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Effects of 5HT-1A agonists on locomotor and investigatory behaviors in rats differ from those of hallucinogens.

Behavioral profiles composed of both locomotor activity and investigatory behavior were established for the 5HT-1A agonists 8OHDPAT, buspirone, gepirone, and ipsapirone using rats tested in a Behavioral Pattern Monitor. Typically these compounds dose-relatedly decreased horizontal locomotion and investigatory activity during the first half of the 1-h test session. Time-course studies revealed that the time interval between injection and placement of the animal in the testing chamber made no difference in the temporal distribution of locomotor activity following most 5HT-1A agonists. These results were compared and contrasted to the behavioral profiles previously established for hallucinogenic compounds such as LSD and DOM, the psychoactive properties of which have been suggested to be mediated by 5HT-2 binding sites. Examination of ipsapirone and 8OHDPAT in a familiar environment paradigm revealed that both drugs decreased behavioral responding independently of the animals' familiarity with the test environment, in contrast to the behaviorally suppressive effects of hallucinogenic 5HT-2 antagonists which disappear in a familiar environment. Additionally, d,l-propranolol was used as a 5HT-1 antagonist and was found to block the behavioral effects of the 5HT-1A agonists ipsapirone and buspirone without having significant effects by itself. Propranolol was also used to identify the contribution of the 5HT-1 binding site to the behavioral effects of LSD. Even at relatively high doses, propranolol only partially antagonized the effects of LSD, supporting the hypothesis that the behavioral effects of LSD reflect the activation of both 5HT-1 and 5HT-2 receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Evolutionary relationships between laboratory mice and subspecies of Mus musculus based on the restriction fragment length variants of the chymotrypsin gene at the Prt-2 locus.

Restriction endonuclease fragment length variants in mice were compared by Southern blot analysis using the cDNA probe pcXP33 for the chymotrypsin gene. The variants were detected in the restriction patterns generated by fragments from digestions with BglII, EcoRI, HindIII, Pstl, SacI, and XbaI. The set of protein phenotypes and the restriction patterns of chymotrypsin gene were examined in many laboratory strains and wild subspecies. Most laboratory strains (26 strains) are grouped into a set defined as Set 1, but only a few laboratory strains (AU/SsJ and five BALB/c sublines) are classified as belonging to Set 2. Of wild subspecies, only BRV-MPL (M. brevirostris) can be placed in Set 1, while DOM-BLG and SK/Cam (M. domesticus) belong in Set 2. The assignment of an appropriate set defined by the characteristics of the chymotrypsin gene has also been investigated in M. musculus, two Chinese subspecies, M. yamashinai, M. molossinus, and M. castaneus, and the evolutionary relationship between laboratory mice and various subspecies of Mus has been examined.

Animals↗

Effects of Linalool on glutamatergic system in the rat cerebral cortex.

Linalool is a monoterpene compound reported to be a major component of essential oils in various aromatic species. Several Linalool-producing species are used in traditional medical systems, including Aeolanthus suaveolens G. Dom (Labiatae) used as anticonvulsant in the Brazilian Amazon. Psychopharmacological in vivo evaluation of Linalool showed that this compound have dose-dependent marked sedative effects at the Central Nervous System, including hypnotic, anticonvulsant and hypothermic properties. The present study reports an inhibitory effect of Linalool on Glutamate binding in rat cortex. It is suggested that this neurochemical effect might be underlining Linalool psychopharmacological effects. These findings provide a rational basis for many of the traditional medical use of Linalool producing plant species.

Acyclic Monoterpenes↗

Suppression of domoic acid induced seizures by 8-(OH)-DPAT.

Microinjections of the neuroexcitotoxin, domoic acid (DOM), in the ipsilateral rat hippocampal CA-3 region, induced generalized electrical seizure discharge activity, characterized by spikes and waves, followed by intermittent burst discharges. Computerized EEG analysis exhibited relative dominance of delta and theta and reductions in alpha and beta activities during domoic acid epileptogenesis. Seizure discharge activity was attenuated by the microinjection of the 5-HT1A agonist, 8-hydroxy-2-(di-N-propylamino)tetralin(8-(OH)-DPAT) and augmented by the specific 5-HT1A antagonist, spiroxatrine in the contralateral hippocampal CA-3 region. Neuronal recovery following 8-(OH)-DPAT was associated with significant reductions in the relative dominance of delta and theta and increases in the alpha and beta activities. The results suggest that activation of serotonergic 5-HT1A receptor in the hippocampus has a neuroprotective action.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Breast cancer screening, with particular reference to the concept of 'high risk' groups.

