Decrease In Total Protein Content Research Articles

PHARMACOLOGICAL SCREENING OF EUPHORBIA NERIIFOLIA LEAF HYDROALCOHOLIC EXTRACT

Pharmacological screening of Euphorbia neriifolia Linn. (Euphorbiaceous) leaf hydro alcoholic extract were performed to explore the analgesic, anti-inflammatory, diuretic, antidiarrhoeal and antiulcer activities. All tests were conducted on rats using 100, 200 and 400 mg/kg dose as LD50 of extract was found to 2779.71 mg/kg. Study revealed strong analgesic effect of E. neriifolia against thermal (P<0.001), and in mechanical and chemical (p < 0.01) noxious stimuli and anti-inflammatory activity (P<0.001 to 0.01) at the 1000 mg/kg dose. In carrageenan-induced paw edema and cotton pellet induced granuloma model E. neriifolia extract showed significant (P<0.001 to 0.01) antiinflammatory activity. Extract considerably increases urine volume as an effective hypernatraemic and hyperchloraemic diuretic. E. neriifolia showed laxative property by increasing wet defecation along with castor oil. Extract showed very prominent protection against ethanol-induced ulceration as well as on pyloric ligated ulceration in dose dependent manner. Extract increases total hexoses (P<0.001), hexosamine (P<0.05), sialic acid and total carbohydrate content (P<0.001) with a decrease in total protein content (P<0.001) of gastric mucosa at 400 mg/kg dose. Presence of phytoconstituents like tannins, flavonoids, alkaloids and triterpenoidal saponins may be responsible for the found pharmacological activities.

Hydrogen Enhances Nickel Tolerance in the Purple Sulfur Bacterium Thiocapsa roseopersicina

A common microbial strategy for detoxifying metals involves redox transformation which often results in metal precipitation and/or immobilization. In the present study, the influence of ionic nickel [Ni(II)] on growth of the purple sulfur bacterium Thiocapsa roseopersicina was investigated. The results suggest that Ni(II) in the bulk medium at micromolar concentrations results in growth inhibition, specifically an increase in the lag phase of growth, a decrease in the specific growth rate, and a decrease in total protein concentration when compared to growth controls containing no added Ni(II). The inhibitory effects of Ni(II) on the growth of T. roseopersicina could be partially overcome by the addition of hydrogen (H(2)) gas. However, the inhibitory effects of Ni(II) on the growth of T. roseopersicina were not alleviated by H(2) in a strain containing deletions in all hydrogenase-encoding genes. Transmission electron micrographs of wild-type T. roseopersicina grown in the presence of Ni(II) and H(2) revealed a significantly greater number of dense nanoparticulates associated with the cells when compared to wild-type cells grown in the absence of H(2) and hydrogenase mutant strains grown in the presence of H(2). X-ray diffraction and vibrating sample magnetometry of the dense nanoparticles indicated the presence of zerovalent Ni, suggesting Ni(II) reduction. Purified T. roseopersicina hyn-encoded hydrogenase catalyzed the formation of zerovalent Ni particles in vitro, suggesting a role for this hydrogenase in Ni(II) reduction in vivo. Collectively, these results suggest a link among H(2) metabolism, Ni(II) tolerance, and Ni(II) reduction in T. roseopersicina .

Transcriptional activation of a 37 kDa ethylene responsive cysteine protease gene, RbCP1, is associated with protein degradation during petal abscission in rose.

Cysteine proteases play an important role in several developmental processes in plants, particularly those related to senescence and cell death. A cysteine protease gene, RbCP1, has been identified that encodes a putative protein of 357 amino acids and is expressed in the abscission zone (AZ) of petals in rose. The gene was responsive to ethylene in petals, petal abscission zones, leaves, and thalamus. The expression of RbCP1 increased during both ethylene-induced as well as natural abscission and was inhibited by 1-MCP. Transcript accumulation of RbCP1 was accompanied by the appearance of a 37 kDa cysteine protease, a concomitant increase in protease activity and a substantial decrease in total protein content in the AZ of petals. Agro-injection of rose petals with a 2.0 kb region upstream of the RbCP1 gene could drive GUS expression in an abscission zone-specific manner and was blocked by 1-MCP. It is concluded that petal abscission is associated with a decrease in total protein content resulting from rapid transcription of RbCP1 and the expression of a 37 kDa protease.

