bluuebunny/crossref_metadata_2025_split huggingface dataset:
Various members of the Toll‐like receptor family contribute to the innate immune response of human epidermal keratinocytes
Authors: Köllisch Gabriele , Kalali Behnam Naderi , Voelcker Verena , Wallich Reinhard , Behrendt Heidrun , Ring Johannes , Bauer Stefan , Jakob Thilo , Mempel Martin , Ollert Markus 
Published: March 2005

SummaryToll‐like receptors (TLRs) are important pattern recognition molecules that activate the nuclear factor (NF)‐κB pathway leading to the production of antimicrobial immune mediators. As keratinocytes represent the first barrier against exogenous pathogens in human skin, we investigated their complete functional TLR1–10 expression profile. First, reverse transcription–polymerase chain reaction (PCR) analysis revealed a very similar pattern of TLR mRNA expression when comparing freshly isolated human epidermis and cultured primary human keratinocytes. Thus, further experiments were carried out with primary keratinocytes in comparison with the spontaneously immortalized human keratinocyte cell line HaCaT. The quantitative expression of TLR1–10 mRNA in real‐time PCR of primary human keratinocytes and HaCaT cells was analysed. Both cell types constitutively expressed TLR2, TLR3, TLR5, and to a lesser extent TLR10. TLR4 was only found in HaCaT cells, TLR1 to a higher degree in primary keratinocytes. In line with this, LPS induced mRNA expression of CD14 and TLR4 only in HaCaT cells. After stimulation with various TLR ligands, the NF‐κB‐activated chemokine interleukin‐8 (IL‐8) was measured. In primary keratinocytes and HaCaT cells the TLR3 ligand poly (I:C) was the most potent stimulator of IL‐8 secretion. The TLR ligands peptidoglycan, Pam3Cys and flagellin which bind to TLR2, TLR1/TLR2 heterodimer, and TLR5, respectively, also induced IL‐8 secretion, whereas no IL‐8 was induced by LPS, R‐848, loxoribine and cytosine guanine dinucleotide‐containing oligodeoxynucleotide. A corresponding pattern was found in the RelA NF‐κB translocation assay after ligand stimulation of primary keratinocytes. These studies provide substantial evidence for a functional TLR expression and signalling profile of normal human keratinocytes contributing to the antimicrobial defence barrier of human skin.
Reduced susceptibility to dextran sulphate sodium‐induced colitis in the interleukin‐2 heterozygous (IL‐2+/–) mouse
Authors: Sund Malin , Xu Li Li , Rahman Arman , Qian Bi‐Feng , Hammarström Marie‐Louise , Danielsson Åke 
Published: March 2005

SummaryMice homozygous for an inactivation of the interleukin‐2 (IL‐2) gene develop a T‐cell dependent colitis. Heterozygous (IL‐2+/–) mice are clinically healthy but have been shown to express reduced levels of IL‐2 in the colon. Splenocytes from the IL‐2+/– mice had a poorer proliferative response to polyclonal T‐cell activation and these mice have reduced numbers of intestinal regulatory T cells (CD4+ CD25+ cells) when compared to wild type mice. When exposed to dextran sulphate sodium (DSS) IL‐2+/– mice showed a markedly reduced susceptibility to DSS‐induced colitis. While DSS treatment caused a marked increase in both CD4+ and CD8+ colonic T cells expressing increased levels of IL‐2, IL‐4, and IL‐10 in wild type mice none of these changes were seen in IL‐2+/– mice. On the contrary, cytokine expression in intestinal T cells of IL‐2+/– mice was actually reduced after DSS treatment. These results suggest that reduced levels of IL‐2 leads to attenuated activation and function of intestinal T cells in IL‐2+/– mice and a failure to react adequately to DSS exposure.
Identification and characterization of a functional, alternatively spliced Toll‐like receptor 7 (TLR7) and genomic disruption of TLR8 in chickens
Authors: Philbin Victoria J. , Iqbal Muhammad , Boyd Yvonne , Goodchild Marianne J. , Beal Richard K. , Bumstead Nat , Young John , Smith Adrian L. 
Published: March 2005

