Conducting Informal Discovery of a Party\u27s Former Employees: Legal and Ethical Concerns and Constraints

This Article identifies and critiques existing sources of confusion in the law and proposes revised and alternative discovery procedures to provide equal access to information possessed by ex-employees, while simultaneously safeguarding the integrity of that information. Its primary emphasis is on federal jurisprudence, although important points of consensus and departure between state and federal law are noted, as appropriate. Part I explains the issues that arise in informal discovery, and the difficulties with clearly resolving those issues given the conflicting state of the law. Part II discusses application of the attorney-client privilege to communications between corporate counsel and former employees, concluding that the privilege should not shield the content of such communications from discovery by opposing counsel. Because the attorney-client privilege issue and the debate over ex parte contact both turn on whether a former employee is a party to the litigation, an exposition of the ethical concerns of ex parte contact follows in Part III, which concludes that ex parte contact by opposing counsel should be allowed. Part IV examines the applicability of the work product doctrine in shielding from discovery certain tangible materials related to the interview. This Article advocates absolute immunity from discovery for attorney notes and memoranda, and for collections of documents selected by counsel for discussion with former employees; limited discovery of signed witness statements generated pursuant to the interview; and fairly broad discovery of the content of counsel\u27s questions and statements during the interview. The proposed discovery guidelines in the Conclusion are offered with the ambitious, if not elusive, goal of reconciling the competing interests and policies that rouse litigants to battle when informal discovery is conducted of former employees, while simultaneously giving due regard to the pragmatic impact that new or revised rules may have on the litigants and the adversarial process. The overriding objective is to provide equal access by all parties to the information possessed by former employees, while at the same time providing mechanisms to deter, and if necessary to reveal, inappropriate manipulation of these potential witnesses by counsel

Production of Monoclonal Antibody Against Salmonella H: g,m Flagellar Antigen and Potential Diagnostic Application

WOS: 000283852200006PubMed ID: 21050043In this study, the flagella antigen was detached at high speed by shaking vigorously with glass pearl beads, from Salmonella enteritidis in a yield of 3.9 mg/mL(-1) after being concentrated with polyethylene glycol (PEG). A monoclonal antibody (MAb), designated D7 clone, was generated from Balb/c mice immunized with Salmonella enteritidis flagella using the conventional hybridoma method. The D7 clone secreting MAb was classified as IgG2a isotype. ELISA analyses of D7 MAb to Salmonella-specific flagella demonstrated that the antibody reacted with H: m. However, Western immunoblot analyses of D7 clone appear to be secreting heavy chain of IgG2a antibody, which was eligible for the diagnosis of Salmonella enteritidis

Patterned adhesion of cells

Symposium on Ion-Beam-Based Nanofabrication held at the 2007 MRS Spring Meeting -- APR 10-12, 2007 -- San Francisco, CAWOS: 000250475100010It is well known that silver deposition avoids bacterial growth and inhibits the natural process of attachment of connective tissue to biocompatible materials in vivo. We have completed a five year investigation of the precise spatial control of cell growth on glassy polymeric carbon implanted with silver using ion beam techniques, and the persistence of the inhibitory effect on cell growth. Long term inhibition of cell growth on GPC is a desirable improvement on current cardiac implants and other biocompatible materials placed in the blood stream. We have used implanted silver ions near the surface of GPC to completely inhibit cell attachment and adhesion. Cells attach and strongly adhere to areas close to the silver implanted surfaces. Patterned ion implantation permits precise control of tissue growth on GPC and other biocompatible substrates. Cell growth limited to micrometric patterns on a substrate may be useful for in vitro studies of associated biological processes in an otherwise identical environment. The patterned inhibition of cell attachment persists for periods of time significant relative to typical implant lifetimes.Center for Irradiation of Materials, Alabama AM University; National Science FoundationNational Science Foundation (NSF) [EPS-0447675]Research sponsored by the Center for Irradiation of Materials, Alabama A&M University and by National Science Foundation under Grant No. EPS-0447675

