8 and 6.9 after 72 h of fermentation. S. xylosusused as single starter culture also showed a good growth potential with however, a higher level of microbial load, 12.5 Log cfu/g and a pH value of 7.2. Lanhouin produced with the mixture culture of B. subtilis, B. licheniformis and S. xylosus gave a sauce with a desirable taste and aroma preferred by 100% of the panellists. Key words: Fish, starter, inoculation, Bacillus licheniformis, Bacillus subtilis,Staphylococcus xylosus.

Bacterial biofilms resist oxidising agents due to the presence of organic matter

This study evaluated the susceptibility of planktonic and biofilm cells of Staphylococcus spp. (n = 87), Klebsiella spp. (n = 30), and Escherichia coli (n = 74) isolates originating from food contact surfaces of milk and meat processing plants to benzalkonium chloride (BAC), sodium hypochlorite (NaClO), chloramine B (CAB), and peracetic acid (PAA). Bacterial growth and reduction of viable cells in the presence of disinfectants were determined in tryptone soya broth (TSB) and water, respectively. Biofilm positive isolates (n = 73) were tested for the presence of selected qac genes. Unlike BAC, chlorine‑based disinfectants and PAA were poorly efficient in TSB, especially in the case of biofilms. However, when tested in water, the efficacy of NaClO, CAB and PAA substantially increased, which was particularly evident in biofilms. In water, staphylococcal biofilms were even more susceptible to CAB than planktonic cells. A part (23.3%) of the biofilm positive staphylococci carried the qac genes but did not express an increased resistance to BAC. This study showed that bacterial biofilms protected with organic matter could be one of the main reasons for disinfection failure.  

Repeated open fermentative production of optically pure l-lactic acid using a thermophilic Bacillus sp. strain

A thermophilic Bacillus sp. strain 2–6 was used in completely open repeated batch fermentation for producing optically pure l-lactic acid. Up to 107 g l −1 l-lactic acid of optical purity 99.8% was obtained with NaOH as pH regulator. Unexpectedly, accumulated cells did not necessarily lead to increased l-lactic acid production. Kinetic and viable cells analyses revealed that l-lactic acid production by Bacillus sp. 2–6 was closely related to the growth of viable cells. Accumulated unviable cells had a slight or negative effect on production efficiency. Therefore, the number of repeated batches should be limited to avoid the accumulation of unviable cells. In this study, repeated open operation was shown to be feasible for optically pure l-lactic acid production, and new light shed on the roles of viable and unviable cells in repeated batch fermentation.

Pro-oxidative activities and dose–response relationship of (−)-epigallocatechin-3-gallate in the inhibition of lung cancer cell growth: a comparative study in vivo and in vitro

(-)-Epigallocatechin-3-gallate (EGCG), the major polyphenol in green tea, has been shown to inhibit tumorigenesis and cancer cell growth in animal models. Nevertheless, the dose-response relationship of the inhibitory activity in vivo has not been systematically characterized. The present studies were conducted to address these issues, as well as the involvement of reactive oxygen species (ROS), in the inhibitory action of EGCG in vivo and in vitro. We characterized the inhibitory actions of EGCG against human lung cancer H1299 cells in culture and in xenograft tumors. The growth of tumors was dose dependently inhibited by EGCG at doses of 0.1, 0.3 and 0.5% in the diet. Tumor cell apoptosis and oxidative DNA damage, assessed by the formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG) and phosphorylated histone 2A variant X (gamma-H2AX), were dose dependently increased by EGCG treatment. However, the levels of 8-OHdG and gamma-H2AX were not changed by the EGCG treatment in host organs. In culture, the growth of viable H1299 cells was dose dependently reduced by EGCG; the estimated concentration that causes 50% inhibition (IC(50)) (20 microM) was much higher than the IC(50) (0.15 microM) observed in vivo. The action of EGCG was mostly abolished by the presence of superoxide dismutase (SOD) and catalase, which decompose the ROS formed in the culture medium. Treatment with EGCG also caused the generation of intracellular ROS and mitochondrial ROS. Although EGCG is generally considered to be an antioxidant, the present study demonstrates the pro-oxidative activities of EGCG in vivo and in vitro in the described experimental system.

