Use of an in vitro model of tissue-engineered human skin to study keratinocyte attachment and migration in the process of reepithelialization

To produce a stable epidermis, keratinocytes need to be firmly attached to the basement membrane. However, following wounding, keratinocytes are required to develop a migratory phenotype in order to reepithelialize the wound. To investigate some of the issues underlying reepithelialization, we have developed a three-dimensional in vitro model of tissue-engineered skin, comprising sterilized human dermis seeded with human keratinocytes and dermal fibroblasts. Using this model, we have shown that the inclusion of fibroblasts within the model increases the stability of keratinocyte attachment. We have also demonstrated that keratinocyte migration occurs most effectively in the absence of a basement membrane and following the inclusion of fibroblasts in the model. In addition, subjecting the keratinocyte layer to mechanical trauma induces a migratory phenotype. We conclude that this three-dimensional in vitro wound model can be used to increase our understanding of the factors that enhance keratinocyte migration and hence wound healing in vivo.     

Use of ambulance dispatch data as an early warning system for communitywide influenzalike illness,New York City

In 1998, the New York City Department of Health and the Mayor’s Office of Emergency Management began monitoring the volume of ambulance dispatch calls as a surveillance tool for biologic terrorism. We adapted statistical techniques designed to measure excess influenza mortality and applied them to outbreak detection using ambulance dispatch data. Since 1999, we have been performing serial daily regressions to determine the alarm threshold for the current day. In this article, we evaluate this approach by simulating a series of 2,200 daily regressions. In the influenza detection implementation of this model, there were 71 (3.2%) alarms at the 99% level. Of these alarms, 64 (90%) occurred shortly before or during a period of peak influenza in each of six influenza seasons. In the bioterrorism detection implementation of this methodology, after accounting for current influenza activity, there were 24 (1.1%) alarms at the 99% level. Two occurred during a large snowstorm, 1 is unexplained, and 21 occurred shortly before or during a period of peak influenza activity in each of six influenza seasons. Our findings suggest that this surveillance system is sensitive to communitywide respiratory outbreaks with relatively few false alarms. More work needs to be done to evaluate the sensitivity of this approach for detecting nonrespiratory illness and more localized outbreaks.     

Physiological neutrophilia of pregnancy is not associated with a rise in plasma granulocyte colony-stimulating factor (G-CSF)

A patient with neutropenia and life-threatening infections secondary to T-γ lymphoproliferative disease, who did not respond to treatment with recombinant human G-CSF (filgrastim), was treated with filgrastim plus cyclosporine A (CyA). The patient achieved a good response in the absolute neutrophil count and subsequently required a dose reduction in the filgrastim. The patient was eventually discontinued from the CyA but continues on filgrastim alone. While on therapy, the large granular lymphocytes disappeared from the circulation and the beta-TCR rearrangement, which was present prior to beginning therapy, became undetectable. The patient had no significant toxicity to the CyA or the filgrastim and he has not experienced any serious infections or required hospitalization. Filgrastim has proven to be relatively nontoxic and of some benefit to patients with this disease and should probably be utilized first when treatment is necessary. However, if improvement is not observed, these findings suggest that a trial of the combination of CyA plus filgrastim may be beneficial.     

Exploring the nature of the relationship between child sexual abuse and substance use among women

Aims. This study investigated whether child sexual abuse (CSA) was associated with earlier substance use and greater severity of substance dependence and what aspects of CSA might predict substance abuse. Design. The study compared (a) drug and alcohol treatment clients with and without a history of CSA and (b) CSA survivors outside drug and alcohol treatment who did or did not have current substance abuse. Settings. Semi-structured interviews took place at participants' homes, treatment agencies or the research centre. Participants. Volunteer participants included 100 women recruited from drug and alcohol treatment programmes and 80 CSA survivors recruited through CSA counselling services and media advertising. Measurements. The results focus on data from the Opiate Treatment Index, Severity of Alcohol Dependence Questionnaire, Substance Dependence Scale, Self-Esteem Inventory and self-reported histories of CSA. Findings. There were no differences between CSA survivors and other drug and alcohol treatment clients in their severity of dependence. Women with a history of CSA more frequently identified stimulants as their main problem drug and reported an earlier age of first intoxication and earlier use of inhalants. Among CSA survivors outside drug and alcohol treatment, women with current substance abuse had typically been abused during adolescence by someone outside the family, whereas those without current substance abuse were typically abused by family members before adolescence. Conclusions. The results suggest that adolescence is a crucial time for the influence of CSA experiences on substance abuse.     

