bluuebunny/crossref_metadata_2025_split huggingface dataset:
Toll‐like receptor‐9 induced by physical trauma mediates release of cytokines following exposure to CpG motif in mouse skin
Authors: Liu Ling , Zhou Xiaohui , Shi Jianying , Xie Xin , Yuan Zhenghong 
Published: October 2003

SummaryThe skin plays a crucial role in defence against microbial infection via the innate immune system, but the exact cellular mechanisms of this defence are not well understood. Toll‐like receptors (TLRs), a newly recognized 10‐member family of vertebrate pattern recognition receptors (PRRs), have been identified as crucial mediators of innate immune recognition. Although both TLR2 and TLR4 have been detected in normal human skin, little is known about the expression and function of TLR9, a CpG motif receptor, in skin. In this study, reverse transcription–polymerase chain reaction and in situ hybridization analysis were used to identify TLR9 mRNA expression in mouse skin. Results showed that TLR9 mRNA was not detected in normal mouse skin, but its presence in skin could be induced by intradermal injection of either normal saline, or the bacteria‐based CpG motif in a time‐ and volume‐dependent manner. Furthermore, intradermal injection of CpG motif induced increased expression of mRNAs for proinflammatory cytokines such as interleukin (IL)‐1, IL‐6, IL‐12 and tumour necrosis factor α. This suggests that TLR9, while not present basally in skin, can be induced by physical trauma and then mediate responses to CpG motif. In conclusion, TLR9 is involved in the innate immune response in skin and that it may have a role in secondary inflammation following physical trauma such as epidermal damage or microbial infection. This role of TLR9 may help explain the previously identified enhancement of DNA immunization by CpG ODN.
Lymphocyte populations in human lymph nodes. Alterations in CD4+ CD25+ T regulatory cell phenotype and T‐cell receptor Vβ repertoire
Authors: Battaglia Alessandra , Ferrandina Gabriella , Buzzonetti Alessia , Malinconico Paolo , Legge Francesco , Salutari Vanda , Scambia Giovanni , Fattorossi Andrea 
Published: October 2003

SummaryHere we provide a description of lymphocyte populations in human lymph nodes (LN) with a special emphasis on the CD4+ lymphocyte population constitutively expressing CD25 at a high level and endowed with immunoregulatory properties [T regulatory (Treg) cells]. Lymph nodes were analysed by multicolour flow cytometry in parallel with correspondent peripheral blood (PB). Immunomagnetically purified Treg cells were tested for anergy and suppressive activity in a CD3/T‐cell receptor (TCR)‐driven proliferation assay. Compared to PB, there was a reduced T/B lymphocyte ratio in LN. Both LN and PB contained a similar proportion of CD4+ lymphocytes but, conversely, CD8+ lymphocytes were less represented in PB, with a consequent increase in the ratio of CD4+/CD8+ natural killer cells were < 2% (PB range 6–22%). No significant differences existed in the frequency of the other lymphocyte subpopulations examined (naïve‐type CD4+ and CD8+ lymphocytes, activated B and CD4+ lymphocytes, and effector‐type CD8+ lymphocytes). LN and PB contained similar percentages of CD4+ lymphocytes constitutively expressing intermediate or high levels of CD25. CD4+ CD25++ cells constitutively coexpressed high levels of CD152 and were therefore identified as Treg cells. Treg cells in LN and PB differed in terms of CD45RB, HLA‐DR, CD45RO, and CD62L expression. Also the TCRVβ repertoire diverged between Treg cells from LN and PB. Similar to Treg cells from PB, Treg cells from LN were anergic and efficiently inhibited other CD4+ and CD8+ lymphocyte proliferation. This study extends the information on the diversities in lymphocyte composition between human LN and PB, and reports for the first time a description of the phenotypic and functional characteristics of Treg cells in human LN, highlighting the importance of the LN microenvironment in shaping the surface phenotype of Treg cells.
Embryonic age influences the capacity for cytokine induction in chicken thymocytes
Authors: Peters Michelle A. , Browning Glenn F. , Washington Elizabeth A. , Crabb Brendan S. , Kaiser Pete 
Published: October 2003

