Effector molecules in expression of the antimicrobial activity of macrophages against Mycobacterium avium complex: roles of reactive nitrogen intermediates, reactive oxygen intermediates, and free fatty acids.

We studied microbicidal activities of reactive nitrogen intermediates (RNI), free fatty acids (FFA), and reactive oxygen intermediates (ROI) against Mycobacterium avium complex (MAC) and the mode of macrophage (mphi) production of these effectors. (1) Intracellular growth of MAC in murine peritoneal mphis was accelerated by scavengers for ROI or RNI and inhibitors of nitric oxide synthase or phospholipase A2, indicating roles of ROI, RNI, and FFA in mphi anti-MAC functions. (2) Acidified NaNO2-derived RNI, FFA (linolenic and arachidonic acids), and the H2O2-mediated halogenation system exhibited a significant anti-MAC bactericidal activity. The combination of RNI with FFA showed a synergistic effect. However, the H2O2-halogenation system in combination with either RNI or FFA showed an antagonism. When Listeria monocytogenes (Lm) was used as a target organism, the combinations of RNI + FFA and RNI + H2O2-halogenation gave a synergistic effect, whereas FFA + H2O2-halogenation showed an antagonism in exerting bactericidal activity. In addition, when ROI generated by the xanthine oxidase-acetaldehyde system was combined with RNI, anti-Lm but not anti-MAC activity was potentiated. (3) ROI production by murine peritoneal mphis was observed immediately after contact with MAC organisms (MAC stimulation) and ceased within 2 h. FFA release was seen 1-24 h after MAC stimulation. RNI production was initiated from 3 h and increased during the first 36 h and continued at least for 4 days. These findings suggest that RNI and FFA rather than ROI are important effectors of anti-MAC functions of mphis, and the collaborating action of RNI with FFA temporarily participates in mphi-mediated killing of MAC in the relatively early phase after MAC stimulation.

Keratin type intermediate filaments in sweat gland myoepithelial cells.

A study was undertaken to clarify the origin of sweat gland myoepithelial cells using monoclonal antibodies EKH1, EKH4, and AN3. EKH1 recognizes all classes of intermediate filaments. EKH4 and AN3 recognize keratin type intermediate filaments. Since within the skin, only epithelial cells of ectodermal origin contain keratin, EKH4 and AN3 could be used as ectodermal markers within the skin. Sweat gland myoepithelial cells were labeled by all three antibodies. In contrast, arrector pili muscle and vascular smooth muscle were recognized only by EKH1, but not by EKH4 and AN3. This study demonstrated that myoepithelial cells of sweat glands contain keratin type intermediate filaments and suggested their ectodermal origin. On the other hand, arrector pili muscle and vascular smooth muscle did not contain keratin type intermediate filaments, despite their ultrastructural similarity to myoepithelial cells. Electron microscopic studies using human fetal and adult skin revealed that myoepithelial cells are developed from basal cells of the coiled tip of fetal gland and not from mesenchymal cells. In order to determine the time of appearance of myoepithelial cells during fetal development, embryonic and newborn mouse skin was also examined. It was found that sweat gland myoepithelial cells first appear around 20 weeks of gestation in humans and after birth in mice.

Monoclonal nonspecific suppressor factor beta (MNSF beta) inhibits the production of TNF-alpha by lipopolysaccharide-activated macrophages.

The monoclonal nonspecific suppressor factor (MNSF), a lymphokine produced by a murine T cell hybridoma, shows a pleiotropic antigen-nonspecific suppressive function. Most recently, a cDNA encoding a subunit of MNSF (MNSF beta) has been isolated and characterized. Recombinant form of MNSF beta (rMNSF beta) inhibits lymphokine functions, as does native MNSF. In this study, we investigated whether rMNSF beta also affects macrophage function in terms of LPS-induced TNF-alpha production by a mouse macrophage cell line, J774. rMNSF beta suppressed the TNF-alpha production in a dose-dependent manner. This suppressive effect was remarkably reduced when rMNSF beta was added after 6 h of LPS stimulation. In addition, enhancement of TNF-alpha production by IFN-gamma was also suppressed by rMNSF beta. The suppressive effect was partly neutralized by the addition of the serine/threonine phosphatase inhibitor, okadaic acid. This finding suggests that serine/threonine protein phosphatases type 1 and/or 2A may be implicated in the mechanism of action of MNSF.

Skin mast cell promotion of matrix remodeling in burn wound healing in mice: relevance of chymase.

