Eeffect of pH on growth in a respiration-deficient mutant of Saccharomyces cerevisiae

The effects of pH on the budding cycle of a respiration-deficient mutant of Saccharomyces cerevisiae were investigation by monitoring the time course of bud development on single cells. The volume of each bud was measured at various time intervals between the inception of its development and inception of development of the next bud on the mother cell. A previous report that the budding cycle consisted of two phases (a rapid-growth phase and a slow-growth phase) was confirmed. With increase in pH from 3.8 to 6.0 the budding cycle shortened as a result of both increase in rate of the rapid-growth phase and decrease in the duration of the slow-growth phase. Although further increase in pH to 7.4 further increased the rate of the rapid-growth phase, the budding cycle lengthened as a result of an increase in time lag and increase in duration of the slow-growth phase. The growth rate, in terms of bud volume, conformed with the expression: (1/V)(dV/dτ) = ξ exp(–V/η), where the values of ξ and η were dependent on pH. The cell volume distribution in a batch culture was compared with the cell volume distribution calculated from the growth curve of a single bud. Similarities in the curves suggested that the growth pattern of a whole culture reflected the growth pattern of a single cell.     

Floral ontogeny in tribe Dalbergieae (Leguminosae: Papilionoideae):Dalbergia brasiliensis, Machaerium villosum s. l.Platymiscium floribundum, andPterocarpus rotundifolius

Floral organogenesis and development of the tropical legume treesDalbergia brasiliensis, Machaerium villosum, Platymiscium floribundum, andPterocarpus rotundifolius were studied using scanning electron microscopy. The aims were to compare ontogenies and to elucidate if floral ontogenetic data will provide new character states diagnostic of the tribe Dalbergieae, which is considered a basal papilionoid tribe and primarily defined on fruit characters. Organ inception is principally acropetal in all taxa studied. Carpel inception is, however, consistently precocious. InD. brasiliensis sepals are initiated in an order not previously reported in Papilionoideae. It may be considered modified helical. InP. rotundifolius the inner whorl of stamens initiate in an unusual way, this is lateral two stamens first, then the two abaxial ones, and last the adaxial one, opposed to the unidirectional order usually seen in Papilionoideae. Generally the differences in flower development among the studied genera appear at initiation and late stage in ontogenesis, rather than at mid-stage.     

Changs in pyrimidine nucleotide biosynthesis during germination of white spruce (picea glauca) somatic embryos

Summary Changes in pyrimidine metabolism were investigated in germinating white spruce somatic embryos by following the metabolic fate of [2-¹⁴C]uracil and [2-¹⁴C]uridine, intermediate metabolites of the salvage pathway and [6-¹⁴C]orotic acid, a central metabolite of the de novo. nucleotide biosynthesis. An active uridine salvage was found to be responsible for the enlargement of the nucleotide pool at the inception of germination. Uridine kinase, which catalyzes the conversion of uridine to uridine monophosphate (UMP), was found to be very active in partially dried embryos and during the early phases of imbibition. The contribution of uracil to the nucleotide pool was negligible since a large amount of radioactivity from [2-¹⁴C]uracil was recovered in degradation products. As germination progressed, the decline of the uridine salvage pathway was concomitant with an increase of the de novo biosynthetic pathway. The central enzyme of the de novo pathway, orotate phosphoribosyltransferase, showed increased activity and contributed to the larger amount of orotate being anabolized. These results suggest that although both the salvage and de novo pathways operate in germinating white spruce somatic embryos, their contribution to the enlargement of the nucleotide pool appears tightly regulated as germination progresses.     

Duration of the cell cycle phases in mutants for the tumor suppressor <Emphasis Type="Italic">Merlin</Emphasis> in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The cell cycle duration was estimated in Drosophila melanogaster mutants for the tumor suppressor Merlin with the use of different approaches. Experiments on induction of mosaic clones in tissues of the larval wing imaginal disc showed that the cell cycle in mutant discs is shorter than that in control. Flow fluorescence cytometry revealed no differences between mutant and normal animals in the relative duration of the cell cycle phases, which suggests proportional shortening of the cell cycle phases. The study with pulselabeled mitoses confirmed these results and showed that the length of the cell cycle is 7 h (S phase duration 3 h) in control individuals and 5 h (S phase duration 2 h) in Merlin gene mutants.     

