Enhanced immunity in intradermal vaccination by novel hollow microneedles.

BACKGROUND: The intradermal (ID) route for vaccination represents an effective alternative to subcutaneous (SC)/intramuscular administration to induce protective immunity. However, a critical issue associated with ID vaccination is the precise delivery of solution in the upper dermis, which ensures enhanced immunity. METHODS: We fabricated a hollow microneedle unit made of poly-glycolic acid by injection molding and bonding, and created a dedicated prototype injector. To ensure ID delivery of solution, the injected site was macroscopically and microscopically examined. Serum immunoglobulin G antibody production was measured by enzyme immunoassay and compared in groups of rats following either ID delivery with microneedles or SC administration with a 27-G stainless needle of graded vaccine doses. RESULTS: The unit used a tandem array of six microneedles, each with a side delivery hole, and a conduit inside for solution. Microneedles installed in the injector punctured the skin with the aid of a spring. Injection of solution formed a wheal due to ID distribution. Histologically, a wedge-shaped skin defect in the upper skin corresponded to each puncture site. Antibody titers following vaccinations on days 1 and 8 were significantly higher with ID injection than with SC delivery on day 15 and every 7 days thereafter until day 36 with mumps vaccination, and until day 36 with varicella vaccination. CONCLUSIONS: The microneedle unit presented here delivered solution intradermally without any difficulty and evoked antibody responses against viruses even with the reduced vaccine volume. Our findings confirm promising results of ID delivery as an immunogenic option to enhance vaccination efficacy.

Primary mucinous carcinoma of the skin.

BACKGROUND: Primary mucinous carcinoma is a rare primary adnexal neoplasm of the skin with a high recurrence rate. OBJECTIVE: To improve the recognition of this clinical entity and aid in establishing the origin of this tumor. METHODS: Two cases of this tumor were reported and investigated histopathologically and histochemically. Furthermore, some mucinous carcinomas arising from the breast or gastrointestinal tract were compared with this tumor. RESULTS: Both cases showed no evidence of primary neoplasms as a source of metastasis and had characteristic findings of primary mucinous carcinoma of elongated cords and small lobules of epithelial cells floating in lakes of sialomucin. The origin of this tumor seemed to be an eccrine sweat gland. CONCLUSION: This tumor should be resected widely and, at least, the breasts and gastrointestinal tract should be investigated for primary lesions.

Systematized keratotic nevus with finger contracture.

A case of systematized keratotic nevus which is a peculiar form of epidermal nevus is reported. The present case is rare because of the dysfunction of incomplete flexion of fingers by systematized keratotic nevus. Furthermore, it involved mixed clinical and histological changes of epidermal nevus from fingers to elbow. An operation with full-thickness skin graft was undertaken.

Spatial organization of extracellular matrix and fibroblast activity: effects of serum, transforming growth factor beta, and fibronectin.

The goal of our research is to understand reciprocal relationships between cell function and tissue organization. We studied the regulation of fibroblast activity in an in vitro culture model that recapitulates in continuous fashion the cycle of events occurring during connective tissue repair. We present evidence that concomitant with spatial reorganization of the extracellular matrix, there was a dramatic decline in extracellular matrix synthesis and cell proliferation. Therefore, spatial reorganization was a crucial turning point for fibroblast activity. Factors that regulated the timing of spatial reorganization included serum, transforming growth factor beta, and fibronectin. By accelerating spatial reorganization of the cultures, transforming growth factor beta led to a relative decrease in cell proliferation and extracellular matrix synthesis. By retarding spatial reorganization of the cultures, fibronectin led to a relative increase in cell proliferation and extracellular matrix synthesis. The results indicate that spatial information in the three-dimensional cell-matrix interaction permits higher order, tissue-level regulation of fibroblast function.

Collagen processing, crosslinking, and fibril bundle assembly in matrix produced by fibroblasts in long-term cultures supplemented with ascorbic acid.

Human foreskin fibroblasts were cultured for up to 6 weeks in medium supplemented with ascorbic acid. During this time, the cells produced an extensive new connective tissue matrix in which the accumulated collagen (mostly type I) amounted to about 0.25 mg/10(6) cells. The matrix was highly differentiated as shown by complete processing of procollagen to collagen alpha-chains and covalent crosslinking of the collagen. Alignment of collagen fibrils occurred as the fibrils were deposited between cells, and binding of adjacent fibrils to the cell surface appeared to hold the fibrils in register. Groups of aligned fibrils were subdivided into bundles by cell-surface folds. If beta-aminopropionitrile was added to the medium, collagen crosslinking was inhibited, but not collagen synthesis or fibril bundle organization. If ascorbic acid was omitted from the culture medium, the extensive new connective tissue matrix was not produced. Our results indicate that fibroblasts in long-term cultures supplemented with ascorbic acid produce a connective tissue matrix with many in vivo-like properties including supermolecular organization of collagen.

