Cryopreservation of in vitro Shoot Tips of Ulluco (Ullucus tuberosus Cal.) Using D Cryo-Plate Method.

　　BACKGROUND: Maintenance of in vitro collections of ulluco (Ullucus tuberosus Cal.) is cumbersome and costly in an ex-situ genebank. An alternative method for long term preservation which is safe and cost-effective is required. OBJECTIVE: To apply a novel cryopreservation procedure using the cryo-plate system to improve the long-term conservation of ulluco. MATERIALS AND METHODS: Initially V and D cryo-plate methods were tested, subsequently the D cryo-plate method was selected for ulluco cryopreservation. The D cryo-plate procedures were optimized for post-LN regrowth procedures including cold-hardening, sucrose addition in alginate gel, and duration of LS treatment. Optimized procedures were tested with 11 ulluco lines. RESULTS: Shoot tips were isolated from cold-hardened shoots for 3-4 weeks at 5 degree C were excised to 1.0-1.5 mm long and 0.5 mm wide and precultured for 16h at 25 degree C on MS with 0.3 M sucrose. The shoot tips were attached on the cryo-plates by alginate gel with 0.4M sucrose. The cryo-plates with attached shoot tips were treated with 2.0 M glycerol and 1.0 M sucrose solution for 90 min at 25 degree C and dehydrated on filter paper in a Petri dish by air current flow at 25 degree C for 45 min before direct immersion in LN. This optimized procedure was applied to shoot tips of 11 ulluco lines, resulting regrowth ranging from 73 % to 97 %, with an average of 90 % post-LN regrowth. CONCLUSION: D cryo-plate is a practical and simple procedure for cryo-storage of in vitro grown ulluco shoot tips in an ex situ genebank.

Epidermodysplasia verruciformis: viral particles in early malignant lesions.

Two cases of epidermodysplasia verruciformis were studied histologically and electron microscopically. Four lesions examined were histologically benign, and had viral particles morphologically similar to that of common human warts. Two lesions on the forehead and the face were histologically in the stages of malignant transformation, the intraepidermal epithelioma and the early invasive squamous cell carcinoma. Similar viral particles were also observed in the upper layers of these 2 lesions. These results suggest that at least some of the virus-induced lesions of epidermodysplasia verruciformis actually become malignant.

Prolidase deficiency: its dermatological manifestations and some additional biochemical studies.

Prolidase deficiency occurred in a 13-year-old girl. Determinations were made of prolidase and prolinase activities in cultured fibroblasts, and thin layer chromatographic studies of skin prolidase were performed. The patient had chronic, recurrent ulcers on the legs and feet, diffuse telangiectasia, shallow scar-like atrophic lesions on the face and arms, soft and thin abdominal skin, and premature gray hairs. Prolidase in the patient's skin fibroblasts was absent. Greatly reduced prolidase activity was demonstrated in the patient's skin. A review of hitherto reported cases of this disease showed such skin manifestations as (1) skin ulceration or skin fragility with scar formation, (2) purpuric lesions, (3) telangiectasia and/or photosensitivity, and (4) thickening of the skin with lymphedema.

Stationary positron emission tomography and its image reconstruction.

Feasibility of stationary positron emission tomography (PET) using discrete detectors has been investigated by simulation studies. To enable full utilization of detector resolution, a "bank array" of detectors is proposed and an EM algorithm is adopted for image reconstruction. The bank array consists of an odd number of detector banks arranged on a circular ring with a gap equal to one half the detector width. The EM algorithm [11] is used with some modifications for reducing the quantity of computation, improving the convergence speed, and suppressing statistical noise, so as to meet the present purpose. Simulation studies involving several phantoms show that the stationary PET with the new detector array provides image quality which is good enough for clinical applications. For fast dynamic studies with low spatial resolution, the convolution-backprojection method is efficient, but for high-resolution static imaging, resolution enhancement by an iterative method is required. Problems arising in the corrections for attenuation of photons and detector sensitivity, etc., are also discussed. A totally stationary PET avoids the mechanical problems associated with accurate movement of heavy assemblies and is particularly advantageous in gated cardiac imaging or in fast dynamic studies. Elimination of a scan along the detector plane allows a quick scan in the axial direction to achieve three-dimensional imaging with a small number of detector rings.