Inflammatory Lesions of Cochlea in Murine Cytomegalovirus-Infected Mice with Hearing Loss

We sought to determine the distribution of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and lymphocytes in the cochlea of mice infected with murine cytomegalovirus (MCMV). For this purpose, 16 newborn mice were divided equally into model and control (uninfected) groups. In model group, 10 μl of MCMV was injected into the brain of each mouse whereas in control group, 10 μl of physiological saline was injected. Fourteen days after the injection of MCMV, the auditory brainstem response audiometry was performed. Later, the mice were killed and the acoustic capsule samples were collected for polymerase chain reaction analysis, histopathological and immunohistochemical studies. Compared with control mice, the incubation period was longer (F = 13.797; P = 0.003) and the amplitude was lower (F = 5.095; P = 0.043) in model group mice. It indicated that the intracerebral injection of MCMV caused hearing loss in mice. Histopathological examination of cochleas revealed increased levels of lymphocytic infiltration in the membrana vestibularis. Higher levels of TNF-α and IL-6 in scala tympani were detected by immunohistochemical staining. Taken together, the hearing loss in mice could be related with the inflammatory changes occurring in cochlea after inception of MCMV infection.     

FLORAL DEVELOPMENT OF COCHLOSPERMUM TINCTORIUM AND BIXA ORELLANA WITH SPECIAL EMPHASIS ON THE ANDROECIUM

The initiation and development of the flower of Bixa orellana L. and Cochlospermum tinctorium A. Rich, were investigated using the scanning electron microscope to elucidate the nature of the androecial development and the relationships of both taxa. Initiation of floral buds starts with the inception of five sepals in a 2/5 sequence between two bracteoles. The petals are formed successively on the irregular pentagonal apex in a phyllotactic pattern different from that of the calyx. Stamen development proceeds centrifugally on a broad circular primordium or ringwall, which attains its final size at the time of initiation of individual stamens. Stamen primordia arise in successive whorls without connection to the perianth. The residue of the apex is differentiated into a gynoecial circular primordium. In Cochlospermum 3 to 4 carpels are initiated; in Bixa no individual carpels are visible. The origin of the androecial circular primordium is discussed in relation to other types of androecial development. A comparison is made with existing theories of the evolution of multistaminate androecia. Few differences were found in the floral development of Bixa and Cochlospermum, except in the gynoecium. It is proposed to retain them in a single family Bixaceae for a number of reasons. The floral development supports a thealean, dillenialean, or violalean affinity rather than a malvalean. The inception of a broad androecial circular primordium is highly different from the development in Malvales, where more space for stamens is provided by the continuous growth of a tube.     

BREEDING SYSTEM IN FICUS CARICA,THE COMMON FIG. I. FLORAL DIVERSITY

Coevolution in Ficus carica (Moraceae) and the fig wasp (Blastophaga psenes, Agaonidae, Chalcidoidea) has resulted in a complex breeding system involving two tree morphs (Caprifig and Edible fig), three floral forms (long-styled female, short-styled female, and male flowers) and the insect pollinator. The two female floral forms have been reported to differ only in style length and stigma shape. In the present study, we demonstrate that the two female flowers differ from inception—short-styled flower primordia are smaller and exhibit significantly greater individual variation than do those of the long-styled flower, and the relative growth rate of each flower type differs. Mature forms exhibit disparity in style length, in stigma characteristics, and in degree of fusion of stylar lobes. Female flowers of both tree morphs are unisexual from inception. Male flowers of the Caprifig tree morph are initiated as hermaphrodites and gynoecium abortion occurs before megaspore mother cell stage. A single inflorescence therefore expresses two pathways to unisexuality. Hermaphrodite flower primordia were repeatedly found in the supposedly unisexual female syconium of the Edible fig tree morph. Based on its developmental morphology, Ficus carica appears to be of gynomonoecious ancestry.     