Enhanced expression of serine proteases during floral senescence in Gladiolus

Programmed cell death during senescence in plants is associated with proteolysis that helps in remobilization of nitrogen to other growing tissues. In this paper, we provide one of the few reports for the expression of specific serine proteases during senescence associated proteolysis in Gladiolus grandiflorus flowers. Senescence in tepals, stamens and carpels results in an increase in total protease activity and a decrease in total protein content. Of the total protease activity, serine proteases account for about 67–70% while cysteine proteases account for only 23–25%. In-gel assays using gelatin as a substrate and specific protease inhibitors reveal the enhanced activity of two trypsin-type serine proteases of sizes 75 kDa and 125 kDa during the course of senescence. The activity of the 125 kDa protease increases not only during tepal senescence but also during stamen and carpel senescence indicating that it is responsive to general senescence signals.

Influência do hipotireoidismo induzido por propiltiouracil na mucosa gengival do rato

This work aimed at verifying the influence of propyl-thiouracil (PTU)-induced thyroid hormone deficiency on gingival mucosa of young male rats, measuring total protein concentration, collagen content and DNA concentration as indices of cellular population. Fifty Sprague-Dawley rats were used. The animals were grouped in: PTU-treated (i.p. 10 mg/d) and control rats (C). The experience was maintained for a period of 10 weeks. Total protein content of gingival mucosa tissue was determined by the Lowry method; hydroxyprolin rate, as prototype amino acid of collagen, was determined using the Newman method, and DNA concentration was measured by Burton's methodology. The results showed decreased amounts of PTU-treated rats gingival total protein content (PTU= 41.23 +/- 24.05 vs. C= 63.36 +/- 18.05); no alterations were seen in hydroxyprolin concentration neither in DNA content of PTU treated rats, respectively (PTU= 2.18 +/- 1.48 vs. C= 2.29 +/- 1.51) and (PTU= 0.33 +/- 0.19 vs. C= 0.46 +/- 0.41). Thus, PTU treatment promotes a decrease in total protein content of rat gingival mucosa that may be interpreted as a decrease in protein synthesis induced by the hypothyroid condition, but with no alteration either in collagen or nucleic acid rates.

Determination of the Grape Invertase Content (Using PTA−ELISA) following Various Fining Treatments versus Changes in the Total Protein Content of Wine. Relationships with Wine Foamability

Proteins have proven to play a major role in the stabilization of foam in Champagne wines despite their low concentration that ranges from 4 to 20 mg/L. The aim of this study was to evaluate the effect of fining on total protein and grape invertase contents of champenois base wines and their foaming properties. Data showed that fining and especially the use of bentonite at doses ranging from 10 to 50 g/hL leads to a significant decrease in the total protein content of wines together with that of the grape invertase content, with such a decrease being very detrimental to the foaming properties of the treated wines in terms of foam height (HM) and foam stability (HS). Only a slight decrease in the total protein content, in the grape invertase concentration, and in the foam quality of wines was observed when using casein (10 and 20 g/hL) or bentonite combined with casein (both at 20 g/hL). Our study thus clearly establishes the good correlation existing between the wine protein concentration and its foaming properties. A remarkable correlation was observed between the decrease in the grape invertase content and the total protein content of wines, following bentonite treatments, suggesting that the grape invertase (which represents at least 10-20% of the wine proteins) follows a similar behavior upon fining to other proteins of Champagne wines, despite the high molecular mass and the highly glycosylated structure of this particular protein. Moreover, the decrease in total protein and grape invertase contents of wine after fining with bentonite was found to be correlated with a decrease in the foaming properties of the corresponding wines (with respectively R(2) = 0.89 and 0.95).

