bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunocytochemical Investigation of Nuclear Progestin Receptor Expression within Dopaminergic Neurones of the Female Rat Brain
Authors: Lonstein J. S. , Blaustein J. D.
Published: May 2004

Progesterone influences most processes involved in female reproduction, including ovulation, sexual behaviour, pregnancy, parturition, lactation and maternal behaviour. One neurotransmitter through which progesterone might regulate many of these functions is dopamine. To determine where in the brain progesterone might alter dopaminergic activity necessary for these and other processes in rats via cell nuclear progestin receptors, ovariectomized rats were injected subcutaneously with either 4 µg oestradiol benzoate to induce high levels of hypothalamic progestin receptor expression, or oil, and perfused 48 h later. Dual‐label immunocytochemistry was used to visualize cells having immunoreactivity (ir) for progestin receptors and tyrosine hydroxylase, a rate‐limiting enzyme for dopamine synthesis. Many cells containing both progestin receptor‐ir and tyrosine hydroxylase‐ir were found throughout the periventricular hypothalamus of oestradiol‐treated females. Conversely, very few cells in the hypothalamus of oil‐treated controls contained progestin receptor‐ir and, consequently, few dual‐labelled cells were found in this group. The greatest percentage of tyrosine hydroxylase immunoreactive cells expressing progestin receptors in oestradiol‐treated females was in, or near, the arcuate nucleus (A12 group), where up to 55% of tyrosine hydroxylase‐expressing cells coexpressed progestin receptors. Notably, dual‐labelled cells in oestradiol‐treated females were also found more rostrally than previously reported, with approximately 15–20% of tyrosine hydroxylase‐ir cells in the preoptic area/anterior hypothalamus (A14 group) also containing progestin receptor‐ir. No dual‐labelled cells were found for either group in the posterodorsal hypothalamus (A11 group), zona incerta (A13 group), retrorubral field (A8 group), ventral tegmental area (A10 group) or substantia nigra (A9 group) because little or no progestin receptor‐ir was found in these sites. These data provide new information about the neural substrate where progesterone might regulate dopamine release in the preoptic area/anterior hypothalamus. Using more sensitive techniques than those used previously, they also confirm the relationship between progestin receptor and tyrosine hydroxylase in the arcuate nucleus, which could be important for the regulation of prolactin release throughout the female reproductive cycle. Additionally, although progesterone alters mesolimbic and nigrostriatal dopamine release, and the numerous behaviours that these pathways influence, these data again suggest that it does not do so via nuclear progestin receptor in dopaminergic cells of the ventral tegmental area and substantia nigra.
Cellular Expression of V1a Vasopressin Receptor mRNA in the Female Rat Preoptic Area: Effects of Oestrogen
Authors: Kalamatianos T. , Kalló I. , Goubillon M‐L. , Coen C. W.
Published: May 2004

The preovulatory luteinizing hormone (LH) surge in female rats is dependent upon signals from the suprachiasmatic nucleus (SCN), the site of a dominant circadian pacemaker. Various lines of evidence indicate that arginine‐vasopressin (AVP)‐containing projections from the SCN to the preoptic area (POA) contribute to the production of the surge of LH‐releasing hormone (LHRH). These actions may be mediated by V1a because the transcript for this AVP receptor subtype is predominant within the POA of the female rat. In this study, in situ hybridization histochemistry was used to examine V1a mRNA expression, either by itself or together with LHRH or glutamic acid decarboxylase 65 (GAD65) mRNA, within the POA of ovariectomized rats in the presence or absence of oestrogen. V1a mRNA was found in cells across the rostro‐caudal axis of the POA; some were in close proximity to cells expressing LHRH mRNA. Coexpression of V1a and LHRH mRNAs was detected only very rarely. By contrast, cells with V1a mRNA commonly displayed GAD65 mRNA. The density of V1a mRNA‐expressing cells was particularly high within the anteroventral periventricular nucleus; at this site, V1a mRNA expression was elevated following oestrogen treatment. The present results indicate that V1a‐mediated AVP actions may influence LHRH release via cells in the immediate vicinity of LHRH neurones and/or via oestrogen‐regulated cells in the anteroventral periventricular nucleus, which is a site that lacks LHRH neurones but plays an essential role in initiating the preovulatory LH surge.
Chronic Glucocorticoid Deficiency‐Induced Abnormal Aggression, Autonomic Hypoarousal, and Social Deficit in Rats
Authors: Haller J. , Halász J. , Mikics É. , Kruk M. R.
Published: May 2004

Certain aggression‐related psychopathologies are associated with decreased glucocorticoid production and autonomic functions in humans. We have previously shown that experimentally‐induced chronic glucocorticoid deficiency leads to abnormal forms of attack in rats. Here, we compared the effects of acute and chronic glucocorticoid deficiency on aggressive behaviour, autonomic responses to challenges, and anxiety. Glucocorticoid synthesis was blocked acutely by the glucocorticoid synthesis blocker metyrapone or chronically by adrenalectomy and low glucocorticoid replacement (ADXr). As shown previously, chronic glucocorticoid deficiency facilitated aberrant attacks directed towards the most vulnerable parts of the opponent's body. The acute inhibition of glucocorticoid synthesis lowered aggressive behaviour without affecting attack targeting. In a different experiment, ADXr rats and their sham‐operated controls were exposed to different challenges whereas their heart rate and locomotion were telemetrically recorded. Autonomic responses to social challenges were lowered by chronic, but not by acute glucocorticoid deficiency. Autonomic responses to the elevated plus‐maze were only slightly affected by chronic glucocorticoid deficiency. Locomotor behaviour was not affected in either challenge; thus, the altered autonomic reactions were not due to interference from workload. The behaviour of ADXr rats was similar to that of sham‐operated controls in the elevated plus‐maze, but ADXr rats showed reduced social interactions in the social interaction test. Our data demonstrate that, in rats, chronic but not acute glucocorticoid deficiency induces abnormal attack patterns, deviant cardiovascular responses and social deficits that are similar to those seen in abnormally violent humans. Thus, the similar correlations found in humans probably cover a causal relationship. Experimentally‐induced glucocorticoid deficiency may be used to assess the mechanisms underlying glucocorticoid deficiency‐induced abnormal forms of aggressiveness.
Stimulation of Luteinizing Hormone Subunit Gene Expression by Pulsatile Intracerebroventricular Microinjection of Galanin in Female Rats
Authors: Gajewska A. , Zwierzchowski L. , Kochman K.
Published: May 2004

