Acidification of lakes in Šumava (Bohemia) and in the High Tatra Mountains (Slovakia)

Acidification of lakes takes place when pH of rainwater is less than 4.5 and the catchments lie on sensitive geology. Both conditions are met for most lakes in Bohemia and Slovakia. Since 1978 we have studied mountain lakes in the Sumava and in the High Tatra Mountains. In Šumava the three lakes under study are of glacial origin. The catchments are small, with steep sides covered by spruce. The bedrocks are biotite-rich paragneiss, together with gneiss, quartzite and granite. In summer 1936 surface pH was 5.7–6.9 in the Lake Čertovo and 6.9–7.0 in the Lake Černé. Now the pH values are 4.3–4.8 in the two lakes and in the Lake Prášilské as well. Old reports on zooplankton are from the years 1871, 1892–96, 1935–37, 1947 and 1960. Since 1979 we have not found any planktonic Crustacea in the lakes Černé and Čertovo. Lake Prášilské is inhabited by Daphnia longispina and Cyclops abyssorum. In July 1989 the pH values were 4.4, 4.7 and 4.7, concentrations of labile monomeric Al were 0.83, 0.68 and 0.24 mg l^-1 in the lakes Čertovo, Černé and Prašilské, respectively. High levels of toxic Al compounds might be responsible for the extinction of planktonic Crustacea in the lakes Čertovo and Černé. All the three lakes are void of fish at present. In the High Tatra Mts. we examined more than 40 lakes above timberline in altitudes 1612–2145 m. They are all clearwater, naturally fishless lakes. The bedrock is granite. Owing to different levels of calcium the lakes are now in different stages of acidification. According to recent changes in the zooplankton they can be divided into three groups: (1) Species composition of planktonic Crustacea has not changed. (2) Planktonic Crustacea were present until 1973 but are absent now. (3) From the original species of Crustacea only Chydorus sphaericus is present. The three groups are well separated along the gradients of calcium, ANC and pH. They can be identified with the Henriksen's bicarbonate (our group 1), intermediate (our group 2) and acid (our group 3) lakes. We suppose that in the process of acidification the lakes of the group (2) have been shifted from oligotrophy to ultraoligotrophy.     

Isolation of Sporothrix schenckii From the Claws of Domestic Cats (Indoor and Outdoor) and in Captivity in São Paulo (Brazil)

Sporotrichosis is a subcutaneous mycosis and is also a zoonosis (sapro- and anthropozoonosis). The objective of the present study was to determine the occurrence of sporotrichosis in domestic cats and in wild or exotic felines in captivity through the isolation of Sporothrix spp. from claw impressions in a culture medium. The samples included 132 felines, of which 120 (91.0 %) were domestic cats, 11 (8.3 %) were wild felines, and one (0.7 %) was an exotic felid. Twenty-one (17.5 %) were outdoor cats. Of the total, 89 (67.4 %) had contact with other animals of the same species. It was possible to isolate Sporothrix schenckii from the claws of one (0.7 %) of the felids probed; this animal exhibited generalised sporotrichosis and had infected a female veterinarian. The potential pathogenic agents Microsporum canis and Malassezia pachydermatis were isolated in 12.1 and 5.3 % of the animals, respectively. The following anemophilous fungi, which were considered to be contaminants, were also isolated: Penicillium sp. (28 or 21.2 %), Aspergillus sp. (13 or 9.8 %), Rhodotorula sp. (5 or 3.8 %), Candida sp. (5 or 3.8 %), Trichoderma sp. (1 or 0.7 %), and Acremonium sp. (1 or 0.7 %). Due to the low magnitude of occurrence (0.7 %) of Sporothrix in feline claws, the potential of the cats evaluated in this study to be sources of infection in the city of São Paulo is considerably low.     

