Distribution In Different Organs Research Articles

In silico structure-based screening of versatile P-glycoprotein inhibitors using polynomial empirical scoring functions

P-glycoprotein (P-gp) is an ATP (adenosine triphosphate)-binding cassette transporter that causes multidrug resistance of various chemotherapeutic substances by active efflux from mammalian cells. P-gp plays a pivotal role in limiting drug absorption and distribution in different organs, including the intestines and brain. Thus, the prediction of P-gp–drug interactions is of vital importance in assessing drug pharmacokinetic and pharmacodynamic properties. To find the strongest P-gp blockers, we performed an in silico structure-based screening of P-gp inhibitor library (1,300 molecules) by the gradient optimization method, using polynomial empirical scoring (POLSCORE) functions. We report a strong correlation (r2=0.80, F=16.27, n=6, P<0.0157) of inhibition constants (Kiexp or pKiexp; experimental Ki or negative decimal logarithm of Kiexp) converted from experimental IC50 (half maximal inhibitory concentration) values with POLSCORE-predicted constants (KiPOLSCORE or pKiPOLSCORE), using a linear regression fitting technique. The hydrophobic interactions between P-gp and selected drug substances were detected as the main forces responsible for the inhibition effect. The results showed that this scoring technique might be useful in the virtual screening and filtering of databases of drug-like compounds at the early stage of drug development processes.

Effects of soil C:N on growth and distribution of nitrogen and carbon of Malus hupehensis seedlings

Aims Soil C:N is a sensitive indicator of soil quality and an indicator for assessing carbon and nitrogen nutrition balance of soils. Our objective was to explore the effects of soil C:N on growth and distribution of nitrogen and carbon of Malus hupehensis seedlings.Methods Using the track technology of C and N double mark, we investigated growth parameters(height, stem diameter and dry matter of different organs),15N parameters(absorption, N derived from fertilizer, distribution and utilization) and13C parameters(distribution in different organs) of two-year old M. hupehensis seedlings under six different soil C:N treatments(T1–T6 were 4.70, 9.78, 14.70, 19.96, 25.60 and 28.83, respectively).Important findings With increase of soil C:N, dry matter of roots increased significantly, while the height, stem diameter and dry matter of above ground parts and total plant increased at first and then decreased. The highest value appeared in the T4 treatment. There were significant differences in15N utilization efficiency among the six different treatments.15N utilization efficiency increased from T1 to T4, and the rate of T4(18.46%) was 1.73 times than that of T1. But, the15N utilization efficiency decreased with a further increase of soil C:N, as the rate of T5and T6 reduced that of T4 by 1.59% and 2.58%, respectively. In the two lower soil C:N treatments(T1 and T2),the order of Ndff value(15N derived from fertilizer) in the organs was roots leaves stems. With increase of soil C:N, Ndff value in leaves was the highest, followed by roots and stems.15N distribution ratio in leaves increased,while13C distribution decreased with increase of soil C:N, but the changes were the opposite in roots. Considering plant growth and utilization of nitrogen, the suitable soil C:N was 21–23 under these experimental conditions.

Assessing pathogenicity of Gallibacterium anatis in a natural infection model: the respiratory and reproductive tracts of chickens are targets for bacterial colonization

Two separate bird trials were performed to establish a reliable route of infection for Gallibacterium anatis in chickens, comparing intranasal (i.n.) and intravenous (i.v.) applications. Additionally, three mutually divergent isolates from three geographical locations, as shown by MALDI-TOF-MS and partial rpoB gene sequence analysis, were compared. In the first trial, birds were infected with one of the selected isolates by the i.v. or i.n. route. Subsequently, birds were killed 3, 12 and 24 h post infection following i.v. infection while at 3, 7 and 10 days post infection (dpi) in the case of i.n. infection along with birds of the control group. As a result, i.n. infection showed prominent and consistent bacterial tissue distribution in different organs persisting until 10 dpi, which was a striking contrast to the i.v. infection route. Likewise, histopathology revealed mild to severe tracheal lesions following i.n. infection. The second trial was set up to confirm both the achieved results and the robustness of i.n. infection but with an extended observation period, until 28 dpi In agreement with the preceding trial, identical results for bacteriological and histopathological examinations were obtained with persistency of bacteria until 28 dpi Comparing the three different isolates from Mexico, China and Austria, the Mexican isolate showed a somewhat higher pathogenicity than the other strains. Consequently, pathogenesis of G. anatis strains was studied in chickens elucidating i.n. infection as the most reliable route characterized by a long-lasting bacteraemia, targeting the respiratory and reproductive tract.