A semiquantitative measurement of anti-nuclear antibody using immuno-microfluorimetry.

1. Using microfluorimetry, the strength of fluorescence was measured on fluoresceinated anti-nuclear antibody of SLE sera. The indirect "Sandwich" method was applied using human peripheral lymphocytes as substrates tissue. 2. The results of using three FITC-labelled anti-human IgG conjugates of different types were compared with each other. 3. More specific and more consistent results were obtained with conjugates with a lower F/P ratio and lower antibody and protein concentration that those in the case of conjugates with a higher F/P molar ratio and higher antibody and protein concentration. 4. The immuno-microfluorimetric technique is very useful for the objective determination of the results of fluorescence microscopy and for the comparison of conjugates of different qualities.

Persistence of foam cells in rabbit xanthoma after normalization of serum cholesterol level.

Xanthoma was produced in diet-induced hypercholesterolemic rabbits by intradermal dextran sulfate injections. The serum cholesterol level returned to the normal range at about 10 weeks after ending the cholesterol diet. Gross observations after cessation of the cholesterol diet revealed a decrease in xanthomatous infiltrations. However, the dense foam cell infiltrations and cholesterol accumulations showed no signs of regression at even 9 months after ending the cholesterol diet. Signs of foam cell migration into the blood stream were not observed. The persistence of the xanthoma may be due to a lack of acceptors, such as high-density lipoproteins, that remove the cholesterol from the foam cells. During our 9-month observation period, some foam cells were degenerated and a few were fused with each other to transform into Touton-type giant cells. Nonfoamy histiocytes were infiltrated around these degenerating foam cells. The histiocytes may have transformed into foam cells by incorporating the lipids of the degenerated foam cells.

The local noise property in positron volume imaging and optimal conditions for the signal-to-noise ratio of the 3D reconstructed image.

The local noise property of a 3D PET reconstructed image is investigated for a uniform-activity sphere distributed in a constantly attenuating spherical object. The positional dependence of the statistical noise is approximately derived and calculated for some special cases. It is suggested that a larger diameter of the activity sphere causes noise amplification, and the noise property for the large attenuating sphere is close to that for a non-attenuating object with the same total number of measuring counts. By considering noise propagation of two spherical activity distributions, we suggest that the signal-to-noise ratio of the image depends on a set of projection directions and the sizes and intensities of the activity distributions. In a simple case, we derive an optimal value of the maximum acceptance angle for the projection directions to improve the signal-to-noise ratio of the image.

Marked increase in the rate of ocular lens regeneration in the newt, Cynops pyrrhogaster, following partial body exposure to low dose X-rays.

In recent years, concern over the stimulating effects of low-dose X-rays has been growing. Therefore, the effects of low-dose X-irradiation on lens regeneration in the newt were examined. Newts were subjected to sham or whole-body X-ray exposure at a dose of 0.2 Gy or 0.4 Gy, delivered at a rate of 0.43 Gy min(-1). The eyeballs were fixed in formalin solution, embedded in paraffin and assessed histologically. On day 14 after lens removal, unexposed animals showed the formation of a hollow epithelial vesicle of depigmented cells continuous with the laminae of the iris corresponding to the expected regeneration stage (Reyer's regeneration stage II). In contrast, lenses from newts exposed to a 0.2 Gy dose of X-rays showed some formation of the primary lens fibre nucleus corresponding to the fibre differentiation stage (Reyer's regeneration stage III-early). Thus, low-dose X-irradiation induced regeneration compared with the unexposed groups. An acceleration from Reyer's stage II to III-early was also found on day 14 following irradiation of only the upper belly, including the spleen. The effects of low-dose X-irradiation on lens regeneration may be mediated by changes in immune activity.