Antinociceptive interaction of (±)-CPP and propentofylline in monoarthritic rats

Introduction

Multiple studies have shown that glial cells of the spinal cord, such as astrocytes and microglia, have close contact with neurons, suggesting the term tripartite synapse. In these synapses, astrocytes surrounding neurons contribute to neuronal excitability and synaptic transmission, thereby increasing nociception and thus the persistence of chronic pain. Conversely, the N-methyl-D-aspartate (NMDA) receptor is crucial in the generation and maintenance of chronic pain. It has multiple sites of modulation. One is the site of recognition of extracellular neurotransmitter (glutamate), which can be blocked by competitive antagonists such as (3-(2-carboxipiperazin-4)1-propyl phosphonic acid), (±)-CPP, resulting in a blockade of the calcium current and thus the intracellular transduction process. In the present study, we investigated whether the potential antinociceptive effect of glial inhibition produced by propentofylline (PPF) can be enhanced when combined with an NMDA-receptor inhibitor such as (±)-CPP.

Methods

We used Sprague-Dawley monoarthritic rats. The monoarthritis was induced by injection of complete Freund adjuvant in the right tibiotarsal joint. Four weeks later, rats were treated with PPF (1, 10, 30, and 100 μg/10 μl) intrathecally (i.t.) for 10 days, injected once with (±)-CPP (2.5, 5, 12.5, 25, 50, and 100 μg/10 μl, i.t.), or both treatments combined. The antinociceptive effect was evaluated on day 11 for PPF and immediately to (±)-CPP, by assessing the vocalization threshold to mechanical stimulation of the arthritic paw.

Results

The data indicate that intrathecal administration of increasing concentrations of (±)-CPP or PPF produced a significant dose-dependent antinociceptive effect with respect to monoarthritic rats receiving saline. The linear regression analysis showed that the dose that produces 30% of maximal effect (ED₃₀) for i.t. (±)-CPP was 3.97 μg, and 1.42 μg for i.t. PPF. The administration of the PPF and (±)-CPP combination in fixed proportions of ED₃₀ produced a dose-dependent antinociceptive effect, showing an interaction of the supraadditive type.

Conclusions

The results suggest that glia inhibitors can synergically potentiate the effect of glutamate blockers for the treatment of chronic inflammatory pain.     

Characterization of Nuclear Localization Signals (NLSs) and Function of NLSs and Phosphorylation of Serine Residues in Subcellular and Subnuclear Localization of Transformer-2β (Tra2β)

The serine/arginine-rich (SR) proteins are one type of major actors in regulation of pre-mRNA splicing. Their functions are closely related to the intracellular spatial organization. The RS domain and phosphorylation status of SR proteins are two critical factors in determining the subcellular distribution. Mammalian Transformer-2β (Tra2β) protein, a member of SR proteins, is known to play multiple important roles in development and diseases. In the present study, we characterized the subcellular and subnuclear localization of Tra2β protein and its related mechanisms. The results demonstrated that in the brain the nuclear and cytoplasmic localization of Tra2β were correlated with its phosphorylation status. Using deletional mutation analysis, we showed that the nuclear localization of Tra2β was determined by multiple nuclear localization signals (NLSs) in the RS domains. The point-mutation analysis disclosed that phosphorylation of serine residues in the NLSs inhibited the function of NLS in directing Tra2β to the nucleus. In addition, we identified at least two nuclear speckle localization signals within the RS1 domain, but not in the RS2 domain. The nuclear speckle localization signals determined the localization of RS1 domain-contained proteins to the nuclear speckle. The function of the signals did not depend on the presence of serine residues. The results provide new insight into the mechanisms by which the subcellular and subnuclear localization of Tra2β proteins are regulated.     

Vitellogenesis and changes in lipid and protein content of oocytes of Trophon geversianus (Neogastropoda: Muricidae) in Golfo San José (Chubut,Argentina)