Effects of Winter Ryegrass Planting on Soil Heavy Metal Content and Accumulation and Distribution in Different Organs of Late Rice in Six Paddy Soils

Investigation of the effects of winter ryegrass planting on soil heavy metal content and its accumulation and distribution in different organs of late rice in double-rice cropping paddy field with different paddy soil types will play an important role on rice quality safety and environmental security assessment of paddy field.Based on seven years long-term located experiments in paddy fields,we analyzed the effects of winter ryegrass planting on soil heavy metal content and its accumulation and distribution in rice in double-rice cropping paddy field with different paddy soil types including granitic sandy soil(GSS),grey clayey soil(GCS),purple clayey soil(PCS),red yellow clayey soil(RCS),yellow clayey soil(YCS) and alluvial sandy soil(ASS).The results showed that the Cd content in GCS was significantly higher than that in other treatments,while the Pd content in YCS was significantly higher than that in other treatments.RCS had the highest Hg content with 0.029 mg kg-1,and the highest As contents with 29.5 mg kg-1 in YCS.The contents of Cd,Pb and As in late rice showed an order as root stem and leaves brown rice.Cd contents in root,stem and leaves,and brown rice planting in RCS were significantly higher than those in other treatments.And the highest Pb and As contents in root of rice planting in YCS was 33.0 and 119.5 mg kg-1,respectively.Meanwhile,the Hg and As contents in rice stem and leaves were significantly higher in ASS than in other soil types.Our results clearly demonstrated that the heavy metal content of brown rice was decreased,and the quality of late rice was ultimately improved under YCS condition.

Comparison of cadmium subcellular distribution in different organs of two water spinach (Ipomoea aquatica Forsk.) cultivars

Mechanisms of low cadmium (Cd) accumulations in cultivars of water spinach are poorly investigated. We aimed to improve understanding of the subcellular biochemical properties of the mechanisms involved. A pot experiment was conducted to investigate the subcellular distributions of Cd in lateral and main roots, stems, and young and old leaves of a high-Cd (T308) and a low-Cd cultivar (QLQ). The ratio of main root:lateral roots Cd concentration in QLQ was lower (0.34–0.35) than that in T308 (0.39–0.55). The ratio of stem:main root Cd concentration in QLQ was much lower (0.60–0.73) than that in T308 (1.19–1.58). QLQ has higher capacity to sequester Cd in cell wall fractions of main and lateral roots than T308. The difference in shoot Cd concentration between QLQ and T308 is attributable to the difference in Cd translocation from lateral to main roots and from roots to the stem. Fixation of large amounts of Cd in old leaves is beneficial to protect young leaves from Cd toxicity. Cadmium immobilization by the cell wall is important in Cd detoxification, especially in main and lateral roots of QLQ and the shoot of T308.

Selenium Treatment Mitigates the Effect of Lead Exposure in Coleus Blumei Benth

We examined the effects of selenium (Se) on coleus (Coleus blumei Benth.) under lead (Pb) stress, to determine possible mitigating mechanisms of Se. Coleus plants were exposed to 1.0 mM Pb(NO3)2 and varying concentrations of Na2SeO3 for 21 d. Application of 1.0 mM Se enhanced biomass allocation and Pb distribution in different organs, decreased superoxide dismutase (SOD) and peroxidase (POD) activity in the roots, and acted as an antioxidant by inhibiting lipid peroxidation via increasing glutathione levels. Root catalase (CAT) and glutathione peroxidase (GSH-Px) activities increased with Se concentration, and changes to root and leaf particles were observed by scanning electron microscopy and X-ray diffraction. Our results indicate Pb is tolerated by coleus plants through allocation plasticity, activation of antioxidant systems, and improvements in particle size and mineralogical effects. C. blumei can be useful in phytoremediation of aquatic systems contaminated with Pb, especially with addition of low concentrations of Se.