Although galanin, which exerts its effects both at the hypothalamic and pituitary level, has been implicated as an important neuroendocrine regulator of hypothalamic‐pituitary‐gonadal axis activity, there is a lack of data concerning its involvement in the regulation of gonadotropin subunit gene expression. To elucidate whether galanin can influence luteinizing hormone (LH) subunit mRNA content, as well as affect gonadotropin‐releasing hormone (GnRH) receptor activity, a model based on pulsatile (one pulse per hour over 5 h) galanin (1 nM) microinjections directly into the third cerebral ventricle of ovariectomized (OVX) and/or oestrogen/progesterone‐pretreated rats was used. Furthermore, to determine galanin effects on GnRH‐induced LH subunit mRNA synthesis, a cocktail of 1 nM GnRH and 1 nM galanin was coadministered in a pulsatile manner to OVX/steroid primed rats. Subsequently, to obtain data concerning the role of galanin receptors in the regulation of pituitary α (common to LH, follicle‐stimulating hormone, thyroid‐stimulating hormone) and LHβ subunit gene expression, OVX/oestrogen/progesterone rats received microinjections of 1 nM of the receptor antagonist galantide and 1 nM of galanin. In this case, both substances were administered separately, with a 30 min lag, according to which each galantide pulse always preceded a galanin pulse. Northern‐blot analysis revealed that intracerebroventricular pulsatile galanin injections were effective in stimulation of both α and LHβ subunit mRNA levels and that this effect was apparently steroid‐dependent. Moreover, galanin also up‐regulated GnRH receptor functional parameters (affinity and maximum binding capacity) but was ineffective in potentiating GnRH‐induced accumulation of both subunit mRNAs. The results from the study also indicate that galanin acts through its own receptor(s) because a receptor antagonist, galantide, significantly reduced the stimulatory effect exerted by galanin on the expression of both LH subunit genes in vivo.
Differential modulation of murine host immune response by salivary gland extracts from the mosquitoes Aedes aegypti and Culex quinquefasciatus
Authors: Wanasen N. , Nussenzveig R. H. , Champagne D. E. , Soong L. , Higgs S.
Published: June 2004

Abstract.  Mosquitoes (Diptera: Culicidae) are major vectors of numerous infectious agents. Compounds in mosquito saliva not only facilitate blood‐feeding, but may also have an impact upon the immune system of vertebrate hosts. Consequently, the exposure to mosquito saliva may influence pathogen transmission, establishment and disease development. Using two medically important vector mosquitoes, Aedes aegypti (L.) and Culex quinquefasciatus Say, we examined the effects of mosquito saliva on immune cells of host mice. After antigen‐specific or non‐specific stimulation, murine splenocyte proliferation and production of both Th1 and Th2 cytokines were significantly reduced in the presence of salivary gland extract (SGE) from Ae. aegypti, but not SGE from Cx. quinquefasciatus. T cell populations were highly susceptible to this suppression, showing increased mortality and reduced division rates – judged by flow cytometric analyses. Evidently these two culicine mosquitoes differ in their host immunomodulatory activities.
Endolysosomal Processing of Exogenous Antigen into Major Histocompatibility Complex Class I‐Binding Peptides
Authors: Chen L. , Jondal M.
Published: June 2004

An alternative endolysosomal pathway has recently been suggested for the processing of MHC‐I‐binding peptides, and peptide/MHC‐I complexes have been demonstrated in this compartment. However, it remains unclear where in the antigen‐presenting cells such peptides are processed, in the endolysosomes themselves or in the proteasomal complex. Here, we have investigated this using monoclonal antibodies specific for the immunodominant SIINFEKL/Kb complex (25‐D1) or for the carbohydrate part of Db‐ or Kb‐binding glycopeptides in combination with inhibitors for classical and endolysosomal MHC‐I‐processing pathways. Alternative processing was detected in both wt and TAP1–/– immature DC (iDC) as the expression of SIINFEKL/Kb complexes on the surface of OVA‐treated cells in the presence of Brefeldin A (BFA) or lactacystin and their absence in the presence of the lysosomotropic amines ammonium chloride, chloroquine and methylamine. Internalized Db‐ and Kb‐binding glycopeptides, detected with high specificity using an anti‐galabiose (Gal2) monoclonal antibody, were found to appear on the cell surface of BFA‐treated cells after intracellular MHC‐I‐binding. Peptide exchange in Kb was demonstrated as the gradual appearance of SIINFEKL/Kb complexes on BFA‐treated cells which earlier had been saturated with another Kb‐binding peptide. Our data support the presence of a fully functional endolysosomal processing pathway in iDC guided by the chaperone function of MHC‐I molecules.
Red Blood Cells Inhibit Proliferation and Stimulate Apoptosis in Human Lung Fibroblasts In Vitro
Authors: Fredriksson K. , Stridh H. , Lundahl J. , Rennard S. I. , Skold C. M.
Published: June 2004