The reproductive cycle of Trophon geversianus (Pallas 1774) population from Golfo San José (Chubut, Argentina, 42°33′S, 64°33′W) was studied using histochemical methods and digital image analysis. For such purpose, ovary samples were taken monthly between July 2006 and August 2007 and stained with hematoxylin and eosin, Sudan black B (to identify lipids) or Schiff’s ninhydrin (to identify proteins). Four different gametogenic stages were described: oogonias, previtellogenic oocytes, early vitellogenic oocytes, and late vitellogenic oocytes. Two spawning events were registered; one between September and October 2006, and a second one between February and March 2007. Oocyte quality was determined by the changes in lipid and protein composition during gametogenesis. Through digital image analysis, a lipid and a protein indexes (LI and PI) were calculated. Significant differences were observed in LI over month sampled, contrarily to what PI showed. A significant and positive correlation was found between lipid and protein content and oocytes areas, indicating that these nutrients accumulate during the entire vitellogenesis. Also, when dividing the oocytes into two size groups, analysis indicates a positive correlation between LI and oocyte area for smaller area oocytes. This demonstrates that while proteins accumulate linearly throughout vitellogenesis process, lipids accumulate in two steps: first at a growing rate, secondly at constant rate. This information is essential to determine the nutritional requirements of brood stock individuals at hatcheries in this potential fishery resource that inhabits patagonian waters.     

Speciation of dimethyltin(IV) – and trimethyltin(IV) – carbocysteinate and – glutamate systems in aqueous media

Abstract

The formation of complex species in the dimethyltin(IV) and trimethyltin(IV)-carboxymethyl-L-cysteinate (carbocysteinate) systems in NaCl_aq, at different ionic strengths, and in a multicomponent Na⁺, K⁺, Ca²⁺ ,Mg²⁺, Cl^? and SO₄^2-? medium representative of the seawater major composition, is discussed. Experimental results give evidence for the formation of the following species (L = carbocysteinate): [(CH₃)₂Sn(L)]⁰, [(CH₃)₂Sn(HL)]⁺, [(CH₃)₂Sn(OH)(L)]^?, [(CH₃)₂Sn(OH)₂(L)]^2? in the DMT–CCYS system, and [(CH₃)₃Sn(HL)]⁰, [(CH₃)₃Sn(L)]^? and [(CH₃)₃Sn(OH)(L)]^2? in the TMT-CCYS system. The ionic strength dependence of formation constants was taken into account by an extended Debye Hückel type equation and by the SIT (Specific ion Interaction Theory). Measurements were carried out also on the dimethyltin(IV)-glutamate and trimethyltin(IV)-glutamate systems in NaCl_aq, owing the strict similarity of glutamate and carbocysteinate. Results obtained show the formation of complex species having the same stoichiometry as those formed in the DMT- and TMT-carbocysteinate systems, with very similar stability, confirming that carbocysteinate behaves as a dicarboxylic amino acid without involving the sulfur-bridge potential binding site in metal coordination.     

Numerical increases and distributional shifts of Chrysaora quinquecirrha (Desor) and Aurelia aurita (Linné) (Cnidaria: Scyphozoa) in the northern Gulf of Mexico

Fisheries resource trawl survey data from the National Marine Fisheries Service from a 11–13-year period to 1997 were examined to quantify numerical and distributional changes of two species of northern Gulf of Mexico scyphomedusae: the Atlantic sea nettle, Chrysaora quinquecirrha (Desor), and the moon jelly, Aurelia aurita (Linné). Trawl surveys were grouped into 10 statistical regions from Mobile Bay, Alabama to the southern extent of Texas, and extended seaward to the shelf break. Records of summertime C. quinquecirrha medusa populations show both an overall numerical increase and a distributional expansion away from shore in the down-stream productivity field of two major river system outflows: Mobile Bay and the Mississippi-Atchafalaya Rivers. In addition, there is a significant overlap between summer C. quinquecirrha and lower water column hypoxia on the Louisiana shelf. In trawl surveys from the fall, A. aurita medusae showed significant trends of numerical increase in over half of the regions analyzed. For both species, there were statistical regions of no significant change, but there were no regions that showed significant decrease in number or distribution. The relationships between natural and human-induced (e.g. coastal eutrophication, fishing activity and hard substrate supplementation) ecosystem modifications are very complex in the Gulf of Mexico, and the potential impact of increased jellyfish populations in one of North America's most valuable fishing grounds is a most critical issue. Several hypotheses are developed and discussed to guide future research efforts in the Gulf of Mexico.     

