Nerve growth factor in normal and psoriatic skin equivalent models.

Our objective was to determine the pattern and time course of nerve growth factor expression in an established skin equivalent model that we have used in the past to study wound healing and psoriasis phenotypes. Skin equivalents were constructed in triplicate using normal neonatal foreskin keratinocytes plated on collagen gels containing fibroblast lines. These lines were derived from five specimens of psoriatic lesions, three specimens of normal skin from patients with psoriasis, and three specimens of eyelid skin from normal donors. Immunohistochemistry and a monoclonal nerve growth factor-b antibody were used to determine the pattern of protein staining over 2 weeks. We looked at the wound healing phenotype using the skin equivalent model for 7-14 days. When keratinocytes invaginate into the dermis of skin equivalents (beginning at around 7 days of growth), dark staining of nerve growth factor was seen under the basal membrane zone, suggesting that nerve growth factor serves in the development of the basal membrane zone and the epidermis, and may influence the migration of nerves through the basal membrane zone into the regenerated skin.

Artificial skin equivalent differentiation depends on fibroblast donor site: use of eyelid fibroblasts.

Our objective was to construct and explore human skin equivalents from several normal and pathologic donor skin sites to determine if the fibroblast origin influences epidermal differentiation. Also, we wanted to find out if fibroblasts from some donor sites produced epidermis of superior quality for plastic surgery repairs. Skin equivalents were constructed from 15 normal skin specimens, 9 normal eyelid specimens, 15 lesional skin specimens taken from patients with psoriasis, and 4 specimens from keloid scars. Results show that the tissue origin of the donor fibroblasts determines epidermal differentiation and the time period for regeneration. Eyelid fibroblasts were very dependable in establishing well-differentiated skin equivalents in all nine specimens. Our findings contribute to the accumulated knowledge of wound healing and should also be of value in skin grafting, especially when large areas are denuded as in burns, severe trauma, or cancer ablation.

Interleukin-8 is induced in skin equivalents and is highest in those derived from psoriatic fibroblasts.

Interleukin-8 (IL-8) may be important in psoriasis as it is expressed in the stratum granulosum, attracts polymorphonuclear cells, and stimulates angiogenesis and keratinocyte mitogenesis. To study intrinsic cutaneous factors in psoriasis, we constructed skin equivalents from psoriatic or adult control fibroblasts with normal foreskin keratinocytes. IL-8 levels were measured in supernatants by enzyme-linked immunosorbent assay and in skin equivalents by immunohistochemistry and in situ hybridization. IL-8 was highly induced in skin equivalents compared to cells grown alone. Epidermal stratification varied among fibroblast lines and was correlated with IL-8 levels, but lesional and nonlesional psoriatic skin equivalents from the same donor were similar. Six fibroblast lines (two psoriasis lesion and four normal) supported only monolayers, while 12 lines (seven psoriasis lesion and five normal) produced stratification. Mean IL-8 levels were significantly lower in dermal equivalents of the first group than the second (0.78 +/- 0.40 vs 3.93 +/- 2.83 ng per ml, mean +/- SD, p = 0.01, analysis of variance). Significantly more IL-8 was secreted by psoriatic than normal fibroblast skin equivalents over 14 d (p = 0.015) with greatest differences at 1 and 4 d. Psoriatic IL-8 levels peaked first and remained increased. IL-8 protein and mRNA were initially strongest in dermal fibroblasts, and at the dermal-epidermal interface. Diffuse epidermal expression was replaced by accentuation in the stratum granulosum. Psoriatic skin equivalents were thicker, had more intense IL-8 staining, and developed invagination. We hypothesize that an IL-8 paracrine loop between fibroblasts and keratinocytes may play a key role in epidermal regeneration in the skin equivalent, in normal wound healing, and in the determination of an intrinsic psoriatic wound-healing phenotype.

[Diagnosis of supravalvular stenosis of the aorta]. / K voprosu diagnostiki nadklapannogo stenoza aorty.

