Application of the names Crataegus calycina Peterm. and C. oxyacantha L.

Authentic herbarium material of Crataegus calycina Petermann (1849) supports Hrabetová-Uhrová's contention in 1969 that this is conspecific with C. macrocarpa Hegetschweiler; the name C. calycina , for which a Petermann specimen in Musée Botanique Cantonal, Lausanne is designated as lectotype, had been incorrectly applied in Flora Europaea.
Examination of the specimens in the Linnaean Herbarium has led to acceptance of Dandy's proposal in 1946 that sheet 643.12 should be designated as the lectotype of C. oxyacantha L. (1753). This specimen is not, as frequently assumed, C. laevigata (Poiret) DC. (C oxyacanthoides Thuill.) but the species described in Flora Europaea as "C. calycina Peterm. subsp. curvisepala (Lindman) Franco". It is suggested that the name C. oxyacantha L. is a source of confusion and should be rejected under Article 69 of the International Code of Botanical Nomenclature. This gives priority to the name C. curvisepala Lindman, which should replace C. calycina Peterm. in Flora Europaea.     

Lecithinase-negative variants of Clostridium perfringens; the identity of C. plagarum with C. perfringens.

Identity of Clostridium plagarum (Prévot) com. nov. (1938) with C. perfringens was demonstrated in the tests for the cultural and biochemical properties, DNA-DNA homology, susceptibility to C. perfringens-specific lysin, complementary synthesis of C. perfringens theta-toxin between C. plagarum and theta-toxin-negative mutants of C. perfringens, as well as the tests for gas-chromatographic patterns.     

Cryptosporidium canis n. sp. from domestic dogs.

Oocysts of Cryptosporidium, from the feces of a naturally infected dog and from an HIV-infected human, were identified as the previously reported canine genotype of Cryptosporidium parvum, hereafter referred to as Cryptosporidium canis n. sp. Also among the oocysts from the dog, a trace amount of C. parvum bovine genotype was detected. Cryptosporidium canis oocysts from both the dog and human were infectious for calves. Oocysts excreted by calf 1 (dog source) were approximately 90% C. canis and 10% C. parvum, whereas those excreted by calf 3 (human source) were 100% C. canis. Oocysts from calf 1 infected calf 2 resulting in excretion by calf 2 of oocysts approximately 90% C. parvum and 10% C. canis. Oocysts of C. canis were not infectious for BALB/c neonatal mice or immunosuppressed C57 juvenile mice, although all control mice became infected with the C. parvum Beltsville isolate. Oocysts of C. canis from calf 1 and the human were structurally indistinguishable from oocysts of the C. parvum Beltsville isolate (bovine). However, C. canis oocysts differed markedly at the molecular level from all known species of Cryptosporidium based on sequence data for the 18S rDNA and the HSP 70 gene. The differences in genetics and host specificity clearly differentiate C. canis as a new species.     

13C n.m.r. study of L-alanine peptides

The di-, tri-, and tetrapeptides of L-alanine have been studied in aqueous solution by 13C n.m.r. spectroscopy at 25 and 50 MHz. By using selectively 13C enriched analogs containing either 90% 13C methyl or carbonyl carbons and measurements as a function of pH, assignment of the chemical shifts of the peptides has been made. T1 and NOE measurements of the peptides in their cationic, anionic, and zwitterionic states have been recorded as a function of concentration. The results show considerable segmental motion along the backbone carbons of the peptides, with only small changes occurring in the dynamic motions of the peptides as their charge states are altered. The lack of concentration dependence of the chemical shift and T1 values, as well as the similarity of T1 values for individual peptides in the three charge states, indicate that the peptides do not self-associate in aqueous solution.     

Changes in heated and autoclaved forest soils of S.E. Australia. I. Carbon and nitrogen