SummaryThymocyte responses to functional activation are of relevance to the evaluation of the efficacy of in ovo immunotherapies and vaccines in chickens. In this study we have demonstrated differences in chicken thymocyte responses according to developmental age. RNA samples from stimulated and unstimulated chicken thymocytes were assayed for messenger RNA encoding the cytokines interleukin‐1β (IL‐1β), IL‐2, interferon‐α (IFN‐α), IFN‐β, IFN‐γ and transforming growth factor‐β4 (TGF‐β4), and also components of the major histocompatibility complex (MHC), β2‐microglobulin (β2M) and the MHC class I α‐chain (MHC IA). At embryonic day 14 thymocytes were least responsive to functional activation and differences existed even between thymocyte populations at embryonic day 18 and day 1 post‐hatch. The duration of proliferation in response to stimulation was found to increase with increasing embryonic age. Mitogen stimulation of embryonic day 18 and day 1 post‐hatch thymocytes induced up‐regulation of IFN‐γ, IL‐1β and TGF‐β transcripts, and down‐regulation of IFN‐α, IFN‐β and IL‐2 transcripts, with a higher induction of IFN‐γ, IL‐1β and TGF‐β transcripts in more immature T‐cell‐receptor‐negative (TCR−) than TCR+ (TCR1+, TCR2+, or TCR3+) subsets. In contrast, in the mouse and human, both mature and immature thymocytes respond to mitogen stimulation with up‐regulation of IL‐2. Thymocytes from embryonic day 14 chicks responded to mitogen with a short burst of unsustained proliferation, and transcriptional down‐regulation of the cytokines IL‐2, IL‐1β, IFN‐α, IFN‐β and IFN‐γ. These results suggest that embryonic day 14 thymocytes are largely unresponsive to mitogen. Transcripts encoding TGF‐β and type I interferons (IFN‐α and IFN‐β) were constitutively expressed at high levels in very early thymocytes at embryonic day 14. Thymocytes at embryonic days 14 and 18 and day 1 post‐hatch responded to mitogen stimulation with up‐regulation of MHC IA transcript. The pattern of β2M transcription following mitogen stimulation was distinct from that of the globally up‐regulated MHC IA transcript, with up‐regulation of β2M transcription observed at embryonic day 18 and day 1 post‐hatch but not at embryonic day 14. In thymocyte subsets, up‐regulation of β2M transcription was found to be specific to the CD8+ TCR+ population. The balance of responses in the embryonic thymus suggests that at all stages thymocytes have a reduced capacity for activation in comparison to mature thymocyte populations.
In vivo expression of interleukin‐8, and regulated on activation, normal, T‐cell expressed, and secreted, by human germinal centre B lymphocytes
Authors: Sims‐Mourtada Jennifer C. , Guzman‐Rojas Liliana , Rangel Roberto , Nghiem Dat X. , Ullrich Stephen E. , Guret Christiane , Cain Kelly , Martinez‐Valdez Hector 
Published: October 2003

SummaryT‐cell homing within germinal centres (GCs) is required for humoral B‐cell responses. However, the mechanisms implicated in the recruitment of T cells into the GC are not completely understood. Here we show, by immunohistology, and Northern and Western blots, that in vivo human GC B lymphocytes can express CxC and CC chemokines. Moreover, B‐cell subset‐specific experiments reveal that interleukin (IL)‐8 and regulated on activation, normal, T‐cell expressed, and secreted (RANTES) are predominantly expressed by GC centroblast and centrocytes, suggesting that chemokine expression is essential at stages in which B‐lymphocytes engage in active antigen‐dependent interactions with T lymphocytes. In keeping with this hypothesis, we show that the T cells recruited into the GC correlatively express the receptors for IL‐8 and RANTES. We propose that chemokine expression is a key B‐cell function that facilitates T‐lymphocyte recruitment into the GCs and supports cognate B‐cell : T‐cell encounters. Moreover, our data are consistent with the impaired homing of T cells to secondary lymphoid organs in mice that are either deficient in CC and CxC chemokines or their receptors.
Induction of protective immunity against a Chlamydia trachomatis genital infection in three genetically distinct strains of mice
Authors: Pal Sukumar , Peterson Ellena M. , De La Maza Luis M. 
Published: October 2003