SummaryBased upon the recognition of antiviral compounds and single stranded viral RNA the Toll‐like receptors TLR7 and TLR8 are suggested to play a significant role in initiating antiviral immune responses. Here we report the molecular characterization of the chicken TLR7/8 loci which revealed an intact TLR7 gene and fragments of a TLR8‐like gene with a 6‐kilobase insertion containing chicken repeat 1 (CR1) retroviral‐like insertion elements. The chicken TLR7 gene encodes a 1047‐amino‐acid protein with 62% identity to human TLR7 and a conserved pattern of predicted leucine‐rich repeats. Highest levels of chicken TLR7 mRNA were detected in immune‐related tissues and cells, especially the spleen, caecal, tonsil and splenic B cells. Alternative spliced forms of TLR7 mRNA were identified in chicken, mouse and human and expressed in similar tissues and cell types to the major form of chicken TLR7. The chicken TLR7+ HD11 cell line and fresh splenocytes produced elevated levels of interleukin‐1β (IL‐1β) mRNA after exposure to the agonists R848 and loxoribine. Interestingly, none of the TLR7 agonists stimulated increased type I interferon (IFN) mRNA whereas poly(I:C) (a TLR3 agonist) up‐regulated both chicken IFN‐α and chicken IFN‐β mRNA. In contrast, TLR7 agonists, particularly R848 and poly(U) stimulated up‐regulation of chicken IL‐1β, and chicken IL‐8 mRNAs more effectively than poly(I:C). Stimulation of chicken TLR7 with R848 was chloroquine sensitive, suggesting signalling within an endosomal compartment, as for mammalian TLR7. The deletion of TLR8 in galliforms, accompanied with the differential response after exposure to TLR7 agonists, offers insight into the evolution of vertebrate TLR function.
Tracking the dynamics of T‐cell activation in response to Salmonella infection
Authors: Ravindran Rajesh , McSorley Stephen J. 
Published: March 2005

SummaryDespite the current availability of Salmonella vaccines, typhoid fever remains a significant public health problem in developing countries. A greater understanding of T‐cell activation and the development of immunological memory during Salmonella infection should lead to the development of more effective prophylactic intervention. Here, we review recent literature on the initiation, expansion and memory development of T‐cell responses using the mouse model of typhoid. We pay particular attention to strategies for tracking T‐cell responses in vivo and ex vivo, and suggest models to integrate some these studies.
Mutation C677T in the methylenetetrahydrofolate reductase gene is associated with male infertility in an Indian population1
Authors: SINGH KIRAN , SINGH S. K. , SAH R. , SINGH INDU , RAMAN RAJIVA 
Published: March 2005

SummaryA mutation (C677T) in the gene, MTHFR, is known to increase susceptibility to various multifactorial disorders. In order to assess this single nucleotide polymorphism (SNP) as risk factor for idiopathic male infertility, a case–control study was done on an Indian population. DNA from 151 cases of non‐obstruction, idiopathic oligo‐/azoospermia and 200 fertile males (controls) was polymerase chain reaction amplified using site‐specific primers, and analysed for the mutation following HinfI‐digestion. Our results show a significantly increased frequency of CT heterozygotes among infertile patients (p value <0.04). More importantly, while there were no T homozygotes in the control population, six of 151 infertile cases were T homozygous. Considering that T allele occurs in very low frequency in the control population, 677T is clearly a risk factor for infertility in the Indian population. We contend that the same could also be true for African and Southeast Asian populations where the frequency of 677T is very low. The lack of similar association in western populations could be because of the overall dietary enrichment of folates, which could nullify or minimize the effect of this polymorphism.
Improvement of semen quality by nocturnal scrotal cooling in oligozoospermic men with a history of testicular maldescent
Authors: JUNG A. , SCHILL W. ‐B. , SCHUPPE H. ‐C. 
Published: March 2005

SummaryFor infertile men with a history of testicular maldescent only few therapeutic options exist beside assisted reproduction. The aim of our study was to evaluate the influence of nocturnal scrotal cooling on semen quality in such patients presenting with oligozoospermia. Twenty infertile men with a history of testicular maldescent and oligozoospermia were included for nocturnal scrotal cooling over 12 weeks for every night. To increase nocturnal periscrotal air circulation we used a membrane pump connected via plastic tubes to receptacles placed in both groins. Semen analysis was performed at the beginning of the cooling period and at weeks 4, 8 and 12. Another 20 infertile patients with a history of testicular maldescent and oligozoospermia were followed without specific treatment and served as a retrospectively built control group. Scrotal cooling at night by means of a perigenital air stream resulted in a scrotal temperature drop by 0.8 °C (median). A significant increase in sperm concentration and total sperm count was achieved by nocturnal cooling after 8 weeks (p < 0.01; p < 0.05; respectively) and 12 weeks (p < 0.01; p < 0.01; respectively). The improvement of sperm motility and sperm morphology was statistically insignificant. The present study suggests nocturnal scrotal cooling as a therapeutic option to improve semen quality. In a further controlled prospective study the influence on pregnancy rates should be evaluated.
Characterization of a critical region in the hormone binding domain of sperm progesterone receptor
Authors: SACHDEVA GEETANJALI , GADKAR SUSHAMA , SHAH CHIRAG ASHWINKUMAR , KHOLKUTE SANJIVA DATTATRAYA , PURI CHANDER PRAKASH 
Published: March 2005

