General Growth Inhibition Research Articles

Antagonistic and additive effect when combining biopesticides against the fall armyworm, Spodoptera frugiperda

Fall armyworm, Spodoptera frugiperda (FAW) is a cosmopolitan crop pest species that has recently become established in sub-Saharan Africa and Southeast Asia. Current FAW control is almost entirely dependent on synthetic pesticides. Biopesticides offer a more sustainable alternative but have limitations. For example, pyrethrum is an effective botanical insecticide with low mammalian toxicity but is highly UV labile, resulting in a rapid loss of efficacy in the field. Beauveria bassiana is an entomopathogenic fungus that is more persistent, but there is a time lag of several days before it causes insect mortality and leads to effective control. The combination of these biopesticides could mitigate their drawbacks for FAW control. Here we evaluated the efficacy of pyrethrum and B. bassiana as individual treatments and in combination against 3rd instar FAW. Four different combinations of these two biopesticides were tested, resulting in an antagonistic relationship at the lowest concentrations of B. bassiana and pyrethrum (1 × 104 conidia mL−1 with 25 ppm) and an additive effect for the other 3 combined treatments (1 × 104 conidia mL−1 with 100 ppm and 1 × 105 conidia mL−1 with 25 ppm and 100 ppm pyrethrum). Additionally, a delay in efficacy from B. bassiana was observed when combined with pyrethrum as well as a general inhibition of growth on agar plates. These results appear to show that this particular combination of biopesticides is not universally beneficial or detrimental to pest control strategies and is dependent on the doses of each biopesticide applied. However, the additive effect shown here at specific concentrations does indicate that combining biopesticides could help overcome the challenges of persistence seen in botanical pesticides and the slow establishment of EPF, with the potential to improve effectiveness of biopesticides for IPM.

Biochemical, biological characteristics and antibacterial activity of glycoprotein extracted from the epidermal mucus of African catfish (Clarias gariepinus)

Catfish glycoprotein (CFG) was extracted from the cutaneous mucus of Egyptian catfish by ammonium sulphate precipitation and purified on gel filtration column (sephadex G-50). After purification, CFG produced one band on SDS-PAGE (22 kDa). Urea-PAGE and the pH-solubility of CFG indicated its positive charge (IEP 8). CFG contained 12 saccharides. FTIR spectrum shows 3 groups of bands at 1800–2900, 1100–1700 and 700–1100 cm−1. CFG exhibited antibacterial activity against 9 pathogenic bacteria with low MIC (50 μg/mL), where two Gram+ bacteria, i.e.; Streptococcus pyogenes (St. pyogenes) and Listeria ivanovii (L. ivanovii) were the most sensitive. The growth curves of the bacteria subjected to 1 MIC of CFG during 30 h showed general growth inhibition, particularly in case of Gram− bacteria such as E. coli. TEM images showed evidently reduced relative content of the intact cells and clear incurred cellular malformations. Combining CFG with specific antibiotic at equal ratios induced synergistic antibacterial actions, amounting to 40% of the mathematical sum of the combination. Substituting the antibiotic chloramphenicol with gradual increasing ratios of CFG of its starting concentration (30 μg/mL), produced proportionally bigger antibacterial actions against St. pyogenes growth and increasing synergistic effect up to 37% at 80% of CFG substitution.

Temporal kinetics of the transcriptional response to carbon depletion and sucrose readdition in Arabidopsis seedlings.

To investigate whether the transcriptional response to carbon (C) depletion and sucrose resupply depends on the duration and severity of the C depletion, Arabidopsis seedlings were grown in liquid culture and harvested 3, 6, 12, 24, 48 and 72 h after removing sucrose from the medium and 30 min after resupplying sucrose at each time. Expression profiling revealed early transcriptional inhibition of cell wall synthesis and remodelling of signalling, followed by induction of C recycling and photosynthesis and general inhibition of growth. The temporal sequence differed from the published response to progressive exhaustion of C during a night and extended night in vegetatively growing plants. The response to sucrose readdition was conserved across the C-depletion time course. Intriguingly, the vast majority of rapidly responding transcripts decreased rather than increased. The majority of transcripts that respond rapidly to sucrose and many transcripts that respond during C depletion also decrease after treating seedlings with the transcriptional inhibitor cordycepin A. Comparison with published responses to overexpression of otsA, AKIN10 and bZIP11 revealed that many genes that respond to C depletion, and especially sucrose resupply, respond to one or more of these C-signalling components. Thus, multiple factors contribute to C responsiveness, including many signalling components, transcriptional regulation and transcript turnover.

