Dynamic alteration of human β-defensin 2 localization from cytoplasm to intercellular space in psoriatic skin

Defensins are cationic antimicrobial peptides with a broad spectrum. Recently human beta-defensin 2 (hBD-2) has been isolated from psoriatic skin; however, its exact localization and fate have not been fully understood. We studied the distribution pattern of hBD-2 in skin tissues of psoriasis and other inflammatory skin diseases. In the upper spinous and granular layer of psoriasis vulgaris hBD-2 was present in the cytoplasm. In the horny layer the positive signals were in a basket-weave pattern, indicating possible accumulation of hBD-2 in the intercellular space. The similar pattern of hBD-2 distribution was observed in the lesions of nummular eczema and atopic dermatitis. hBD-2 was not detected in the section of normal elbow and knee skin. When isolated psoriatic scales were stained, hBD-2 was detected in a wrapping paper-like distribution pattern surrounding the corneocytes. In horny layer of psoriatic skin hBD-2 was closely associated or colocalized with elafin, which is known to be in extracellular space, as demonstrated by double staining. Western blot analysis using cultured human keratinocytes detected hBD-2 with an expected size in the conditioned medium and in the cell lysates when stimulated with 5% FCS or IL-alpha. These results indicate that hBD-2 was synthesized and remained in cytoplasm in the upper spinous and granular layer, and then secreted into intercellular space in the horny layer. This dynamic change in hBD-2 distribution in epidermis is certainly relevant to function as an innate host defense mechanism against invading micro-organisms.     

Purification of fully activated Clostridium botulinum serotype B toxin for treatment of patients with dystonia

Clostridium botulinum serotype B toxins 12S and 16S were separated by using a beta-lactose gel column at pH 6.0; toxin 12S passed through the column, whereas toxin 16S bound to the column and eluted with lactose. The fully activated neurotoxin was obtained by applying the trypsin-treated 16S toxin on the same column at pH 8.0; the neurotoxin passed through the column, whereas remaining nontoxic components bound to the column. The toxicity of this purified fully activated neurotoxin was retained for a long period by addition of albumin in the preparation.     

Stress-induced impairment of inhibitory avoidance learning in female neuromedin B receptor-deficient mice

Neuromedin B (NMB) is a mammalian bombesin (BN)-like peptide that exerts its function via the neuromedin B receptor (NMB-R). The NMB/NMB-R system is involved in stress response, and therefore we examined behavioral properties in female mice lacking NMB-R using a restraint-induced stress paradigm. Thirty minutes of restraint in a wire mesh cage constituted a sufficient stress stimulus for mice as evidenced by elevated blood glucose concentrations in stressed wild-type and NMB-R-deficient mice. Using a one-trial passive avoidance test, stressed NMB-R-deficient mice exhibited a marked reduction in memory performance. NMB-R-deficient mice exhibited elevated spontaneous activity in a novel environment compared to non-stressed mutant mice after 30-min stress, and a similar difference was also observed between stressed/non-stressed wild-type mice. An elevated plus maze test showed that the stress stimulus had no effect on anxiety in either wild-type or NMB-R-deficient mice. Furthermore, pain response of wild-type and NMB-R-deficient mice induced by electric foot shock was not affected under either stressed or non-stressed conditions. These results indicate that impaired memory performance in stressed NMB-R-deficient mice is not a consequence of changes in spontaneous activity, anxiety, or pain response, and suggest that the NMB/NMB-R pathway may play a role in regulating the stress response via the neural system that controls learning and memory.     

Release of non-glycosylated polymeric immunoglobulin receptor protein

Using a recombinant vaccinia virus containing the T7 RNA polymerase, we have established a system for the transient expression of human polymeric immunoglobulin receptor (pIgR) in baby hamster kidney cells, a baby hamster-derived fibroblastic cell line. This transfection system resulted in the successful expression of pIgR in these cells, and Western blot analysis showed that human pIgR was expressed as two different molecular weight forms of 92 and 107 kDa. Treatment with endoglycosidase H showed that the difference between these two forms was due to the glycosylation status of the protein. In order to examine the functional role of glycosylation, we treated the transfected cells with tunicamycin, which prevents a core glycosylation step in the endoplasmic reticulum. Non-glycosylated pIgR was released into the culture medium of the transfected cells, albeit with extremely low efficiency. Taking these results together, we conclude that the glycosylation of pIgR may play a positive role in the efficient transport or release of free pIgR.     

Uncommon Type of Meningiomas with Conspicuous Plasmo-lymphocytic Infiltration: An Immunohistochemical and Histochemical Study

Two cases of meningioma revealing conspicuous plasmo lymphocytic tissue and hyalinized fibrous tissue components are reported. Histopathological examination of the plasmo lymphocytic infiltration was performed. Both lesions showed polyclonality of plasma cells as revealed by positive reactions for 1gG and paraimmunoglobulin χ- and λ light chains, and amyloid infiltration into the fibrous stroma and blood vessel walls. The histochemical and immunohistochemical characteristics of the lesion in relation to its etiology are briefly discussed. Acta Pathol. Jpn. 32: 190∼194, 1989.     