An overview is presented of the salient aspects of the DOM-project of breast cancer screening carried out in the city of Utrecht, the Netherlands. Results on stage at diagnosis, case-fatality, and mortality rates suggest that the program has been effective. Attention is devoted to the issue whether knowledge about risk factors for breast cancer is helpful in making a screening program more effective or efficient. It is concluded that such knowledge is of limited importance.

Breast Neoplasms↗

Single primer amplification reaction methods reveal exotic and indigenous mulberry varieties are similarly diverse.

Mulberry is the sole food source for mulberry silkworm and a number of indigenous and exotic varieties are used in sericulture. Studies on assessment of genetic diversity have been done amongst a few mulberry varieties using one or at the most two methods. However, no comprehensive study on a large number of varieties has been carried out. In present study, single primer amplification reaction (SPAR) methods have been used for determination of diversity in 27 mulberry varieties (exotic as well as indigenous), using four minisatellite core sequence primers for directed amplification of minisatellite DNA (DAMD), three simple sequence repeat (SSR) motifs as primers for inter simple sequence repeat (ISSR) and 20 arbitrary sequence decamer primers for ran-dom amplified polymorphic DNA (RAPD) reactions. The Jaccard coefficients were determined for the DAMD, ISSR and RAPD band data (total of 58, 39 and 235 bands respectively). All three methods revealed wide range of distances supporting a wide range of mulberry genetic diversity. A cumulative analysis of the data generated by three methods resulted in a neighbour-joining (NJ) tree that gave a better reflection of the relatedness and affinities of the varieties to each other. Comparison of the three methods by marker indices and the Mantel test of correlation indicated that though all methods were useful for the assessment of diversity in mulberry, the DAMD method was better. When considered as two groups (10 exotic and 17 indigenous varieties), the mulberry varieties in the exotic group were found to have slightly greater diversity than the indigenous ones. These results support the concept of naturalization of mulberry varieties at locales distant from their origins.

Cluster Analysis↗

Low doses of domoic acid during postnatal development produce permanent changes in rat behaviour and hippocampal morphology.

It is well established that the developing brain is a highly dynamic environment that is susceptible to toxicity produced by a number of pharmacological, chemical and environmental insults. We report herein on permanent behavioural and morphological changes produced by exposing newborn rats to very low (subconvulsive) doses of kainate receptor agonists during a critical window of brain development. Daily treatment of SD rat pups with either 5 or 20 microg/kg of domoic acid (DOM) from postnatal day 8-14 resulted in a permanent and reproducible seizure-like syndrome when animals were exposed to different tests of spatial cognition as adults. Similar results were obtained when animals were treated with equi-efficacious doses of kainic acid (KA; 25 or 100 microg/kg). Treated rats had significant increases in hippocampal mossy fiber staining and reductions in hippocampal cell counts consistent with effects seen in adult rats following acute injections of high doses of kainic acid. In situ hybridization also revealed an elevation in hippocampal brain derived neurotrophic factor (BDNF) mRNA in region CA1 without a corresponding increase in neuropeptide Y (NPY) mRNA. These results provide evidence of long-lasting behavioural and histochemical consequences arising from relatively subtle changes in glutamatergic activity during development, that may be relevant to understanding the aetiology of seizure disorders and other forms of neurological disease.

Animals↗

Speciation of heavy metals in environmental water by ion chromatography coupled to ICP-MS.