The authors analyzed the potentialities of combined radiodiagnosis of supravalvular stenosis (SS) and concomitant diseases of the heart and major vessels (Williams-Beuren syndrome) in 7 patients aged 7 mos. to 24 yrs. Polypositional chest and heart x-ray procedure, catheterization of the cardiac cavities, pulmonary artery and aorta, left ventriculography (6), right ventriculography (4) and angiopulmonography were used. The diagnostic potentialities of each method were defined. It was proved that SS practically in all patients was accompanied by major vascular changes in the pulmonary, vertebral, coronary, carotid, subclavicular, renal and other arteries, heart failure (atrial septal defect, and mitral incompetence).

[Isolation and identification of the antibiotics amicetin and nebramycin formed by a Streptomyces coeruleoaurantiacus 4009 culture]. / Vydelenie i identifikatsiia antibiotikov amitsetina i nebramitsina, obrazuemykh kul'turoi Streptomyces coeruleoaurantiacus 4009.

It was found that Str. coeruleoaurantiacus, strain 4009 produced antibiotics 4009-A and 4009-B belonging to different groups. Antibiotic 4009-A was identified as amicetin belonging to the group of pyrimidine bases and antibiotic 4009-B as the nebramycin complex belonging to the group of aminoglycosides. The identity of the antibiotics was confirmed by the physico-chemical constants, spectral and chromatographic characteristics and their chemotherapeutic activity.

The structure of nocamycin, a new antitumor antibiotic.

The structure of nocamycin, a new antitumor antibiotic, has been elucidated with the aid of mass- and PMR-spectroscopic investigation of the antibiotic and its various chemical transformation products. Nocamycin is structurally related to tirandamycins.

[Physicochmemical characteristics of the new antineoplastic antibiotic, nocamycin]. / Fiziko-khimicheskaia kharakteristika novogo protivoopukholevogo antibiotika nokamitsina.

Nocamycin is produced by Nocardiopsis syringae. It is recovered from the culture fluid by extraction with chloroform. The molecular weight of the crystalline antibiotic is 503, its melting point is 147--149 degrees, [alpha]20 degrees D = --50 degrees (c. 0.21, chloroform), lambdamax235 and 348 nm(E1%sm= = 150 and 420), the summation formula is C26 H33NO9, the biological activity is 100000 Units/mg with respect to Bacillus mycoides. Nocamycin forms salts with alkalies soluble in water. On hydrolysis with an alkali it forms carbonic acid having no ester bond (IP spectrum) and methoxylic group present in the antibiotic molecule. Nocamycin is a new natural substance.

[Extraction of rifamycin B from native and aqueous solutions]. / Ekstraktsiia rifamitsina B iz nativnogo i vodnogo rastvorov

Optimal conditions for extraction of rifamycin B from aqueous solutions and fermentation broth filtrates at pH values within 2.0-7.0 were determined. When the antibiotic was extracted from the aqueous solutions, the highest yield was obtained at pH 2.0. When the antibiotic was extracted from the fermentation broth filtrates, it was found that chloroform was the most selective solvent with respect to rifamycin B, the chloroform selectivity being increased at pH 3.5-4.0. It was shown that rifamicin B passed from the buffer solutions with a concentration of 3-20 mg/ml to chloroform in amounts of 6-7 mg/ml and to ethylacetate and butanol in amounts of 20 mg/ml. Such conditions of chloroform and butanol (9 : 1) increased the rifamycin B contents in the extract up to 40 mg/ml.

[Spontaneous and induced variability in Actinomyces rubiginosohelvolus, a new producer of the antibiotic rubomycin]. / Spontannaia i indutsirovannaia izmenchivost' Actinomyces rubiginosohelvolus--novogo produtsenta antibiotika rubomitsina.

Mutants possessing 3 times higher activity as compared to the initial culture were obtained as a result of selection of active variants of Act. rubiginosonelvolus, a new organism producing rubomycin against the background of variation induced by N-methyl-N1-nitro-N-nitrozoguinidine, diethylsulphate and their combinations. Further selection among the mutants stable to 6-mercaptopurine and riboflavine resulted in obtaining an active culture No. 3912 at least 7 times more active with respect to rubomycin production than the initial soil culture.