Haemagglutination Inhibition Antibodies, Rectal Temperature and Total Protein of Chickens Infected with a Local Nigerian Isolate of Velogenic Newcastle Disease Virus

Changes in values of haemagglutination inhibition (HI) antibody titre, rectal temperature (RT) and total protein (TP) were determined for Shaver Brown chickens infected with Newcastle disease virus (NDV) Kudu 113. The infected chickens came down with Newcastle disease by day 3 post infection (PI). The major clinical signs were depression, greenish diarrhoea, paralysis of legs and wings, opisthotonus and torticolis. Mortality and morbidity were 52% and 1000%, respectively. There were haemorrhagic lesions in the wall of the intestine, proventricular mucosa and caecal tonsils. There were necrosis and mononuclear cell infiltration of the liver, kidney and spleen. There was a significant increase in daily mean HI antibody titres from days 3 to 9 PI. Similarly, significant rise in daily mean RTs were noticed in the infected chickens from days 1 to 13 PI. On the other hand, there was a decrease in daily mean TP concentrations of infected chickens, beginning from day 3 PI, and the lowest concentration of 2.60 +/- 0.15 g/dl was obtained by days 7 and 11 PI. The values of HI, RT and TP for the control chickens were relatively constant during the experiment. The correlation coefficient (r) between HI and RT was positive and highly significant (r = 0.725, p<0.001), while the relationship between HI and TP was negative but highly significant (r = -0.712, p<0.001). It was concluded that NDV Kudu 113 induced increases in values of HI and RT, which occurred concurrently with a decrease in TP concentrations of infected chickens.

Short-term response of benthic bacteria and nanoflagellates to sediment resuspension: an experimental study

We investigated benthic bacterial abundance, biomass and activities and heterotrophic nanoflagellate (HNF) abundance and biomass during sediment resuspension and after a resilience period through short-term laboratory experiments. The strong sediment resuspension increased bacterial carbon production only slightly (Tukey's test, P < 0.05) but somehow interfered with microbial growth. During sediment resuspension, after an initial increase (at 48 and 72 h) bacterial abundance and biomass decreased until the end of resuspension. Sediment resuspension also altered the relationship between HNFs and bacteria (increase in the ratios of bacterium to HNF abundance and biomass after sediment resuspension; Tukey's test, P < 0.01). After sediment resuspension, the fraction of bacterial C production removed by flagellates decreased, whereas bacterial turnover did not change significantly, suggesting that HNF pressure on bacteria decreased. These results are consistent with the significant decrease in both HNF abundance and biomass (Tukey's test, P < 0.01) and suggest that this small benthic component is less resilient to sediment resuspension than bacteria. Sediment resuspension also determined significant changes in sediment organic matter pools (proteins and carbohydrates). Sediment resuspension determined a decrease in total protein concentrations, whereas total carbohydrate concentrations did not change significantly. Sediment resuspension had significant effects on extracellular enzymatic activities, as aminopeptidase increased and β-glucosidase activity decreased. Based on total protein and carbohydrate concentrations and enzymatic activities, we observed that sediment resuspension resulted in an increase in protein turnover and a decrease in carbohydrate turnover. These results indicate that sediment resuspension plays a major role in the early diagenesis of sediment organic matter.

The toxicity of opiates and their metabolites in HepG2 cells

The toxicity of codeine (C), codeinone (CO), morphine (M), oxycodone (OC), pholcodine (P) and pholcodine-N-oxide (P-NOX) was assessed in HepG2 cells by determining cell viability via the measurement of lactate dehydrogenase (LDH) leakage through the membrane, depletion of reduced glutathione (GSH) and measurement of total protein content. Incubation of C, M, OC, P or P-NOX with HepG2 cells resulted in no significant loss of cell viability, depletion of GSH or decreased total protein content. In contrast, with CO there was a marked depletion of GSH with significant differences from control cells (P<0.05) being detected after as little as 5 min. This effect preceded the loss of cell viability and the decrease in total protein content. To identify the cause of GSH depletion during incubations with CO, the incubation solutions were analysed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). Analysis showed that a codeinone–glutathione conjugate (CO–SG) had been formed. This adduct was synthesised and characterised by LC/MS/MS and by nuclear magnetic resonance spectroscopy (NMR). CO–SG was quantified in the incubation solutions using the synthesised standard substance. Results obtained in this study support the hypothesis that the toxicity of CO may be partly due to GSH depletion. The absence of LDH leakage and GSH depletion in the incubations containing C or OC suggests, that the presence of both a double bond at Δ 7 and an adjoining keto-group in the 6-position are necessary to elicit the toxicity of M analogues with regard to GSH depletion.