SummaryOur previous studies have demonstrated that the ligand binding characteristics of the non‐genomic sperm progesterone receptor (sPR) are different from those of the conventional progesterone receptor (PR). Unlike PR, sPR does not bind efficiently to progesterone antagonists i.e. mifepristone (RU38486), onapristone (ZK98.299) etc. The present study was undertaken to determine the molecular composition of a region that plays a critical role in the interaction of the hormone‐binding domain (HBD) of sPR with progesterone and its antagonists. Detection, cloning and sequencing of the HBD region of sPR revealed its complete sequence homology to the corresponding region in the conventional PR. No nucleotide substitution/mutation/deletion, which could account for the differential antihormone binding, was observed in the HBD of sPR. The critical codon (nucleotide 3216–3218) in all three clones for the HBD of sPR was found to encode for glycine. This ruled out the possibility of steric hindrance because of the placement of amino acids with side chains at a critical position in the HBD, which may interfere in binding of sPR with antiprogestins. It is likely that the post‐transcriptional modifications contribute to the differential binding characteristics of sPR. This warrants future investigations to focus more on the characterization of the mature sPR protein.
A new and simple method to evaluate early membrane changes in frozen–thawed boar spermatozoa
Authors: PEÑA F. J. , SARAVIA F. , JOHANNISSON A. , WALGREN M. , RODRÍGUEZ‐MARTÍNEZ H. 
Published: March 2005

SummaryDetection of early changes in the sperm plasma membrane during cryopreservation is of utmost importance when designing freezing protocols and has previously been studied in the pig species using annexin‐V detection of phosphatidylserine translocation. In the present study we designed a new assay to detect these changes in boar spermatozoa, based on the slight increase of sperm membrane permeability occurring during the early stages of cryoinjury, using the combination of three fluorescent probes, SNARF‐1, YO‐PRO‐1 and ethidium homodimer. Four ejaculates from five different boars were frozen–thawed and flow cytometrically (FC) evaluated as paired samples. One of the samples was assayed using the annexin‐V/propidium iodide staining and the other sample was evaluated using the new triple staining. Using this combination of probes, four sperm subpopulations were easily detected: viable, with stable membranes (SNARF‐1 positive cells), and three with compromised membranes, one of YO‐PRO‐1+/Eth− cells, one ethidium homodimer+ spermatozoa and, finally spermatozoa stained both with YO‐PRO‐1 and ethidium homodimer (YO‐PRO‐1+/Eth+). The latter three categories corresponded to dead spermatozoa, but with different degree of membrane damage, being YO‐PRO+/Eth− an earlier stage of membrane destabilization, (manifested by an increase in membrane permeability, while still maintaining membrane integrity) than YO‐PRO+/Eth+. A method agreement analysis between both methods was performed revealing good agreement, although the percentage of live cells was 9.44% larger for the triple stain than the annexin‐V assay. The new assay stained all sperm sub‐populations present in the sample, making it especially suitable for both fluorescence microscopy and flow cytometry, facilitating the exclusion of debris and egg‐yolk particles when using FC.
The impact of mitochondrial genetics on male infertility
Authors: JOHN JUSTIN C. ST. , JOKHI ROOBIN P. , BARRATT CHRISTOPHER L. R. 
Published: March 2005

SummaryHuman mitochondrial DNA (mtDNA) encodes 13 of the polypeptides associated with the process of oxidative phosphorylation (OXPHOS), the cells most important ATP generating pathway. Until recently, the effects of mtDNA rearrangements on male fertility have been largely ignored. However, it is becoming increasingly evident that both point mutations and large‐scale deletions may have an impact on sperm motility and morphology. We discuss the implications of these rearrangements in the context of the clinical setting. We further discuss the possible consequences resulting from the transmission of sperm mtDNA deletions to the offspring. The role of nucleo‐cytoplasmic interaction is investigated in the context of nuclear transcription and replication factors that regulate mtDNA transcription and replication.
Ontogenetic profile and thyroid hormone regulation of type‐1 and type‐8 glucose transporters in rat Sertoli cells
Authors: CAROSA ELEONORA , RADICO CARLA , GIANSANTE NADIA , ROSSI SIMONA , D'ADAMO FABIO , DI STASI SAVINO M. , LENZI ANDREA , JANNINI EMMANUELE A. 
Published: March 2005