Characterization of junctional sarcoplasmic reticulum Ca2+ content and release by short-term mechanical restitution in cardiac muscle

To study Ca2+ handling by the junctional sarcoplasmic reticulum (JSR), the time course of short-term mechanical restitution after varying magnitudes of twitch contractions was assessed in rat papillary muscle. Mechanical restitution consisted of a pretwitch latency period followed by a rapid and a subsequent much slower restitution of twitch force. The rate of rapid restitution was independent of the magnitude of the preceding twitch, which suggests that the rate of JSR Ca2+ repletion was dependent on the amount of Ca2+ remaining in the JSR after a twitch contraction. Based on this finding, the functions Gt and Ht, representing the time courses of JSR Ca2+ repletion and release, respectively, were derived graphically from a family of the mechanical restitution curves. Gt increased monotonically with time at a decreasing rate, while Ht increased with time in a sigmoid manner. The mechanical alternans were simulated by using experimental values and mathematically predicted values of Gt and Ht. A substitution of extracellular Na+ with Li+ to inhibit Na+/Ca2+ exchange resulted in an augmentation of Gt by approximately 10%, presumably by increasing the tubular SR Ca2+ uptake. The inhibition of tubular SR Ca2+ uptake by thapsigargin (10 microM) reduced mechanical restitution by approximately 13% of the maximal twitch force, independent of the phase of mechanical restitution; the effect was greater at an earlier time point in the mechanical restitution. These results suggest that early JSR Ca2+ replenishment results mainly from the movement of Ca2+ from the tubular SR.     

BASAL CELL ADENOMA WITH ACINIC DIFFERENTIATION

A case of basal cell adenoma in the left parotid region of a 45 years old male was reported. The tumor measured 1.1 cm × 0.9 cm, was spherical and covered with a fibrous capsule. Histologically, it was a monomorphic adenoma, forming solid, trabecular, tubular and acinic structure. The tumor cells secreted PAS-positive substance. Electron microscopically, the tumor consisted of three kinds of cells - secretory cells containing electron-dense secretory granules, non-secretory cells, and myoepithelial cells. The acinus was formed of single-layered secretory cells, in the base of which myoepithelial cells were observed. The tubulus was formed of both secretory and non-secretory cells, and myoepithelial cells were found in the base of the tubules. The interstitium was narrow, and was composed of a small amount of collagen fibers, myoepithelial cells, and basement membrane-like substance.     

Detection of Mason-Pfizer monkey virus infection by syncytia formation of human cells doubly transformed by Rous sarcoma virus and simian virus 40

Summary Human cells doubly transformed by Rous sarcoma virus and SV40 (RSb cells) formed syncytia by cocultivation with Mason-Pfizer monkey virus (MPMV)-producing cells. This cell fusion was blocked by anti-MPMV serum indicating that the phenomenon is MPMV specific. The RSb cells were successfully used for MPMV infectivity assay in the same manner as KC cells.With 1 Figure     

Studies on complement-potentiated neutralizing antibodies (C'-PNAb) induced in rabbits inoculated with japanese encephalitis virus (JEV). I. The nature of C'-PNAb

The C'-PNAb induced by JEV grown in porcine kidney stable (PS) cells [JEV(PS)] inactivate not only the corresponding virus, hut also Western equine encephalitis (WEE), Eastern equine encephalitis (EEE), vesicular stomatitis (VS) and Sindbis viruses grown in PS cells or primary hamster kidney (HK) cell cultures in the presence of complement. The degree of complement-potentiated neutralizing (C'-PN) ability varies for each virus. The C'-PNAb do not, inactivate these viruses grown in mouse brain, even JEV. The C'-PN activity against viruses other than JEV(PS) is completely removed by absorption with the microsomal fraction of PS or HK cells, but not of mouse brain. The antibodies in fraction IgM induced by the microsomal fraction of PS or HK cells inactivate the viruses grown in PS cells to a different degree in the presence of complement, but not viruses grown in mouse brain. The activity of C'-PNAb against JEV(PS) is reduced to 2% of the original activity by absorption with sheep red cells. After absorption, the remaining C'-PNAb are not further reduced by absorption with the microsomal fraction of PS cells, nor do they inactivate the other viruses grown on PS cells. The early rabbit hemagglutination-inhibition (HI) antibodies in fraction IgM induced by JEV(PS) could not only inhibit hemagglutination with JE, WEE, EEE, and Sindbis viruses grown on PS cells in the absence of complement, but could also facilitate HI in the presence of complement. However, they could not inhibit hemagglutination with these viruses grown in mouse brain, in the presence or absence of complement. This activity of HI could also be removed by absorption with the microsomal fraction of PS cells. These findings suggest that C'-PNAb are induced by host cell components associated with the virus, and that the early HI antibodies in fraction IgM are the same entities as C'-PNAb.