Investigation of Possible Genotoxic and Cytotoxic Effects of Differential Boron Compounds in CCL 62 (Hela Contaminant) Human Amniotic Ephitelial Cell Line

Epidemiological and in vitro studies have showed that boron may have anti-carcinogenic properties. Chromosome aberrations assay and sister chromatid exchange assays have showed that boron has no genotoxic effects on cytogenetically stable cell lines. Aim of this investigation is determine to effects of different boron compounds which are boric acid (BA), borax pentahydrate (BP) and disodium pentaborate decahydrate (DPD) ) on exist cytogenetic disruption with variable doses (250, 500 and 1000µM ) on cytogenitically unstabel CCL 62 (HeLa Contaminant) cell line. In order to test this hypothesis, following the boron treatment on the cell lines for cytotoxcity was performed using MTT that cell viability assay. For genotoxicity was used chromosome aberrations assay (CAs) and micronucleus assay (MN), CAs and MN frequency calculated in each boron dose. Boron compaunds effected proliferation of CCL 62 (human amniotic ephitelial) cell lines in a time, species and dose dependent manner. According to data obtained from CAs and MN assays, no significant difference was found between control groups which were not treated any of boron compaund with boron treated groups (p>0,05). In conclusion, we established that BA, BP, and DPD effected proliferation of CCL 62 cell lines in a time, compaund and dose dependent manner, however no evidence was observed suggesting these compounds cause an increase or decrease in the level of existing cytogenetic defects in these cell lines. [Med-Science 2013; 2(1.000): 454-68

Metabolic Activity and Monoclonal Antibody Production of Salmonella Enteritidis O and H Antigen Specific Hybridoma Cells in Static Culture

WOS: 000290030800012PubMed ID: 21529293The aim of the present work was to study the kinetics of two hybridomas that produce monoclonal antibody against Salmonella Enteritidis O (5A8) and H (D7) antigen. The hybridomas originated from the Ag8x653 (5A8) and Sp2/O (D7) myeloma cell lines. The relationship between the uptake of glucose and glutamine and the release of the lactate and ammonia and monoclonal antibody production into hybridoma growth were investigated in static culture with serum-containing DMEM/F:12 medium for the determination of pilot-production strategies of the hybridomas. Results showed that glucose and glutamine concentrations were reduced, with an increase in ammonia and lactate concentration in the culture medium. The hybridoma cell line 5A8 has shown lower metabolic activities compared with D7, whereas its monoclonal antibody productivity was found to be two-fold higher than the D7. MAb production by the hybridoma cell line 5A8 seems promising, considering the moderate level of productivity compared to that found in the literature.Ege UniversityEge University [06MUH05]This work was supported in part by a grant from the Research Fund of Ege University (06MUH05). We thank Melis Kuban for helpful discussions and a critical review of this manuscript

Production of Monoclonal Antibodies in a Mouse Model via Lipopolysaccharide Conjugates with Synthetic Polymers Specific to Salmonella Enteritidis O Antigen

WOS: 000284837300011PubMed ID: 20707736Monoclonal antibodies (Mabs) specific for lipopolysaccharide (LPS) of Salmonella Enteritidis were evaluated in a model of LPS conjugated synthetic polymer immunization of Balb/c mice by conventional hybridoma method. Nine hybridoma cell lines were determined as antibody positive against LPS. The clone 5A8 secreting the highest antibody was selected for further characterization. Evaluated results indicate that the synthetic polymer can be used as an effective adjuvant in immunization with LPS, because the 5A8 Mab were obtained using synthetic polymer as an adjuvant. 5A8 Mab was classified as IgG2a isotype by antibody capture enzyme-linked immunosorbent assay. The reactivity of the Mab against lipid A and different LPS of Salmonella were investigated using an indirect enzyme-linked immunosorbent assay. Mab presented a wide spectrum of reactivity, coupling with antigens against Salmonella. The hybridoma 5A8 determined in this study has a great potential to be used in the development of diagnostic, prophylactic, and therapeutic agents specific for Salmonella Enteritidis and Salmonella LPS.Ege UniversityEge University [2002FEN09]This work was financially supported in part by a grant from Research Fund of Ege University (2002FEN09)

