Search

Ricinus communis agglutinin (RCA)-binding sites were visualized by transmission electron microscopy (TEM) in mice kidney, epididymis and other tissue cells, using the horseradish peroxidase (HRP) as a marker. RCA, a galactose-binding lectin, was conjugated covalently with HRP by the periodate method. Positive reaction with RCA was found at the plasma membrane, Golgi apparatus and multivesicular bodies. Control experiments (e.g. omission of the conjugate, addition of lactose to the incubating medium) established the specificity of the reaction. Plasma membrane staining is in agreement with previously published studies. The staining of the Golgi apparatus was for the first time demonstrated in this study. The positive sites were the cisternal surface of the vesicles and of the saccules of the Golgi apparatus. RCA-HRP staining can be used as a new Golgi marker in cytochemistry.

When a 13-day-old chick embryonic tarsometatarsal skin was cultured for 4 days in medium containing hydrocortisone (20 nM) and 5% delipidized fetal calf serum (FCS), epidermal growth factor (EGF, 100 ng/ml) decreased epidermal DNA content 42% and inhibited epidermal DNA synthesis 87%. Tonofilament bundles within the basal and intermediate cells of the EGF-treated epidermis were not as conspicuous as those in the glucocorticoid-induced keratinized epidermis, and the upper region of the EGF-treated epidermis did not form either the filament bundles or the electron-dense amorphous masses seen in the cytoplasm of the glucocorticoid-induced keratinized layer. EGF stimulated degradation of glucocorticoid-induced α-keratin. Furthermore, EGF caused a twofold increase in glucocorticoid-induced epidermal transglutaminase activity and in the amount of epidermal glucocorticoid receptor. In the absence of FCS, however, EGF did not inhibit steroid-induced α-type keratinization and did not affect either steroid-induced epidermal transglutaminase activity or amount of epidermal glucocorticoid receptor. Hence, the effect of EGF on glucocorticoid-induced epidermal transglutaminase activity was observed only in the presence of delipidized FCS and might be supported by an increase in the amount of glucocorticoid receptor.

Reactions of 4-(substituted benzylidene)-3, 5-dimethylisopyrazoles (1a-1c) with acetic anhydride, hydrochloric acid in methanol, bromine in acetic acid, dimethyl sulfate, and acyl chlorides (acetyl chloride, benzoyl chloride, ethyl chloroformate, and p-toluenesulfonyl chloride) in pyridine gave 1, 4-addition products (2-10) in fairly good yield. On the other hand, 4-(o-nitrobenzylidene) isopyrazole (1a) was converted to 5-chloro-3-(1-substituted 3, 5-dimethylpyrazolyl) anthranils (13, 14, and 15) by treatment with acetyl chloride, benzoyl chloride, or ethyl chloroformate without pyridine.

Changes in lectin binding of developing fetal mouse testes and ovaries were examined by light and electron microscopy, with much attention paid particularly to those in carbohydrates of germ cells. Characteristic binding patterns were observed with three lectins (BPA, GS-I, and GS-II) in the germ cells and the somatic cells during the process of testicular and ovarian development. GS-I and BPA, which showed similar binding patterns, preferentially bound to the plasma membrane and small dense bodies (SDB) of germ cells in both testes and ovaries during the 12th to 14th day post coitum (p.c.). In the fetal testes on day 16 p.c., the reaction with both GS-I and BPA completely disappeared. While, in the ovaries, a weak reaction with these lectins was retained as it was in germ cells until the 16th day p.c. The reaction with GS-II was restricted to Sertoli cells in the fetal testes during the 12th to 14th day p.c., and thereafter disappeared on day 16 p.c. The distribution of GS-II binding sites was in agreement with that of the glycogen granules. No positive staining with GS-II was seen in the ovaries throughout their development. These results indicate that certain glycoconjugates containing d-galactose and N-acetyl-d-galactosamine residues are expressed on the cell surface and in the SDB of germ cells during the period of the 12th to 14th day p.c., and that striking changes in function as well as in structure may take place in both germ cells and somatic cells during the 14th to 16th day p.c. in association with testicular and ovarjan development.