Biogenic (e.g. phytochelatins, porphyrins, DOM) as well as anthropogenic (e.g. NTA, EDTA, phosphonates) chelators affect the mobility and cycling of heavy metals in environmental waters. Since such chelators can form strongly bound anionic heavy metal complexes that are stable and highly mobile, anion-exchange chromatography coupled to ICP-MS was investigated. A narrow bore HPLC system was connected to a micro concentric nebuliser for in-line sample introduction. A new chromatographic procedure based on a synthetic hydrophilic quaternary ammonium anion exchanger in combination with nitrate as a strong eluent anion, and gradient elution, provided high separation selectivity and a large analytical window. Low detection limits (nmol L(-1)) were achieved by on-column matrix removal and sample preconcentration. This allowed the method to be successfully applied to different environmental research areas. In ecotoxicological studies of heavy metal effects on algae low concentrations of metal EDTA complexes were determined in nutrient solutions without interference from high (buffer) salt concentrations. In groundwater, infiltrated by a polluted river, mobile metal EDTA species were observed. In river water of different pollution levels beside CuEDTA other anionic Cu-complexes were found in nmol L(-1) concentrations.

Animals↗

Optimization and application of SPME for the gas chromatographic determination of endosulfan and its major metabolites in the ng L(-1) range in aqueous solutions.

In the present study an analytical method was optimized for the determination of alpha-endosulfan, beta-endosulfan, endosulfan sulfate, endosulfan ether and endosulfan lactone in small volumes of environmental aqueous samples using solid-phase microextraction (SPME) and gas chromatography-electron capture detection (GC-ECD). A 100 micro m polydimethylsiloxane (PDMS) phase was used for the extraction. The limit of detection (LOD) for the analytes varied between 0.01 and 0.03 micro g L(-1) with a relative standard deviation of 3 to 11%. The influence of the ionic strength on the extraction efficiency was investigated for the individual compounds. alpha-Endosulfan, beta-endosulfan, endosulfan sulfate and endosulfan ether were extracted successfully without salt addition. The extraction efficiency of endosulfan lactone was improved with 30% NaCl content. A general decrease in extraction efficiency for alpha-endosulfan, beta-endosulfan, endosulfan sulfate and endosulfan ether with high NaCl content (20-30%) in the solution was observed due to glass surface adsorption. No effect of dissolved organic material (DOM) on the extraction efficiency was observed. The extraction coefficients changed between Log K=2.17 and 3.33. A sample from the Antarctic region was analyzed using the optimized GC-ECD/SPME method. To confirm the results obtained for the real sample a GC with a mass spectrometer (MS) was used. Endosulfan sulfate, the most toxic metabolite of endosulfan, was found in the sample at a concentration of 0.3 micro g L(-1).

Antarctic Regions↗

Transformations of metal species in ageing humic hydrocolloids studied by competitive ligand and metal exchange.

Transformations of metal species (particularly Al, Ca, Fe, Mg, Mn, Zn) in ageing humic hydrocolloids were studied, applying a competitive ligand and metal exchange approach. For this purpose, metal-containing hydrocolloids, freshly collected from humic-rich German bog lake waters (Hohlohsee (HO), Black Forest; Venner Moor (VM), Muensterland; Arnsberger Wald (AW), Northrhine-Westfalia) and conventionally pre-filtered through 0.45 microm membranes, were subjected on-site to an exchange with EDTA and Cu(II) ions, respectively, as a function of time. EDTA complexes gradually formed, metal fractions exchanged by Cu(II) (as well as free Cu(II) concentrations) were operationally discriminated by means of a small time-controlled tangential-flow ultrafiltration unit (nominal cutoff: 1 kDa). Metal and DOM (dissolved organic matter) fractions obtained this way were determined off-site using instrumental methods (AAS, ICP-OES, carbon analyzer). After weeks of storage, the collected hydrocolloids were studied again by this approach. The EDTA availability of colloid-bound metals (particularly Al and Fe) exhibited different ageing trends, dependent on the sample (VM: decrease of Fe availability (98-76%), HO: increase of Fe availability (76-82%)). In contrast, the Cu(II) exchange equilibria of colloid-bound metals revealed merely low availability of Al (16-38%) and Fe (5-11%) towards Cu(II) ions, also dependent on ageing effects. In particular, the conditional copper exchange constants Kex obtained from the exchange between Cu(II) ions and available metal species (such as Ca, Mg, Mn, Zn) exhibited a strong decrease (by a factor of 2-100) during sample storage, indicating considerable non-equilibria complexation of these metal ions in the original bogwaters studied on-site.