Vibrio salmonicida pathogenesis analyzed by experimental challenge of Atlantic salmon (Salmo salar)

Cold-water vibriosis (CV) is a bacterial septicemia of farmed salmonid fish and cod caused by the Gram-negative bacterium Vibrio (Aliivibrio) salmonicida. To study the pathogenesis of this marine pathogen, Atlantic salmon was experimentally infected by immersion challenge with wild type V. salmonicida and the bacterial distribution in different organs was investigated at different time points. V. salmonicida was identified in the blood as early as 2 h after challenge demonstrating a rapid establishment of bacteremia without an initial period of colonization of the host. Two days after immersion challenge, only a few V. salmonicida were identified in the intestines, but the amount increased with time. In prolonged CV cases, V. salmonicida was the dominating bacterium of the gut microbiota causing a release of the pathogen to the water. We hypothesize that V. salmonicida uses the blood volume for proliferation during the infection of the fish and the salmonid intestine as a reservoir that favors survival and transmission. In addition, a motility-deficient V. salmonicida strain led us to investigate the impact of motility in the CV pathogenesis by comparing the virulence properties of the mutant with the wild type LFI1238 strain in both i.p. and immersion challenge experiments. V. salmonicida was shown to be highly dependent on motility to gain access to the fish host. After invasion, motility was no longer required for virulence, but the absence of normal flagellation delayed the disease development.

CD8+ T Cells and IFN-γ Mediate the Time-Dependent Accumulation of Infected Red Blood Cells in Deep Organs during Experimental Cerebral Malaria

BackgroundInfection with Plasmodium berghei ANKA (PbA) in susceptible mice induces a syndrome called experimental cerebral malaria (ECM) with severe pathologies occurring in various mouse organs. Immune mediators such as T cells or cytokines have been implicated in the pathogenesis of ECM. Red blood cells infected with PbA parasites have been shown to accumulate in the brain and other tissues during infection. This accumulation is thought to be involved in PbA–induced pathologies, which mechanisms are poorly understood.Methods and FindingsUsing transgenic PbA parasites expressing the luciferase protein, we have assessed by real-time in vivo imaging the dynamic and temporal contribution of different immune factors in infected red blood cell (IRBC) accumulation and distribution in different organs during PbA infection. Using deficient mice or depleting antibodies, we observed that CD8+ T cells and IFN-γ drive the rapid increase in total parasite biomass and accumulation of IRBC in the brain and in different organs 6–12 days post-infection, at a time when mice develop ECM. Other cells types like CD4+ T cells, monocytes or neutrophils or cytokines such as IL-12 and TNF-α did not influence the early increase of total parasite biomass and IRBC accumulation in different organs.ConclusionsCD8+ T cells and IFN-γ are the major immune mediators controlling the time-dependent accumulation of P. berghei-infected red blood cells in tissues.

Bioavailability and tissular distribution of docetaxel, a P-glycoprotein substrate, are modified by interferon-α in rats