Stress reactions in Vitis vinifera L. following soil application of the herbicide flumioxazin

In order to evaluate the stress effects of flumioxazin (fmx) on grapevine, a non-target plant ( Vitis vinifera L.), physiological parameters such as carbohydrate content, water status or nitrogenous metabolites were investigated on fruiting cuttings and plants grown in vineyard. In the leaves of cuttings, the soil-applied herbicide induced stress manifestations including a decrease of the dry weight percentage and the soluble carbohydrate content during the first week after treatment. Thereafter, a decrease of the osmotic potential was observed, as well as a decrease of total protein content and a parallel accumulation of free amino acids, including proline. Altogether, these results suggest that soil-applied fmx induced a stress in grapevines, leading to leaf proteolysis. However, this stress was partially recovered 3 weeks after herbicide application, suggesting that the cuttings were capable to adapt to the fmx exposure. In the vineyard, the flumioxazin effects were still significant 5 months after the treatment, particularly in the CH cv. They included a decrease of the leaf dry weight percentage and soluble carbohydrate content, as well as an increase of the osmotic potential. The decrease of leaf soluble carbohydrates may have dramatic consequences for the berry growth and the reserve constitution. Moreover, treated plants were characterized by a decrease of the free amino acid content and an accumulation of ammonium, while the protein level did not significantly increase, suggesting a degradation of amino acids. The alteration of carbon and nitrogen status after herbicide treatment may affect the grapevine vigour in a long term.

24-h Langendorff-perfused neonatal rat heart used to study the impact of adenoviral gene transfer.

The human genome project has increased the demand for simple experimental systems that allow the impact of gene manipulations to be studied under controlled ex vivo conditions. We hypothesized that, in contrast to adult hearts, neonatal hearts allow long-term perfusion and efficient gene transfer ex vivo. A Langendorff perfusion system was modified to allow perfusion for >24 h with particular emphasis on uncompromised contractile activity, sterility, online measurement of force of contraction, inotropic response to beta-adrenergic stimulation, and efficient gene transfer. The hearts were perfused with serum-free medium (DMEM + medium 199, 4 + 1) supplemented with hydrocortisone, triiodothyronine, ascorbic acid, insulin, pyruvate, l-carnitine, creatine, taurine, l-glutamine, mannitol, and antibiotics recirculating (500 ml/2 hearts) at 1 ml/min. Hearts from 2 day-old rats beat constantly at 135-155 beats/min and developed active force of 1-2 mN. During 24 h of perfusion, twitch tension increased to approximately 165% of initial values (P < 0.05), whereas the inotropic response to isoprenaline remained constant. A decrease in total protein content of 10% and histological examination indicated moderate edema, but actin and calsequestrin concentration remained unchanged and perfusion pressure remained constant at 7-11 mmHg. Perfusion with a LacZ-encoding adenovirus at 3 x 108 active virus particles yielded homogeneous transfection of approximately 80% throughout the heart and did not affect heart rate, force of contraction, or response to isoprenaline compared with uninfected controls (n = 7 each). Taken together, the 24-h Langendorff-perfused neonatal rat heart is a relatively simple, inexpensive, and robust new heart model that appears feasible as a test bed for functional genomics.

Metabolic depression and whole-body response to enforced starvation by Crassostrea gigas postlarvae

Physiological and biochemical measurements were performed on six oyster ( Crassostrea gigas) cohorts, in order to: (a) investigate the whole-body response (growth, energy content, metabolic and excretion rates) of 2-week-old postlarvae (spat) to enforced (0–8 days) starvation, and (b) test the potential use of three aerobic enzyme systems as indices of physiological condition. Starvation resulted in exponential reduction of postlarval metabolic and excretion rates, as well as a linear decrease in enzyme activity. These response mechanisms effectively limited the loss of endogenous reserves after 2 days of starvation and maintained the oyster's functional integrity over prolonged (8 days) starvation. Proteins appeared to be selectively conserved during short-term (2 days) starvation, as suggested by a decrease in total protein content, while maintaining constant weight-specific enzyme activity. Postlarvae starved for 2 days exhibited relatively higher lipid losses, lower mortality and lower metabolism than metamorphosing stages, thus suggesting a greater buffering capacity to starvation in the former. The activity of the electron transport system may be a useful indicator of long-term stress or developmental condition of oyster postlarvae, while citrate synthase and cytochrome oxidase could be used as indicators of growth rate. None of these enzyme systems is recommended as an index of aerobic metabolism during short-term starvation.