The effect of heating and autoclaving on extractable nitrogen, N mineralisation and C metabolism was studied by heating five forest soils in the laboratory, simulating the range of effects of heat due to bushfire. Top soil (0–5 cm) was heated to 60 °C, 120 °C and 250 °C for 30 minutes; unheated soil was taken as a control. Samples of the soil heated to 250 °C were also inoculated with fresh soil to accelerate the recovery of the microbial population. Soil autoclaving was carried out as another heat treatment (moist heat). Soils were analysed immediately after heating and 3 times during seven months of incubation to assess immediate and longer-term effects of heating.Extractable N (organic and mineral forms) increased after heating to 120 °C, but decreased with further heating to 250 °C suggesting the volatilisation of N. N associated with microbial biomass diminished with heating and was barely detectable after the 250 °C treatment. Microbial biomass was an important source of soluble N in heated soils, and only partly recovered during subsequent long incubation. The amount of N mineralised during incubation depended on both soil and temperature. Nitrification did not occur when soils were heated to 250 °C (with or without inoculum), or after autoclaving, demonstrating the high sensitivity of nitrifiers to heat. At the beginning of soil incubation, respiration was enhanced in heated soils (250 °C, 250 °C inoculated) and autoclaved soils, but after 30 days of incubation respiration decreased to values either similar to or lower than those in control. This respiration pattern indicated that a fraction of labile C was released by heating, which was quickly mineralised within 30 days of incubation. These results demonstrate some effects of soil heating on C and N dynamics in forest soils.     

Determination of ploidy levels in Ipheion uniflorum (R. C. Graham) Rafin (Liliaceae)

In this study, chromosome number and ploidy levels of Ipheion uniflorum cv. "Wisley Blue" (spring starflower) were determined. In meristematic root tip cells, chromosome number was found as 2n = 12 and 4n = 24. The ratios of diploid and tetraploid cells were found as 80.74% and 19.26%, respectively. In differentiated root tissues and mature leaf tissues ploidy levels were analysed by flow cytometry and polysomaty were found in both organs. In differentiated root tissues, ploidy levels were found as 2C, 4C, 8C and 16C DNA. In root tissues percentages of 2C, 4C, 8C and 16C nuclear DNA content were observed as 57.2%, 33.1%, 2.47% and 7.23%, respectively. In mature leaf tissues, ploidy levels were determined 2C, 4C, 8C and 16C DNA. In this tissue the frequency of 4C DNA was found very higher (74.3%) and 2C DNA content was determined as 19.2%. In mature leaf tissue, 8C and 16C nuclear DNA contents were observed as 2.72% and 3.78%, respectively. When nuclear DNA contents in leaves and roots were compared, an apparent difference in 2C and 4C DNA contents was found.     

Branchiobdellida (Oligochaeta) from the farthest South-East of the U.S.S.R.

Eight species of the genera Branchiobdella and Cirrodrilus are reported from the Soviet Far East. Six of them from the Komarovka Stream in the Ussuriiski State Nature Reserve: Branchiobdella cheni (Liu, 1964), B. domina sp.n., Cirrodrilus chosen (Yamaguchi, 1934), C. suzukii (Yamaguchi, 1934), C. pugnax sp.n., and C. fimbriatus sp.n. Branchiobdellidans are abundant in this stream, occurring on the crayfishes Cambaroides dauricus wladiwostokensis as well as in benthic samples. Two from the northern Amur River basin near Khabarovsk: Branchiobdella minuta Pierantoni, 1912, and Cirrodrilus quadritentacularis (Liu, 1984), living on the crayfishes Cambaroides dauricus dauricus and C. schrencki . The genus Cirrodrilus demonstrates several trends of parallel variation on the Asian continent and the Japanese Islands; the more apomorphic genus Branchiobdella seems to be absent in Japan.     

Visualization of freezing damage. II. Structural alterations during warming

There is a growing amount of indirect evidence which suggests that the loss in viability of rapidly cooled cells is due to recrystallization of intracellular ice. This possibility was tested by an evaluation of the formation of morphological artifacts in rapidly cooled cells to determine whether this process can account for the loss in viability. Samples of the common yeast Saccharomyces cerevisiae were frozen at 1.8 or 1500 °C/min, and the structure of the frozen cells was examined by the use of freeze-fracturing techniques. Other cells cooled at the same rate were warmed to temperatures ranging from ?20 ° to ?50 °C and then rapidly cooled to ?196 °C, a procedure that should cause small ice crystals to coalesce by the process of migratory recrystallization. Cells cooled at 1500 °C/min and then warmed to temperatures above ?40 °C formed large intracellular ice crystals within 30 min, and appreciable recrystallization occurred at temperatures as low as ?45 °C. Cells cooled at 1.8 °C/min and warmed to temperatures as high as ?20 °C underwent little structural alteration. These results demonstrate that intracellular ice can cause morphological artifacts. The correlation between the temperature at which rapid recrystallization begins and the temperature at which the cells are inactivated indicates that recrystallization is responsible for the death of rapidly cooled cells.     

Studies on nitrate reductase of Clostridium perfringens. II. Purification and some properties of ferredoxin.