Interferon-alpha (IFN-alpha) inhibits intestinal P-glycoprotein (P-gp) expression in rats. In the present study, the effects of repeated pre-treatment with recombinant human INF-alpha (rhIFN-alpha) on oral and intravenous pharmacokinetics of a P-gp substrate, docetaxel (DTX; Taxotere) were investigated in a rat model. The bioavailability and distribution in different organs were also studied. Sprague-Dawley rats were subcutaneously pre-treated with either rhIFN-alpha for 8 days (4MIU kg(-1), once daily) or with pegylated-IFN-alpha (ViraferonPeg; 60 microg kg(-1), Days 1, 4 and 7). The rats were then distributed into sub-groups (n = 5-6) according to the pre-treatment type, and received one dose of [(14)C]DTX (20 mgkg(-1)) either orally or intravenously. Pharmacokinetics studies were then performed over 240 min, at the end of which tissues (intestine, liver, kidneys, lung, heart and brain) were immediately removed for radioactivity quantitation. Non-pegylated and pegylated IFN-alpha both increased DTX oral bioavailability parameters: C(max) (17.0+/-4.0 microg L(-1) (P < 0.02) and 18+/-5.5 microg L(-1) (P < 0.05), respectively, vs 7.4+/-2.5 microg L(-1) for the control) and AUC (0.036+/-0.010 microg h mL(-1) (P < 0.01) and 0.033+/-0.009 microg h mL(-1) (P < 0.01), respectively, versus 0.012+/-0.004 microg h mL(-1) for the control). IFN-alpha also delayed DTX absorption from 60 min in controls to about 95 min and 80 min in non-pegylated and pegylated treated animals, respectively. However, IFN-alpha did not affect intravenous DTX pharmacokinetics and it had a limited effect on tissue distribution at 240 min. [(14)C]DTX was decreased in intestine and enhanced in brain in both pre-treated groups. rhIFN-alpha modified the P-gp-dependent pharmacokinetics of DTX, limited its intestinal efflux and markedly enhanced its oral bioavailability.

Visible Human Utilization to Render Induced Electric Field and Current Density Images Inside the Human

The external and internal exposure to power frequency electromagnetic field, generated by high-voltage power transmission lines, raises serious safety concerns. Since we cannot measure the induced electric fields and current densities inside the human body, we used the Visible Human (VH) to investigate the induced electric fields and currents in human body tissues and organs of a worker standing 2 m away from conductor phase C of a double-circuit 132 kV transmission line. The double circuit 132 kV 60 Hz transmission line has a power rating of 293 MVA and a maximum recorded peak load current of 603 A. Charge simulation method and the Biot-Savart law have been used for computation of external electric and magnetic fields. Finite-difference time-domain technique was used to calculate the organs' internal induced electric field and circulating current densities in more than 40 different tissues of the VH with 3 mm voxel size. The simulation indicates that, at 2 m away from a 132 kV transmission line, the computed external electric field is 6.485 kV/m and the external magnetic field is 66.4 muT, which are below the limits set by the IEEE standards for external exposure for live-line workers. The maximum induced electric fields in the brain and heart are 23 and 14 mV/m, respectively. These values are below the IEEE standard recommended limits of 53 mV/m for the brain and 943 mV/m for the heart. The VH data allowed us to obtain two- and three-dimensional images of the induced electric field and current density distribution in different organs, tissues, and cross-sections of the human body.

Evaluation of the Experimental Inoculation of Cryptococcus albidus and Cryptococcus laurentii in Normal Mice: Virulence Factors and Molecular Profile Before and After Animal Passage

The genus Cryptococcus includes free-developing species, a few of which are of medical importance. Some, such as C. neoformans and C. gattii, cause infections in man frequently and C. albidus and C. laurentii cause less so. The aims of this study were to evaluate organ colonization after inoculation of C. albidus and C. laurentii isolates in normal BALB/c mice, the virulence factors (growth at 37 degrees C, capsule, melanin, proteinase, and phospholipase production) and the molecular profile (PCR-fingerprinting) of the yeasts before and after infection. The importance of different profiles (virulence and molecular) was considered in relation to the distribution in different organs and to the time intervals of isolation from organs. C. albidus was isolated from animal organs 2 to 10 days after inoculation and C. laurentii from 2 to 120 days. Most isolates of the two species kept the virulence factors showed before inoculation. The high homogeneity of the molecular profile of C. albidus and the high heterogeneity of C. laurentii were kept through the passages in animals. It is concluded that most isolates of both species were recovered from the animal organs after 5 or more days, and phenotypes were not altered by inoculation. No molecular alteration was detected and the virulence factors were not related to the time intervals before isolation from organs.