Growth responses and ion accumulation in the halophytic legume Prosopis strombulifera are determined by Na2SO4 and NaCl

Halophytes are potential gene sources for genetic manipulation of economically important crop species. This study addresses the physiological responses of a widespread halophyte, Prosopis strombulifera (Lam.) Benth to salinity. We hypothesised that increasing concentrations of the two major salts present in soils of central Argentina (Na2SO4, NaCl, or their iso-osmotic mixture) would produce distinct physiological responses. We used hydroponically grown P. strombulifera to test this hypothesis, analysing growth parameters, water relations, photosynthetic pigments, cations and anions. These plants showed a halophytic response to NaCl, but strong general inhibition of growth in response to iso-osmotic solutions containing Na2SO4. The explanation for the adaptive success of P. strombulifera in high NaCl conditions seems to be related to a delicate balance between Na(+) accumulation (and its use for osmotic adjustment) and efficient compartmentalisation in vacuoles, the ability of the whole plant to ensure sufficient K(+) supply by maintaining high K(+)/Na(+) discrimination, and maintenance of normal Ca(2+) levels in leaves. The three salt treatments had different effects on the accumulation of ions. Findings in bi-saline-treated plants were of particular interest, where most of the physiological parameters studied showed partial alleviation of SO4(2-)-induced toxicity by Cl(-). Thus, discussions on physiological responses to salinity could be further expanded in a way that more closely mimics natural salt environments.

In vitro activity of extracts and isolated polyphenols from West African medicinal plants against Plasmodium falciparum

The aim of the study was to screen 11 selected traditional medicinal plants from West Africa for their in vitro antiplasmodial activity in order to determine the activity of single and of combination of plant extracts and to examine the activity of isolated pure compounds. Ethanolic and aqueous extracts of the 11 selected plants and pure compounds from Phyllanthus muellerianus and Anogeissus leiocarpus were tested in vitro against Plasmodium falciparum 3D7. Proliferation inhibitory effects were monitored after 48 h. Among the plants and pure compounds investigated in this study, geraniin from P. muellerianus, ellagic, gentisic, and gallic acids from A. leiocarpus, and extracts from A. leiocarpus, P. muellerianus and combination of A. leiocarpus with P. muellerianus affected the proliferation of P. falciparum most potently. Significant inhibitory activity was observed in combination of A. leiocarpus with P. muellerianus (IC(50) = 10.8 μg/ml), in combination of A. leiocarpus with Khaya senegalensis (IC(50) = 12.5 μg/ml), ellagic acid (IC(50) = 2.88 μM), and geraniin (IC(50) = 11.74 μM). In general growth inhibition was concentration-dependent revealing IC(50) values ranging between 10.8 and -40.1 μg/ml and 2.88 and 11.74 μM for plant extracts and pure substances respectively. Comparison with literature sources of in vivo and in vitro toxicity data revealed that thresholds are up to two times higher than the determined IC(50) values. Thus, the present study suggests that geraniin from P. muellerianus; ellagic acid, gallic acid, and gentisic acid from A. leiocarpus; and combination of extracts from A. leiocarpus with either P. muellerianus or K. senegalensis could be a potential option for malaria treatment.

A case of an extremely large accessory bone with unusual sutures and foramina parietalia permagna in multiple premature craniosynostoses

BackgroundWithin the entity of craniofacial malformations premature craniosynostoses represent the majority of cases. They originate from disturbances in the ossification process, resulting in an osseous closure of cranial sutures ahead of time. Depending on severity and number of the affected sutures, a local or general growth inhibition of the skull follows. In the rare instance of accessory bones along these affected sutures, they may interfere with diagnostics and therapy. PatientThis clinical report describes the case of a seven-month-old male infant with multiple craniosynostoses, an extraordinary large accessory median calvarial bone, two foramina parietalia permagna and a submucosal cleft palate. Chromosomal and genetic analysis did not reveal potential mutations. ResultsThe osseous abnormalities were diagnosed and displayed by three-dimensional computed tomography. The sequential surgical treatment consisted of occipital remodelling at seven months of age and frontoorbital advancement at fourteen months. ConclusionThis rare anatomical variation of the cranial bones – superimposed by multiple premature craniosynostoses – demonstrates the necessity of accurately conducted preoperative diagnostics for appropriate surgical planning. Knowledge, and precise medical examination of potential anatomical variations facilitate the planning and secure performance of surgery as well as its outcome.