CENTRIFUGAL STAMEN INITIATION IN PHYTELEPHANTOID PALMS

The phytelephantoid group, considered very highly specialized among the 15 major groups of palms, includes only three dioecious genera which are distinguished by multipartite flowers. Staminate flowers of these genera have from 120 to over 900 stamens, the largest number known in palms. Developmental material shows centrifugal inception of stamens in Palandra and Phytelephas, and the course of the trunk bundles indicates centrifugal initiation of stamens in Ammandra. Stamens of all three genera are supplied by large trunk bundles that develop acropetally following the different centrifugal expansion of the floral apex in each genus. Branches of the trunk bundles form later to supply stamen primordia as they develop. Polyandric androecia occur in ten of the major groups of palms. Other multistaminate genera show expansion and change in shape of the floral apex before stamen initiation. Centrifugal development appears to represent a different method of expansion of the floral apex to accommodate increase in stamen number in this group. Centrifugal inception of stamens in monocotyledons has previously been established only for two genera of Butomaceae.     

Different instability of nuclear DNA at acid hydrolysis in cancerous and noncancerous cells as revealed by fluorescent staining with acridine orange

Summary Ehrlich cancer cells and inflammatory cells in mouse ascitic fluid were hydrolyzed and stained with acridine orange (AO). The AO hydrolysis curves for G₁/G₂+M phase cancer cells and inflammatory cells were differentially determined using flow cytometry by monitoring the metachromatic red-shifted fluorescence of the fluorochrome bound to the single-stranded DNA produced by acid hydrolysis. By computer fitting of the Bateman function to the hydrolysis curves, the kinetic parameters k ₁ (rate constant for the degradation of the produced single-stranded DNA), and y ₀ (theoretical value of the single-stranded DNA present initially) were determined. It was found that the k ₂ value, which reflects the degree of DNA instability, was much higher for cancer cells in both the G₁ and G₂+M phases than for inflammatory cells. This finding led us to develop a method for the differential AO staining of cancer cells and non-cancerous cells utilizing the different degree of DNA instability at acid hydrolysis. AO staining after hydrolysis with 2N HCl at 30°C for 8.5 min was found to be the optimal method. In the 60 cases of human malignant epithelial and nonepithelial tumors tested, all of the malignant tumor cells emitted metachromatic red fluorescence, while all of the nonmalignant tumor cells (5 cases of benign tumor) and normal cells emitted orthochromatic green fluorescence when observed with a violet excitation light under a fluorescence microscope. This new technique can be a useful tool for the screening of malignancy in exfoliative cytology and also for basic cancer research.In honour of Prof. P. van Duijn     

Spatial and temporal dynamics of the steady-state phytoplankton assemblages in a temperate shallow hypertrophic lake (Lake Manyas,Turkey)

Spatial and temporal dynamics of phytoplankton biomass and species composition in the shallow hypertrophic Lake Manyas, Turkey, were studied biweekly from January 2003 to December 2004 to determine steady-state phases in phytoplankton assemblages. Steady-state phases were defined when one, two or three coexisting species contributed to at least 80% of the standing biomass for at least 2 weeks and during that time the total biomass did not change significantly. Ten steady-state phases were identified throughout the study peiod. During those periods, Achnanthes microcephala (Kützing) Cleve twice dominated the phytoplankton biomass alone and contributed to more than 50% of the total biomass in seven phases. Microcystis aeruginosa (Kützing) Kützing, Anabaena spiroides Klebahn, Cyclotella stylorum Brightwell, Pediastrum boryanum (Turpin) Meneghini and Phacus pusillus Lemmermann were also represented once in steady-state phytoplankton assemblages. A. microcephala was dominant usually during cold periods of the year, while M. aeruginosa and A. spiroides were usually dominant in warm seasons. The total number of species showed a clear decrease during steady-state phases at all stations. All stations were significantly different in terms of the measured physical and chemical parameters (P < 0.05) and phytoplankton biomass (F = 117, P < 0.05).     