SummaryThe glucose transporters (GLUTs) gene encode glycoproteins responsible for facilitating transfer of glucose across plasma membrane. In testis, different members of this family are present. In particular the main GLUT mRNA expression within the adult testis is the type 8, while type 1 is more expressed in prepubertal testis. Thyroid hormone, which receptors and function have been characterized in the testis, plays a crucial role in the cellular energetic metabolism. In fact, in the immature Sertoli cells, GLUT1 is up regulated by l‐triiodothyronine (T3). The aim of this paper is to investigate the expression profile of GLUT1 and GLUT8 in the testis during development and in adulthood and analyse the role of T3 on their expression. To analyse the expression of GLUT8 and GLUT1 we performed Northern blot and RT‐PCR experiments in the whole testis and in Sertoli cells from rats of different ages. Treatments in vivo and in vitro with T3 were used to study the effect of thyroid hormones on GLUT1 and GLUT8 expression. The activity of the rat GLUT1 promoter and its regulation by T3 was studied with transient transfections in gonadal and non‐gonadal cell lines and in primary Sertoli cell cultures. GLUT8 is expressed at a low level in the prepubertal testis and Sertoli cells and does not appear to be under T3 control. GLUT1 is the predominant form in immature Sertoli cells. The effect of T3 on its mRNA accumulation was quantified and confirmed by RT‐PCR (control: 0.65 ± 0.17; T3: 1.23 ± 0.04, arbitrary units, p < 0.05). However, transfection experiments showed that T3 does not directly regulate GLUT1 promoter in any cell line tested. This is confirmed by the evidence that, upon extensive analysis, the rat GLUT1 promoter and the first intron sequence do not shows any thyroid responsive elements. Our data demonstrate that GLUT1 and GLUT8 are both expressed in prepubertal testis, but only GLUT1 is regulated by T3. In addition, we found that the effect of T3 cannot be attributed to its action on GLUT1 promoter.
Control of Phytophthora cryptogea in the hydroponic forcing of witloof chicory with the rhamnolipid‐based biosurfactant formulation PRO1
Authors: De Jonghe K. , De Dobbelaere I. , Sarrazyn R. , Höfte M. 
Published: March 2005

Rhamnolipids, extracellular metabolites of Pseudomonas aeruginosa with surfactant properties, proved to be very effective in controlling the spread of brown root rot disease caused by Phytophthora cryptogea in the hydroponic forcing system of witloof chicory (Cichorium intybus var. foliosum). The biosurfactant was applied as the product PRO1, a formulation of 25% rhamnolipids in oil. Both an in vitro screening and in vivo experiments in a mini‐hydroponic system demonstrated the ability of PRO1 to control brown root rot. A 25 µg mL−1 rhamnolipids nutrient solution was enough to obtain good control of an artificial infection with a zoospore suspension of P. cryptogea. The biosurfactant PRO1 performed well in a semicommercial system under growers’ conditions. A treatment of 25 µg mL−1 rhamnolipids (100 µg mL−1 PRO1) reduced the disease incidence significantly in two independent experiments. However, PRO1 was not effective when a mycelial suspension was used as inoculum. Rhamnolipids have good potential to limit the spread of P. cryptogea in the hydroponic forcing system of witloof chicory, and can be used as a preventive measure against brown root rot.
Comparison of populations of the wilt pathogen Ceratocystis albifundus in South Africa and Uganda
Authors: Barnes I. , Nakabonge G. , Roux J. , Wingfield B. D. , Wingfield M. J. 
Published: March 2005