SummaryTo establish the feasibility of inducing a protective immune response against a chlamydial genital infection in animals with different genetic backgrounds, groups of C3H/HeN (H‐2k), BALB/c (H‐2d) and C57BL/6 (H‐2b) mice, were immunized intranasally with elementary bodies (EB) of the Chlamydia trachomatis mouse pneumonitis biovar. Following the intranasal immunization strong Chlamydia‐specific humoral and cell‐mediated immune (CMI) responses were detected in the three strains of mice. Eight weeks following immunization the animals were challenged with C. trachomatis in the genital tract. Vaginal cultures showed that the three strains of mice immunized with EB were significantly protected in comparison to the sham immunized animals. To determine the ability of this immunization protocol to protect against infertility six weeks after the genital challenge the animals were mated. Mice of the three strains immunized with EB showed significant protection as demonstrated by the number of animals that were fertile, and the number of embryos present in their uterine horns, in comparison to the sham immunized mice.
Resting CD8 T cells recognize β‐galactosidase expressed in the immune‐privileged retina and mediate autoimmune disease when activated
Authors: McPherson Scott W. , Yang Jing , Chan Chi‐Chao , Dou Chunzhi , Gregerson Dale S. 
Published: October 2003

SummaryAlthough the expression of class II major histocompatibility complex (MHC) in retina is extremely low, it is an established fact that activated CD4 T cells, specific for retinal antigens (Ags), mediate experimental autoimmune uveoretinitis (EAU). Conversely, CD8 T cells have not been shown to recognize Ag in the retina. This study investigated whether retinal‐specific Ags are detected by class I MHC‐restricted CD8 T cells. Using a CD8 T‐cell clone (β3) specific for an immunodominant epitope of β‐galactosidase (β‐gal), local Ag recognition was shown by transfer of activated β3 cells into β‐gal transgenic (Tg) mice expressing β‐gal in the retina (hi‐arr‐β‐gal mice), or in the brain and eye (GFAP‐β‐gal mice). β‐gal‐positive photoreceptor cells were damaged in the retina of hi‐arr‐β‐gal mice, and anterior segment disease was found in the eyes of GFAP‐β‐gal mice. Ag recognition by resting CD8 T cells was also evaluated. Recovery of 5(6)‐carboxyfluorescein diacetate N‐succinimidyl ester (CFSE)‐labelled β3 cells from hi‐arr‐β‐gal mice was slightly decreased compared to recovery from B10.A mice, while recovery from GFAP‐β‐gal mice was transiently increased. Conversely, recovery of CFSE− cells increased in hi‐arr‐β‐gal mice, consistent with an Ag‐dependent response. The CFSE content of the CFSE+ population was unchanged relative to β3 cells recovered from controls. Intracellular cytokine responses of β3 cells recovered from hi‐arr‐β‐gal and GFAP‐β‐gal mice correlated with the number of cells recovered, regardless of CFSE content. Even though their production of interferon‐γ and tumour necrosis factor‐α was affected little by transfer into hi‐arr‐β‐gal recipients, the ability of β3 cells to mediate delayed‐type hypersesitivity was inhibited in hi‐arr‐β‐gal mice. These results show that resting CD8 T cells are affected by the presence of Ag that originates in retina and, when activated prior to transfer, mediate pathogenic autoimmunity against retinal and other ocular targets.
Disorganization of actin bundles of ectoplasmic specialization
Authors: Sakai Yasuhiro , Yamashina Shohei 
Published: October 2003

The degradation of ectoplasmic specialization consisting of bundles of actin sandwiched between the plasma membrane and endoplasmic reticulum of the Sertoli cell, occurs just before spermiation. For elucidation of the processes involved in this degradation, changes in fibrous actin of the rat testis were analyzed using BODIPY‐phalloidin by fluorescence and electron microscopy.Before step 17, the fluorescence of BODIPY‐phalloidin was evenly distributed around the spermatid head. When the spermatids became positioned at the luminal surface, the fluorescence had condensed on the concave side of the spermatid head. At step 19, lines of fluorescence distributed at regular intervals projected at right angles from the head. Ultrastructural observation showed that the tubulobulbar complex was formed at step 19 and electron‐dense material accumulated around thin tubules of the tubulobulbar complex. Immunohistochemical examination of BODIPY‐phalloidin showed that the electron dense materials around the thin tubules of the tubulobulbar complex had the capacity to bind to phallotoxin. Therefore the pattern of fluorescence in the spermatid at step 19 corresponds to that of the tubulobulbar complex.Actin bundles of the ectoplasmic specialization would thus appear to de‐polymerize into actin monomers via electron dense materials around the thin tubules of the tubulobulbar complex. The tubulobulbar complex may contribute to the disorganization of actin bundles.
Characterization and localization of tropomyosin proteins in Xenopus embryos with specific antibodies
Authors: Tanaka Tetsuya S. , Tatsuta Takashi , Ikenishi Kohji 
Published: October 2003