Demonstration of immunoglobulin G in normal human epidermis by peroxidase-labeled antibody.

Cytoplasmic immunoglobulin G (IgG) in normal human epidermis was defined by a peroxidase-labeled antibody method. A correlation between cytoplasmic staining and the serum level of IgG was found. Epidermal cells containing IgG were not present when the serum level of IgG was less than 1000 microgram/ml.

Cutaneous mucinosis associated with SLE--a case provoked by PUVA.

A patient with SLE which was complicated by a characteristic cutaneous mucinosis is reported. The skin lesions were composed of intradermal soft nodules, purple-red plaques, and alopecia of the scalp. Acid mucopolysaccharides accumulated in each lesion. PUVA provoked an infiltrated skin lesion in which acid mucopolysaccharides were deposited in the dermis. Deposition of immunoglobulins and complements were also observed at the dermo-epidermal junction and on the vascular walls. Some immunological processes which may stimulate the synthesis of acid mucopolysaccharides in the dermis were suggested.

[Comparative clinical study of roxithromycin and josamycin for suppurative skin and soft tissue infections by a double-blind method].

The clinical efficacy and safety of Roxithromycin (RU 28965, RU), a new macrolide preparation, were compared with those of Josamycin (JM) in superficial suppurative skin infections. The study was designed as double-blind controlled trial with daily dosages of 300 mg in RU group and 1200 mg in JM group. A total of 209 cases (RU:105; JM:104) was analyzed and the final global improvement rating was 82.9% in the RU group and 80.8% in the JM group; there was no significant difference between the two groups. Slight adverse reactions were observed in 3.6% (4 cases) of the RU group and in 4.6% (5 cases) of the JM group. In conclusion, RU at daily doses of 300 mg is as effective as JM at daily doses of 1200 mg in superficial suppurative skin infections.

[Experimental studies of cefmetazole in the field of dermatology].

Cefmetazole was studied experimentally in the field of dermatology. The following results were obtained. 1. The in vitro antibacterial activities of cefmetazole against S. aureus and S. epidermidis were studied. Most strains of both staphylococci were inhibited by 1.56 micrograms/ml or less of cefmetazole. 2. Serum and skin concentrations of cefmetazole in rats were studied after intramuscular administration of 20 mg/kg of cefmetazole. Mean serum concentrations were 40.3, 21.5, 7.1, 0.56 micrograms/ml respectively at 0.25, 0.5, 1 and 2 hours, and the corresponding skin concentrations were 12.5, 6.1, 0.62, 0.61 micrograms/g (n = 4).

[Clindamycin-2-phosphate in the field of dermatology (author's transl)].

1. MIC against 20 strains of staphylococcus isolated from skin infections was studied. MIC was less than or equal to 0.05 mug/ml in 11 strains and greater than 100 mug/ml in 9 strains. 2. Clindamycin-2-phosphate was injected intramuscularly to rats at the dosis of 10 mg/kg in solution of 10 mg/ml. The serum and skin levels of this drug were determined at 0.5, 1, 2, 4 and 7 hours. Mean serum levels of 4 rats were 0.893, 0.578, 0.463, 0.268 and 0.167 mug/ml respectively at 0.5, 1, 2, 4 and 7 hours. The corresponding skin levels were 0.500, 0.707, 0.431, 0.313 and 0.269 mug/g. 3. Clindamycin-2-phosphate was used clinically in 7 patients of skin infection with the following result: excellent in 2 cases, fair in 3 cases and poor in 2 cases. Abdominal pain was complained in one case and loose stool in one case.

[Clinical use of amoxicillin preparation with prolonged activity in dermatology (author's transl)].

C-AMOX was administered to adult patients of acute bacterial skin infection at the dosage of 500 mg 2 times a day after meals in the morning and in the evening. Evaluation was done in 4 grades, i.e. excellent, good, fair and poor. Of 42 cases treated, excellent was marked in 11 cases and good was in 14 cases. Percentage of excellent and good results was 59.5%. Fair results being included, the percentage rises to 83.3%. As subjective side effects, stomach trouble (including stomachache) were observed in 2 cases and diarrhea, perleche, rash, nausea and loss of appetite were each 1 case.