Cell proliferation and apoptosis are both important mechanisms for the regulation of tissue homeostasis. For instance, proliferation is crucial in wound repair, whereas apoptosis is important for removal of damaged cells and resolution of inflammation. Imbalance between cell proliferation and apoptosis can therefore lead to pathological conditions and disease. In inflammatory and fibrotic lung disorders, red blood cells (RBCs) can interact with fibroblasts and connective tissue. In the present study, we therefore hypothesized that the presence of RBCs can affect fibroblast proliferation and apoptosis. Human foetal lung fibroblasts (HFL‐1) were cultured in the presence or absence of purified whole RBCs and RBC‐conditioned media. RBC significantly decreased fibroblast proliferation as determined both by DNA content analysis (Hoechst 33258 staining, P < 0.01; WST‐1, P < 0.001) and BrdU incorporation. After treatment with staurosporine (STS) for 48 h, apoptosis was determined by TUNEL and propidium iodide staining followed by flow cytometry analysis. RBCs augmented STS‐induced apoptosis (median: 46.4%; range 12.0–90.4) compared to control cells (median 26.2%; range 7.1–45.5). Thus, our data indicate that the presence of RBCs affects both fibroblast proliferation and susceptibility to undergo apoptosis. Our findings therefore suggest a role for RBCs in regulating fibroblast homeostasis after tissue injury.
The study of the participation of basement membrane zone antibodies in the formation of the lupus band in systemic lupus erythematosus
Authors: Yang Sen , Gao Yu , Song Yingxue , Liu Jiangbo , Yang Chunjun , Liu Jinli , Wang Zaixing , Zhang Xuejun
Published: May 2004

Background  The lupus band test (LBT) is an important auxiliary method in the diagnosis of systemic lupus erythematosus (SLE), but the mechanism of its formation is still unknown. There are many kinds of autoantibodies, such as basement membrane zone autoantibodies (BMZ‐Abs), in patients with SLE.

Aim  To detect whether skin BMZ‐Abs participated in the formation of the lupus band in SLE.

Methods  Immunoglobulin G (IgG)‐type BMZ‐Abs in 15 SLE patients were detected by means of immunofluorescence (IF), immunoblotting (IB), and immunoelectron microscopy (IEM).

Results  Direct immunofluorescence (DIF) on salt‐split skin showed epidermal fluorescence in 12 of the 15 SLE patients. Two of the 12 patients also showed dermal fluorescence. Indirect immunofluorescence (IIF) on salt‐split skin revealed that, in 12 of the 15 (80%) sera, antibodies were bound to the epidermal roof of the salt‐split skin. IB showed that, in 14 SLE sera, autoantibodies reacted to 230, 200, 180, 130, and 97 kDa epidermal extracts and 75 kDa dermal extracts. Direct (DIEM) and indirect (IIEM) immunoelectron microscopy showed that gold particles were directed to every region of the BMZ, including hemidesmosomes, lamina lucida, lamina densa, and sublamina densa.

Conclusions  BMZ‐Abs in SLE sera may participate in the formation of the lupus band.
Broad‐band UVB fiber‐optic comb for the treatment of scalp psoriasis: a pilot study
Authors: Taneja Atul , Racette Ashley , Gourgouliatos Zafirios , Taylor Charles R.
Published: May 2004

Background  Scalp psoriasis is a frustrating condition with limited treatment options. Our objective was to test a new fiber‐optic broad‐band UVB comb device for the treatment of this condition.

Methods  Scalp psoriasis on adult subjects was treated three times a week for up to 12 weeks with a fiber‐optic UVB comb. A minimum 50‐cm2 scalp area served as the control. Starting doses were based on the standard minimal erythema dose (MED) with increments up to 20% for subsequent treatments.

Results  Fourteen subjects completed the study without sunburn reactions or blisters. Treated sites showed a mean improvement in the modified PASI score of 3.6. The difference in the mean modified PASI scores between the control and treated sites was 3.9 (< 0.0001).

Conclusions  This fiber‐optic broad‐band UVB comb can safely and effectively treat scalp psoriasis.
Melanoma epidemic: True or false?
Authors: Flórez Ángeles , Cruces Manuel
Published: May 2004

Melanoma incidence has increased dramatically over the last decades in most industrial countries, mainly as a result of the large numbers of early melanomas being diagnosed. Simultaneously, a lack of commensurate change in mortality has been reported, raising the possibility that skin melanoma may have modified its aggressiveness as a result of the increased diagnosis of biologically benign lesions. The main data and controversies arising from the melanoma epidemic are reviewed.
Celecoxib‐induced photoallergic drug eruption
Authors: Yazici Ayca Cordan , Baz Kiymet , Ikizoglu Guliz , Kokturk Aysin , Uzumlu Hale , Tataroglu Canten
Published: May 2004

Photoallergic dermatitis is caused by a photosensitizing substance plus sunlight exposure in a sensitized person. If the photosensitizer is delivered internally, it is called a photoallergic drug reaction. Celecoxib is a new generation non‐steroidal anti‐inflammatory drug and sulfonamide derivative. We report a photoallergic drug eruption associated with the introduction of celecoxib. To our knowledge, this is the first report of photoallergic drug reaction associated with celecoxib.
Tinea faciei, an often deceptive facial eruption
Authors: Lin Richie L. , Szepietowski Jacek C. , Schwartz Robert A.
Published: May 2004

Tinea faciei is a relatively uncommon superficial dermatophyte infection limited to the glabrous skin of the face. It is usually seen in pediatric patients. This fungus can be found worldwide, but has a predilection for tropical humid climates. Although the infecting organisms and treatments are the same as for other superficial fungal infections, it is frequently misdiagnosed clinically and histologically.
Templated assembly of the pH‐sensitive membrane‐lytic peptide GALA
Authors: Haas D.H. , Murphy R.M.
Published: May 2004