Ceratocystis albifundus is an important fungal pathogen of Acacia mearnsii trees in South Africa. In a previous study, a high level of gene diversity was demonstrated in a South African population of C. albifundus. This, together with the occurrence of the pathogen on native Protea species and its exclusive occurrence in South Africa, led to the hypothesis that C. albifundus is probably native to that country. More recently, C. albifundus has been reported from A. mearnsii in south‐western Uganda. The aim of this study was to compare the populations of C. albifundus from Uganda and South Africa based on genetic diversity, population structure and possible gene flow. This was achieved using codominant microsatellite markers developed for the closely related species Ceratocystis fimbriata. Available isolates for comparison were from six different areas of South Africa and six jungle stands in Uganda. Eight of the 11 available markers amplified loci in C. albifundus. Gene diversity was higher in the Ugandan population, but genotypic diversity was greater for the South African isolates. There were no common genotypes between the two populations and they shared only 22% of the total alleles. The populations were genetically isolated from each other and highly substructured within. There was no association between isolates collected from the same geographic locations, and gene flow between the two populations was low. Results suggest that C. albifundus was probably not introduced into Uganda from South Africa but rather that an ancestral population, yet to be discovered, is the source of both populations.
Ustilospores of Tilletia ehrhartae, a smut of Ehrharta calycina, are common contaminants of Australian wheat grain, and a potential source of confusion with Tilletia indica, the cause of Karnal bunt of wheat
Authors: Pascoe I. G. , Priest M. J. , Shivas R. G. , Cunnington J. H. 
Published: March 2005

Australian wheat consigned for export from Australian ports was surveyed in March 2004 using a national diagnostic protocol for detection and identification of Tilletia indica. No ustilospores of T. indica were detected, confirming previous surveys which have failed to detect T. indica in Australia. However, the survey detected moderate levels of the common smuts Tilletia caries (syn. Tilletia tritici), Tilletia laevis and Urocystis agropyri, and very low levels (average fewer than six ustilospores per 150 g sample) of an unidentified dark, tuberculate‐spored Tilletia in ≈ 60% of samples tested. Comparison with herbarium specimens enabled identification of the majority of the tuberculate ustilospores as Tilletia ehrhartae, a smut fungus known to infect only Ehrharta calycina (perennial veldt grass) and which is common in southern Australia. A smaller number of tuberculate smut ustilospores were identified as Tilletia walkeri, a smut of Lolium spp. recorded in Australia but apparently uncommon. Both T. ehrhartae and T. walkeri bear sufficient resemblance to T. indica for misidentifications to be possible where only a very few ustilospores are seen, although T. ehrhartae ustilospores are always <25 µm in diameter. The frequent presence of ustilospores of both T. ehrhartae and T. walkeri as contaminants of Australian wheat grain exports has significance for diagnosticians testing Australian export wheat, as it demonstrates the potential for tuberculate ustilospores of species other than those covered in existing diagnostic protocols to be misidentified as T. indica. This paper describes T. ehrhartae in detail, and provides criteria for its differentiation from T. indica, T. walkeri and some other species.
Erwinia amylovora populations resistant to oxolinic acid in Israel: prevalence, persistence and fitness
Authors: Kleitman F. , Shtienberg D. , Blachinsky D. , Oppenheim D. , Zilberstaine M. , Dror O. , Manulis S. 
Published: March 2005

Oxolinic acid (OA) has been the only bactericide used against fire blight in pear and quince orchards in Israel since 1998. OA‐resistant Erwinia amylovora strains (Ea‐OAR) were detected in several orchards in two restricted areas in the northern Galilee region during 1999–2001. In the following years, resistant strains could not be detected in some of these locations. Documenting the fate of Ea‐OAR strains in commercial orchards at eight sites in northern Israel during 2000/03 revealed that the resistant population appeared irrespective of the number of sprays applied and the severity of the disease. The persistence of the Ea‐OAR populations varied from site to site, ranging from 4 to 20 months; these differences could be attributed to the fire blight management activities of growers. Comparative studies on the fitness of Ea‐OAR and E. amylovora strains sensitive to OA (Ea‐OAS) were conducted in vitro and in planta using two strains of each group. In four of the six comparisons, disease incidence on detached blossoms inoculated with Ea‐OAS was significantly higher than that on blossoms inoculated with Ea‐OAR. In two experiments conducted on 8‐year‐old pear trees grown under netting, the colonization of Ea‐OAS in blossoms, annual shoots and perennial spurs was significantly higher than that of the Ea‐OAR. In two experiments conducted on 2‐year‐old trees grown under netting in an experimental station, the incidence of shoots exhibiting fire blight symptoms and the rate of symptom progress within the branches were significantly higher in trees inoculated with Ea‐OAS than in those inoculated with Ea‐OAR. The results of this study suggest that OA‐resistant E. amylovora strains have lower fitness than wild‐type strains. These findings may have implications for fire blight management.