A ferredoxin was purified from Clostridium perfringens by DEAE-cellulose chromatography and Sephadex G-50 gel filtration. It had absorption maxima at 390 and 280 nm. The molecular weight was estimated to be 6,000 by Sephadex gel filtration and from the results of amino acid analysis. The isoelectric point was 3.0. It contained four atoms of iron, four atoms of labile sulfur, and six cysteine residues. This ferredoxin as well as ferredoxin from C. pasteurianum acted as an electron donor for nitrate reductase from C. perfringens. The ferredoxin could also act as an electron donor for the hydrogenase from C. pasteurianum in hydrogen evolution.     

Antibody-independent activation of C1. II. Evidence for two classes of nonimmune activators of the classical pathway of complement

Nonimmune activation of the first component of complement (C1) by cardiolipin (CL) vesicles present specific features which were not demonstrated on immune complexes. CL vesicles which activate C1 in the presence of C1-inhibitor (C1-INH) were found to bind C1s in the absence of C1r, and to induce a specific C1r-independent cleavage of C1q-bound C1s. Therefore, several known natural nonimmune activators were analyzed by comparing their ability to activate C1 in the presence of C1-INH and to mediate a C1r-independent cleavage of C1s. Freshly isolated human heart mitochondria (HHM) activated C1 only in the absence of C1-INH. However, mitoplasts derived from HHM (HHMP) activated C1 regardless of the presence of C1-INH, and induced a specific cleavage of C1q-bound C1s. The same pattern was observed in the case of smooth E. coli and a semi-rough E. coli strain. DNA, known to activate C1 only in the absence of C1-INH, does not induce C1s cleavage in the absence of C1r. Thus, nonimmune activators can be classified into two distinct categories. "Strong" activators, such as CL vesicles, HHMP, or the semi-rough E. coli strain J5 can activate C1 in the presence of C1-INH. By using C1qs2 as a probe, they exhibit a specific, C1r-independent cleavage of C1s. C1s-binding to C1q is a critical factor for the activation process in this group. In the case of "weak" activators, such as E. coli smooth strains, DNA, or HHM, no C1s-binding to activator-bound C1q was detected, and C1r-independent C1s cleavage and C1 activation in the presence of C1-INH were not observed. As in the case of immune complexes, C1r activation appears to play a key role in the C1 activation by "weak" activators.     

Vitamin E analogue Trolox C. E.s.r. and pulse-radiolysis studies of free-radical reactions.

The reactions between Trolox C, a water-soluble vitamin E analogue, and several oxidizing free radicals including the hydroxyl radical and various peroxy radicals were examined by using the pulse-radiolysis technique. The results demonstrate that Trolox C may undergo rapid one-electron-transfer reactions as well as hydrogen-transfer processes; the resulting phenoxyl radical is shown to be relatively stable, in common with the phenoxyl radical derived from vitamin E. The reactions between the Trolox C phenoxyl radical and a variety of biologically relevant reducing compounds were examined by using both pulse radiolysis and e.s.r. The results demonstrate that the Trolox C phenoxyl radical is readily repaired by ascorbate (k = 8.3 x 10(6) dm3.mol-1.s-1) and certain thiols (k less than 10(5) dm3.mol-1.s-1) but not by urate, NADH or propyl gallate. Evidence from e.s.r. studies indicates that thiol-containing compounds may also enter into similar repair reactions with the alpha-tocopherol phenoxyl radical. Kinetic evidence is presented that suggests that Trolox C may 'repair' proteins that have been oxidized by free radicals.     

The action of caffeine on X-irradiated HeLa cells. IX. Hypothermic effects

Hypothermic enhancement of the lethal effect of 3.5 Gy of 220-kV X rays in the absence of caffeine as well as in its presence (4 mM) was examined at temperatures between 10 and 34 degrees C in monolayer cultures in the G1 phase of the cell cycle. Correction has been made for the toxicity of low temperatures, and of caffeine at low temperatures, by concomitantly measuring cell killing in unirradiated cells. In the absence of caffeine, incubation of irradiated cells for up to 34 h at temperatures in the range 15 to 30 degrees C (or possibly 34 degrees C) enhances killing compared to that observed at 38 degrees C; the amount of enhancement is about the same throughout this range, but is nil at 10 degrees C. The enhanced killing induced by caffeine at 38 degrees C decreases as the temperature is lowered to 15 degrees C; there is no enhancement at 10 degrees C. Less killing is manifested in the range 15 to 25 degrees C in the presence of caffeine than in its absence. Recovery (loss of sensitivity to caffeine) and fixation of potentially lethal damage were studied in late-S/G2-phase cells at reduced temperatures by delaying treatment with caffeine for increasing times after irradiation. As the temperature is progressively lowered to 20 degrees C, less recovery is manifested after 5 h of incubation; no recovery is detected in the range 10 to 20 degrees C. Despite extensive recovery at 34 degrees C, no fixation is observed at that (or any lower) temperature in G2-phase cells: the cells are able to recover essentially fully when returned to 38 degrees C. In addition, responses of unirradiated control series to incubation at low temperatures appear to differ from those reported by others for longer treatment times of different cell systems.     