Influence of refining steps on trace allergenic protein content in sunflower oil

Background: Although allergy to sunflower seed and oil is a relatively rare occurrence, several cases of sunflower seed allergy have been observed, and we have already described one case of anaphylaxis after eating sunflower oil and margarine. Objective: The aim of our study was to determine and characterize the allergens from sunflower oil at the different steps of the refining process: crude pressed oil (step A), acidification and neutralization (step B), pregumming by centrifugation (step C), washing (step D), bleaching (step E), gumming by filtration (step F), and deodorization (step G). Methods: A sample of oil from each step of the process (steps A to G) was heat extracted with PBS. The protein concentration of each extract was evaluated by using the micro-Bradford assay. Samples were run on SDS-PAGE. The immunoblot was performed with the serum of a patient sensitized to sunflower seed and oil. Results: The extracts obtained after each step reveal a decrease in total protein concentration from 13.6 μg/mL to 0.22 μg/mL. The result of SDS-PAGE shows 5 bands, from 67 kd to 145 kd, with the most abundant being the 67-kd protein. The amount of this protein decreases after each step of the process. It is, however, still present in trace amounts in the refined oil. The 67-kd protein, which is mainly present in the crude oil and slightly in the refined oil, has been shown to be allergenic. Conclusion: Because of the presence of allergenic proteins, refined sunflower oil may pose a threat to people highly sensitized to sunflower seeds. (J Allergy Clin Immunol 2000;106:962-7.)

140 Ultrastructural Changes during the Senescence of Petals in Hibiscus syriacus L.

Five distinctive developmental stages were chronologically suggested. Cells at Stage I and II were essentially free of cytoplasmic or vacuolar abnormalities and the cytoplasm contained numerous electron-dense mitochondria with well-developed cristae. At Stage III, there were a localized dilation of mitochondria matrix and a partial-diluted cytoplasm in mesophyll cells. At Stage IV, characterized by high levels of fresh weight and osmolality, most mesophyll cells were seen to be ruptured, resulting in a general mixing of cell contents and diluting cytoplasm. It can be explained as an irreversible senescence phenomena that tonoplast in mesophyll cell was ruptured partly, corresponding to rapid increase in petal cell size and turgidity. Petal turgidity was due to an increase of content in soluble sugar. At Stage V, there was a loss of petal fresh weight. With a loss of turgidity, most mesophyll cells have collapsed completely. There were a notable plasmolysis in vasculature. The activity of protease in petals was found to increase between Stage II and III, and then decreased rapidly at Stage IV, resulting in the decrease of total protein content before senescence. Unexpectedly, there were stomata in hibiscus petals. Ultrastructural disorganization, like as a broken tonoplast, was observed in mesophyll cells at Stage IV. ABA and the stomata on petal might promote the disorganization. The final stages of senescence involved breakdown of cellular organization leading to hydrolysis of previously separated compartments. The cellular disorganization triggered during the flowers are still in the process of opening may be one of the earliest physiological signal that senescence is under way.

Effects of UV-B radiation on Olea europaea: comparisons between a greenhouse and a field experiment.

We compared growth, morphology, anatomy and physiology of field-grown Olea europaea L. plants after one year with or without supplemental UV-B radiation equivalent to the increase at ground level resulting from a 15% depletion in stratospheric ozone concentration over Patras, Greece (38.3 degrees N, 29.1 degrees E). In a parallel greenhouse experiment, plants were exposed for four months to either zero UV-B radiation or UV-B radiation equivalent to that of the supplemented outdoor treatment. In the field, the only significant changes in response to supplemental UV-B radiation were an increase in adaxial epidermal thickness and a decrease in total protein concentration. In the greenhouse-grown plants, UV-B radiation caused significant increases in abaxial cuticle thickness and trichome UV-B absorbing compounds. We conclude that Olea europaea exhibits high UV-B tolerance and will not be affected by the predicted increases in UV-B radiation.