Low concentration of ethylenediaminetetraacetic acid (EDTA) affects biofilm formation of Listeria monocytogenes by inhibiting its initial adherence

The distribution and survival of the food-borne pathogen Listeria monocytogenes is associated with its biofilm formation ability, which is affected by various environmental factors. Here we present the first evidence that EDTA at low concentration levels inhibits the biofilm formation of L. monocytogenes. This effect of EDTA is not caused by a general growth inhibition, as 0.1 mM EDTA efficiently reduced the biofilm formation of L. monocytogenes without affecting the planktonic growth. Adding 0.1 mM of EDTA at the starting time of biofilm formation had the strongest biofilm inhibitory effect, while the addition of EDTA after 8 h had no biofilm inhibitory effects. EDTA was shown to inhibit cell-to-surface interactions and cell-to-cell interactions, which at least partially contributed to the repressed initial adherence. The addition of sufficient amounts of cations to saturate EDTA did not restore the biofilm formation, indicating the biofilm inhibition was not due to the chelating properties of EDTA. The study suggests that EDTA functions in the early stage of biofilm process by affecting the initial adherence of L. monocytogenes cells onto abiotic surfaces.

Metabolite Profiling Uncovers Plasmid-Induced Cobalt Limitation under Methylotrophic Growth Conditions

BackgroundThe introduction and maintenance of plasmids in cells is often associated with a reduction of growth rate. The reason for this growth reduction is unclear in many cases.Methodology/Principal FindingsWe observed a surprisingly large reduction in growth rate of about 50% of Methylobacterium extorquens AM1 during methylotrophic growth in the presence of a plasmid, pCM80 expressing the tetA gene, relative to the wild-type. A less pronounced growth delay during growth under non-methylotrophic growth conditions was observed; this suggested an inhibition of one-carbon metabolism rather than a general growth inhibition or metabolic burden. Metabolome analyses revealed an increase in pool sizes of ethylmalonyl-CoA and methylmalonyl-CoA of more than 6- and 35-fold, respectively, relative to wild type, suggesting a strongly reduced conversion of these central intermediates, which are essential for glyoxylate regeneration in this model methylotroph. Similar results were found for M. extorquens AM1 pCM160 which confers kanamycin resistance. These intermediates of the ethylmalonyl-CoA pathway have in common their conversion by coenzyme B12-dependent mutases, which have cobalt as a central ligand. The one-carbon metabolism-related growth delay was restored by providing higher cobalt concentrations, by heterologous expression of isocitrate lyase as an alternative path for glyoxylate regeneration, or by identification and overproduction of proteins involved in cobalt import.Conclusions/SignificanceThis study demonstrates that the introduction of the plasmids leads to an apparent inhibition of the cobalt-dependent enzymes of the ethylmalonyl-CoA pathway. Possible explanations are presented and point to a limited cobalt concentration in the cell as a consequence of the antibiotic stress.

The toxic potential of an industrial effluent determined with the Saccharomyces cerevisiae-based assay

Increasing levels of environmental pollution and the continuous monitoring of water quality both request specific and sensitive methods for the detection of detrimental water contents. On a regulatory basis genotoxicity is assessed by the standard umu-test (ISO 13829) that responds to DNA damage induced by chemicals. The focus of this study was the examination of the toxic potential of samples taken from the wastewater treatment plant of a refinery factory to explore the applicability of the Saccharomyces cerevisiae (bakers yeast) test for the detection of bio-available genotoxic activity in complex matrices. The toxic potential of samples without pre-treatment and following centrifugation was determined with the eukaryotic Saccharomyces cerevisiae bioassay based on the transcriptional activation of the green fluorescent protein (gfp) fused to the DNA damage inducible RAD54 promoter and general growth inhibition. Primary effluent samples were taken as qualified sterile spot samples from the final effluent of the purification plant. The Saccharomyces cerevisiae assay yielded geno- and cytotoxic responses in all complex untreated and centrifuged samples with high reproducibility. The obtained results suggest that the yeast assay is suited as a screening tool to monitor genotoxic potential of wastewater.