Binding of protein S to C4b-binding protein. Mutagenesis of protein S.

Protein S and C4b-binding protein (C4BP) form a tight complex (Kd approximately 0.6 nM) the physiologic purpose of which is unknown. The participation of protein S in this complex was investigated using site-specific mutagenesis. Normal recombinant human protein S (rHPS) and five specifically mutated protein S analogs were expressed in transformed human kidney 293 cells and the following properties were characterized: solution-phase C4BP binding, ability to be cleaved by thrombin, ability to act as a cofactor in the activated protein C-catalyzed inactivation of factor Va, and gamma-carboxyglutamic acid content. In some cases, beta-hydroxyaspartic acid plus beta-hydroxyasparagine content was also determined. Binding studies indicated that while clearly important for a high affinity interaction, the amino acid sequence Gly605-Ile614 identified by Walker (Walker, F J. (1989) J. Biol. Chem. 264, 17645-17648) does not account for all the binding energy of the HPS-C4BP interaction. All mutants perturbed in this region or lacking it altogether displayed reduced C4BP binding, and some retained anticoagulant cofactor function. Neither human factor X nor human steroid-binding protein had any measurable ability to compete with plasma HPS for C4BP binding. Furthermore, bovine protein S and a rHPS analog with bovine sequence from Gly597-Trp629 bound to human C4BP with the same affinity as did HPS, and both proteins substituted effectively for HPS as a cofactor for activated protein C in an otherwise human anticoagulation system. Together these results suggest that optimal binding of protein S to C4BP requires the putative alpha-helix Gly605-Ile614, as well as other undetermined regions of protein S, and that the regions of HPS responsible for C4BP binding and activated protein C cofactor function are structurally isolated.     

Studies on the sporulation of Phytophthora parasitica Dastur var. piperina Dast. Responsible for Leaf and Foot-rot of Pan,Piper betle. L.

Summary Sporangial formation was found restricted within the range of temperature from 20–31° C. Atmospheric humidity of 100 % did not favour very much in developing sporangia. Free water appeared to be essential in sporangial formation. The range of temperature within which the liberation of zoospores took place was even narrower, 19 to 25° C. Temperature from 21 to 23° C appeared to be best condition for maximum liberation of zoospores. Similarly as with sporangial development atmospheric humidity as high as 100 % had very little effect on the liberation of zoospores unless there was condensation of water. The liberated zoospores in free water were found to germinate only within 20–24° C. At about 22° C the germination rate was found optimum.     

Immunodiagnosis of Candida-infections. I. Sensitivity of the antigens

Attemps were made to prepare a sensitive antigen from C. albicans suitable for detecting humoral antibodies and hypersensitivity in deep-seated candidiasis, in patients at risk of invasive candidiasis and in allergic states caused-by Candida.5343 persons suffering from systemic, bronchial, vaginal candidiasis, bronchial asthma, chronic bronchitis, polyarthritis nodosa, ulcus cruris, malignancy, rhinitis pollinosa, vasomotorica, and non infected miners, farmers and blood donors were investigated on the presence of antibodies and hypersensitivity against 8 different antigen preparations.The extracellular protein and mannan- protein isolated from the cultivation medium of C. albicans proved the most sensitive for specific anticandida antibodies. The mannan, especially the mannan isolated from the cell surface of C. albicans determined best for the allergy.Comparison made of commercial Candidine showed similar activity. The whole cell C. albicans antigen as well as the mixed Candida antigen reacted much weakly.Comparison made of autoantigen, C. albicans and mixed C. albicans antigen proved the highest sensitivity of the autoantigen.     

Cyanate as an inactivator of complement proteins.