Evaluation of weed eradication programs: the delimitation of extent

Eradication is a management strategy that can provide substantial ecological and economic benefits by eliminating incursions of pest organisms. In contrast to eradication efforts that target other pests, weed eradication programs can be very protracted owing to the presence of persistent seed banks and difficulties in detecting the target. Hence there is a need to develop criteria to assist in the evaluation of progress towards eradication. Knowledge of the extent of a weed incursion (the ‘delimitation’ criterion) is considered fundamental for eradication success, as an incursion will progress from any infestations that remain undetected and thus uncontrolled. This criterion is examined with regard to eradication programs targeting Bassia scoparia L. A.J. Scott [= Kochia scoparia L. Schrader], Chondrilla juncea L. (both in Western Australia) and Orobanche ramosa L. in South Australia. The B. scoparia incursion, which has been eradicated, was largely delimited within 12 months of the inception of its eradication program. In contrast, the Western Australian C. juncea incursion has never been delimited, owing to insufficient investment in surveillance during an eradication program spanning 30 years. An exponential decrease in the detection ratio (infested area detected/area searched) over time suggests that delimitation has been approached within 6 years of the inception of the eradication program for O. ramosa. An effective surveillance program is essential for achieving delimitation of a weed incursion.     

Mechanism of growth retardation of the adenocarcinoma EO 771

Summary Growth retardation of tumors has been predominantly described by an increase of the cell loss factor. However, this cell loss factor alone merely reflects the growth deceleration without giving information on the mechanism that causes growth retardation. In the present study a quantitative analysis of the mechanism causing growth retardation of the adenocarcinoma EO 771 has been carried out by determining separately the components of the cell loss factor, namely the cell production rate and the cell loss rate of the tumor cell population. For this purpose the alteration of the histology of the tumor (proportion of necrotic tumor tissue, tumor cell density) and the proliferative capacity of the tumor cell population as a function of the tumor size was studied by applying morphometric and cell kinetic methods. The results show that growth deceleration is due to a decrease of the cell production ratek _p and a simultaneous increase of the cell loss ratek _l . Both processes contribute to about the same extent to the growth deceleration of the tumor cell population. In early tumor growth deceleration is mainly due to a prolongation of the cycle time of the tumor cells, in later phases of tumor growth to an increasing probability of the tumor cells to decycle leading to a decrease of the growth fraction GF and an increase of the cell loss ratek _l .Dedicated to Prof. L.E. Feinendegen on the occasion of his 60th birthday     

Centrifugal ovule inception

Nine samples from Sattler (1973) were found to exhibit a remarkable correlation between the sequence of ovule inception and ovary structure. In every case, the sequence of ovule inception is centrifugal from its supposed final position of the floral apex. Thus, it is hypothesized that ovules are initiated centrifugally from their floral apex and that this is regulated in some mechanical way. To prove this hypothesis, ontogenetical development of the ovary was reexamined inSilene cucubalus, where the sequence of ovule inception was expected to be not simply basipetal but basipetal on the central column and acropetal on the septal protuberances. The results appear to support the hypothesis. A basipetal sequence is clearly visible on the central column. Although the ovule number is very small on the septal placenta, an acropetal sequence can be recognized, i.e., the distal primordia are obviously initiated later than the proximal ones. Two possible mechanisms for centrifugal ovule inception are suggested: (1) that it is regulated by chemical information diffusing from the floral apex or (2) that it is dependent on gradation of cell age in the placenta. The second mechanism is discussed and supporting evidence is suggested in the epidermal cell arrangement of the septal wall. Contribution from the Osaka Museum of Natural History No. 275.     

Mast cells dysregulate apoptotic and cell cycle genes in mucosal squamous cell carcinoma

Background  

Mucosal squamous cell carcinoma of the head and neck is a disease of high mortality and morbidity. Interactions between the squamous cell carcinoma and the host's local immunity, and how the latter contributes to the biological behavior of the tumor are unclear. In vivo studies have demonstrated sequential mast cell infiltration and degranulation during squamous cell carcinogenesis. The degree of mast cell activation correlates closely with distinct phases of hyperkeratosis, dysplasia, carcinoma in-situ and invasive carcinoma. However, the role of mast cells in carcinogenesis is unclear.     