Effects of polyamines on K+, Na+ and Cl− content and distribution in different organs of cucumber (Cucumis sativus L.) seedlings under NaCl stress

Seedlings from the salt-sensitive cucumber cultivar Jinchun No. 2 and the salt-tolerant cucumber cultivar Changchun Mici were exposed for 8 days to 50 mmol/L NaCl in the absence or in the presence of exogenous foliar spraying PAs [putrescine (Put), spermidine (Spd), and spermine (Spm) 1 mmol/L] to compare the effects of different kinds of polyamines (PAs) on plant tolerance to salinity. This paper studied the effects of exogenous PAs on K+, Na+ and Cl− in different organs of cucumber seedlings. The results showed that K+ content as well as the ratios of K/Na and Cl/Na decreased, while Na+ and Cl− concentrations increased in salt-treated cucumber seedlings. The differences in K+, Na+ and Cl− content and the K/Na and Cl/Na ratios were greater for the salt sensitive cultivar Jinchun No. 2 than for the salt-tolerant cultivar Changchun Mici. Cucumber seedlings treated with exogenous polyamines and combined with salinity exhibited a higher level of K+ accumulation and lower levels of Na+ and Cl− accumulation compared with the seedlings treated only with salt stress. Among the three kinds of polyamines, Spd and Spm were more effective in inhibiting the accumulation of Na+ and reduction of K+. However, Put was more effective in reducing Cl− accumulation. Furthermore, all of the three kinds of exogenous polyamines could increase the ratio of K/Na, improving the absorption and transport selectivities of K+ and Na+ from stems to leaves for both cultivars. In conclusion, exogenous polyamines could alleviate salt damage to some extent and enhance the accumulation of biomass. Among the three kinds of polyamines, spermidine was most effective. Exogenous polyamines could improve tolerance of cucumber seedlings under salt stress by regulating the absorption and distribution of ions in different organs.

2,3,7,8‐tetrachlorodibenzo‐p‐dioxin increases bovine herpesvirus type‐1 (BHV‐1) replication in madin‐darby bovine kidney (MDBK) cells in vitro

Dioxin-2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a common environmental toxin of current interest. In the last years, higher levels of TCDD than those permitted in UE [European Commission. 2002. European Commission Recommendation 2002/201/CE. Official Gazette, L 67/69] were detected in milk samples from cow, water buffalo, goat, and sheep raised on some areas of Campania Region (South Italy). Dioxin often causes immunosuppression and might render the animal liable to viral infections. In addition, viral infections are able to alter the pattern of dioxin distribution in different organs of the exposed animals. Bovine Herpesvirus type-1 (BHV-1) is a widespread pathogen, which causes infectious rhinotracheitis and infectious pustular vulvovaginitis in cattle. Herein, we have studied the effects of TCDD and BHV-1 infection, in Madin-Darby Bovine Kidney (MDBK) cells, alone as well as in association, so as cellular proliferation, apoptosis, and virus replication. We have observed an increase in cell viability of confluent monolayers at low TCDD concentrations. TCDD treated cells demonstrated increased viability compared to controls as evaluated by MTT test. TCDD exposure increased cell proliferation but induced no changes on apoptosis. Cells exposed to TCDD along with BHV-1 showed a dose-dependent increase in cytopathy, represented by ample syncytia formation with the elimination of the cellular sheets and increased viral titer. These results suggest that TCDD increases viral replication in MDBK cells while BHV-1 further decreases viability of TCDD exposed cells. Since very low concentrations (0.01 pg/ml) are sufficient to augment BHV-1 titer, TCDD may contribute to reactivate BHV-1 from latency, leading to recurrent disease and increase virus transmission.

Variability of Flavonol Contents during Floral Morphogenesis in the Poppy Papaver somniferum L.

We studied the contents of flavonols (kaempferol and quercetin) in the meristem of vegetative and generative apices of the main plant shoot in floral Papaver somniferum L. mutants, as well as in the normal plants at successive stages of flower development. Five stages of flower development were distinguished. Flavonols (kaempferol and quercetin) were present in all flower organs at all stages of floral morphogenesis we studied. However, their contents and distribution in different organs and at different stages of flower development markedly varied. No significant differences were found in the contents of flavonols in the meristems of vegetative and generative apices of the main shoot in the lines of floral mutants, as well as between the lines with different amounts of vegetative phytomeres. In the plants with normal flower structure, the contents of flavonols (kaempferol + quercetin) sharply increased with the beginning of differentiation of flower organs, i.e. from stage 3, to reach a maximum in the open flower, when gametogenesis is terminated and fertilization takes place. The level of flavonol contents in the petals (upper part) and stamen was at a maximum at all stages of flower development, while that in the gynaecium was at a minimum. The kaempferol : quercetin ratio was shifted towards quercetin at successive stages of flower development, most significantly in the stamens. The involvement of flavonols in the regulation of floral morphogenesis at stages of flower organs differentiation and functioning is discussed.