Colloids↗

Effects of dissolved organic material on binding and toxicokinetics of pyrene in the waterflea Daphnia magna.

The binding and bioavailability of pyrene was studied in the laboratory in two humic fresh waters and in a reference water without dissolved organic material (DOM), measured as dissolved organic carbon (DOC). The uptake of pyrene by Daphnia magna in short-term (24 h) accumulation experiments was fitted to a first-order rate-kinetic equation to calculate simultaneous uptake and elimination rates. The partition coefficients of pyrene to DOC (KDOC) were 37.1 x 103 in Pielisjoki River (9.4 mg DOC L(-1)), and 34.9 x 103 in Lake Kontiolampi (17.4 mg DOC L(-1)) waters, indicating similar binding affinities of pyrene for both humic waters. The uptake clearance of pyrene (ku) in the DOC-rich Lake Kontiolampi water was 290.4 ml g wet weight(-1) h(-1), which was lower than those in Pielisjoki River and DOC-free waters (395.8 and 346.0 ml g wet weight(-1) h(-1), respectively). Pyrene elimination (ke) was in average 0.2 h(-1) with no significant differences between the waters. The observed bioconcentration factors of pyrene in Pielisjoki River and Lake Kontiolampi waters were 84% and 46% of that in the DOC-free reference water, respectively, thus indicating that a high concentration of DOC is needed to decrease the bioavailability of waterborne pyrene to D. magna.

Animals↗

Exploring bacteriplankton growth and protein synthesis to determine conversion factors across a gradient of dissolved organic matter.

The effect of bacterial specific growth rates of abundance (micro) and protein synthesis (b) on conversion factor (CF) variability was explored in order to provide an alternative approach to the controversial application of just one universal CF to field data. Nine regrowth cultures (RCs) were set up from very diverse aquatic ecosystems, controlling temperature and adding N and P to avoid mineral limitation and force organic carbon limitation. The values of micro varied one order of magnitude from 0.26 to 3.34 d(-1), whereas b values varied two orders of magnitude from 0.28 to 34.87 d(-1). We found no relationships between micro or b values and the dissolved organic carbon (DOC) concentration or the dissolved organic matter (DOM) quality indexes assayed. Abundance and protein synthesis increased exponentially and synchronously in four RCs, leading to balanced growth (micro = b). In contrast, abundance and protein synthesis increased logistically in the other five RCs and b values were significantly higher than g values, leading to unbalanced growth (micro not equal b). CFs ranged from 0.0062 to 0.0576 x 10(18) cells mol leucine(-1) with an average of 0.0305 x 10(18) cells mol leucine(-1). CFs obtained in RCs with balanced growth were generally higher than CFs obtained in RCs with unbalanced growth and were not alike, impeding the establishment of an upper limit for CFs. A positive and significant relationship (n = 8, p < 0.0 1, r2 = 0.71) was found between CFs and DOC concentration (CF (x10(18) cells mol leucine(-1)) = 0.0104 + 0.0094 DOC (mM)) when the value for the most productive system was excluded. This function permits the estimation of site-specific CFs based on DOC concentration instead of the controversial use of a single CF for different systems.

Bacteria↗

Effects of small-scale turbulence on bacteria: a matter of size.

We examined the influence of small-scale turbulence and its associated shear on bacterioplankton abundance and cell size. We incubated natural microbial assemblages and bacteria-only fractions and subjected them to treatments with turbulence and additions of mineral nutrients and/or organic carbon. Bacterial abundance was not affected directly by turbulence in bacteria-only incubations. In natural microbial assemblage incubations, bacterial concentrations were higher under turbulence than in still-water controls when nutrients were added. In general, in the turbulence treatments bacteria increased significantly in size, mainly due to elongation of cells. The addition of inorganic nutrients had a negative effect on bacterial size, but a significantly positive effect on abundance independently of other factors such as turbulence and the presence of predators. Flagellate grazing did not trigger an increase in bacterial size as a grazing resistance response in unmixed containers. With the addition of organic carbon, bacteria elongated and partly settled to the bottom of the containers, in both the turbulent and still treatment, but bacterial abundance did not further increase. Furthermore, bacteria aggregated in the turbulence treatments after the second day of incubation even in the absence of other components of the microbial community. We found that turbulence and the associated shear increase bacterial size and change bacterial morphology, at least under certain nutrient conditions. This might be due to a physiological response (enhanced growth rate and/or unbalanced growth) or due to the selection of opportunistic strains when organic carbon is in excess compared to mineral nutrients. We suggest that shear associated with turbulent flow enhances the DOM flux to bacteria directly as well as indirectly through enhanced grazing activity and photosynthetic release. The formation of bacterial aggregates and filaments under turbulence might give selective advantage to bacteria in terms of nutrient uptake and grazing resistance.

