The h-Current in Periglomerular Dopaminergic Neurons of the Mouse Olfactory Bulb

The properties of the hyperpolarization-activated cation current (I_h) were investigated in rat periglomerular dopaminergic neurons using patch-clamp recordings in thin slices. A reliable identification of single dopaminergic neurons was made possible by use of a transgenic line of mice expressing eGFP under the tyrosine hydroxylase promoter. At 37 °C and minimizing the disturbance of the intracellular milieu with perforated patches, this current shows a midpoint of activation around −82.7 mV, with a significant level of opening already at rest, thereby giving a substantial contribution to the resting potential, and ultimately playing a relevant function in the control of the cell excitability. The blockage of I_h has a profound influence on the spontaneous firing of these neurons, which result as strongly depressed. However the effect is not due to a direct role of the current in the pacemaker process, but to the I_h influence on the resting membrane potential. I_h kinetics is sensitive to the intracellular levels of cAMP, whose increase promotes a shift of the activation curve towards more positive potentials. The direct application of DA and 5-HT neurotransmitters, physiologically released onto bulbar dopaminergic neurons and known to act on metabotropic receptors coupled to the cAMP pathway, do not modifythe I_h amplitude. On the contrary, noradrenaline almost halves the I_h amplitude. Our data indicate that the HCN channels do not participate directly to the pacemaker activity of periglomerular dopaminergic neurons, but influence their resting membrane potential by controlling the excitability profile of these cells, and possibly affecting the processing of sensory information taking place at the entry of the bulbar circuitry.     

Effect of the endosymbiotic pea crab Calyptraeotheres garthi on the metabolic rate and oxidative status of the slipper limpet Crepidula cachimilla

Parasites may induce metabolic changes and imbalances in the redox status of hosts. This study tested the effect of parasites on the O₂ consumption rate (O₂‐CR) of hosts, and explored the link between O₂‐CR and oxidative stress in parasitized hosts. We used the symbiotic pea crab Calyptraeotheres garthi and its slipper limpet host Crepidula cachimilla as models. The O₂‐CR of long‐term (3 months) infested limpets was 2.5 times greater than that of long‐term uninfested limpets. Also, the O₂‐CR of limpets stripped of crabs 24 h before measurements was intermediate between that of long‐term infested and uninfested limpets. These results indicate a parasitic relationship between C. garthi and Cr. cachimilla, and suggest that the effect of the parasite on the metabolic rate of limpets is reversible. Lastly, the activity of two antioxidant enzymes (CAT and GST) as well as lipid peroxidation did not vary between infested and uninfested limpets. Thus, increased O₂‐CR is not necessarily coupled with oxidative stress in pea crab‐parasitized slipper limpets.     

Oligosaccharide transfer from lipid sugar intermediates to endogenous protein(s) of rat liver microsomal subfractions

Summary The subcellular localization and characterization of some of the components involved in the glycosylation of asparagine type glycoproteins was attempted using dolichyl diphosphate [¹⁴c]mannose oligosaccharide as precursor of the glycosylation reaction in vitro. Isolated rough and smooth microsomel fractions were able to carry out the transfer of the carbohydrate moiety from lipid oligosaccharide to endogenous protein acceptors. The protein glycosylating activity remained practically the same after stripping the vesicles from their ribosomes or partially releasing their vesicular content. Isolation of polysomes from rough microsomes after glycosylation has taken place, reveals that a large proportion of mannose labeled glycoproteins is in the membranous fraction. The remaining labeled glycoproteins co-sediment with the polysomal fraction. If the isolation is carried out before glycosylation only the membranous fraction shows enzyme activity, whereas the polysomes alone are not able to carry out glycosylation. All these results taken together indicate that the protein glycosylating enzyme is a structural component of the rough and smooth microsomes of rat liver.     

Cadmium-induced responses in duckweed <Emphasis Type="Italic">Lemna minor</Emphasis> L.

Although duckweed Lemna minor L. is a known accumulator of cadmium, detailed studies on its physiological and/or defense responses to this metal are still lacking. In this study, the effects of 10 μM CdCl₂ on Lemna minor were monitored after 6 and 12 days of treatment, while growth was estimated every 2 days. Cadmium treatment resulted in progressive accumulation of the metal in the plants and led to a decrease in the growth rate to 54% of the control value. The metal also considerably impaired chloroplast ultrastructure and caused a significant reduction in pigment content, i.e., at day 12, by 30 and 34% for chlorophylls a and b, and by 25% for carotenoids. During cadmium treatment, the contents of malondialdehyde and endogenous H₂O₂ progressively increased (rising 77 and 46% above the controls by day 12), indicating that cadmium induced considerable oxidative stress. On the other hand, higher activities of pyrogallol peroxidase (PPX), ascorbate peroxidase (APX) and catalase (CAT), as well as the induction of a new APX isoform, in cadmium-treated plants, clearly showed activation of an antioxidative response. At day 6, only PPX activity was significantly above the controls (15%), while, at day 12, PPX, APX and CAT activities were increased (74, 78 and 63%). Cadmium also led to accumulation of the heat shock protein 70 (HSP70) and induced an additional isoform of this protein. The obtained results suggest that cadmium (10 μM) is phytotoxic to Lemna minor, inducing oxidative stress, and that antioxidative enzymes and HSP70 play important roles in the defense against cadmium toxicity. M. Tkalec and T. Prebeg contributed equally to this work     

Updated checklist of the mosquitoes (Diptera: Culicidae) of Croatia

Improvement of morphological and molecular identification methods allows the detection of new species of mosquitoes. The mosquito fauna of Croatia currently includes 52 species, belonging to eight genera, including Anopheles (12 species), Aedes (24 species), Coquillettidia (one species), Culex (seven species), Culiseta (six species), Orthopodomyia (one species), and Uranotaenia (one species). This is an updated checklist, which includes five new species found in Croatian mosquito fauna. Two of these are invasive mosquito species, Aedes albopictus (Skuse, 1895) and Aedes japonicus (Theobald 1901), which are spreading across Europe and Croatia. The other three species, Culex laticinctus (Edwards 1913), Culex torrentium (Martini 1925), and Anopheles daciae (Linton, Nicolescu & Harbach 2004) are autochthonous species which haven't been recorded so far. Since there are several more invasive species spreading across Europe, we assume that this is not the final list.     