Influence of Catecholamines on Migration and Differentiation of GnRH Neurons in Rats

We studied the contents of flavonols (kaempferol and quercetin) in the meristem of vegetative and generative apices of the main plant shoot in floral Papaver somniferum mutants, as well as in the normal plants at successive stages of flower development. Five stages of flower development were distinguished. Flavonols (kaempferol and quercetin) were present in all flower organs at all stages of floral morphogenesis we studied. However, their contents and distribution in different organs and at different stages of flower development markedly varied. No significant differences were found in the contents of flavonols in the meristems of vegetative and generative apices of the main shoot in the lines of floral mutants, as well as between the lines with different amounts of vegetative phytomeres. In the plants with normal flower structure, the contents of flavonols (kaempferol + quercetin) sharply increased with the beginning of differentiation of flower organs, i.e. from stage 3, to reach a maximum in the open flower, when gametogenesis is terminated and fertilization takes place. The level of flavonol contents in the petals (upper part) and stamen was at a maximum at all stages of flower development, while that in the gynaecium was at a minimum. The kaempferol: quercetin ratio shifted towards quercetin at successive stages of flower development, most significantly in the stamens. The involvement of flavonols in the regulation of floral morphogenesis at stages of flower organs differentiation and functioning is discussed.

Hyperoxia and local organ blood flow in the developing chick embryo.

1. Hyperoxia can cause local vasoconstriction in adult animal organs as a protective mechanism against hyperoxia-induced toxicity. It is not known at what time during development this vasoconstrictor capacity is present. Therefore, we measured the cardiac output (CO) distribution in different organs during a period of acute hyperoxia (100 % O2) in the developing chick embryo. 2. Fertile eggs were divided into five incubation time groups (10 and 11, 12 and 13, 14 and 15, 16 and 17, and 18 and 19 days of a normal incubation time of 21 days). Eggs were opened at the air cell and a catheter was inserted into a branch of the chorioallantoic vein for injections of 15 microm fluorescent microspheres during normoxia and at the end of 5 min (test group 1; n = 39) or 20 min (test group 2; n = 21) of hyperoxia exposure (100 % O2). The fraction of CO to an organ was calculated as the fluorescence of the organ sample divided by the sum of the fluorescence of all organs. 3. Only in 18- and 19-day-old embryos did hyperoxia cause a decrease in the fractions of CO to the heart and carcass, and an increase in those to the yolk-sac and chorioallantoic membrane. This response was more pronounced after 20 min (test group 2) than after 5 min (test group 1) of hyperoxia with an additional decrease in the fractions of CO to the brain, intestine and liver (test group 2). 4. These data indicate that local mechanisms for hyperoxia-induced vasoconstriction in the heart, brain, liver, intestine and carcass develop late, during the final 15 % of the incubation period, in the developing chick embryo.

Inhalation of substance P and thiorphan: acute toxicity and effects on respiration in conscious guinea pigs.

Substance P is a tachykinin and a biologically active neuropeptide. The peptide produces salivation, neuronal excitation, vasodilatation, increased vascular permeability and contraction of smooth muscles in the respiratory tract. The study was designed to evaluate the acute effects in guinea pigs of inhaled aerosolized Substance P (SP). Apart from the acute toxic effect of the peptide, the distribution in different organs was also investigated. The acute inhalation toxicity of SP (LC50, 15 min) when co-administrated with the neutral endopeptidase inhibitor thiorphan was 368 microg m(-3). The peptide caused an increase in respiratory rate proceeding a decrease in tidal volume. As the exposure proceeded, a decrease in both respiratory rate and further decreases in tidal volume were observed until either the animal died or the exposure was terminated. The decreases in respiratory rate and tidal volume were probably due to bronchoconstriction caused by SP. Eighteen per cent of the inhaled amount of radioactive SP was retained in the body, and the highest concentrations of radioactivity were found in the kidney, lung and liver. Substance P in combination with thiorphan administered as an aerosol is extremely toxic and highly potent. Exposure to the substance at extremely low air concentrations may result in incapacitation in humans.