Abstract:  Delivery of protein or nucleic acid therapeutics into intracellular compartments may require facilitation to allow these macromolecules to cross otherwise impermeant cellular membranes. Peptides capable of forming membrane‐spanning channels hold promise as just such facilitators, although the requirement for peptide oligomerization to form these channels may limit their effectiveness. Synthetic molecules containing multiple copies of membrane‐active peptides attached to a template molecule in a pre‐oligomerized form have attracted interest for drug‐delivery applications. Using three template designs, we synthesized multimeric versions of the pH‐sensitive lytic peptide GALA and compared their performance to monomeric GALA. Template assembly stabilized helix formation: templated GALA retained α‐helical structure even at neutral pH, unlike monomeric GALA. In membrane leakage assays, templated GALA retained the pH sensitivity of the monomer, with improved leakage for dimeric GALA. Surprisingly, trimeric GALA was less effective, particularly when synthesized with a larger and more flexible spacer. Surface plasmon resonance analysis indicated that reversible binding of templated GALA to lipid surfaces at acidic conditions was greatly reduced compared with monomeric GALA, but that the amount of irreversibly bound material was similar. We interpreted these results to indicate that templated peptides may cyclize into ‘self‐satisfied’ oligomeric structures, incapable of further aggregation and subsequent pore formation. Future design of templated peptides must be carefully performed to avoid this unwanted consequence.
Nonpeptide/peptide chimeric ligands for the nociceptin/orphanin FQ receptor: design, synthesis and in vitro pharmacological activity
Authors: Guerrini R. , Carra’ G. , Calo’ G. , Trapella C. , Marzola E. , Rizzi D. , Regoli D. , Salvadori S.
Published: May 2004

Abstract:  Nociceptin/orphanin FQ (N/OFQ) is the endogenous ligand for the G‐protein coupled receptor referred to as N/OFQ peptide (NOP) receptor. NOP receptor activation by N/OFQ modulates several biological functions both at central and peripheral level. Structure activity relationship (SAR) studies demonstrated that the N/OFQ sequence can be divided into a N‐terminal tetrapeptide ‘message’ crucial for receptor activation and a C‐terminal ‘address’ important for receptor binding. On the basis of this message/address concept we synthesized some chimeric compounds in which we substituted the natural message domain with the nonselective nonpeptide NOP ligand (8‐Naphthalen‐1‐yl‐methyl‐4‐oxo‐1‐phenyl‐1,3,8‐triaza‐spiro[4,5]dec‐3‐yl)‐aceticacid methyl ester (NNC 63‐0532) and used as address domain the peptide sequences Thr‐NH2, N/OFQ(5‐9)‐NH2, N/OFQ(5‐13)‐NH2 and N/OFQ(5‐17)‐NH2. All the compounds were pharmacologically evaluated in the electrically stimulated guinea‐pig ileum. NNC 63‐0532 produced a concentration‐dependent inhibition of the electrically induced twitches showing, in comparison with N/OFQ, lower potency and higher maximal effects. In addition, contrary to N/OFQ, the effects of NNC 63‐0532 were insensitive to the NOP selective antagonist [Nphe1, Arg14, Lys15]N/OFQ‐NH2 (UFP‐101) while prevented by naloxone. Similar results were obtained with NNC 63‐0532/Thr‐NH2 and NNC 63‐0532/N/OFQ(1‐9)‐NH2. On the contrary, the inhibitory effects of NNC 63‐0532/N/OFQ(5‐13)‐NH2 and NNC 63‐0532/N/OFQ(5‐17)‐NH2 were slightly antagonized by UFP‐101 while naloxone prevented the effects of the high but not of the low concentrations of the two ligands. These data indicate that it is possible to functionalize with the N/OFQ address sequence a nonpeptide NOP ligand for increasing its binding to the NOP receptor. Moreover, these results corroborate the idea that the 5–13 sequence represents the crucial core of the N/OFQ address domain.
Effect of feeding intensity and time on feed on intramuscular fatty acid composition of Simmental bulls
Authors: Sami A. S. , Augustini C. , Schwarz F. J.
Published: May 2004

Seventy‐two Simmental bulls, weighing 489 kg initially and approximately 15 months old, were divided into four groups to determine the effects of feeding intensity and time on feed on intramuscular fatty acid (FA) composition. Two groups of 18 bulls each were extensively (E) or intensively (I) fed on maize silage and concentrates with a daily gain of 943 g (E) or 1371 g (I). Half of each group were slaughtered after 100 days (S) or 138 days (L) on feed. In addition to carcass fatness parameters, intramuscular FA composition was also measured. Only small differences in the sum of saturated FA (SFA) percentages were found with 47.7 and 47.5% FA methyl esters (FAME) for SE and LE, respectively, and 48.7% FAME for each of SI and LI. Monounsaturated fatty acids (MUFA) significantly increased with longer, and higher feeding intensity to 44.2% FAME (LI), whereas the other groups had similar contents of 41.9, 42.2 and 42.0% FAME (SE, LE and SI respectively). Polyunsaturated FA (PUFA) decreased with higher feeding intensity to 8.39% (SI) and 6.71% (LI) FAME (p < 0.05) in comparison with 9.48% (SE) and 9.54% (LE). Intensive feeding decreased the ratio of PUFA : SFA to 0.17 (SI) and 0.14 (LI, p < 0.05) in comparison with 0.20 (SE and LE). The mean conjugated linoleic acid (CLA) concentration was 0.41% FAME. Time on feed had only a small effect on the FA composition compared with feeding intensity. No significant relationships were detected between meat quality attributes and the pattern of FA.
Inhibition of ruminal microbial methane production by β‐cyclodextrin iodopropane, malate and their combination in vitro
Authors: Mohammed N. , Lila Z. A. , Ajisaka N. , Hara K. , Mikuni K. , Hara K. , Kanda S. , Itabashi H.
Published: May 2004