Sodium cyanate added to normal human serum or serum from patients with sickle-cell disease resulted in the functional inactivation of C3, C5, C6, C7, and the C3b inactivator, but not C8 and C9. Final concentrations as low as 0.5 mM in serum caused inactivation of 12 to 64% of the C3 after 8 hr at 37 degrees C. The activity of the inactivated C3, C5, and C3b inactivator was not restored by dialysis. Most of the functional activity of C3 in cyanate-treated sera was destroyed by very small quantities of 14C-labeled cyanate that was bound to the protein. C3 inactivation by cyanate occurred in heated sera (50 degrees C, 30 min) and sera treated with EDTA, probably indicating that one mechanism for inactivation was by a direct carbamylation reaction. Both C3 and C5 showed two anodal-migrating forms in two dimensional antigen-antibody crossed electrophoresis in some sera treated with low concentrations of cyanate. Measurements of circular dichroism of highly purified carbamylated C3 showed no detectable changes in structure even though most of the functional activity was destroyed. Purified, inactive C3 that was carbamylated with 14C-labeled cyanate was capable of binding to EAC142, but the resulting EAC1423 was weakly positive for immune adherence and negative for agglutination with anti-C3 antiserum. Unlabeled, cell-bound C3b on EAC142 was not susceptible to cyanate action as shown by no loss in immune adherence and positive agglutination with anti-C3 antiserum. The C3b inactivator was more susceptible to cyanate than C3 in a short time period, whereas both were inactivated after 8 hr. Since cyanate is currently being evaluated as a treatment for sickle-cell disease, the inactivation of C3 by the drug is an important consideration for such patients who are already deficient in C3 dependent heat-labile opsonins that aid in host defense.     

Stressors and pain sensitivity in CFW mice. Role of opioid peptides.

Effects of several environmental situations on pain threshold were studied in CFW male mice. Immobilization induced significant and naloxone reversible analgesia. Isolation produced analgesia which was partially reversed by naloxone. One minute swimming in + 4 degrees C or + 42 degrees C water increased naloxone reversible analgesia. Isolation produced analgesia which was partially reversed by naloxone. One minute swimming in 4 degrees C or + 42 degrees C water increased naloxone irreversible pain threshold. Other situations: drinking 2% NaCl solution, disturbance of light-dark cycle or social aggregation did not produce analgesia. The role of these situations as stress-inducers, as well as the role of endogenous opioid peptides in stress-induced analgesia, were discussed.     

Isolation of Chromobacterium spp. from foods, soil, and water.

Chromobacterium violaceum, a soil and water inhabitant, has been implicated in human disease with a high mortality rate, particularly in the southeastern United States. The psychrotrophic Chromobacterium lividum has been isolated from foods, water, and soil, but is not considered pathogenic. To determine the distribution of Chromobacterium spp. in soil, water, and foods in the Gainesville area, we evaluated Bennett, Ryalls and Moss, and Aeromonas membrane agars for their ability to recover these organisms from various samples when incubated at 25 or 35 degrees C. Bennett agar was best for the isolation of both species when incubated at 25 degrees C; however, at 35 degrees C, Aeromonas membrane agar gave the highest recoveries of C. violaceum. C. violaceum was recovered only from soil and water, whereas C. lividum was frequently recovered from foods as well as soil and water.     

Molecular characterization of Calocedrus rupestris Aver., H.T. Nguyen & L.K. Phan, 2008 (Cupressaceae) based on ITS1 partial sequence

Calocedrus rupestris Aver., H.T. Nguyen & L.K. Phan was described in 2008 based on some morphological characters that were not sufficiently significant to discriminate it as a species distinct from C. macrolepis Kurz. We applied a new approach to resolve these conflicting views by using sequence data from DNA (ITS) to elucidate phylogenetic relationships between the two species. Analyses of a partial ITS1 sequence in 5 individuals of 2 subpopulations of C. macrolepis and 18 individuals of 8 subpopulations of C. rupestris collected in Vietnam were done. Molecular characterization of the two species showed its low divergence with the lack of autapomorphic characters. In addition, the ITS1 partial sequences of some C. rupestris individuals were identical with C. macrolepis. Due to the less distinctive morphology between C. rupestris and C. macrolepis, the divergence between them does not exceed the interspecific levels, and therefore, C. rupestris could not be regarded as an independent species in relation to C. macrolepis but only as one of its varieties, C. macrolepis var. rupestris (Aver., H.T. Nguyen & L.K. Phan) L.K. Phan, Long K. Phan & Aver.