Publisher Summary This chapter discusses the preparation of a number of thiamine homologs and analogs. For the preparation of pyrimidine, 4-amino-5-bromomethyl-2-methylpyrimidine hydrobromide, the methods of Andersag and Grewe are excellent. And for the preparation of the thiazole, 4-methyl-5-(2-hydroxyethyl)thiazole, the method of Buchmann is outstanding. These methods can be applied to the preparation of a number of thiamine homologs and analogs. A practical method for the preparation of the 4,5-dimethyl compound (V: R = CH 3 ) as well as pyrithiamine is presented in the chapter. The intermediate, thiothiamine, formed by the condensation of 4-amino-5-aminomethyl-2-methylpyrimidine, CS 2 , and 3-acetyl-3-chloro-1-propanol is a water-insoluble crystalline compound and is easily converted by oxidation into thiamine in good yield. This method is very useful not only in the laboratory but also in industry and can be applied to the preparation of thiamine homologs and analogs by allowing the various 5-aminomethylpyrimidines to react with various α-chloroketones.

There are three isomers of dihydrothiamine, i.e., normal, iso and pseudo forms. In the present paper normal- and iso-dihydrothiamine were proved to have perhydro-furothiazole structure by means of infrared absorption spectral measurements and syntheses of their related compounds. It is assumed that the ring fusion is trans in the normal compound and cis in the iso compound.

The copper ferrocyanide method recently developed in our laboratory for the ultracytochemical demonstration of oxidoreductases was applied in the rat cerebrum. It was found that ferricyanide was reduced in synaptic membranes of both vesiculated and non-vesiculated bags, the synaptic cleft and membranes covering non-vesiculated bags in synapses, and in membranes of the neuronal perikaryon as well as processes including axons, when sodium succinate and cytochrome c were used as substrate. Some mitochondria in non-vesiculated bags of synapses and in the neuronal perikayron as well as processes also showed the reaction product. Based on control studies performed it was postulated that the oxidoreductase activity seen in membranes was due mainly to the cytochrome c-ferricyanide reductase.

Ferritin conjugates of two plant agglutinins, concanavalin A and ricin, have been used as specific electron microscopic stains for covalently-bound saccharide residues on membrane fragments from a myeloma-cell homogenate. The results indicate that different saccharide residues are uniformly localized to a single surface of each membrane fragment. In particular, the ferritin-concanavalin A conjugate binds exclusively to the cisternal side of membrane fragments of the rough endoplasmic reticulum. If it is postulated that the biogenesis of eukaryotic plasma membranes involves an assembly-line process from precursor intracellular membranes, these observed asymmetric distributions of saccharides on cell membranes can be explained.

Background Exogenous lipoid pneumonia is a rare disease caused by aspiration or inhalation of oily substances. Case presentation A 66-year-old male with dry cough (Case 1) and a 38-year-old female with shortness of breath (Case 2) demonstrated ground-glass opacities on chest computed tomography and were diagnosed with lipoid pneumonia based on the confirmation of lipid-laden alveolar macrophages. Both patients habitually performed sesame oil pulling via nasal or mouth washing for several months prior to the diagnosis. Conclusion Steroid therapy and bronchoalveolar lavage resulted in improvement in Case 1, and no intensive therapy was required for Case 2. Sesame oil pulling has been rarely been reported to cause lipoid pneumonia.

Background Uterine sarcoma is a rare tumor that is often difficult to classify based on morphological and immunohistochemical analysis alone. Limited access to molecular biological analysis in routine practice would hinder making a definitive diagnosis. Case Presentation In this report, we describe a case of a mesenchymal tumor arising from the uterine cervix in a 52-year-old woman. From microscopic morphology of the resected specimen, epithelioid leiomyosarcoma, high-grade endometrial stromal sarcoma, or uterine gastrointestinal stromal tumor (GIST) were considered as differential diagnoses. The immunophenotype of the tumor featured smooth muscle differentiation and hormone receptor expression. The cell membrane and cytoplasm were positive for c-kit, although no mutation was found in the c-kit or PDGFRA gene. Fluorescence in situ hybridization (FISH) analysis revealed a relatively low frequency of YWHAE rearrangement, whereas there were few NUTM2A and NUTM2B split signals. Conclusions In this case, the tumor was not typical of any three of the differential diagnoses mentioned above. However, insufficient frequency of YWHAE, NUTM2A, and NUTM2B gene rearrangement and absence of mutation in both the c-kit and PDGFRA genes suggested that this tumor should be categorized as epithelioid leiomyosarcoma. This is an instructive case showing a potential diagnostic pitfall of uterine sarcoma. Comprehensive approaches including molecular biological techniques are required for definitive diagnosis.