Methods for the separation of lactate dehydrogenases and clinical significance of the enzyme.

Lactate dehydrogenase (LDH), an ubiquitous enzyme among vertebrates, invertebrates, plants and microbes was discovered in the early period of enzymology. The enzyme has been dissolved in several distinguishable molecular forms. In mammals, three types of subunits encoded by the genes Ldh-A, Ldh-B and Ldh-C give rise to a selected number of tetrameric isoenzymes. LDH-A4, LDH-B4 and the mixed hybrid forms of the A- and B-subunits are present in many tissues but with certain distribution patterns. LDH-C4 is confined in mammals to testes and sperm. Numerous techniques have been employed to purify, characterize and separate the different forms of the enzyme. This report deals with the main protocols and procedures of purification of LDH and its isoenzymes including chromatographic and electrophoretic methods, partitioning in aqueous two-phase systems and precipitation approaches. In particular, affinity separation techniques based on natural and pseudo-biospecific ligands are described in detail. In addition, basic physico-chemical and kinetic properties of the enzyme from different sources are summarized in a second part, the clinical significance of the determination of LDH in diverse body fluids in respect to the total activity and the isoenzyme distribution in different organs is discussed.

Distribution of Externally Applied 15NH4 + or 15NO3 − in White Lupins at Different Developmental Stages

Abstract White lupins (Lupinus albus L., var. Kievsky mutant) were grown in Mitscherlich pots. 15NH4NO3 or NH4 15NO3 (10 or 95 at% 15N exc.) was applied at the late vegetative stage (14–16 leaves) or at the generative stage (end of bloom), respectively. Dry weight increase (ΔDW) of the plants as well as the 15N distribution in different organs and N fractions (NO3 −-N, NH2-N, protein N) were investigated after 5 days and 50 days (maturity). The following results were obtained: 1. In comparison with the 15N derived from NO3 −, the 15N derived from NH4 + was more strongly retained in the roots. When transported into the shoot, it was more strongly translocated into the tip. 2. More 15N derived from NH4 + was incorporated into proteins and reduced soluble N compounds than 15N derived from NO4 −. Even in the above-ground plant tissues, a considerable proportion of the latter was found as NO3 −. 3. In the long-term experiment (harvest at maturity 50 days after application), no difference in translocation and i...

Desiccation leads to the rapid accumulation of both cytosolic and chloroplastic proteins in the resurrection plant Craterostigma plantagineum Hochst

A number of desiccation-related and abscisic-acid (ABA)-inducible transcripts have been isolated from the resurrection plant Craterostigma plantagineum (Scrophulariaceae). They have been analysed at the transcriptional level (D. Bartels et al., 1990, Planta 181, 27–34) and their nucleotide sequences determined (D. Piatkowski et al., 1990, Plant Physiol. 94, 1682–1688). Three such genes encoded polypeptides with substantial homologies to proteins abundantly expressed during late embryogenesis in many higher plants; two other genes encoded novel transcripts. The temporal expression patterns of these gene products and their distribution in different organs of the plant and in callus tissues have now been analysed immunologically. For this, in-situ RNA hybridizations and immunocytochemical studies using tissue sections were carried out at both the light and electron microscope level. All of the products were found to be present in leaf tissue, and some were also found in roots and in seeds. Three desiccation-related proteins were localized in the cytosol, while two others, one associated with the thylakoid membranes, the other soluble in the stroma, were detected in the chloroplast. In C. plantagineum the severe ultrastructural changes observed during the desiccation-rehydration process indicate the need for protectants: the gene products characterized in this publication may be good candidates for this role.