Use of microbial peptide inhibitors for characterization of the substrate specificity of thermitase,a thermostable serine protease fromThermoactinomyces vulgaris

Seven microbial peptide inhibitors—chymostatin, antipain, elastatinal, leupeptin, pepstatin, bestatin, and phosphoramidon—were tested for their efficiency to inhibit thermitase, a thermostable serine protease fromThermoactinomyces vulgaris. Chymostatin and antipain were the most effective inhibitors, with K_i values of 7×10^–8 M and 2×10^–7 M, respectively. Except for leupeptin, all inhibitors resist hydrolysis by thermitase. Leupeptin, however, is cleaved by thermitase between the two leucylresidues. Further, a close relationship in specificity between thermitase and subtilisin BPN and their distinct discrimination from elastase specificity was demonstrated by using these inhibitors.     

Ultrastruktur und Differenzierung der prostatoiden Organe von Polystyliphora filum (Plathelminthes,Proseriata)

Zusammenfassung Polystyliphora filum besitzt neben einem dem männlichen Begattungsorgan angeschlossenen Stilett eine Vielzahl gleichartiger prostatoider Organe, stets caudal des Begattungsorgans serial angeordnet. Jedes dieser prostatoiden Organe besteht aus einem Stilett, das in ein Atrium reicht, und einem caudal anschließenden Bulbus. Das Stilett hat die Form eines gebogenen Trichterrohres mit einem plattenförmigen Fortsatz in der Mitte; Trichterrohr und Fortsatz werden zusammen in einer einzelnen Zelle ausdifferenziert. In einer frühen Bildungsphase wird in der basalen Hälfte zunächst ein Gerüst aus Mikrotubuli angelegt, an das sich elektronendichtes Material anlagert. In einem späteren Bildungsstadium werden teilweise die Zwischenräume zwischen der entstehenden Hartstruktur und der Außenmembran der Bildungszelle mit elektronendichtem Material ausgefüllt. Die Spitze des Stiletts wird durch Anlagerung elektronendichten Materials an die Außenmembran gebildet. Die Differenzierung der gesamten Hartstruktur erfolgt simultan und intrazellulär. Gleichzeitig wird auch die gesamte Muskulatur des prostatoiden Organs ausgebildet. Die vollständig ausdifferenzierten prostatoiden Organe enthalten keine Spermien, sondern nur große Mengen eines grobscholligen Sekretes.

---

Ultrastructure and differentiation of the prostatoid organs of Polystyliphora filum (Plathelminthes, Proseriata)

Summary In addition to a male copulatory organ containing a stylet, Polystyliphora filum has numerous uniform prostatoid organs which are arranged in series caudally to the copulatory organ. Each of these prostatoid organs consists of a stylet, extending into an atrium, and caudal to this a bulb. The stylet is funnel-shaped with a curved distal part and a flattened projection in the middle; funnel and projection are differentiated together in a single cell. In an early phase of differentiation, a framework of microtubules is built in the basal part, and this becomes enveloped by electron-dense material. In a later phase, the space between the formed hard structure and the outer membrane of the style building cell is partially filled up with electrondense material. The distal part of the stylet if formed by electron-dense material taken up to the outer membrane. The whole hard structure is differentiated simultaneously and intracellularly. At the same time the whole muscular system of the prostatoid organ is formed. The completed prostatoid organs do not contain sperm, but much coarsegrained medium electron-dense secretion.

---

Abkürzungen am Atriummuskulatur - bl Basallamina - bm Bulbusmuskulatur - bmk Kernregion einer Bulbusmuskelzelle - bz Stilettbildungszelle - cw Cilienwurzel - ep Epidermis - fz Füllzelle - hz Hüllzelle - mv Mikrovilli - n Nerv - p Protraktor - pa Protraktoransatz - r ciliärer Rezeptor - s Stilett - sd Septatdesmosomen - sm Schließmuskel - sz Sekretzelleg     

Proteinaceous crystals in cells of virus infected plants

Crystal-containing organelles in cells of virus infected plants lying at chloroplasts and mitochondria are identical with single membrane-bound microbodies containing crystals of catalase described in healthy plants. Massive complex inclusions caused by turnip mosaic virus very frequently contain the same microbodies with crystal inclusions; that phenomenon may be related to some pathophysiological changes of virus infected plants. Comparable proteinaceous crystals, but not lying within microbodies limited by a membrane, may also be found in cytoplasm of infected cells. These crystals are sometimes surrounded by a substance resembling the microbody matrix. Disintegrated cytoplasm of virus infected cells may also contain the same crystals lying free in “empty spaces”. Cytopathological effects responsible for this phenomenon and possible artifacts as well are discussed.     