The objective of this study was to evaluate the effects of different concentrations of l‐malate (0, 5, 10 and 20 mm), 2‐iodopropane‐β‐cyclodextrin complex (CD‐IP) (0, 0.1, 0.2 and 0.4 mm) and a combination of malate (10 and 20 mm) plus CD‐IP (0.2 and 0.4 mm) on methane production from corn starch. Ruminal fluid was collected from dairy cows, mixed with phosphate buffer (1 : 2) and incubated (30 ml) anaerobically at 38 °C for 6 h with or without additives. Fermentation of corn starch in the presence of malate resulted in an increase (p < 0.05) in pH of the medium, total volatile fatty acid (VFA), total gas production and molar proportion of propionate. Acetate and ammonia‐N concentration were unchanged. Methane production was decreased (p < 0.05) (15.5 to 20.4%). Addition of CD‐IP in corn starch resulted in an increase (p < 0.05) in total VFA and molar proportion of propionate. Acetate, pH and ammonia‐N concentration of the medium were decreased (p < 0.05). Total gas production was unchanged. Methane production was decreased (p < 0.05) (25.2 to 97.1%) and hydrogen production was increased (p < 0.05). Addition of l‐malate to CD‐IP resulted in an increase (p < 0.05) in total VFA, total gas production and molar proportion of propionate. Acetate and ammonia‐N concentration were decreased (p < 0.05). No effects were observed on medium pH. Methane production was decreased (p < 0.05) (49.5 to 97.1%). Hydrogen production was also decreased (p < 0.05) (54.5 to 64.1%) compared with those of CD‐IP alone. Therefore, these additives may be used as supplements to inhibit methane production as well as to improve rumen fermentation and animal performance.
Influence of dietary linseed, fish and coconut oil on growth performance of growing pigs kept on small holdings in central Vietnam
Authors: Nguyen L. Q. , Everts H. , Beynen A. C.
Published: May 2004

The aim of the study was to investigate the effect on growth performance of the addition of 5% of either linseed, fish or coconut oil to the diet of growing pigs kept on small holdings in central Vietnam. The diets were fed on six different farms; there were three animals per treatment per farm. The farmers fed a base diet according to personal choice, but were instructed to use linseed, fish and coconut oil. The diets were fed to the pigs from 70 to 130 days of age. The diets with 5% linseed oil on average contained 2.4 g α‐linolenic acid/MJ of metabolizable energy (ME) and the diets with 5% fish oil on average contained 0.06 and 0.75 g eicosapentaenoic and docosahexaenoic acid/MJ ME, respectively. The relative percentages of docosahexaenoic, eicosapentaenoic, α‐linolenic and linoleic acid in adipose tissue were determined by the intake of the corresponding fatty acids. Dietary linseed oil vs. coconut oil significantly enhanced daily weight gain and vs. fish oil it also stimulated growth, but this effect just failed to reach statistical significance. It is concluded that extra intake of α‐linolenic acid may stimulate growth in growing‐finishing pigs, this effect being independent of the conversion of α‐linolenic acid into eicosapentaenoic and docosahexaenoic acid.
Effects of supplemental L‐carnitine in drinking water on performance and egg quality of laying hens exposed to a high ambient temperature
Authors: Çelik L. B. , Tekeli A. , Öztürkcan O.
Published: May 2004

The present study was conducted to investigate effects of l‐carnitine supplied with drinking water on performance and egg quality of laying hens under high environmental temperature. In the study, 47‐week‐old laying hens (Brown hisex) were divided into two groups (control and treatment) and fed with a standard layer diet. Treatment group was received 50 p.p.m l‐carnitine with drinking water for an 8‐week period. Throughout the study, 8 h hot (35–37 °C) and 16 h thermoneutral (20–22 °C) environmental temperature regime was employed daily. The results showed that l‐carnitine supplementation affected some egg quality characteristics of layers under high environmental temperature. Relative albumen weight and height were increased (p < 0.05) by supplemental l‐carnitine. Live weight gain, feed intake, egg mass, egg weight, yolk weight, shell weight, yolk index, egg‐shape index, yolk colour score and shell thickness were not affected (p < 0.05) by l‐carnitine. It is concluded that l‐carnitine supplementation in laying hens could have potential to improve albumen quality under high environmental temperature.
The study of introgressive lines of Triticum aestivum × Aegilops speltoides by in situ and SSR analyses
Authors: Adonina I. G. , Salina E. A. , Efremova T. T. , Pshenichnikova T. A.
Published: June 2004

Fluorescence in situ hybridization (FISH) with a genome‐specific repeat, Spelt1, and wheat simple sequence repeat (SSR) markers were used to analyse the chromosome constitution of two Triticum aestivum×Aegilops speltoides introgressive lines. The lines 170/98i and 178/98i carried one and two subtelomeric regions of Ae. speltoides (per haploid genome), respectively, marked by Spelt1 repeats according to FISH data. SSR analysis detected homoeologous substitution of wheat chromosome 7D with Ae. speltoides chromosome 7S in the lines 178/98i and 170/98i as well as the assumed terminal translocation in the short arm of chromosome 3A in the line 178/98i. Anthocyanin pigmentation of the coleoptiles was found in the lines 170/98i and 178/98i and resulted from the 7S (7D) substitution. It was demonstrated that Spelt1 could be effectively used for the rapid identification (without DNA isolation) of terminal translocations of T. aestivum×Ae. speltoides introgressive lines as well as for further analysis of the stability of the hybrid plants.
Inheritance of resistance to spot blotch caused by Bipolaris sorokiniana in spring wheat
Authors: Joshi A. K. , Kumar S. , Chand R. , Ortiz‐Ferrara G.
Published: June 2004