Inflammation, granulation, and collagen accumulation, which are observed in the wound healing process, occasionally lead to hypertrophic scarring. Several in vitro reports have suggested that skin mast cells (MCs) and their major protease, chymase, participate in the healing process as well as in fibrotic skin diseases. The present study examined the potential involvement of MCs and MC chymase in the healing of burns in mouse dorsal skin. The size of the burn wounds, density of the capillaries, collagen accumulation, MC number, and chymase activity were measured before and 1, 3, 7, and 14 days after burning. The healing process corresponded strongly with MC density and chymase activity in both acute and subacute phases. The maximum decrease in MC number and chymase activity occurred on day 3 when tissue loss due to necrosis was maximal. From day 7 to 14, the burn wounds retracted rapidly accompanied by increases in capillaries and collagen fibers, in correspondence with fast increments in MC numbers and chymase activity at the wound edges. The present results combined with previous in vitro results strongly support the contention that skin MC chymase plays a role in the normal wound healing process, and presumably in dermal fibrotic disorders.

Inhibitory effect of lead on tube formation by cultured human vascular endothelial cells.

The effect of lead acetate (Pb) on the formation of capillary-like structures (tube formation) by cultured human umbilical vascular endothelial cells (HUVECs) was examined. HUVECs were seeded on a gelled basement membrane matrix (Matrigel). Treatment of HUVECs with 0.3-30.0 microM Pb for 24 hours inhibited the tube formation dose-dependently. The length of tube formation decreased time-dependently with 3.0-10.0 microM Pb. To elucidate the main target factor of Pb for this inhibition, the effects of Pb on the activity of protein kinase C (PKC) and Matrigel were examined. The addition of beta-phorbol 12-myristate 13-acetate (PMA, 50 nM), an activator of PKC, and isoquinolinesulfonamide derivative (H-7, 30 microM), an inhibitor of PKC, showed an increase and decrease in the tube formation, respectively. However, the results of simultaneous addition of Pb and either PMA or H-7 to HUVECs indicated that PMA and H-7 acted not synergistically but additively. When PKC activities in HUVECs were measured by a colorimetric assay after treatments with 3.0-10.0 microM Pb for 24 hours, there was no significant change in PKC activity in the cells. The Pb-inhibition of tube formation was suggested to be independent of PKC activity. Pretreatment of Matrigel with 3.0-10.0 microM Pb for different periods decreased the tube formation dose- and time-dependently. These findings suggest that Pb can inhibit the tube formation by HUVECs dose- and time-dependently and that the inhibitory effect of Pb could be dependent on the degeneration of Matrigel, not on PKC activity.

Expression of cathepsin B-like enzyme activity in cell lysate from cultured human normal keratinocytes.

In the present study, we have found that the cell lysate from cultured human normal keratinocytes from foreskin (HFKs) hydrolyzed alpha-N-benzoyl-DL-arginine beta-naphthylamide (BANA), and the BANA hydrolysis occurred most under conditions of 37 degrees C and pH 6.0. This activity was strongly inhibited by leupeptin, which is an inhibitor to cathepsin B. These results suggested that the cell lysate from cultured HFKs contained cathepsin B-like enzyme activity. This is the first report to demonstrate that cathepsin B-like enzyme activity was expressed in the cell lysate from human normal keratinocytes.

Effects of cycloheximide and actinomycin D on production of acidic nuclear proteins in human lymphocytes activated by phytohemagglutinin.

Previous studies have implicated a role for acidic nuclear proteins in the PHA stimulated activation of human lymphocytes. This study examines the mechanism of synthesis of these proteins. Cycloheximide inhibited their production, while Actinomycin D did not. It is suggested that synthesis proceeds by translation of dormant messenger RNA rather than of newly synthesized messenger RNA.

Ringworm-like cutaneous candidiasis: report of a case of localized type.

A 52-year-old Japanese female patient with ringworm-like cutaneous candidiasis on the right axillary area is described. It appeared to have developed in relationship to previous local steroid therapy in this area for a long time.

Tinea corporis due to Trichophyton verrucosum: report of a patient from the San'in District.

A 70-year-old Japanese housewife, a resident of Shimane Prefecture, developed an erythematous, annular skin lesion on her right forearm. Mycological examinations revealed that it was tinea corporis caused by Trichophyton verrucosum (T. verrucosum). This fungus was then shown to have been transmitted to her from young dairy cattle introduced from the Hokkaido District to the ranch of her son. To the best of our knowledge, she is the first reported patient with tinea due to T. verrucosum who was infected in the San'in District.

Two-dimensional polyacrylamide gel electrophoresis of human hair matrix proteins using isoelectric focusing electrophoresis as the first dimension.