2-β-d-Glucopyranosyloxy-2-methylpropanol in Acacia sieberana var. woodii

A new diol glucoside, 2-β-d-glucopyranosyloxy-2-methylpropanol, the first reported naturally occurring monoglucoside of an aliphatic dihydric alcohol, was isolated from pods of Acacia sieberana var. woodii. Structure elucidation was based on ¹ H and ¹³C NMR spectroscopy, and enzymatic analyses. The compound was hydrolysed very slowly by almond β-glucosidase, but cleaved by a β-glucuronidase enzyme complex from Helix pomatia.     

Stimulation of renal tubular transport by ethacrynic acid in rats of different ages.

The transport system for organic acids in the kidney is not fully developed in the neonatal period. The effect of repeated administrations of ethacrynic acid on the renal excretion of p-aminohippurate (PAH) was studied in rats of different ages. Pretreatment with ethacrynic acid was followed by an increase in the renal excretion of PAH in 33-, 55-, 105- and 240-day-old rats but not in newborn rats. In 55-day-old rats the increase in renal excretion of PAH after pretreatment with ethacrynic acid was not associated with any consistent change of the glomerular filtration rate. It is concluded from these results that the stimulation of transport processes in the kidney by ethacrynic acid and some other drugs is linked with their affinity to tissue proteins.     

L-3-Phosphoglyceric acid,formed by ribulose-1,5-bisphosphate carboxylase,is the primary substrate for photorespiration: Experimental test of a hypothesis

Preparations of RuBP carboxylase are shown to carry out an oxygen dependent decarboxylation of L-3-phosphoglyceric acid. The product of this reaction is probably phosphoglycollate. L-3-phosphoglyceric acid, formed by RuBP carboxylase is therefore proposed to be the primary substrate for photorespiration.     

The spread of rhinoviruses in the Soviet Union (USSR) and in Czechoslovakia (CSSR) and the role of their pathology.

Virological, serological and immunofluorescence studies revealed circulation of rhinoviruses of the strains 1A, 1B, 2, 3, 7, 9, 10, 12, 13, 14, 16-23, 27, 29-33, 42, 48, 53, 56, 60 and 69 on the territory of Czechoslovakia and the Soviet Union. According to virological results, type 48 predominated and was followed in frequency of occurrence by types 27, 14 and 16 in the USSR and 30, 1A and 31 in the CSSR. RV infection in adults with ARD diseases was the aetiology in 28.5% of cases. Among the causative agents of acute rhinitis, rhinosinusitis and otitis, rhinovirus infection makes up a large proportion. Interrelation has been found between rhinovirus infection and chronic inflammation of the mucosa of the nose, the accessory sinuses and the ears. Prolonged course of rhinovirus infection and isolation of the virus in the late stages of disease (1-4 months after its onset) support the presumption that rhinoviruses promote chronicity of inflammation. RV was isolated from patients with chronic pneumonia from the lower respiratory passages (bronchial secretion). Rhinoviraemia was found in a child who died of ARD.     

X-Linked mental retardation with fragile X. a pedigree showing transmission by apparently unaffected males and partial expression in female carriers

Summary A large family is reported in which mental retardation associated with the fragile site at Xq28 was found. Three normal males seemed to have transmitted the trait through their daughters to affected grandchildren.A total of 19 family members were investigated cytogenetically. Mentally retarded males showed macroorchidism and the fragile X. Three mentally retarded females were found, with the fragile X in a high percentage of cells; in contrast, the obligate carriers showed no or only few cells with the fragile X.     

The metabolism of benzyl isothiocyanate and its cysteine conjugate.

1. The corresponding cysteine conjugate was formed when the GSH (reduced glutathione) or cysteinylglycine conjugates of benzyl isothiocyanate were incubated with rat liver or kidney homogenates. When the cysteine conjugate of benzyl isothiocyanate was similarly incubated in the presence of acetyl-CoA, the corresponding N-acetylcysteine conjugate (mercapturic acid) was formed. 2. The non-enzymic reaction of GSH with benzyl isothiocyanate was rapid and was catalysed by rat liver cytosol. 3. The mercapturic acid was excreted in the urine of rats dosed with benzyl isothiocyanate or its GSH, cysteinyl-glycine or cysteine conjugate, and was isolated as the dicyclohexylamine salt. 4. An oral dose of the cysteine conjugate of [14C]benzyl isothiocyanate was rapidly absorbed and excreted by rats and dogs. After 3 days, rats had excreted a mean of 92.4 and 5.6% of the dose in the urine and faeces respectively, and dogs had excreted a mean of 86.3 and 13.2% respectively. 5. After an oral dose of the cystein conjugate of [C]benzyl isothiocyanate, the major 14C-labelled metabolite in rat urine was the corresponding mercapturic acid (62% of the dose), whereas in dog urine it was hippuric acid (40% of the dose). 5. Mercapturic acid biosynthesis may be an important route of metabolism of certain isothiocyanates in some mammalian species.