Aerobic degradation of lindane (γ-hexachlorocyclohexane) in bacteria and its biochemical and molecular basis

γ-Hexachlorocyclohexane (γ-HCH, also called γ-BHC and lindane) is a halogenated organic insecticide that causes serious environmental problems. The aerobic degradation pathway of γ-HCH was extensively revealed in bacterial strain Sphingobium japonicum (formerly Sphingomonas paucimobilis) UT26. γ-HCH is transformed to 2,5-dichlorohydroquinone through sequential reactions catalyzed by LinA, LinB, and LinC, and then 2,5-dichlorohydroquinone is further metabolized by LinD, LinE, LinF, LinGH, and LinJ to succinyl-CoA and acetyl-CoA, which are metabolized in the citrate/tricarboxylic acid cycle. In addition to these catalytic enzymes, a putative ABC-type transporter system encoded by linKLMN is also essential for the γ-HCH utilization in UT26. Preliminary examination of the complete genome sequence of UT26 clearly demonstrated that lin genes for the γ-HCH utilization are dispersed on three large circular replicons with sizes of 3.5 Mb, 682 kb, and 191 kb. Nearly identical lin genes were also found in other HCH-degrading bacterial strains, and it has been suggested that the distribution of lin genes is mainly mediated by insertion sequence IS6100 and plasmids. Recently, it was revealed that two dehalogenases, LinA and LinB, have variants with small number of amino acid differences, and they showed dramatic functional differences for the degradation of HCH isomers, indicating these enzymes are still evolving at high speed.     

DFT study of (17)O, (1)H and (13)C NMR chemical shifts in two forms of native cellulose, I(α) and I(β)

This computational study is intended to shed light on the crystalline and molecular structure, together with the hydrogen bonding (H-bonding) differences between two forms of native cellulose. DFT calculations were carried out to characterize the ¹⁷O, ¹H and ¹³C nuclear magnetic resonance (NMR) parameters in cellulose I_α and I_β with the B3LYP functional employing the 6–311++G7 and 6–31+G1 basis sets. Geometry optimization revealed that the average HB length is shortened by 0.01–0.08 Å when the chains are aligned, whereas the average bond angle increases by about 4–8° exhibiting the enhancement of HB strength. For the isolated cellotetramer chains, the isotropic ¹⁷O–H chemical shifts were plotted as a function of HB length. Our results indicated that as the HB length in cellotetramer I_α increases, the ¹⁷O–H chemical shift isotropy increases, but this parameter changes in the opposite direction for the other structure. Moreover, B3LYP/6–311++G7 calculations reveal that there is an acceptable correlation between the calculated ¹³C chemical shifts of the two structures and their experimental values.     

Synthesis of (±)-Homosarkomycin and (±)-Rosaprostol

(±)-Homosarkomycin (2) and (±)-rosaprostol (3) were synthesized from (±)-methyl 2-oxo-bicyclo[3.1.0]hexane-1-carboxylate (1) by using the nucleophilic ring opening reaction on the double-activated cyclopropane ring as the key step.     

Synthesis of (±)-Lamprolobine and (±)-Epilamprolobine

(±)-Lamprolobine, the (+)-enatiomer of which was isolated from the leaves of Lamprolobium fruticosum, and (±)-epilamprolobine were synthesized from δ-valerolactam.     

Promoter hypermethylation and Up-regulation of thyroid-stimulating-hormone-alpha (TSH-α) in thyroid cancer

Thyroid-stimulating-hormone-alpha (TSH-α) is the common subunit of the heterodimeric hormone TSH and also of other glycoprotein hormones. Although both expression and promoter-methylation profiles of the gene have been observed in the pituitary gland and placenta, no observation has been reported in the thyroid gland. We examined TSH-α expression in normal and cancer thyroid tissues. Real-time RT-PCR and immunohistochemistry indicated that TSH-a was repressed in normal tissues while activated in cancer tissues. To identify the epigenetic mechanism of upregulation of TSH-α, the methylation status of the seven CpG sites in the TSH-a promoter was examined in sixty thyroid cancer tissues. Two CpG sites showed remarkably higher levels of methylation in cancer (46 and 45%) than in normal tissues (24 and 23%) (p=0.010 and 0.003). These findings indicate that TSH-α is expressed in the thyroid cancer tissue per se and that its expression can be affected by promoter methylation.     