In the process of monoclonal antibody (mAb) production against the 38kDa protein which is lacking in the gastrula arrested mutant embryos in Xenopus we incidentally obtained two kinds of mAb (designated as B11 and 2D10 antibodies, respectively) recognizing tropomyosin (TM) proteins in Xenopus embryos. The characterization of the corresponding antigens to those mAb was performed by immunoblotting and silver staining for two‐dimensional (2‐D) gels in the present study. The localization of the antigens in Xenopus embryos was also investigated by fluorescent microscopy.By 2‐D immunoblots with those mAb, three distinct protein spots or TM isoforms were recognized in Xenopus embryos; a 38 kDa spot with a pl of approximately 4.8 reacted with both antibodies in embryos at stages later than the mid‐tailbud (stage 28) and two 30 kDa spots, which are probably isomers, with a pl of approximately 4.8 were detected with 2D10 antibody in embryos at stages extending from the fertilized to the mid‐neurula (stage 20). By immunofluorescent microscopy, B11 antibody was shown to react mainly with muscle cells and their precursor cells. In contrast, 2D10 antibody stained the cytoplasm of almost all cells in embryos at stages from the fertilized to the tadpole.Judging from the results obtained with immunoblotting and fluorescent microscopy, it is likely that the 38 kDa spot is a skeletal muscle TM isoform and the two 30 kDa spots are non‐muscle TM isoforms.
Timing of initiation of muscle‐specific gene expression in the ascidian embryo precedes that of developmental fate restriction in lineage cells
Authors: Satou Yutaka , Kusakabe Takehiro , Araki Lsato , Satoh Noriyuki 
Published: October 2003

The lineage of muscle cells in the ascidian embryo is well documented. Of the B‐line blastomeres, B7.4 cells become restricted to give rise to muscle at the 44‐cell stage, whereas B7.8 and B7.5 cells become restricted to muscle at the 64‐cell stage. In this study we addressed three issues concerning the timing of initiation of muscle‐specific structural gene transcription: (i) whether the initiation of the transcription of muscle‐specific genes is correlated to that of developmental fate restriction; (ii) if there is a relationship in the timing, whether it is applicable to B7.4‐, B7.5‐ and B7.8‐sublineages; and (iii) whether two muscle‐specific genes, one for actin and the other for the myosin heavy chain, show the same pattern of temporal expression.Whole‐mount in situ hybridization revealed that hybridization signal of actin mRNA was first evident in B6.2 (the progenitor of B7.4) at the 32‐cell stage, in B7.8 at the 64‐cell stage, and in B7.5 around the 76‐cell stage, respectively, suggesting that the transcription of the gene is initiated prior to the developmental fate restriction in the B7.4‐sublineage. Presence of actin transcripts in the 32‐cell embryos was confirmed by means of reverse transcription‐polymerase chain reaction (RT‐PCR). The myosin heavy chain gene was expressed in the same manner as the actin gene. Therefore, we concluded that the timing of initiation of transcription of the muscle‐specific genes is not always associated with that of the fate restriction and that the temporal profile of the gene expression differs among the sublineages, even though two different genes have the same expression profile.
Possible involvement of an acylation mechanism in thalidomide‐induced teratogenesis of the newt (Pleurodeles waltl.)
Authors: Audit C. O. , Eger K. , Aimar C. 
Published: October 2003