Three F1 progenies and their families in the segregating generations (F3, F4, F5 and F6), obtained after crossing resistant × susceptible wheat genotypes were studied in the field to determine the genetics of resistance to spot blotch caused by Bipolaris sorokiniana. Spot blotch scores in the F1 generation showed absence of dominance. Individually threshed F2 plants were used to advance the generations. Progenies (200‐250) of resistant genotypes Acc. No. 8226, Mon/Ald, Suzhoe#8 crossed with susceptible ‘Sonalika’ were evaluated in the F3, F4, F5 and F6 generations under induced epiphytotic conditions. Based on disease score distribution in individual progeny rows, F3 progenies were grouped into four classes: homozygous resistant, homozygous susceptible, segregating resistant and segregating susceptible. Resistance appeared to be under the control of three additive genes. The presence of three genes was also noted in the distribution of F4 and F5 lines. In the case of F6 progeny rows, both quantitative and qualitative models were used to estimate the number of segregating genes based on a 2‐year trial. It appeared that resistance to spot blotch was controlled by the additive interaction of more than two genes, possibly only three.
Molecular mapping of a dominant genic male sterility gene Ms in rapeseed (Brassica napus)
Authors: Lu G. Y. , Yang G. S. , Fu T. D.
Published: June 2004

Rs1046AB is a genic male sterile two‐type line in rapeseed that has great potential for hybrid seed production. The sterility of this line is conditioned by the interaction of two genes, i.e. the dominant genic male sterility gene (Ms) and the suppressor gene (Rf). The present study was undertaken to identify DNA markers for the Ms locus in a BC1 population developed from a cross between a male‐sterile plant in Rs1046AB and the fertile canola‐type cultivar ‘Samourai’. Bulked segregant analysis was performed using the amplified fragment length polymorphism (AFLP) methodology. From the survey of 480 AFLP primer combinations, five AFLP markers (P10M13350, P13M8400, P6M6410, E7M1230 and E3M15100) tightly linked to the target gene were identified. Two of them, E3M15100 and P6M6410, located the closest, at either side of Ms at a distance of 3.7 and 5.9 cM, respectively. The Ms locus was subsequently mapped on linkage group LG10 in the map developed in this laboratory, adding two additional markers weakly linked to it. This suite of markers will be valuable in designing a marker‐assisted genic male sterility three‐line breeding programme.
Effect of genetic background on the target fatty acids of acyl‐ACP thioesterase transgenes in Brassica napus
Authors: Tang J. , Scarth R.
Published: June 2004

Acyl‐acyl carrier protein (ACP) thioesterase (TE) is involved in the biosynthetic fatty acid pathway of plants. Conventional canola lines transformed individually with the bay‐TE (Uc FatB1), elm‐TE (Ua FatB1), nutmeg‐TE (Mf FatB1) or Cuphea‐TE transgene (Ch FatB1), produce seed oil with modified fatty acid compositions. This study assessed the effects of genetic background, cytoplasm, maternal parent, and interaction of different TE transgenes, on the target fatty acids using F1 seeds and double haploid (DH) lines. The F1 seeds were produced by crossing four TE transgenic parental lines and three non‐transgenic cultivars with distinct fatty acid compositions. The DH lines were developed from microspores of F1 plants. DH lines from different crosses showed that genetic background does not have an effect on the relative levels of the target fatty acids of the same TE, indicating the stability of the substrate specificity of the TE within canola. However, significant effects of genetic background on the content of the major target fatty acids, lauric acid (C12:0) or palmitic acid (C16:0) depending on the TE, were observed. Expression of the TE in low erucic acid (C22:1) genotypes resulted in higher target fatty acid levels than expression in high C22:1 genotypes. Reciprocal crosses showed maternal effects, but not cytoplasmic effects. In addition, co‐expression of two different TE transgenes in the same seeds was observed. These results indicate the importance of selection for appropriate genetic backgrounds in order to maximize the expression of the target fatty acids of TE transgenes, and also indicate potential uses of TE co‐expression in modifying canola seed oil.
Spanish spelt: a separate gene pool within the spelt germplasm
Authors: Elía M. , Moralejo M. , Rodríguez‐Quijano M. , Molina‐Cano J. L.
Published: June 2004

The renewed interest in spelt (Triticum spelta L.) for wheat improvement programmes requires the study of the available genetic diversity. The purpose of this study was to assess genetic diversity within a Spanish spelt collection. Sixty‐six Spanish spelt accessions, 19 accessions of T. spelta and T. macha from different origins, three bread wheat cultivars (T. aestivum) and one accession of T. dicoccum were screened using simple sequence repeats (SSRs). The diversity observed within the Spanish group was comparable with that observed in the other wheat varieties, despite their broader geographical diversity. Indeed, the highest polymorphic information content value calculated with SSRs for Spanish material (0.90) is similar to that observed for the other wheat varieties (0.98). Principal component analysis explained 46.5% of the cumulative variation and confirmed the Spanish accessions as a separate group. This study showed the Spanish spelt collection to be a variable and unique genetic resource for wheat and spelt breeding programmes.
Single nucleotide polymorphism genotyping of the barley waxy gene by polymerase chain reaction with confronting two‐pair primers
Authors: Domon E. , Yanagisawa T. , Saito A. , Takeda K.
Published: June 2004

A high‐throughput single nucleotide polymorphism (SNP) genotyping procedure was developed to select amylose‐free barley mutants whose waxy genes had a C‐ to T‐base substitution in exon 5, which converted Gln‐89 of the wild‐type gene into a termination codon. An F2 population carrying an amylose‐free waxy gene was checked for segregation. Polymerase chain reaction with confronting two‐pair primers (PCR‐CTPP) produced allele‐specific PCR products that have different sizes and are inherited in a co‐dominant manner. Two alleles of the barley waxy gene with SNP were correctly identified in parental strains using the PCR‐CTPP procedure. Segregation of the SNP as detected by PCR‐CTPP in an F2 population fitted the expected 1:2:1 ratio. The PCR‐CTPP procedure can provide a time saving and cost‐effective alternative to derived cleaved amplified polymorphic sequence in marker‐assisted selection.
Genetics, fertility behaviour and molecular marker analysis of a new TGMS line, TS6, in rice
Authors: Latha R. , Thiyagarajan K. , Senthilvel S.
Published: June 2004