Expression profiling of human renal carcinomas with functional taxonomic analysis

Background  

Molecular characterization has contributed to the understanding of the inception, progression, treatment and prognosis of cancer. Nucleic acid array-based technologies extend molecular characterization of tumors to thousands of gene products. To effectively discriminate between tumor sub-types, reliable laboratory techniques and analytic methods are required.     

THE QUANTITATIVE DETERMINATION OF THE INFECTIVITY OF AGROBACTERIUM TUMEFACIENS

A bioassay relating number of Agrobacterium tumefaciens cells in the inoculum quantitatively to the number of crown-gall tumors initiated on primary pinto bean leaves is described. Variability in estimation of infectious titers by this assay is similar to that observed in comparable plant virus assays, most determinations showing standard errors of 20% of the mean tumor per leaf value. The assay has the advantages of speed and practicality. The efficiency of the system is low, typically requiring between 10⁵ and 10⁶ bacteria for each tumor initiated. Infectivity titers of 10³-10⁴, however, are readily obtained from stationary phase cultures. Statistical analysis of the infectivity titration curve indicates that a single bacterium is the usual infectious unit. The assay is specific within the family Rhizobiaceae to the species Agrobacterium tumefaciens and Agrobacterium rubi. A. tumefaciens strains IIBNV6 and ATCC # 11095 were non-infectious, while strain B6 was the most infectious of the strains tested. The infectivity of the latter strain is shown to decrease about 4-fold between early log and stationary phases of growth. Changes in the growth medium or in the dilution-inoculation medium failed to alter the infectivity of the bacterium.     

<Emphasis Type="Italic">TRAF1/C5</Emphasis>polymorphism is not associated with increased mortality in rheumatoid arthritis: two large longitudinal studies

Introduction  

Recently an association between a genetic variation in TRAF1/C5 and mortality from sepsis or cancer was found in rheumatoid arthritis (RA). The most prevalent cause of death, cardiovascular disease, may have been missed in that study, since patients were enrolled at an advanced disease stage. Therefore, we used an inception cohort of RA patients to investigate the association between TRAF1/C5 and cardiovascular mortality, and replicate the findings on all-cause mortality. As TRAF1/C5 associated mortality may not be restricted to RA, we also studied a large cohort of non-RA patients.     

Quantification of different yeasts associated with the bark beetle,Ips typographus,during its attack on a spruce tree

There were different amounts and types of yeasts associated with individuals ofIps typographus spruce bark beetles during different phases of their attack on a healthy spruce tree. The yeasts were isolated on Sabouraud agar medium in order to identify them and estimate their numbers.Hansenula holstii andCandida diddensii type yeasts were most frequently isolated. The increase in number of these two yeast types probably accounted for most of the total yeast increase found during the later attack phases of the bark beetles. Lesser amounts ofHansenula capsulata, Pichia pinus, Candida nitratophila, and twoCryptococcus type yeasts were also found.     

Candidate proteomic biomarkers for non-alcoholic fatty liver disease (steatosis and non-alcoholic steatohepatitis) discovered with mass-spectrometry: a systematic review

Context: Non-alcoholic fatty liver disease (NAFLD) is characterized by lipid accumulation in the liver which is accompanied by a series of metabolic deregulations. There are sustained research efforts focusing upon biomarker discovery for NAFLD diagnosis and its prognosis in order investigate and follow-up patients as minimally invasive as possible.

Objective: The objective of this study is to critically review proteomic studies that used mass spectrometry techniques and summarize relevant proteomic NAFLD candidate biomarkers.

Methods: Medline and Embase databases were searched from inception to December 2014.

Results: A final number of 22 records were included that identified 251 candidate proteomic biomarkers. Thirty-three biomarkers were confirmed – 14 were found in liver samples, 21 in serum samples, and two from both serum and liver samples.

Conclusion: Some of the biomarkers identified have already been extensively studied regarding their diagnostic and prognostic capacity. However, there are also more potential biomarkers that still need to be addressed in future studies.