An acylation reaction of biological polyamines by thalidomide has been postulated to explain the teratogenic activity of this drug (Fabro et al. 1965). In a further study, thalidomide has been reported to acylate polyamines at physiological pH; the teratogenic activity of this drug appears to be linked to its high acylating power towards polyamines (Audit 1994).In the present study, the action of the thalidomide molecule and its two chemical moieties (phthalimide and glutarimide rings) on Pleurodeles embryonic development has been investigated. The phthalimide moiety, which displays acylating activity, appears to generate Pleurodeles teratogenesis. The occurrence of a correlation between acylating activity and teratogenicity was confirmed using homothalidomide and partially hydrolyzed thalidomide. The glutarimide moiety has been found to act as an enhancer of phthalimide activity and to cause moderate alterations of newt development.As the acylation of polyamines by thalidomide would deprive the embryo of these essential compounds, the effects of polyamine biosynthesis inhibitors have been compared to those of thalidomide. Both thalidomide and polyamine antimetabolites altered the early cleavage process of the Pleurodeles egg and arrested early development.
Transurethral incision of ureteroneocystostomy strictures in kidney transplant recipients
Authors: Katz R. , Pode D. , Gofrit O.N. , Shenfeld O.Z. , Landau E.H. , Golijanin D. , Shapiro A. 
Published: October 2003

OBJECTIVESTo report the treatment of patients who presented with vesico‐ureteric stricture after kidney transplantation, using a minimally invasive endourological approach.PATIENTS AND METHODSPatients (10 men and four women, mean age 34 years, range 22–55) were assessed at presentation by serum creatinine level, ultrasonography and intravenous pyelography when the serum creatinine level was < 200 µmol/L. When there was hydronephrosis of the allograft a percutaneous antegrade pyelogram was taken, followed by inserting a nephrostomy. After decompression a stent nephrostomy was passed into the bladder and the strictures at the vesico‐ureteric junction incised along the stent during cystoscopy.RESULTSAll 14 patients were treated endourologically by an endoscopic incision through the bladder; 13 fared well and one died from sepsis and transplantation problems. The mean follow‐up was 8 months.CONCLUSIONSSimple incision of the stricture via cystoscopy was safe and effective, and succeeded in most patients. The endourological management of ureteric lesions is feasible and is currently our first‐line management of ureteric complications after kidney transplantation.
The association between lower urinary tract symptoms and erectile dysfunction in four centres: the UrEpik study
Authors: Boyle P. , Robertson C. , Mazzetta C. , Keech M. , Hobbs R. , Fourcade R. , Kiemeney L. , Lee C. ,   the UrEpik Study Group
Published: October 2003

OBJECTIVETo report a large‐scale multinational investigation of lower urinary tract symptoms (LUTS) and sexual function, designed to investigate the independent association between them, as recent small‐scale epidemiological studies suggest an association between benign prostatic hyperplasia and sexual dysfunction; both conditions are strongly associated with age and no study has been able to exclude age as a confounding factor in this relationship.SUBJECTS AND METHODSCulturally and linguistically validated versions of standard questionnaires were used to estimate the prevalence of LUTS (using the International Prostate Symptom Score, IPSS) and erectile dysfunction (ED) score (using O’Leary's Sexual Function Inventory) in regions of the UK (Birmingham), the Netherlands (Boxmeer), France (Auxerre) and Korea (Seoul). In each centre, stratified random samples were selected from population registers to provide representative samples of the population of men aged 40–79 years (and their partners) in each community. Direct interviews were held in Seoul and postal questionnaires used in the three European centres. The samples were selected randomly, providing representative samples in each community.RESULTSIn all, 4800 men and 3674 women responded; the response rates among men were 77% in Boxmeer, 21% in Auxerre, 42% in Birmingham and 65% in Seoul. The overall prevalence of ED for men aged 40–79, estimated as an ED score of 0–4, was 21.1%. There was evidence of a linear increase with age (P < 0.001) and the pattern was very similar in the four centres. From the weighted logistic model, there was evidence of an association between sexual dysfunction, other self‐reported diseases and lifestyle. From the ED score, after adjusting for age and country, men with diabetes were more likely to score of 0–4 (odds ratio 1.57, 95% confidence interval 1.09–2.25), as were those with high blood pressure (1.38, 1.09–1.75) and with an IPSS of 8–35 (1.39, 1.10–1.74). For lifestyle, smokers were more likely to score 0–4 (1.54, 1.23–1.92), while physical activity during leisure time was slightly associated with a reduction in the chance of scoring 0–4 (0.87, 0.77–0.99). The analysis gave the same results when repeated using self‐reported ED instead of the dichotomised score.CONCLUSIONSED is clearly age‐related and a problem for a large proportion of men in the community. It can have a profound impact on the quality of life of the man and on his partner. Were all men with this problem to seek medical help there would be a large burden on healthcare systems. There are cultural and age effects on the assessment of this problem.