Bacteria↗

Effects of Resources and Trophic Interactions on Freshwater Bacterioplankton Diversity.

In a study of bacterioplankton in an oligotrophic lake in northern Wisconsin, a community fingerprinting technique, automated ribosomal intergenic spacer analysis (ARISA), was used to determine the effect of resources and trophic interactions on bacterioplankton diversity. Inorganic nitrogen and phosphorus (NP), carbon in the form of glucose (G) or dissolved organic matter extracted from peat (DOM), and carbon and NP in combination were added to two types of experimental systems. Ten-liter mesocosms contained all components of the original aquatic community except for large zooplankton. One-liter dilution cultures were prepared so that the effects of grazers and phytoplankton were removed. During a 3-day incubation, bacterial production showed the greatest response to the carbon plus NP treatment in both experimental systems, but bacterial diversity was strikingly different between them. In the mesocosms, the number of ARISA-PCR fragments averaged 41 per profile, whereas the dilution culture communities were highly reduced in complexity, dominated in most cases by a single PCR fragment. Further analysis of the mesocosm data suggested that whereas the NPDOM addition caused the greatest aggregate bacterial growth response, the addition of NP alone caused the largest shifts in community composition. These results suggest that the measurement of aggregate responses, such as bacterial production, alone in studies of freshwater bacterial communities may mask the effects of resources on bacterioplankton.

Journal Article↗

Biogenic black crusts on buildings in unpolluted environments.

Samples of peeling black crusts from modern and historic buildings in Campeche, Mexico, from a gravestone on the island of Dom Khon, Lao, and from the Anglican cathedral in Belize City were analyzed microbiologically, by scanning electron microscopy plus electron dispersive spectroscopy (EDS) and for pigment composition. In all cases, the surface was covered by a thick mat of cyanobacteria with dark brown sheaths. These were filamentous organisms of the genera Scytonema or Fischerella/Mastigocladus, except for one sample, where coccoid cyanobacteria of Subsection II were predominant. Fungi were not present at all sites and, where seen, were not the major biomass. High scytonemin:chlorophyll a ratios correlated with the dark pigmentation of the cyanobacterial cells and indicated the stressful conditions under which these organisms were living (high temperatures and ultraviolet levels, frequent desiccation). The absence, or low levels, of sulfur in the biofilms confirmed that there was little urban pollution at the sites and the EDS analysis showed that the black coloration was caused solely by cell pigmentation; no dark-colored elements were present at high concentrations. These results demonstrate that, unlike chemically formed thick black crusts found in polluted atmospheres, thin black crusts (which could be called patinas) in clean environments may be predominantly composed of filamentous cyanobacteria.

Biofilms↗

Biarticular and monoarticular muscle activation and injury in human quadriceps muscle.