Neoplastic and Hematological Effects of Endosulfan and Bleomycin in the Swiss Albino Mice Mus musculus

Effects of endosulfan (EN), an insecticide, and bleomycin (BL), an antibiotic, on the body weight in the normal mice, and the in vivo cell growth, tumor weight, and hematological parameters of the Ehrlich ascites carcinoma (EAC) cell-bearing Swiss albino mice Mus musculus were evaluated. EN and BL were respectively administered orally and intraperitoneally to the experimental mice; the control group consisted of EAC cell-bearing untreated mice only. EN reduced the body weight in normal mice, whereas BL resulted in a steady body weight compared to the control. EN increased the EAC cell count significantly by reducing the growth of normal viable cells. In contrast, BL reduced the cell count by increasing the proportion of viable cells in the body. The tumor weights induced by EN were significantly higher than those of the EAC control and the BL-treated animals. In comparisons with the control and the BL mice, hematological parameters such as hemoglobin (%) and the number of RBC and lymphocytes were lowered, while counts of WBC, neutrophils, and monocytes were elevated after EN treatments. These results show that BL is capable of reducing the EN-induced neoplastic and haematological alterations in the mice under laboratory conditions.

Apoptosis and necrosis in developing brain cells due to arsenic toxicity and protection with antioxidants

Epidemiological studies on arsenic contamination in drinking water indicated presence of arsenic in fetal tissues. Experiments on human fetal brain explants on exposure to arsenic in culture showed disturbance in lipid peroxidation, generation of nitric oxide (NO), reactive oxygen species (ROS) and apoptosis. The oxidative stress challenged by antioxidant vitamins C, E or chelator dimercaptosuccinic acid (DMSA) may reverse arsenic toxicity on neuronal development. The concept was tested with the models: (A) human fetal brain explants exposed to arsenic, 0.3 mg/l in culture for 24 h; (B) rat neonatal brain explants from 1-day-old litters exposed to 0.3 mg/l arsenic in drinking water during gestation. Rats ( n=10) were given oral administration of vitamin C, 2.5 mg/kg/day, vitamin E, 148 μg/kg/day during gestation and DMSA, 50 mg/kg for 2 days at the end of gestation. (A) The arsenic induced in human fetal brain explants increase in production of NO, 20% and ROS, 25%, and decrease in DNA, 62% and protein, 54% synthesis. The morphological analyses showed growth of viable cells, neural networking vis-à-vis apoptosis on exposure to arsenic for 24 h and necrosis and loss of ground matrix on arsenic exposure for 18 days. The occurrence of two processes of apoptosis and necrosis in different neurons of same culture indicated existence of a selective cellular defense against arsenic toxicity. (B) The rats exposed to arsenic showed increased generation of NO, 25% and ROS, 22%, loss of glutathione content from 42 to 35 μg/mg protein, 40% increase in lipid peroxidation and decreased superoxide dismutase at 32%. The administration of vitamins C, E and DMSA showed partial reversal of the effects indicating possible protection from arsenic toxicity.

Investigation of induction effect on the steady state performance of a continuous fermentation for recombinant β-ga1actosidase

The performance of continuous fermentation for an induced recombinant protein depends on the concentration of the inducer and on the inflow stream. The latter is characterized by the substrate concentration and the dilution rate, of which the latter is preferred as the manipulated variable for control. This work studies the effects of inducer (isopropyl-β- d-thiogalactopyranoside, IPTG) concentration and dilution rate on the steady state performance of a continuous fermentation for β-galactosidase production by Escherichia coli containing the plasmid pUR2921. A low concentration of IPTG favours the growth of both viable (recombinant) cells and nonviable cells. To promote preferential growth of the former, their mass fraction should be maximized, and this requires a larger, optimal IPTG concentration. The expressed concentration of β-galactosidase passes through maxima with respect to both IPTG concentration and the dilution rate, whereas the viable cell concentration attains a peak only with respect to the dilution rate. While the optimal IPTG concentrations for mass fraction and for β-galactosidase are widely different, the two dilution rates are not far apart. This suggests two-stage fermentation at an intermediate dilution rate and different concentrations of IPTG.