A cost-effective and simple culture method for primary hepatocytes

WOS: 000288661400003Hepatocytes, the major epithelial cells of the liver, maintain their morphology in culture dishes coated with extracellular matrix (ECM) components such as collagen and fibronectin or biodegradable polymers (e.g. chitosan, gelatin). In these coated dishes, survival of cells and maintaining of liver-specific functions may increase. The aim of this study was to determine a suitable, cost-effective and simple system for hepatocyte isolation and culture which may be useful for various applications such as in vitro toxicology studies, hepatocyte transplantation and bioartificial liver (BAL) systems. In order to obtain primary cultures, hepatocytes were isolated from liver by an enzymatic method and cultured on plates coated with collagen, chitosan or gelatin. Collagen, gelatin-sandwich and gelatin-cell mixture methods were also evaluated. Morphology and attachment of the cells were observed by inverted microscope and scanning electron microscope (SEM). An MTT assay was used to determine cell viability and mitochondrial activity.Ege University Science and Technology Society (EBILTET)Ege University [E18/03/06]The authors received partial support from Ege University Science and Technology Society (EBILTET) under the Project No. E18/03/06. In addition, we are grateful to Assoc. Prof. Seda Vatansever, Dr. Feyzan Ozdal Kurt and Dr. Elgin Turkoz from Celal Bayar University for their help in the study

Isolation and purification of O and H antigens from Salmonella Enteritidis as diagnostic tool

WOS: 000285047300025Lipopolysaccharide (LPS) and flagellin are the major antigenic structures of Salmonella Enteritidis. Purified antigens are essential components for the diagnosis of and research on Salmonella Enteritidis. In this study, LPS were isolated and purified with modified hot phenol-water procedure in a yield of 6%. The flagellin isolation and purification were done by strong shaking with glass pearl beads with 3.9 mg/ml protein content. Purity was demonstrated on SDS-PAGE, ELISA and with colorimetric assay for both isolated antigens. The antigen-mediated ELISA results exhibited immunoreactive properties for both LPS and flagellar antigen specific for Salmonella Enteritidis O:9 and H: g, m, respectively. The findings demonstrated low reactivity with Mab O:9 for isolated LPS due to their monoepitopic specificity. However, isolated flagellin reactivity was high with Mab H: g, m related to their cross-reactivity potential between H: g and H: m. Additionally in this study. the results indicated that similar antigenic structures have alternative potential for antigenic isolation of Salmonella Enteritidis, such as Salmonella Gallinarum for LPS antigen and Salmonella Adeyo for flagellar antigen isolation

Isolation and in vitro cultivation of human urine-derived cells: an alternative stem cell source

WOS: 000410005000021PubMed ID: 28861309Objective: For in vitro tissue engineering in urology, stem cells are commonly isolated from tissue specimens obtained during open or endoscopic surgery. Within the context of the present study our aim was to isolate cells from human urine by an alternative and safe technique rather than using the indicated method. Material and methods: After human urine samples had been collected from young and healthy donors via urethral catheterization, cells were precipitated by centrifugation and cultured. Following this isolation procedure, cells were characterized by immunocytochemical method using specific antibodies. Results: When these cells were characterized by immunocytochemical methods using specific antibodies some of them were positive for mesenchymal stem cell marker CD90 while the others were labelled with urothelial marker cytokeratin 7. When all these results were taken into consideration, urothelial cells together with stem cells were observed in the urine-derived cell population. Conclusion: According to the results obtained from this study human urine may be preferred as an alternative stem cell and urothelial cell source in that this method is and easily reproducible non-invasive method.TUBITAK-BIDEB National Scholarship Programme [2211]Secil Erden Tayhan, one of the researchers of this study, was finantially supported by TUBITAK-BIDEB 2211 National Scholarship Programme for PhD Students