Influence of age and gender on secretion of anti-Müllerian hormone in Asian (Elephas maximus) and African (Loxodonta africana) elephants

Anti-Müllerian hormone (AMH) secretion was studied in Asian and African elephants varying in age and reproductive status. Overall mean concentrations did not differ (P > 0.05) between species, but were markedly higher in male than female Asian elephants (31.01 ± 4.22 ng/mL and 0.19 ± 0.02 ng/mL, mean ± SEM) and African elephants (40.27 ± 3.18 ng/mL, 0.17 ± 0.04 ng/mL), respectively. Anti-Müllerian hormone secretion was not significantly affected by ovarian cyclicity status (cycling vs noncycling), but was higher (P < 0.05) in prepubertal (0.40 ± 0.10 ng/mL) than reproductive age (8-35 y; 0.18 ± 0.04 ng/mL) and aged (≥ 36 y; 0.16 ± 0.03 ng/mL) females. In males, AMH secretion was not significantly affected by musth status, but was age-related, with higher concentrations (P > 0.05) in prepubertal (49.08 ± 6.11 ng/mL) as compared to aged (≥36 y; 22.27 ± 5.82 ng/mL) bulls; concentrations in mature bulls (8-35 y; 37.01 ± 3.17 ng/mL) were similar to prepubertal and older bulls. We concluded that circulating AMH concentrations in elephants were similar between species and not affected by reproductive status; however, concentrations were significantly higher in males than females, and in younger animals. The diagnostic value of AMH to assess fertility status of individual elephants remains to be determined.     

Duration of marriages and remarriages in Chiomonte (Turin) in the XVII–XVIII century

This is a preliminary study of the duration of marriages and remarriages by widowed people in Chiomonte (Turin) from 1670 to 1729, a period that saw long periods of war alternating with brief intervals of peace. We analysed 748 marriage acts and evaluated the age at first marriage, the duration of marriage, the frequency of remarriage by widowed people and the number of widowed people who remarried. The data were analysed in five-year intervals. The results show that the population of Chiomonte was essentially permanent, since the end of the marriage could be established in 77,6% of cases. The mean duration of marriages increased from the end of the XVII century and around 51% of all marriages lasted more than 20 years. Historical events had different effects on the matrimonial behaviour. After the mortality crisis in 1690–91, marriages and remarriages increased in 1691–94, a typical response of populations decimated by an epidemic. However, the matrimonial behaviour in the decade 1704–14 was different, in that there was a strong increase in exogamous marriages (56.52% of all marriages). Remarriages, especially of widowed persons, also increased in that period. Although these data are preliminary and from a brief time period, they reveal particular aspects of the marriage and remarriage behaviour, thus helping to clarify the biodemographic evolution of the Chiomonte population.     

Role of 11β-OH-C(19) and C(21) steroids in the coupling of 11β-HSD1 and 17β-HSD3 in regulation of testosterone biosynthesis in rat Leydig cells

Here we describe further experiments to support our hypothesis that bidirectional 11β-HSD1-dehydrogenase in Leydig cells is a NADP(H) regenerating system. In the absence of androstenedione (AD), substrate for 17β-HSD3, incubation of Leydig cells with corticosterone (B) or several C₁₉- and C₂₁-11β-OH-steroids, in the presence of [³H]-11-dehydro-corticosterone (A), stimulated 11β-HSD1-reductase activity. However, in presence of 30 μM AD, testosterone (Teso) synthesis is stimulated from 4 to 197 picomole/25,000 cells/30 min and concomitantly inhibited 11β-HSD1-reductase activity, due to competition for the common cofactor NADPH needed for both reactions. Testo production was further significantly increased (p < 0.05) to 224-267 picomole/25,000 cells/30 min when 10 μM 11β-OH-steroids (in addition to 30 μM AD) were also included. Similar results were obtained in experiments conducted with lower concentrations of AD (5 μM), and B or A (500 nM).Incubations of 0.3-6.0 μM of corticosterone (plus or minus 30 μM AD) were then performed to test the effectiveness of 17β-HSD3 as a possible NADP⁺ regenerating system. In the absence of AD, increasing amounts (3-44 pmol/25,000 cells/30 min) of 11-dehydro-corticosterone were produced with increasing concentrations of corticosterone in the medium. When 30 μM AD was included, the rate of 11-dehydro-corticosterone formation dramatically increased 1.3-5-fold producing 4-210 pmol/25,000 cells/30 min of 11-dehydro-corticosterone. We conclude that 11β-HSD1 is enzymatically coupled to 17β-HSD3, utilizing NADPH and NADP in intermeshed regeneration systems.     