Increased vulnerability of neurones and glial cells to low concentrations of methylmercury in a prooxidant situation.

Using reaggregating rat brain cell cultures at two different stages of differentiation, we examined the biochemical effects of a 10-day treatment with nanomolar concentrations of methylmercuric chloride (monomethylmercury), in the presence or absence of promoters of hydroxyl radical formation (10 microM copper sulphate plus 100 microM ascorbate). A decrease in total protein content accounted for the general cytotoxicity of these compounds, whereas selective effects were assessed by determining the activities of cell type-specific enzymes. Methylmercury, up to 100 nM, as well as the copper ascorbate mixture, when applied separately, induced no general cytotoxicity, and only slight effects on neuronal parameters. However, when applying 100 nM methylmercury and the copper-ascorbate mixture together, a drastic decrease in neuronal and glial parameters was found. Under these conditions, the content of reactive oxygen species, assessed by 2',7'-dichlorofluorescin oxidation, increased greatly, while the activities of antioxidant enzymes decreased. In the presence of copper and ascorbate, differentiated cultures appeared more resistant than immature ones to low methylmercury concentrations (1-10 mM), but did undergo similar changes in both cell type-specific and antioxidant enzyme activities at 100 nM methylmercury. These results suggest that in prooxidant conditions low doses of mercury can become much more deleterious for the central nervous system.

Inhibition by lithium of gentamicin-induced release of N-acetyl- β- d-glucosaminidase in rat submandibular saliva

1. 1. The effects of gentamicin, lithium and concurrent treatment of these drugs on concentration of total protein and activity of the enzyme N-acetyl- β- d-glucosaminidase (NAG) in rat submandibular saliva were studied. 2. 2. Pure submandibular saliva was collected intraorally by micropolyethylene canula from anesthetized rats with the use of pilocarpine as secretagogue. 3. 3. Daily intraperitoneal injection of gentamicin (50 mg kg/day) for 5 consecutive days caused a marked increase in total protein concentration and the NAG activity. 4. 4. Chronic treatment of rats for 10 days with lithium chloride solution (1,200 mg/l) caused a significant decrease in total protein concentration but did not affect the NAG activity. 5. 5. Concurrent treatment of lithium with gentamicin caused the total protein concentration and the NAG activity of submandibular saliva to reach to those of controls. 6. 6. In this regard, the protective effects of lithium may be due to interference of this ion with the phosphoinositide cycle.

Failure of a prolyl 4-hydroxylase inhibitor to alter extracellular matrix deposition during experimental pancreatitis.

In chronic pancreatitis the exocrine pancreatic tissue is replaced by extracellular matrix deposits from fibroblasts. We have stimulated fibrogenesis in the pancreas by inducing a single episode of cerulein pancreatitis (10 microg/kg/h for 12 h). We have used a spectrophotometrical assay to measure the tissue hydroxyproline content and immunohistochemistry to study the transient deposition of extracellular matrix in the pancreas. We have investigated whether a potent prolyl 4-hydroxylase inhibitor (HOE-077) can influence the deposition of extracellular matrix in the pancreas. Three days after the induction of the pancreatitis we found the maximum increase in pancreatic hydroxyproline content (by 63%), the maximum decrease in total protein content and amylase activity (by -39 and -86%, respectively), as well as a significant increase in DNA content and the deposition of interstitial collagen fibers on electron microscopy. By immunohistochemistry the largest expansion of extracellular matrix components was found for fibronectin. HOE 077, regardless of the concentration administered, failed to affect any of these parameters. We conclude that the induction of a single episode of cerulein pancreatitis and the serial determination of pancreatic hydroxyproline content represents a simple method to induce and monitor experimental fibrogenesis in the pancreas. Prolyl 4-hydroxylase inhibition did not affect the course of extracellular matrix deposition in the pancreas.

Effect of Long-term Therapeutic Bath in Malachite Green on Common Carp (Cyprinus carpio L.)