S-carboxymethylated (SCM) matrix proteins from fifteen normal human scalp hair specimens were analyzed using two-dimensional polyacrylamide gel electrophoresis (PAGE) with usual isoelectric focusing electrophoresis as the first dimension and then with usual sodium dodecyl sulfate-PAGE as the second dimension. On the electrophoregrams obtained, one low-mobility smear of SCM proteins embedding one SCM protein band was always seen. Many higher-mobility SCM protein spots were also always seen. However, some variations were seen in these SCM protein compositions; in type I, there were eighteen SCM protein components and, in type II, seventeen components.

Comparison of human hair and nail low-sulfur protein compositions on two-dimensional electrophoresis.

Compositions of human normal hair and nail low-sulfur proteins were compared using two-dimensional electrophoresis of their S-carboxymethylated (SCM) derivatives. Six SCM low-sulfur protein components with molecular weights (MWs) of 76,000, 73,000, 72,000, 64,000, 61,000 and 55,000 were common to the hair and nail. One component with a MW of 61,000 was specific to hair, and two components, both with a MW of 50,000, were specific to nail.

Amounts of fibrous proteins and matrix substances in hairs of different races.

Fibrous proteins (FPs) and matrix substances (MSs) were extracted as S-carboxymethylated (SCM) derivatives from scalp hair of five individuals each of the Mongoloid, Negroid and Caucasoid races. The ratio of the extracted SCM FPs to SCM MSs in dry weight was calculated for each hair specimen. The ratios obtained from the Mongoloid hair were 0.45 +/- 0.03; those from the Negroid, 0.18 +/- 0.02; and those from the Caucasoid, 0.29 +/- 0.02. It has thus been suggested that the ratios of FP to MS contents in the hair may be involved in the determination of the macroscopic racial variations in the form of hair.

Amino acid compositions of human hair fibrous protein components purified with two-dimensional electrophoresis.

Normal human S-carboxymethylated (SCM) hair fibrous protein (HFP) components were purified with two-dimensional electrophoresis, and their amino acid compositions were examined. As previously reported, the SCM cysteine and glycine contents of the crude HFPs were characteristically high and low, respectively, as compared with those reported for the stratum corneum fibrous proteins (SCFPs). However, the SCM cysteine and glycine contents of the purified SCM HFP components were not as high or as low, respectively, but were rather very similar to those of the SCFPs. This suggests that, with respect to cysteine and glycine content, fibrous protein components similar to those of the stratum corneum exist in normal human hair.

Trichilemmal cyst: report of a case in which the cyst was connected with the epidermis by an epidermal canal.

An 87-year-old Japanese woman with a trichilemmal cyst (TC) on the subnasal area is described. Histopathological examination revealed that the cyst was connected with the overlying epidermis by an epidermal canal. This has suggested that the pathogenesis of TCs could be that they originate from the isthmus or lower portion of the hair follicle.

Intravascular papillary endothelial hyperplasia on the penis: report of a case.

An 82-year-old Japanese man with intravascular papillary endothelial hyperplasia on the penis is described. To the best of our knowledge, this is the first report of a patient who developed this tumor on his penis.

Black dot ringworm: the first report of a patient from the San'in District.

A 77-year-old Japanese woman, a resident of Shimane Prefecture, with black dot ringworm caused by a Trichophyton glabrum infection on the scalp is described. To the best of our knowledge, she is the first case of BDR reported from the San'in District.

Proliferating trichilemmal tumor with apocrine sweat glands.

A 43-year-old man with a proliferating trichilemmal tumor is described. Since the tumor had appeared on a pre-existing alopecia and accompanied ectopic apocrine sweat glands, it is supposed that the pathogenesis of the tumor of our patient might be similar to that of an organoid nevus.

Eosinophil-infiltrating folliculitis in childhood--report of a case.

A 5-year-old Japanese girl developed follicular pustules with erythematous plaques on the lower extremities. The histopathology was compatible with eosinophilic pustular folliculitis (EPF). However, the clinical findings were not typical of EPF; the pustules did not appear in crops, the pustules became ulcers, and the skin lesions appeared exclusively on non-seborrheic areas.

Nevus of ota with nevus of Ito--report of a case with cataract.

A 53-year-old Japanese male patient with both nevus of Ota and nevus of Ito developed cataract of the eye ipsilateral to the side of the nevi. In this patient, cataract might have appeared in a close pathogenetic relationship with nevus of Ota.

Tinea of the glans penis: report of a case presenting as a crop of papules.

A 27-year-old Japanese male student with tinea of the glans penis was described. The lesion on the glans penis was unique; it developed as a crop of papules.