Inhibition of TGF-beta-stimulated CTGF gene expression and anchorage-independent growth by cAMP identifies a CTGF-dependent restriction point in the cell cycle.

CTGF is a 38 kDa cysteine-rich peptide whose synthesis and secretion are selectively induced by transforming growth factor beta (TGF-beta) in connective tissue cells. We have investigated the signaling pathways controlling the TGF-beta induction of connective tissue growth factor (CTGF) gene expression. Our studies indicate that inhibitors of tyrosine kinases and protein kinase C do not block the signaling pathway used by TGF-beta to induce CTGF gene expression. In contrast, elevation of cAMP levels within the target cells by a variety of methods blocked the induction of CTGF by TGF-beta. Furthermore, agents that elevate cAMP blocked the induction of anchorage-independent growth (AIG) by TGF-beta. Inhibition of AIG could be overcome by the addition of CTGF, indicating that it was not a general inhibition of growth but a selective inhibition of CTGF synthesis that is responsible for the inhibition of TGF-beta-induced AIG by cAMP. Kinetic studies of the induction of DNA synthesis by CTGF in cells arrested by cAMP indicate that the block occurs in very late G1. These and other studies in monolayer cultures suggest that the CTGF restriction point in the cell cycle is distinct from the adhesion-dependent arrest point.

Cooperation of Calcineurin and Vacuolar H+-ATPase in Intracellular Ca2+Homeostasis of Yeast Cells

Saccharomyces cerevisiae VMA genes, encoding essential components for the expression of vacuolar membrane H(+)-ATPase activity, are involved in intracellular ionic homeostasis and vacuolar biogenesis. We report here that the immunosuppressants FK506 and cyclosporin A cause general growth inhibition of the vma3 mutant. Upon addition of the drugs, the mutant grew neither in the presence of more than 5 mM Ca2+ nor above pH 6.0. The action of the immunosuppressants is dependent on their binding proteins and ascribable to inhibition of calcineurin activity; a mutation of a calcineurin subunit (cnb1) shows synthetic lethal interaction with the vma mutation. The addition of FK506 decreases the cytosolic free concentration of Ca2+ in the vma3 mutant cells. Consequently, FK506 induces an 8.9-fold elevation of a nonexchangeable Ca2+ pool. These results suggest that calcineurin controls calcium homeostasis by repression of Ca2+ flux into a cellular compartment(s) and that the vacuolar H(+)-ATPase is essential for cell growth cooperating with calcineurin to regulate the cytosolic free concentration of Ca2+.

Changes in the cell wall glycoprotein composition of Candida albicans associated to the inhibition of germ tube formation by EDTA.

Hyphal development in Candida albicans was blocked by EDTA. This effect was not due to a general growth inhibition since the chelator did not affect protein and DNA synthesis. Recovery of mycelial growth was observed when EDTA-grown cells were incubated at 37 degrees C in EDTA-free medium. High-molecular-weight mannoproteins (HMWM) that are mycelium-specific wall components, and particularly a 260-kDa species (HMWM-260), were absent in the wall of cells grown under germination conditions in the presence of EDTA. Synthesis of the HMWM-260 species was not inhibited but its incorporation (secretion) into the wall structure seemed to be blocked in EDTA-treated cells.

Toxicity of medium chain fatty acids to Penicillium crustosum Thom and their detoxification to methyl ketones

Conversion of medium chain length fatty acids into methyl ketones by some filamentous fungi is important in the development of flavour in the blue mould‐ripened cheeses and in the promotion of ketonic rancidity in the lauric acid oils. The effect of fatty acid composition of single acid glycerides, pH, temperature and medium chain length fatty acids on conversion of fatty acids into methyl ketones has been studied with two strains of Penicillium crustosum Thom. Medium chain length fatty acids were fungitoxic. The minimum inhibitory concentration was 3776, 2884, 1723 and 3066 mg/l for the C6:0, 8:0, 10:0 and 12:0 acids, respectively, at pH 7·0 in liquid suspension culture. Evidence is provided to suggest that the cellular site for this conversion is the microbody. Growth of P. crustosum on single acid glycerides involves two distinct physiological processes; the first, a general inhibition of growth, and the second, a detoxification. The fatty acid composition and the slip point of the substrate are of paramount importance in these two processes.