Mechanical stretching increases prostaglandin E2 in cultured human amnion cells.

Mechanical stretching increases synthesis and release of prostaglandin E2 (PGE2) in cultured amnion cells. The maximum level of PGE2 in stretched amnion cells is three times higher than that in nonstretched amnion cells. The maximum level of PGE2 in stretched cell medium is nine times higher than the maximal level of PGE2 in nonstretched cell medium. 3H-thymidine incorporation into DNA is 211 +/- 25 cpm/10(5) cells in nonstretched groups; that in stretched groups is 582 +/- 94 cpm/10(5) cells. There is a significant difference between the two groups. These results suggest that stretch stimulation facilitates prostaglandin production in fetal membranes, which may contribute to uterine contraction in labor.

Growth kinetics of fibroblasts derived from normal skin and hypertrophic scar.

In vitro growth kinetics of fibroblasts derived from normal skin and hypertrophic scar were performed using continuous 3H-thymidine labeling method. In fibroblasts derived from normal skin, aging of the donor affects cell growth mainly by growth fraction (GF), but not labeling index (LI) and DNA synthetic time (Ts). When hypertrophic scar-derived fibroblasts are compared with normal fibroblasts, they showed a shorter Ts and lower LI and GF. This result suggests that in hypertrophic scar a small number of fibroblasts proliferate more actively, but most fibroblasts are non-growing cells.

A cell-binding, immunoglobulin-like protein from human plasma. II. Cell-binding activity.

Cell adhesion to plastic surfaces coated with a new high-molecular-mass immunoglobulin-like protein from normal human plasma was studied. Mouse subdermal fibroblasts, hamster kidney cells, human umbilical vein endothelial cells, and human skin fibroblasts were found to become attached to the surface, but cancer cells derived from human stomach cancer and human breast cancer did not. The appearance of the attached cells differed from that of cells attached to surfaces coated with fibronectin or concanavalin A. The cell adhesion to the surfaces coated with the protein was inhibited by goat anti-human IgM. Furthermore, the binding of the protein to the cell surfaces was demonstrated by the indirect immunofluorescence method. It is concluded that this protein is a new cell-binding protein.

A cell-binding, immunoglobulin-like protein from human plasma. I. Isolation and subunit structure.

A protein having Mr of more than 900K (900,000) was isolated by chromatography on a column of Sepharose 4B coupled with solubilized elastin, followed by sucrose density gradient centrifugation. The protein is composed of several disulfide-linked subunits. SDS-polyacrylamide gel electrophoresis and immunoblotting indicated that two of the subunits are identical with the heavy chains of IgM and IgG, and one is immunologically related to the heavy chain of IgA. The results suggested that this protein is a new protein that belongs to the category of, or is closely related to, the immunoglobulins. As described in the succeeding paper, it has cell-binding activity (Fukamizu et al. (1986) J. Biochem. 100, 843-848). We tentatively refer to this protein as "cell-binding immunoglobulin-like protein (CIP)."

Metastatic squamous-cell carcinomas derived from solar keratosis.

Actinic squamous-cell carcinoma has been described to have a low risk of metastasis. Reported here are four Japanese cases of lymphatic metastasis of squamous-cell carcinoma derived form solar keratosis. The primary lesions were on the face in three cases and on the back of the hand in one. Two patients died, and others had recurrent incurable metastatic lymph nodes. Therefore, it is noteworthy that actinic squamous-cell carcinoma in the Japanese can become so aggressive and metastasize.

Tissue cultures of benign and malignant fibrous histiocytomas: SEM observations.

Four dermatofibromas (DF), 2 dermatofibrosarcomata protuberans (DFSP), 2 atypical fibroxanthmas of the skin (AFX), and one malignant fibrous histiocytoma (MFH) were studied by explant culture technique and scanning electron microscopy. Differences in the cellular atypism, phagocytic activity and motility were observed between histiocyte-like cells extending from a DF and DFSP group and an AFX and MFH group. Such cytological characteristics was maintained during in vitro transformation of the cells into fibroblastic cells. It was concluded that culture behaviors of the cells from each tumor group correlated well with in vivo growth and histologic features. We feel that examination of in vitro morphology of fibrous histiocytomas may prove useful in arriving at a correct diagnosis.

Atypical ("pseudosarcomatous") cutaneous histiocytoma.

We report a patient with a cytologically atypical hemosiderotic histiocytoma that histologically suggested a sarcoma. The lesion is differentiated from atypical fibroxanthoma and fibroxanthosarcoma. We wish to emphasize the benign nature of this variant of histiocytoma whose extreme cellular atypicality may simulate a sarcoma.