Further study on the vascular basis for the reimplantation of the hand amputated through the palm

Summary Based on the anatomic data obtained from earlier studies on the vascular anatomy of the hand, the vascular architecture in the palm of the hand was studied on 60 sides of unembalmed adult upper extremities. Each palm was divided into 64 squares by 8 sagittal and 8 transverse sections. The vascular architecture in these squares and the arterial relations between them were observed and measured by angiography, operative microscopic dissection and computerised three-dimensional reconstruction. According to the pattern of the blood-vessels, the amputated palms can be classified into 4 types. The anatomic basis for the vascular anastomosis in each type is defined. There are three key-areas for the blood-supply of the palm and their significance is discussed. Apart from the 4 types of transversely amputated palms, the repair programe of the blood-vessles in 4 types of common obliquely amputated palms are also discussed.

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Etude complémentaire de l'anatomie vasculaire de la main pour la réimplantation des amputations transpalmaires

Résumé Sur la base de données anatomiques obtenues lors de précédents travaux sur l'anatomie vasculaire de la main, l'architecture vasculaire palmaire a été étudiée sur 60 extrémités supérieures de cadavres d'adultes, non embaumés. Chaque paume a été divisée en 64 carrés par 8 sections sagittales et 8 sections transversales. L'architecture vasculaire à l'intérieur des carrés et les relations artérielles entre eux ont été étudiées et mesurées par angiographie, dissection au microscope opérateur et reconstruction computérisée en 3D. Les paumes amputées ont été regroupées en 4 types d'après la distribution des vaisseaux sanguins. Les données anatomiques concernant les anastomoses vasculaires sont précisées. Il existe trois zones clés pour l'irrigation de la paume. Leur importance quant à l'irrigation de la main est exposée. Outre la division des paumes amputées transversalement en 4 types, le programme de réparation de vaisseaux dans les 4 types d'amputations obliques communes de la paume et aussi discuté.

     

Specific antibody-forming B-lymphocyte colonies. I. Distribution and nature of SRBC antibody-forming B-lymphocyte colonies in mouse lyphomyeloid organs.

Wehn normal mouse spleen or lymph node cells are cultured for 7 days in agar-medium containing 2-mercaptoethanol and sheep red blood cells (SRBC), approximately one B lymphocyte colony (BLC) develops per 50-100 cells seeded. Incubation of cultures for 3 h with guinea-pig complement at day 7, demonstrated that 0.05-0.25% of all BLC form specific antibody against SRBC (SRBC-AF-BLC). The SRBC specific colonies appear centrally in lytic plaques of 2-5 mm in diameter and cells recovered from individual SRBC-AF-BLC were shown to produce antibodies of the IgM class against SRBC. Cells forming SRBC-AF-BLC are absent in the new-born and infantile spleen but appear in adult mice with a frequency of 5, 10-25 and 25-70 per 10(6) bone marrow, spleen and lymph node cells respectively. specific immunization in vivo or in vitro does not greatly affect the number of SRBC-AF-BLC-forming cells. Cytolysis of spleen cells with anti-Ig serum plus complement prior to culture reduced the number of total BLC and that of specific SRBC-AF-BLC by 93% and 94% respectively. The peak sedimentation velocity of both SRBC-AF-BLC-forming cells and total BLC-forming cells was 3.5 mm/h. Spleen cells enriched 200-300 times for cells that bind specifically to the hapten NIP were not enriched for cells forming colonies with specific antibody production against NIP. The data indicate that the cells that give rise to specific antibody-forming colonies belong to a mature virgin B cell group of small Ig-positive B lymphocytes.     

An inexpensive restraint for small animals

An animal restraint is described that can be built in the laboratory from a piece of Plexiglas and a few inches of Velcro.     

Reassessment of the rate of fixative diffusion.

The diffusion of fixatives is slow. Early work using plasma gels and animal tissues showed the distance penetrated by a fixative to be a simple function of the fixation time but this relation has not been established in human tissues. The rates of diffusion into whole human spleens were measured for three primary fixatives over periods ranging from one to 25 days. A positive correlation was demonstrated between penetration distance (mm) and fixation time (hours). The diffusion rates were slower than those in previous studies. These results have possible implications for the handling of surgical specimens.