Chromium and Manganese Levels in Biological Samples of Normal and Night Blindness Children of Age Groups (3–7) and (8–12) Years

This study was designed to compare the levels of chromium (Cr) and manganese (Mn) in scalp hair, blood, and urine of night blindness in children age ranged (3–7) and (8–12) years of both genders, comparing them to sex- and age-matched controls. A microwave-assisted wet acid digestion procedure, was developed as a sample pretreatment, for the determination of Cr and Mn in biological samples of night blindness children. The proposed method was validated by using conventional wet digestion and certified reference samples of hair, blood and urine. The digests of all biological samples were analyzed for Cr and Mn by electrothermal atomic absorption spectrometry. The results indicated significantly higher levels of Cr, whilst low level of Mn in the biological samples (blood and scalp hair) of male and female night blindness children, compared with control subjects of both genders. These data present guidance to clinicians and other professional investigating deficiency of Mn and excessive level of Cr in biological samples (scalp hair and blood) of night blindness children.     

Integration of flowering dates in phenology and pollen counts in aerobiology: analysis of their spatial and temporal coherence in Germany (1992–1999)

We studied the possibility of integrating flowering dates in phenology and pollen counts in aerobiology in Germany. Data were analyzed for three pollen types (Betula, Poaceae, Artemisia) at 51 stations with pollen traps, and corresponding phenological flowering dates for 400 adjacent stations (< 25 km) for the years 1992–1993 and 1997–1999. The spatial and temporal coherence of these data sets was investigated by comparing start and peak of the pollen season with local minima and means of plant flowering. Our study revealed that start of birch pollen season occurred on average 5.7 days earlier than local birch flowering. For mugwort and grass, the pollen season started on average after local flowering was observed; mugwort pollen was found 4.8 days later and grass pollen season started almost on the same day (0.6 days later) as local flowering. Whereas the peak of the birch pollen season coincided with the mean flowering dates (0.4 days later), the pollen peaks of the other two species took place much later. On average, the peak of mugwort pollen occurred 15.4 days later than mean local flowering, the peak of grass pollen catches followed 22.6 days after local flowering. The study revealed a great temporal divergence between pollen and flowering dates with an irregular spatial pattern across Germany. Not all pollen catches could be explained by local vegetation flowering. Possible reasons include long-distance transport, pollen contributions of other than phenologically observed species and methodological constraints. The results suggest that further research is needed before using flowering dates in phenology to extrapolate pollen counts.     

Effect of bis(acetato)tetrakis(imidazole) copper(II) in delaying the onset and reducing the mortality rate of strychnine- and thiosemicarbazide—Induced convulsions

The anticonvulsant activity of bis(acetato)tetrakis(imidazole) copper(II), Cu(OAc)₂(Im)₄, was studied in normal mice using chemical convulsions induced by strychnine, thiosemicarbazide, picrotoxin, and pentelenetetrazol. Intraperitoneal administration of Cu(OAc) ₂(Im)₄, 50 mg/kg body mass, has delayed the onset of strychnine (3 mg/kg)-induced convulsion by 204% (p≤0.005) and thiosemicarbazide (20 mg/kg)-induced convulsant by 61% (p≤0.005). The changes in the onset of picrotoxin-(6 mg/kg) and pentelenetetrazol (50 mg/kg)-induced convulsions were not significant. The same dosage of the copper compound was effective in delaying the lethal time and reducing the mortality rate of treated animals. The anticonvulsant activity of Cu(OAc) ₂(Im)₄ complex against strychnine was not related to its constituents because the inorganic form of copper such as copper chloride, copper acetate, and the parent imidazole has no anticonvulsant activity. Other copper(II) complexes like copper(II)aspirinate and bis(acetato)bis(2-methyl imidazole) copper(II) were less effective.