The thermosensitive genic male sterility (TGMS) system has great potential for revolutionizing hybrid rice production through simple, less expensive and more efficient seed production technology. For the successful utilization of this novel male sterility system, knowledge of the breeding and fertility behaviour of a TGMS line is essential. In this study, the fertility transformation behaviour, the critical fertility and sterility temperatures and the mode of inheritance of male sterility were studied for a new TGMS line, TS6, identified at Tamil Nadu Agricultural University, Coimbatore, India. The pollen and spikelet fertilities recorded on plants raised at fortnightly intervals revealed that this line was completely sterile for 78 consecutive days (35/22 to 32/23°C, maximum/minimum temperatures) and reverted to fertile when the temperature was 30/18°C. It remained fertile continuously for 69 days and the maximum pollen and spikelet fertilities recorded were 75 and 70%, respectively. The fertility was highly influenced by daily maximum temperature followed by average and minimum temperatures. It was not influenced by relative humidity, sunshine hours or photoperiod. The critical temperature inducing sterility and fertility was 26.7 and 25.5°C, respectively. The male sterility in TS6 was inherited as a monogenic recessive in the F2 and BC1 populations of TS6 × MRST9 as well as TS6 × IR68281B. Using bulked segregant analysis on an F2 population of TS6 × MRST9, an RAPD marker, OPC052962, was identified to be associated with TGMS in TS6.
Identification of quantitative trait loci underlying milling quality of rice (Oryza sativa) grains
Authors: Li Z. F. , Wan J. M. , Xia J. F. , Zhai H. Q. , Ikehashi H.
Published: June 2004

Milling quality of rice grains is important to both producers and consumers. In this study, quantitative trait loci (QTLs) controlling brown rice rate (BR), milled rice recovery (MR) and head rice recovery (HR) were analysed by composite interval mapping over 2 years using 98 backcross inbred lines (BILs). A total of 12 QTLs for the three traits were detected, of which five were for BR, four for MR and three for HR. The proportion of phenotypic variation explained by individual QTLs ranged from 7.5 to 19.9%, and additive effects contributed by a single QTL accounted for 0.46 to 2.34% of the variation. QTL‐by‐environment interactions were observed by comparing QTL mapping of the same population grown in two consecutive years. Three of five QTLs for BR and two of four QTLs for MR were detected in 2 years, and all three QTLs for HR were detected in 1 year only. BR was significantly correlated with MR, and all four QTLs of MR were located in the same regions as those of BR. This indicated that QTLs for highly correlated traits could often be detected in the same interval.
Optical coherence tomography as a novel tool for non‐destructive measurement of the hull thickness of lupin seeds
Authors: Clements J. C. , Zvyagin A. V. , Silva K. K. M. B. D. , Wanner T. , Sampson D. D. , Cowling W. A.
Published: June 2004

Hull thickness is an important component of seed quality, which effects dehulling ability, feed or food nutritional aspects and cooking times. A breeding objective in Lupinus angustifolius crop improvement is to reduce hull thickness and a rapid screening method is needed to efficiently screen genotypes. Optical coherence tomography (OCT) imaging using infrared illumination at 980 nm was used to compare hull thickness of genotypes of four lupin species. OCT‐derived hull layer thickness correlated highly with actual hull thickness determined by environmental scanning electron microscopy (r = 0.90) and allowed reliable distinction between mutant (thin‐hulled) and parent genotypes of L. angustifolius. The imaging could clearly penetrate lupin seed to a depth of approximately 200 μm. The use of OCT to measure hull thickness has the advantage that it is rapid and non‐destructive and should be very useful in selecting thin hull lines of lupins and other species on a single seed basis in germplasm or progeny from crosses.
EFFECT OF MAGNESIUM ON MECHANICAL PROPERTIES OF PRESSURIZED MESENTERIC SMALL ARTERIES FROM OLD AND ADULT RATS
Authors: Adrian Mark , Laurant Pascal , Berthelot Alain
Published: May 2004

1. The purpose of the present study was to determine the effects of magnesium (Mg) on the mechanical properties of resistance arteries in adult and old rats.

2. Studies were performed in adult (17 weeks) and old (104 weeks) male Wistar rats. The vasodilatory response and the passive mechanical properties of the wall of isolated perfused and pressurized arterial segments of mesenteric small arteries were investigated after Mg and verapamil application, both known for their calcium antagonistic properties.

3. Mesenteric resistance arteries from old rats exhibited an outward hypertrophic remodelling, with enlargment of the lumen, thickening of the media and enlarged media cross‐sectional area.

4. The vasodilatory response induced by the application of increasing extracellular concentrations of Mg and verapamil was significantly smaller in preconstricted mesenteric arteries of old rats than in those of adult rats.

5. Incremental distensibility in response to increasing intravascular pressures did not change. However, the stress–strain curve was shifted to the left in pressurized mesenteric arteries from old rats, indicating arterial wall stiffness. Verapamil (3 µmol/L) did not modify the stress–strain curves in either adult or aged rats. However, Mg (4.8 mmol/L) significantly shifted the curve to the right in mesenteric arteries from adult rats and, to a greater degree, in those from old rats.