Inhibition of the dimorphic transition of Candida albicans by the ornithine decarboxylase inhibitor 1,4-diaminobutanone: alterations in the glycoprotein composition of the cell wall

Hyphal development in Candida albicans was selectively blocked by the ornithine decarboxylase competitive inhibitor 1,4-diaminobutanone (DAB). Inhibition of hyphal development required DAB during both yeast inoculum growth and subsequent incubation at 37 degrees C to induce mycelial growth. This effect was not due to general growth inhibition since DAB did not inhibit yeast growth, and reduced protein synthesis by 30% at most. Moreover, protein synthesis was unaffected by DAB when cells were pre-grown in drug-containing media. Since DAB inhibited dimorphic transition at 37 degrees C, morphology- and temperature-dependent protein synthesis could be distinguished. DAB stimulated the synthesis of several yeast wall-proteins, irrespective of morphology or growth temperature, and two at 37 degrees C only, but it inhibited the synthesis of a single mycelial-specific glycoprotein species.

Plasmodium falciparum gametocyte formation in vitro: its stimulation by phorbol diesters and by 8-bromo cyclic adenosine monophosphate.

Phorbol 12-myristate 13-acetate increased the number of gametocytes by 50 to 100% in well or petri dish cultures of the HB-3 clone of Plasmodium falciparum. Phorbol dibutyrate had a similar effect. The optimal concentration for each of these agents was 20 ng/ml or approximately 30 nM. No effect of forskolin was found, other than a general inhibition of growth at concentrations over 10 microM. An inhibitor of phosphodiesterase, 8-bromo cyclic adenosine monophosphate (at concentrations of 0.1 and 1.0 microM) also significantly increased the number of gametocytes formed by this clone.

Modulation of nuclear proto-oncogene expression and cellular growth in myeloid leukemic cells by human interferon alpha.

To address the mechanisms that regulate expression of specific growth-related nuclear proto-oncogenes, the transcript levels of the c-fos, c-myc, (2'5')-oligoadenylate synthetase, IFN-alpha 1, and IFN-beta 1 genes have been measured in the human leukemic cell lines KG-1, U937, and HL-60 following growth stimulation by serum, induction of differentiation by tumor-promoting agents, and/or treatment of cells with exogenously supplied alpha interferon (rIFN-alpha 2). Production of fos and myc RNA was measured by S1 mapping, using fos DNA probes which identified either primary unspliced transcripts or steady-state-spliced mRNA levels, and using a myc probe which spanned the two major c-myc start sites, P1 and P2. Pretreatment of a quiescent KG-1 cell population with IFN for 18 hours before serum addition decreased the stimulation of both fos and myc RNA production. In HL-60 and U937 cells, IFN pretreatment had no inhibitory effect on serum-induced fos or myc transcription; however, in U937, rIFN-alpha 2 treatment alone stimulated fos mRNA 11-fold. Expression of 2'5'oligoadenylate synthetase was induced in IFN-treated cultures but not in cells stimulated with serum alone. No serum-induced IFN-alpha 1 or IFN-beta 1 gene expression was observed in KG-1 or U937 cells. These results demonstrate that exogenous rIFN-alpha 2 treatment of quiescent KG-1 cells can antagonize the effect of growth factors by altering expression of nuclear proto-oncogenes, but in general growth inhibition is not obligatorily coupled to inhibition of proto-oncogene transcription.

The utilisation of d-galactonate and d-2-oxo-3-deoxygalactonate by Escherichia coli K-12