CXCL12 is a costimulator for CD4+ T cell activation and proliferation in chronic lymphocytic leukemia patients

Activated T cells from patients with chronic lymphocytic leukemia (CLL) provide survival and proliferative signals to the leukemic clone within lymphoid tissues. Recruitment of both, CLL cells and T lymphocytes, to this supportive microenvironment greatly depends on CXCL12 production by stromal and myeloid cells. CXCL12 also supplies survival stimuli to leukemic B cells, but whether it exerts stimulatory effects on T lymphocytes from CLL patients is unknown. In order to evaluate the capacity of CXCL12 to increase CD4⁺ T cell activation and proliferation in CLL patients, peripheral blood mononuclear cells were cultured with or without recombinant human CXCL12 or autologous nurse-like cells, and then T cell activation was induced by anti-CD3 mAb. CXCL12 increases the proliferation and the expression of CD25, CD69, CD154, and IFNγ on CD3-stimulated CD4⁺ T cells from CLL patients, similarly in T cells from ZAP-70⁺ to ZAP-70^? patients. Autologous nurse-like cells establish a close contact with CD4⁺ T cells and increase their activation and proliferation partially through a CXCR4-dependent mechanism. In addition, we found that activated T cells in the presence of CXCL12 enhance the activation and proliferation of the leukemic clone. In conclusion, CXCL12 production by lymphoid tissue microenvironment in CLL patients might play a key dual role on T cell physiology, functioning not only as a chemoattractant but also as a costimulatory factor for activated T cells.     

Side distinct sciatic nerve recovery differences between rats and mice

The Sciatic Functional Index (SFI) is widely used to evaluate functional recovery after sciatic nerve injury, primarily in the rat, and more recently shown useful in the mouse. This quantitative, non-invasive method allows tracking of regeneration capability, visible in the gait of the animal. Using a Martin micro needle holder, carrying a force measured to be 49.2 N, the left sciatic nerve was crushed for 60 s. We accumulated data from walking tracks collected preoperatively and 1, 7, 14, 21, and 28 days after injury. SFI values were first calculated in the traditional manner. Then using the preoperative values as the normal value in the postoperative calculations, SFI was again calculated; this isolated the calculations to either injured or contra lateral leg giving a “split” plot. The traditional SFI calculations resulted in typical shaped graphs for both rats and mice. However, the “split” SFI calculations showed how rats and mice differ in their recovery from sciatic nerve injury. The mouse graph shows the intact leg remaining stable and the injured leg having functional impairment, which then recovers. The rat graph showed functional impairment of the injured leg, however, the intact leg had an increase in SFI values as if to compensate until the injured leg showed recovery.     

The influence of the different morphological changes on gastric mucosa on somatostatin cell number in antrum mucosa and serum somatostatin

The aim of our paper was to investigate the influence of the different morphological changes on gastric mucosa on somatostatin D-cell number in antral mucosa and serum Somatostatin. We analyzed according to Sydney classification to what extent the severity of gastritis affect the observed hormonal values. somatostatin D-cell number in antral mucosa and serum Somatostatin values were compared between three groups of patients; mild, moderate and severe chronic gastritis. The average number of somatostatin cell in biopsy sample of antrum mucosa was 30.41 +/- 35.38 (N = 17) in the case of middle form, 18.69 +/- 26.65 (N = 56) in moderate and in severe case of chronic gastritis 5.23 +/- 5.93 (N = 7) cells in mm2 of mucosa. The level of somatostatin in the serum of middle form gastritis were 26.43 +/- 28.76, moderate 19.95 +/- 35.93 and severe 17.88 +/- 17.66 pg/mL. In order to determine the number of somatostatin cells in antrum mucosa and serum somatostatin with present morphological changes of mucosa, it might helpful to exclude the patients with non-ulcer dyspepsia, but with the higher risk of premalignant and malignant changes.     

Writing and drawing with both hands as indicators of hemispheric dominance

Brain lateralization is a common term used to describe dominance of one brain hemisphere over another for a specific function. The right hand dominance in writing, controlled by the left hemisphere, is preceded by development of communicative gesticulation and followed by development of speech in the same hemisphere. We assumed that some people are not aware of their own capability of using the other hand for tasks involving fine motor sequential movements. To prove this hypothesis, the participants were asked to perform one trained task (writing) and one less-trained task (drawing) with a dominant and a non-dominant hand. The final sample was comprised of 1189 children from 14 elementary schools and 8 high schools in the Osijek area, of which 685 elementary school children were attending 1st to 4th grade and 504 high school children were attending 3rd and 4th grade. The participants were asked to write two words, draw a specific object (a vase with flowers) and fill out a questionnaire with 10 questions concerning the classification of handedness and cerebral hemisphere dominance. The self-reported cerebral lateralization assessed in the questionnaire was compared with the drawing and the writing performance. The self-reported and objectively measured hand dominance deviated in the cases of the ambidextrous who consider themselves right-handers. Given the fact that the number of ambidextrous persons was greater in elementary schools than in high schools, we concluded how training of the right hand decreases the ability of using both hands equally for either of the tested functions - writing and drawing.