Characterization of the purified multifunctional cellulase component of Penicillium funiculosum

Summary The endoglucanase component (CMCase I) ofPenicillium funiculosum cellulase was purified to apparent homogeneity by ultrafiltration and gel chromatography. It consists of a single polypeptide chain with a molecular weight of 56000 and is a glycoprotein. Viscometric and end-product analysis revealed the randomness of enzyme action. Multifunctional characteristic of CMCase I was studied with various carbohydrate substrates.NCL Communication No.: 4307     

ARAGONITE CRYSTALS IN SPIROGYRA SP. (CHLOROPHYTA)

Intracellular crystals of aragonite have been identified by selected area electron diffraction (SAED) in a species of the freshwater filamentous alga Spirogyra from the Thames River, Ontario, Canada. The crystals are 2 to 24 μm in diameter, and characterized by a unique cross-shaped morphology, in which needle-like, or prismatic outgrowths develop from a common axis. Crystals may be dispersed throughout filaments, but tend to cluster as aggregates towards the centre .     

The multicomponent analysis of estrogens in urine by ion exchange chromatography and GC-MS--I. Quantitation of estrogens after initial hydrolysis of conjugates

A method for the multicomponent analysis of estrogens in urine after initial hydrolysis of the conjugates is described. Following protection of the carbonyl functions by ethoximation, estrogen conjugates were extracted on Sep-Pak C18 cartridges and purified on the acetate form of DEAE-Sephadex. The samples were subsequently hydrolysed by Helix pomatia juice and the hydrolysate was purified on the acetate form of QAE-Sephadex. Estrogens with vicinal cis-hydroxyls and diphenolic compounds were fractionated on the borate and bicarbonate form of QAE-Sephadex, respectively. Neutral steroids were removed by the free base form of DEAE-Sephadex after which estrogens were separated into two groups using Lipidex 5000 in a straight phase system. Following trimethylsilyl ether derivatization estrogens were analysed by selected ion monitoring (SIM). The method allows the quantitation of all the important estrogen metabolites including catechol estrogens. It is precise, accurate and sensitive permitting the quantitation of estrogens in urine of males and non-pregnant females.     

Activation of fatty acids by non-glyoxysomal peroxisomes

Ethylene treatment (approx. 20 l ·1^-1 in air for 2 d) of tobacco (Nicotiana tabacum L. cv. Havana 425) plants markedly increases the endo--1,3-glucanase (EC 3.2.1.39) content of leaves. The antigenic form of the enzyme induced is the same one whose production is blocked by treating cultured cells with combinations of auxin (1.1 · 10^-5 M -naphthaleneacetic acid) and cytokinin (1.4 · 10^-6 M kinetin). Evidence is presented that cultured tobacco cells require ethylene for -1,3-glucanase accumulation: i) ethylene treatment increased the accumulation of \-1,3-glucanase in callus tissues >10 d after subculturing and in cell-suspension cultures; ii) callus tissues can produce ethylene; iii) conditions known to inhibit ethylene production (1 mM CoCl₂; 33° C treatment) or ethylene action (approx. 1.6 mmol · 1^-1 norbornadiene in air) inhibited -1,3-glucanase accumulation by callus tissues treated for 4 d following subculturing; and, these inhibitory effects were prevented by exogenous ethylene. Combinations of auxin and cytokinin blocked ethylene-induced accumulation of -1,3-glucanase by cell-suspension cultures. The results favor a model in which ethylene induces results favor a model in which ethylene induces 1,3-glucanase accumulation, and auxin and cytokinin inhibit this induction process.Abbreviations NAA -naphthaleneacetic acid - NDE norbornadiene     

