A novel two-step method for the formation of tissue-engineered cartilage by mature bovine chondrocytes: the alginate-recovered-chondrocyte (ARC) method.

Most attempts to tissue-engineer cartilage have involved seeding of cultured cells into a biological or synthetic scaffold. We have developed a novel two-step culture approach that makes possible the in vitro formation of cartilaginous-like tissue by mature adult bovine chondrocytes without the aid of a synthetic matrix. The first step consists of culturing chondrocytes under conditions that maintain their rounded shape and their molecular phenotype as assessed by type II collagen and aggrecan production. This step was accomplished by culturing the isolated chondrocytes in alginate beads until the cells have reestablished a proteoglycan-rich cell-associated matrix (CM). The second step consists of culturing the cells with their CM, after recovery from the beads, on a tissue culture insert with a porous membrane. In this study, young adult bovine articular chondrocytes were cultured in alginate beads in the presence of 10% or 20% fetal bovine serum (FBS). After 7 days of culture, the alginate beads were dissolved by incubating the beads for 20 min in sodium citrate buffer, a calcium chelator. Following a brief centrifugation, the cells with their CM were recovered, resuspended in medium containing 10% or 20% FBS and seeded onto a tissue culture insert. After 1 week of culture on the insert, the individual cells with their CM progressively became incorporated into a mass of cartilaginous tissue. Culture with 20% FBS resulted in the best formation of tissues. These tissues, easily recovered from the insert, were then subjected to biochemical and histological analyses. The biochemical results showed that the chondrocytes remain phenotypically stable in the tissues. The de novo tissue has a relatively high ratio of PG/collagen. Histological examination of the tissue revealed it contained a cartilage-like matrix strongly stained with toluidine blue. This scaffold-free system appears ideal to study, in vitro, the development of transplantable cartilaginous tissue.     

Status of Women and Infants in Complex Humanitarian Emergencies

Women and children bear the greatest burden in the midst of war and long‐term disasters. Complex humanitarian emergencies are characterized by social disruption, armed conflict, population displacement, collapse of public health infrastructure, and food shortages. Humanitarian assistance for refugees and internally displaced populations requires particular attention to the common issues affecting morbidity and mortality in women and infants. Gender‐based violence and reproductive health concerns are discussed within the context of populations affected by conflict and forced migration. Recommendations for midwives and women's health care providers engaging in care for women and children in complex humanitarian emergencies are discussed.     

Women's Satisfaction with Their Involvement in Health Care Decisions During a High‐Risk Pregnancy

ABSTRACT: Background: Increasingly, women seek involvement in decisions about their health care. The purpose of this study was to examine women's experience of, and satisfaction with, their involvement in health care decisions during a high‐risk pregnancy. Methods: Forty‐seven women with hypertension or threatened preterm delivery (including multiple births) were interviewed after the birth of their child. They received prenatal care at home from nurses in a community program or were hospitalized. The in‐depth interviews were audiotaped and transcribed; data were analyzed using constant comparative methods. Results: Women identified an increased feeling of responsibility for the health of their baby and themselves, but differed in choosing active or passive involvement in health care decisions. Women who wanted active involvement achieved it through one of three processes: struggling for, negotiating, or being encouraged. Women who wanted passive involvement and women facing health crises used the process of trusting in the expertise of nurses and physicians. Women were satisfied if the care from health care professionals was congruent with how they wanted to be involved in decision‐making. Conclusions: Although most women want to be actively involved in health decision‐making during a high‐risk pregnancy, some prefer a passive role. The setting of prenatal care, community‐based or in‐hospital, was less important than the ability of nurses and physicians to support the woman in her preferred role in decision‐making. (BIRTH 30:2 June 2003)     

Morphology of temperate bacteriophage SfV and characterisation of the DNA packaging and capsid genes: the structural genes evolved from two different phage families

The entire genome of SfV, a temperate serotype-converting bacteriophage of Shigella flexneri, has recently been sequenced (Allison, G.E., Angeles, D., Tran-Dinh, N., Verma, N.K. 2002, J. Bacteriol. 184, 1974-1987). Based on the sequence analysis, we further characterised the SfV virion structure and morphogenesis. Electron microscopy indicated that SfV belongs to the Myoviridae morphology family. Analysis of the proteins encoded by orf1, orf2, and orf3 revealed that they were homologous to small and large terminase subunits, and portal proteins, respectively; the protein encoded by orf5 showed homology to capsid proteins. Western immunoblot of the phage with anti-SfV sera revealed two antigenic proteins, and the N-terminal amino acid sequence of the 32-kDa protein corresponded to amino acids 116 to 125 of the ORF5 protein, suggesting that the capsid may be processed. Functional analysis of orf4 showed that it encodes the phage capsid protease. The proteins encoded by orfs1, 2, 3, 4, and 5 are homologous to similar proteins in the Siphoviridae phage family of both gram-positive and gram-negative origin. The capsid and morphogenesis genes are upstream and adjacent to the genes encoding Myoviridae (Mu-like) tail proteins. The organisation of the structural genes of SfV is therefore unique as the head and tail genes originate from different morphology groups.     

Collaborative investigation of the accuracy and reproducibility of Sceptor Breakpoint susceptibility panels.

A combination Sceptor Breakpoint/ID panel (Johnston Laboratories, Inc., Towson, Md.), which determines interpretive susceptibility results (susceptible, moderately susceptible, and resistant) using two to three selected concentrations of antimicrobial agents, was tested in comparison with full-range Sceptor microdilution MIC panels. The inter- and intralaboratory interpretive reproducibilities for 24 control strains tested in three laboratories on three consecutive days were 97.0 and 95.7%, respectively. The equivalency of breakpoint results to category results obtained by the microdilution MIC procedure for 10,368 control organism-antimicrobial agent comparisons was 94.1%. The level of interpretive agreement between breakpoint and MIC category results using 101 fresh clinical isolates was 97.0% for 51 gram-negative and 50 gram-positive bacteria. Among the total 4,872 clinical organism-antimicrobial agent comparisons, major and very major discrepancies were seen in 0.2% of gram-negative bacteria and very major discrepancies were seen in 0.9% of gram-positive bacteria. All very major discrepancies with gram-positive organisms were associated with trailing endpoints using trimethoprim or sulfisoxazole and staphylococci. The breakpoint concept of testing selective antimicrobial agent concentrations was highly reproducible and accurate and allows for placement of more antimicrobial agents into a panel than is possible with full-dilution MIC testing.