6. Although Mg‐induced vasodilatation is impaired in aged rats, increased Mg concentration improved the mechanics of pressurized mesenteric resistance arteries. The fact that Mg decreases arterial stiffness in arteries from old rats suggests that Mg has a beneficial effect on age‐related changes to the vascular wall.
NANOMOLAR LEVEL OF OUABAIN INCREASES INTRACELLULAR CALCIUM TO PRODUCE NITRIC OXIDE IN RAT AORTIC ENDOTHELIAL CELLS
Authors: Dong Xian Hui , Komiyama Yutaka , Nishimura Noriko , Masuda Midori , Takahashi Hakuo
Published: May 2004

1. Changes in [Ca2+]i across the cell membrane and/or the sarcoplasmic reticulum regulate endothelial nitric oxide (NO) synthase activity.

2. In the present study, we investigated the effect of ouabain, a specific inhibitor of Na+/K+‐ATPase, on NO release and [Ca2+]i movements in cultured rat aortic endothelial cells (RAEC) by monitoring NO production continuously using an NO‐specific real‐time sensor and by measuring the change in [Ca2+]i using a fluorescence microscopic imaging technique with high‐speed wavelength switching. The t½ (half‐time of the decline of [Ca2+]i to basal levels after stimulation with 10 µmol/L bradykinin) was used as an index of [Ca2+]i extrusion.

3. A very low concentration of ouabain (10 nmol/L) did not increase the peak of NO production, but decreased the decay of NO release and, accordingly, increased integral NO production by the maximal dose–response concentration induced by bradykinin. The same dose of ouabain affected [Ca2+]i movements across the cell membrane and/or sarcoplasmic reticulum induced by bradykinin with a time‐course similar to that of NO release. Moreover, the t½ was significantly increased.

4. Pretreatment of RAEC with Na+‐free solution, an inhibitor of the Na+/Ca2+ exchanger, and nickel chloride hexahydrate prevented the effects induced by bradykinin and ouabain.

5. These observations using real‐time recording indicate that a small amount of ouabain contributes to the bradykinin‐stimulated increase of NO production through inhibition of plasma membrane Na+/K+‐ATPase activity and an increase in intracellular Na+ concentrations. The membrane was then depolarized, leading to a decline in the bradykinin‐stimulated increase in [Ca2+]i by forward mode Na+/Ca2+ exchange to prolong the Ca2+ signal time.

6. From these results, we suggest that nanomolar levels of ouabain modulate [Ca2+]i movements and NO production in RAEC.
CYTOCHROMES P450 ARE DIFFERENTLY EXPRESSED IN NORMAL AND VARICOSE HUMAN SAPHENOUS VEINS: LINKAGE WITH VARICOSIS
Authors: Bertrand‐Thiebault Céline , Ferrari Luc , Boutherin‐Falson Odile , Kockx Mark , Desquand‐Billiald Stéphanie , Fichelle Jean‐Marc , Nottin Rémi , Renaud Jean‐François , Batt Anne‐Marie , Visvikis Sophie
Published: May 2004

1. The expression of cytochrome P450 (CYP) enzymes and cyclo‐oxygenases (COX) was investigated in human saphenous veins by reverse transcription–polymerase chain reaction analysis. Non‐varicose veins were obtained from patients undergoing aortocoronary bypass grafting, whereas varicose veins were obtained from patients undergoing stripping removal of varicose saphenous veins.

2. In non‐varicose veins, CYP1B1, CYP2C, CYP2E1 and CYP4A11 were detected, whereas CYP2J2, CYP3A5, COX‐1 and COX‐2 were detected almost exclusively in varicose veins. CYP4F2 was not detectable.

3. Except for CYP4A11, the levels of individual CYP mRNA were higher in varicose veins than in control veins.

4. Smooth muscle cell volume, determined by a colour image‐analysis system, was increased approximately 1.5‐fold in varicose veins.

5. Because CYPs and COXs produce various vasoactive compounds, increased expression of these enzymes could be involved in the impairment of vascular tone and may contribute to varicose pathology. Then, CYP or COX modulators may be potentially active in the treatment of chronic venous insufficiency.
CHRONIC DECREASE IN FLOW CONTRIBUTES TO HEART FAILURE‐INDUCED ENDOTHELIAL DYSFUNCTION IN RATS
Authors: Devaux C , Iglarz M , Richard V , Mulder P , Henrion D , Renet S , Henry JP , Thuillez C
Published: May 2004

1. Chronic heart failure (CHF) impairs endothelium‐dependent, nitric oxide (NO)‐mediated dilation. This decreased dilation may be partly secondary to the chronic decrease in blood flow, but this hypothesis has not yet been tested. Thus, we assessed whether a localized, chronic increase in blood flow in vivo reverses endothelial dysfunction of small arteries in rats with CHF.

2. Two months after coronary artery ligation or sham surgery, second‐order side branches of the superior mesenteric artery were ligated in order to obtain persistently elevated blood flow (HF) in the adjacent first‐order side branch compared with normal vessels (NF). One month later, responses to acetylcholine and flow‐mediated vasodilatation (FMD) were assessed in vitro in an arteriograph.

3. Chronic heart failure induced a decrease in mesenteric blood flow (374 ± 25 and 305 ± 27 µL/min for sham and CHF, respectively; P < 0.05). Neither CHF nor the chronic increase in flow affected the responses to acetylcholine.

4. Chronic heart failure decreased FMD (maximal response in sham and control 34 ± 6 and 13 ± 4%, respectively; P < 0.05). Chronic increases in blood flow did not modify FMD in sham, but restored FMD in CHF rats (28 ± 4%; P < 0.05 vs CHF NF).

5. The restored response was abolished by an inhibitor of NO synthesis (NG‐nitro‐l‐arginine). Chronic heart failure did not affect the abundance of mesenteric endothelial NO synthase (eNOS) mRNA.

6. A chronic increase in flow significantly increased the abundance of eNOS mRNA in sham rats, but only moderately and non‐significantly in CHF rats.

7. Thus, endothelial dysfunction of small arteries in CHF appears to be largely the consequence of the chronic decrease in flow.