4-(o-Nitrobenzylidene)-3, 5-dimethylisopyrazole (1) was converted to 1-acyl-4-(α-hydroxy-o-nitrobenzyl)-3, 5-dimethylpyrazoles (2, 3) or 3-(1'-acyl-3', 5'-dimethylpyrazolo)-5-chloroanthranils (6, 7) by treatment with acyl chlorides with or without pyridine.

A musical tone signal is synthesized on the basis of a predetermined modulation operation (e.g. an FM or AM operation) employing a modulating signal and a carrier signal respectively having an audio range frequency. The modulating wave signal and/or carrier signal is derived by reading out a predetermined waveshape signal from a waveshape table in accordance with progressive phase angle data. The value of the waveshape signal read out from the waveshape table is modified in a specified phase section. This modification is effected by applying a simple operation such as gating, shifting or selecting to the phase angle data addressing the waveshape table or to the waveshape signal read out from the waveshape table. The modified waveshape signal is utilized in a predetermined modulation operation as the modulating wave signal and/or carrier signal. The frequency component of the modulating signal and/or carrier signal is controlled by the waveshape modification wherby synthesis of a musical tone having abundant frequency components is realized with a simple circuit construction.

Lectin binding sites of adrenocorticotropic hormone (ACTH) secretory granules of human pituitary adenomas and of nonadenomatous pituitary tissue adjacent to adenomas were studied by postembedding immunocytochemical doublestaining on ultrathin sections followed by electron microscopy. The specific hormones produced by the secretory granules were identified by labeling one side of the section with anti-human pituitary hormone antibodies conjugated to gold particles. Simultaneously, the other side was labeled with horseradish peroxidase-lectin to reveal lectin binding sites. Specimens were obtained from four human ACTH-producing pituitary adenomas and from nonadenomatous pituitary tissue surrounding three other adenomas. The four ACTH-producing adenomas showed either weak or negative reactions with concanavalin A, whereas the nonadenomatous ACTH-producing pituitary cells reacted strongly with concanavalin A. Moreover, ACTH secretory granules were significantly larger in the nonadenomatous cells than in adenoma cells. Differences in biochemical structure and ultrastructure between nonadenomatous (normal) pituitary cells and adenoma cells secreting the same specific hormones were demonstrated, and the clinical implications of the results were discussed.

Structurally different organic Ca antagonists such as D600, diltiazem and nicardipine enhanced the irreversible blockade by phenoxybenzamine (POB) of the histamine response in guinea pig taenia caecum. The isotonic contraction of the muscle to histamine was tested after incubation with POB (5xlO-6M, 10 min) with or without an organic Ca antagonist. The blockade by POB was greater in muscles treated with an organic Ca antagonist at concentrations that singly depressecene contraction to histamine to less than 50% of the control (D600, 5×10-7 M; di 1 tiazem, 1×10−5M;and nicardipine,3xlO-8M), and the effect lasted for more than 8 hrs;but the blockade was unchanged at lower concentrations of Ca antagonists. Mn ion had no such enhancing effect. The response to 40 mM K ion was not affected. The organic Ca antagonists alone at these concentrations did not affect the contraction to histamine after washing for 3 hrs. D557(desmethoxyverapamil) and 1-cis-isomer of diltiazem were less effective in enhancing than D600 & diltiazem, respectively. Antihistamines failed to protect the histamine receptor from the enhanced effect of POB. Only high doses of diphenhydramine which have high membrane stabilizing action could protect only the response to the maximal dose of histamine. Tritiated-POB binding to the intact muscle strips was significantly increased by nicardipine (3×10−°M). As the enhancing effect is parallel with the Ca antagonistic actions of the drugs used, it is concluded that the effect originated from the blockade of the Ca channel. We speculate as follows:The organic Ca antagonists stabilize the Ca channel in resting conformation, which makes some components of the receptor system more susceptible to POB. Eur.J.Pharmacol. 87 319 (1983)