We hypothesized that activation of the quadriceps femoris muscle group during eccentric exercise is related to the increase in magnitude of several markers of muscle injury that developed during the next week. Fourteen male subjects performed six to eight sets of five to ten repetitions of single-leg eccentric-only seated knee extension exercise. Magnetic resonance (MR) images were collected before and immediately after exercise and on days 2-4 and 6 after eccentric exercise. Changes in maximal voluntary contraction (MVC), perceived soreness, muscle volume and muscle transverse relaxation of water protons (T2) were determined for the quadriceps femoris muscle group each day. Changes in muscle volume and T2 were determined every day for each muscle [vastus lateralis (VL), vastus medialis (VM), vastus intermedius (VI), rectus femoris (RF)] of the quadriceps femoris group. Post-exercise T2 was greater than pre-exercise T2 (P < 0.05) for all muscles. The acute deltaT2 (Post-Pre) was similar (P>0.05) among VL, VM, VI, and RF [5.5 (0.3) ms], suggesting that the four muscles were equally activated during eccentric exercise. In the week after eccentric exercise, subjects experienced delayed-onset muscle soreness (DOMS) and all muscles demonstrated a delayed increase in T2 above pre-exercise values (P < 0.05), suggesting that muscle injury had occurred. For the quadriceps femoris muscle group, there was no correlation between acute deltaT2 and delayed (peak T2 during days 2, 3, 4, 6 minus pre-exercise T2) deltaT2 (r=0.04, P>0.05). Similar results were obtained when VL, VM, VI and RF were examined separately. Of the four muscles in quadriceps femoris, the biarticular RF experienced greater muscle injury [delayed deltaT2= 15.2 (2.0) ms] compared to the three monoarticular vasti muscles [delayed deltaT2 = 7.7 (1.3) ms; P< 0.05]. We propose that the disproportionate muscle injury to RF resulted from an ineffective transfer of torque from the knee to hip joint during seated eccentric knee extension exercise, thus causing RF to dissipate greater energy than normal. We conclude that in humans, muscle activation is not a unique determinant of muscle injury.

Adult↗

The SOX10/Sox10 gene from human and mouse: sequence, expression, and transactivation by the encoded HMG domain transcription factor.

The SOX genes form a gene family related by homology to the high-mobility group (HMG) box region of the testis-determining gene SRY. We have cloned and sequenced the SOX10 and Sox10 genes from human and mouse, respectively. Both genes encode proteins of 466 amino acids with 98% sequence identity. Significant expression of the 2.9-kb human SOX10 mRNA is observed in fetal brain and in adult brain, heart, small intestine and colon. Strong expression of Sox10 occurs throughout the peripheral nervous system during mouse embryonic development. SOX10 shows an overall amino acid sequence identity of 59% to SOX9. Like SOX9, SOX10 has a potent transcription activation domain at its C-terminus and is therefore likely to function as a transcription factor. Whereas SOX9 maps to 17q, a SOX10 cosmid has previously been mapped by us to the region 22q13.1. Mutations in SOX10 have recently been identified as one cause of Waardenburg-Hirschsprung disease in humans, while a Sox10 mutation underlies the mouse mutant Dom, a murine Hirschsprung model.

Adult↗

Pain reduction by infrared light-emitting diode irradiation: a pilot study on experimentally induced delayed-onset muscle soreness in humans.

The present pilot study investigated the analgesic efficacy of light-emitting diode (LED). In view of a standardised and controlled pain reduction study design, this in vivo trial was conducted on experimentally induced delayed-onset muscle soreness (DOMS). Thirty-two eligible human volunteers were randomly assigned to either an experimental (n=16) or placebo group (n=16). Immediately following the induction of muscle soreness, perceived pain was measured by means of a visual analog scale (VAS), followed by a more objective mechanical pain threshold (MPT) measurement and finally an eccentric/concentric isokinetic peak torque (IPT) assessment. The experimental group was treated with infrared LED at one of both arms, the other arm served as control. Irradiation lasted 6 min at a continuous power output of 160 mW, resulting in an energy density of 3.2 J/cm(2). The subjects of the placebo group received sham irradiation at both sides. In post-treatment, a second daily assessment of MPT and VAS took place. The treatment and assessment procedure (MPT, VAS and IPT) was performed during 4 consecutive days. Statistical analysis (a general linear model followed by post hoc least significant difference) revealed no apparent significant analgesic effects of LED at the above-described light parameters and treatment procedure for none of the three outcome measures. However, as the means of all VAS and MPT variables disclose a general analgesic effect of LED irradiation in favour of the experimental group, precaution should be taken in view of any clinical decision on LED. Future research should therefore focus on the investigation of the mechanisms of LED action and on the exploration of the analgesic effects of LED in a larger randomised clinical trial and eventually in more clinical settings.

Adult↗