Svobodova Z., L. Groch, M. Flajshans, B. Vykusova. 1. Machova: The Effect of Long-term Therapeutic Bath of Malachite Gteen on Common Carp (C-,prinus carpio L.). Acta vet. Bmo 1997,66: 111-117. The aim of this paper was to determine acute toxicity of malachite green of the Vert Malachite BE SA Crystaux type for common carp and to evaluate the effect of a 6-day therapeutic bath in O.S mg.!-I concentration of this pharmaceutical on values of selected haematological indicators including micronuclei test, as well as on histological composition of tissues of oneto two-year-old carp. The 48hLCSO value of the malachite green tested was 0.84 mg.!-I. After the 6-day bath, changes in red blood picture [decrease in haematocrit value PCV (P<O.OOI), decrease in mean corpuscular volume MCV (P<O.OS) and increase in mean corpuscular haemoglobin concentration MCHC (p<0.0 I) 1 as well as in white blood picture /decrease in both relative and absolute monocyte count (P<O.OS and P<O.OI), respectively/ were recorded in treated fish (n=30) compared to the control group (n=29). Other indices of red and white blood picture in these fish did not differ from the control group. A decrease in total protein concentration (TP, P<O.O I) in blood plasma was found in fish treated with malachite green. By means of a micronuclei test, no effect of long-term bath in malachite green on increase of their frequency in erythrocytes was proved. Histological examination revealed mild regressive changes on gills, mild dystrophic changes in parenchymatous tissues and an increased macrophage activity in treated fish. These changes indicate an increased sensitivity of the fish treated with malachite green to hypoxia and impaired proteosynthesis. Acute toxicity, haematology, micronuclei test, histopathological changes Malachite green is stilI one of the mostly used preparations for inhibition of protozoal ectoparasitoses and mycoses of fish and their eggs (N 0 g a 1995), although it has numerous negative side effects. Its current importance even increased, since it was recommended for therapy ofa proliferative kidney disease PKD (C I i fton H ad ley and AI derm an 1987, 1988; Deb u f 1991). The negative effects of malachite green are as follows: high acute toxicity for fish (Machova and Svobodova 1984; Alderman 1985; Obiekezic and 0 k a for 1995; Van H e e r den et ai. 1995), capacity to accumulate in fish and longterm persistence of residues in tissues (B au e r et ai. 1988; A Ide r man 1988; A Ide r man and Clifton-Hadley 1993: Machova et al. 1996). Furthermore, malachite green is characterized as respiratory poison, teratogen and suspect carcinogen (M eye rand J 0 r g ens e n 1983). The negative effect of therapeutic baths in malachite green on haematological indicators and histopathological status of tissues was reported by numerous authorsinsalmonids(Ross etaI.1985; Pickering and Pottinger 1985; Hlavek and Bulkley 1980; Gerundo etaI.I991)andinsilurids(Srivastava etaI.1995). This paper deals with acute toxicity determination of the malachite green Vert Malachite 13 ESA Crystaux (imported from Italy) and with observation on the effect of a long-term therapeutic bath in this pharmaceutical on haematological indicators and histopathological composition of tissues of common carp. This type of malachite green was succesfully introduced and distributed among Czech fish farms.

Effects of cyclosporine a on the functions of submandibular and parotid glands of rats

1. 1. The present study was designed to investigate the possible effects of long-term (45 days) administration of therapeutic doses of cyclosporine A (25 mg/kg/day), on the functions of submandibular and parotid glands of rats. Pure submandibular and parotid saliva were collected intraorally by microcannulation of the ducts. 2. 2. The weight gains of the treated animals during the study and the weights of the salivary glands at the end of 45 days were reduced significantly as compared with those of controls. 3. 3. Sialochemistry studies revealed a marked decrease in total protein concentration in saliva obtained from submandibular glands (P<0.05). 4. 4. Determination of electrolyte concentrations in saliva of submandibular gland and serum showed considerable differences between treated and control groups. 5. 5. Significant elevation of amylase activity in serum and parotid saliva was observed in the treated rats in comparison with controls (P<0.001). 6. 6. Data presented here indicates that long-term administration of therapeutic doses of cyclosporine A causes significant alterations in salivary output and composition.