The right ventricular working heart preparation

An isolated working rat heart preparation was modified to study right ventricular (RV) performance. All hearts were perfused with a Krebs-Henseleit bicarbonate buffer via a Langendorff column at 90 mm Hg. Right atrial filling (preload) was varied by raising a buffer reservoir from 5 cm below to 10 cm above the right atrium while pulmonary artery outflow resistance remained fixed. RV systolic pressure and the maximum rise and decrease in pressure development (+/- dP/dt) were measured via a catheter in the RV. Cardiac output was collected with a catheter placed in the pulmonary artery. One group of hearts, monitored at a fixed preload (0 cm H2O) for 2 hr, and another group of hearts, in which two ventricular function curves were performed, demonstrated the stability and reproducibility of the preparation. Additionally, the ability of this preparation to measure changes in inotropy was studied. A negative inotropic effect was measured after verapamil (5 X 10(-8) M) treatment. Positive dP/dt showed the greatest depression (30%) and was significantly lower at every preload. A positive inotropic effect was demonstrated by reducing the buffer Ca2+ concentration to 1.9 mM for the first work curve followed by an addition of Ca2+ (2.8 mM final concentration) or ouabain (5 X 10(-5) M) for the second work curve. Again, the greatest effect was found in the dP/dt measurements (elevated by 20 and 30%, respectively). Thus, this preparation manifests qualities similar to those used in studying the left ventricle and allows investigation of various cardiac diseases which may affect RV pump function.     

Hynes pharyngoplasty revisited

The treatment of velopharyngeal incompetence remains unsatisfactory because the causes are many, as are the variations in anatomic and physiologic defects. Therefore, full assessment and investigation are essential in tailoring the surgery to the defect. A modified Hynes pharyngoplasty has been used in 40 patients, aged 4 to 52, over a 4-year period for velopharyngeal incompetence of varying etiologic causes. Speech was assessed before and at least 6 months after pharyngoplasty. At the same time, radiologic and, when possible, nasendoscopic investigations were undertaken. Thirty-eight patients had no or variable nasal escape (variable defined as achieving intermittent closure), whereas 33 had normal or slight hyponasal resonance. There was only one complication, an asymptomatic dehiscence of the "bucket handle" flap from the posterior wall. Thirteen patients had an assortment of side effects, none requiring surgical treatment. We believe that patients who are suitable for the described sphincter pharyngoplasty are those with slight or moderate nasal escape having a mobile palate with an anteroposterior gap of 5 mm or less.     

Aspirin and acetaminophen use by pregnant women and subsequent child IQ and attention decrements

In a longitudinal prospective study of 1,529 women pregnant in 1974-1975, aspirin and acetaminophen were the two medications most frequently taken during the first half of pregnancy (46 and 41%, respectively). In a selected cohort of 421 offspring of these women, examined at 4 years of age, maternal aspirin use during the first half of pregnancy was significantly related to IQ and attention decrements in the exposed children. Multiple regression analyses were used to statistically adjust for a variety of potentially confounding factors including demographic characteristics, child characteristics, other exposures, and lifestyle/environmental variables. Continuous dose-response and step-function parameterizations of aspirin exposure were both statistically significant and not clearly distinguishable from each other. The estimated aspirin effect is significantly greater for girls than boys. Aspirin effects on offspring function were found in the absence of effects on physical size both at birth and at 4 years. Maternal acetaminophen use was not significantly related to child IQ or attention. As this exploratory research originated from observations of a data set gathered for other purposes, it would be desirable to have these findings replicated in other studies. Further follow-up of the children at a later age is planned.     

Autoradiographic, ultrastructural and interferometric analyses of the temperature effect on the synthesis and migration of ribosomes in root meristem cells of Helianthus annuus L. relation to S phase initiation

Experimental model consisted in blocking cells in G1 phase by cold treatment (12 h, 10 degrees C); following 3 h of postincubation at 20 degrees C, cells initiated S phase. In the present studies it has been shown that 2 h postincubation at 20 degrees C of cold-treated young seedlings of Helianthus annuus L. results in transformation of inactive meristematic nucleoli, characterized by small sizes, reduced amount of dry mass and granular component and by the presence of few and large fibrillar centres into large active nucleoli displaying high dry mass and granular component contents, numerous and small fibrillar centres. After 3 h of postincubation at 20 degrees C, nucleoli lose their granular component, decrease in size and dry mass content. At this moment cytoplasm enriches in ribosomes and its dry mass increases. Maximum of nucleolar activity is preceded by an accumulation of proteins in nucleoli. It is concluded that an enhanced transport of ribosomes is one of the conditions of S phase initiation.