[Experimental and clinical studies of fortimicin in the field of dermatology].

1. Fortimicin (KW-1070) was found to be similar to amikacin in vitro activity against S. aureus of 27 strains and S. epidermidis of 17 strains isolated in dermatological field. 2. Serum and skin levels of the drug were determined in rats. Mean serum levels (n = 4) were 4.91 mcg/ml at 1/4 hour, 2.40 mcg/ml at 1/2 hour, 2.40 mcg/ml at 1 hour and 0.51 mcg/ml at 2 hours. The corresponding skin levels were 2.15 mcg/g, 2.27 mcg/g, 0.83 mcg/g and 0.19 mcg/g. 3. Seven cases with dermatological infections were treated with fortimicin. No side effects were observed both in subjective and in laboratory findings.

[Double-blind comparison of cefadroxil and cephalexin granules in the treatment of impetigo].

A double-blind clinical comparison of cefadroxil granules (CDX) and a long-acting preparation of cephalexin granules (L-CEX) was conducted in the treatment of impetigo in children. The long-acting cephalexin preparation was composed of 3 parts of granules soluble in the stomach and 7 parts of those soluble in the intestine. Drugs were administered at 3 different doses depending on the body weight of patients (Group I: less than 10 kg, Group II: between 10 kg and 20 kg, Group III: over 20 kg). Overall clinical evaluation by attending doctors: CDX group scored slightly better but not statistically significant results. Evaluation of usefulness: CDX group scored slightly better but not statistically significant results. Partially standardized evaluation: CDX scored statistically significantly higher points in the evaluation of total cases and Group II on Day 5, and Group II on Day 7. Cumulative points through Day 5 and Day 7 were not statistically significant. Follow-up evaluation by attending doctors: CDX group got a better result in total cases and Group II on day 3, and in total cases on days 5 and 7. Otherwise there was no statistically significant difference. Incidence of side effects was not significantly different between the 2 drug groups. As shown in Fig. 1, CDX was administered on rising in the morning, after lunch, and at bed time whereas L-CEX was given on rising and at bed time when the patients were relatively in fasting condition. This may have reduced the expected merit of L-CEX and some reserve is required in the interpretations of the results shown herein.

[Use of cefroxadine dry syrup in the management of acute skin infections in children (author's transl)].

1. Cefroxadine dry syrup was in principle administered at the dosage of 10 mg per kilogram of body weight 3 times a day. 2. Evaluation was done in 4 grades, i.e. excellent, good, fair and poor. 3. According to subjective judgement by attending doctors, 'excellent' or 'good' was recorded in 90.7%. 4. If the evaluation was partially standardized, 'excellent' or 'good' was obtained in 74.8% of total 163 cases and in 78.7% of 108 impetigo cases. 5. Side effects were observed in 3 cases (diarrhea 1, fever 2). No direct correlation of these complaints with the administration of the present drug was confirmed.

[Response analysis for an approximate 3-D image reconstruction in cone-beam SPECT].

Cone-beam Single Photon Emission Computed Tomography (SPECT) offers the potential for a large increase in sensitivity as compared with parallel hole or fan-beam collimation. Three-dimensional image reconstruction was approximately accomplished by backprojecting filtered projections using a two-dimensional fan-beam algorithm. The cone-beam projection data were formed from mathematical phantoms as analytically derived line integrals of the density. In order to reduce the processing time, the filtered projections were backprojected into each planes parallel to the circle on which the focal point moved. Discrepancy of source position and degradation of resolution were investigated by computer simulation in three-dimensional image space. The obtained results suggest that, the nearer to the central plane or the axis of rotation, the less image degradation is performed. By introducing a parameter of angular difference between the focal point and fixed point in the image space during rotation, degradation of the reconstructed image can be estimated for any cone-beam SPECT system.