1. Escherichia coli K-12 mutants unable to grow on d-galactonate have been isolated and found to be defective in either galactonate dehydratase, 2-oxo-3-deoxygalactonate 6-phosphate aldolase or devoid of both of these enzymes and of 2-oxo-3-deoxygalactonate kinase. 2. 2-Oxo-3-deoxygalactonate kinase and 2-oxo-3-deoxygalactonate 6-phosphate aldolase are still induced by galactonate in mutants lacking galactonate dehydratase, suggesting that galactonate rather than a catabolic product of galactonate is the inducer of the galactonate catabolic enzymes. Synthesis of the enzymes is subject to glucose catabolite repression. 3. Mutants defective in 2-oxo-3-deoxygalactonate 6-phosphate aldolase accumulate 2-oxo-3-deoxygalactonate 6-phosphate when exposed to galactonate and this compound causes general growth inhibition. 4. Secondary mutants that no longer show this inhibition fail to make 2-oxo-3-deoxygalactonate 6-phosphate due to additional defects in galactonate transport, galactonate dehydratase, 2-oxo-3-deoxygalactonate kinase or a putative promoter mutation that prevents formation of these enzymes. 5. A spontaneous mutant capable of growth on 2-oxo-3-deoxygalactonate has been isolated. It has two genetically distinct mutations. One permits constitutive formation of the galactonate catabolic enzymes and the other allows the uptake of 2-oxo-3-deoxygalactonate. Neither mutation on its own permitted growth on 2-oxo-3-deoxygalactonate. 6. Genes specifying the various galactonate catabolic enzymes have been located at min 81.7 on the E. coli K-12 linkage map and probably constitute an operon. The gene sequence in this region was shown to by: pyrE uhp dgo dnaA.

A re-evaluation of the effect of cysteine onCandida albicans

The effect of cysteine on Candida albicans has been re-examined. Concentrations between 0-1 and 10-0 mM were tested on 12 isolates in yeast and hyphal form. The growth of both forms was inhibited by 10 mM cysteine. Though the rate of hyphal elongation was decreased there was no suppression of initiation of hyphal formation. All isolates showed a random variation in their general inhibitory response to cysteine. We found no support for the concept of cysteine-mediated from determination in rapidly growing C. albicans in submerged culture. High concentrations of cystiene cause a general inhibition of growth of C. albicans.

Behavior and brain growth in rats treated with p-chlorophenylalanine in the first weeks of life.

AbstractIntraperitoneal injections of p‐chlorophenylalanine (pCPA) were given to 71 rats in the first 7 weeks of life, decreasing brain 5‐hydroxytryptamine concentrations to 20% of control values. The brain weight was significantly decreased, evident after 2 weeks of treatment, and also 52 days after the last injection. The retarded brain growth was not due to a general growth inhibition only: body weight was decreased initially, but from Day 24 the experimental animals gained weight more rapidly than the controls, resulting in a significantly higher body weight from Day 67.The decrease in brain growth was accompanied by behavioral changes, evident later in life. The first test was given 4 weeks after the last pCPA‐injection. The changes observed were moderate, but consistent in all the tests. There were no learning deficits or clear motivational changes. In all the tests there was evidence of a reduced arousal level in the pCPA‐treated rats: latencies were long, exploration was low, habituation to new environment was fast, and the reactivity was low.

Anomalies of the internal organs and diminished calcification of vertebrae in fetuses after benzhydrylpiperazine treatment of pregnant rats

Indications of the effects of chlorcyclizine treatment of pregnant rats on the internal organs of the rat fetus were sought. Hydrocephalus, hydronephrosis, undescended testes, and the previously described cleft palate occurred at their highest incidences when chlorcyclizine was given late in the period of organogenesis (days 13–16). The same defects occurred when drug was administered for various intervals earlier in gestation (days 7–12), but a higher dose was needed. A low incidence of resorptions, placental effects (enlargment, hemorrhagic and polycystic), and alternate effects occurred following the earlier times of administration. Thus a relatively constant syndrome of higher incidence of internal and external effects is produced, and evidence of a fluid imbalance in the fetus is presented. The dry weights of the whole fetus and of the fetal kidneys and heart were also reduced following relatively low doses of chlorcyclizine, 10 and 20 mg/kg. Dry weight of the brain was reduced by 40 mg/kg. Only with doses above 70 mg/kg was there a marked general inhibition of growth. When the dose was 20 mg/kg, the kidney and heart dry weights were normal 20 days after birth. Few litters survived the first day after birth when the dose was 40 mg/kg. Thus, an organ-specific effect can occur when one-third to one-half of the threshold dose for cleft palate formation is used. Calcification of the central vertebral bodies was reduced by 40 mg/kg. The results are discussed from the standpoint of the relative constancy of the syndrome of effects rather than from that of a changing spectrum, and from the standpoint of the potential persistence of effects at the various dose levels. Some of the findings represent additional evidence and manifestations of fluid imbalance at the lower doses.