Human and mouse perforin are processed in part through cleavage by the lysosomal cysteine proteinase cathepsin L

Summary Infection of mice with the gastrointestinal nematode Trichuris muris represents a valuable tool to investigate and dissect intestinal immune responses. Resistant mouse strains respond to T. muris infection by mounting a T helper type 2 response. Previous results have shown that CD4⁺ T cells play a critical role in protective immunity, and that CD4⁺ T cells localize to the infected large intestinal mucosa to confer protection. Further, transfer of CD4⁺ T cells from immune mice to immunodeficient SCID mice can prevent the development of a chronic infection. In the current study, we characterize the protective CD4⁺ T cells, describe their chemokine receptor expression and explore the functional significance of these receptors in recruitment to the large intestinal mucosa post‐T. muris infection. We show that the ability to mediate expulsion resides within a subpopulation of CD4⁺ T cells marked by down‐regulation of CD62L. These cells can be isolated from intestine‐draining mesenteric lymph nodes (MLN) from day 14 post‐infection, but are rare or absent in MLN before this and in spleen at all times post‐infection. Among CD4⁺ CD62L^low MLN cells, the two most abundantly expressed chemokine receptors were CCR6 and CXCR3. We demonstrate for the first time that CD4⁺ CD62L^low T‐cell migration to the large intestinal mucosa is dependent on the family of Gα_i‐coupled receptors, to which chemokine receptors belong. CCR6 and CXCR3 were however dispensable for this process because neutralization of CCR6 and CXCR3 did not prevent CD4⁺ CD62L^low cell migration to the large intestinal mucosa during T. muris infection.     

Physiological costs and temporo-spatial parameters of walking on a treadmill vary with body weight unloading and speed in both healthy young and older women

Walking on a treadmill with Body Weight Unloading (BWU), which has been successfully used on patients with neurological conditions, may also be used as a training tool to increase walking speed in healthy individuals. We hypothesised that BWU enables individuals to walk at a faster speed on a treadmill than they would do in normal gravity conditions without increasing their effort and with an increase in both stride length (SL) and stride frequency (SF). Oxygen uptake, heart rate (HR), SL and SF of six older women (mean ± SD; 70 ± 4 years) and six young women (26 ± 3 years) were measured during treadmill walking at three self-selected speeds (comfortable, slow and fast) and three different percentages of BWU (0, 20 and 40%). No significant differences were found between the groups in any self-selected walking speeds and any of the other variables. The combined data of the two groups showed that walking energy cost per unit of time (WECt) and HR at fast speed with 40% of BWU (258 ± 60 J kg⁻¹ min⁻¹ and 95 ± 15 beats min⁻¹, respectively) were similar to those measured at comfortable speed with no BWU (273 ± 47 J kg⁻¹ min⁻¹ and 101 ± 16 beats min⁻¹, respectively). Also SL and SF increased significantly with speed (P < 0.017) at any given percentage of BWU. The results suggest that 40% of BWU enables both young and older women to walk at a faster speed on a treadmill without increasing their effort and with an increase in both SL and SF.     

Three cases of pancreatic serous cystadenoma and endocrine tumour

AIMS: To report three cases of serous cystadenoma and endocrine tumour in the same pancreas, to review the literature and to evaluate the clinicopathological features of the tumours. Cases: Three women (71, 57 and 31 years old) were admitted to hospital, two for diseases unrelated to the pancreas and the third for increasing obstructive jaundice in von Hippel-Lindau disease. Preoperative examination showed two distinct lesions in the first patient and only cystic lesions in the other two. RESULTS: Histological examination of the pancreas showed one serous oligocystic adenoma associated with a benign, well-differentiated endocrine tumour, one serous oligocystic adenoma associated with an endocrine microadenoma, and a von Hippel-Lindau-related cystic neoplasm with a well-differentiated endocrine carcinoma. CONCLUSIONS: Serous cystadenoma associated with endocrine tumour shows some clinicopathological differences with respect to the two tumours considered separately, and with respect to von Hippel-Lindau-related cases, although there is no convincing evidence at present to justify considering this association as a separate entity.     

Continuous axial contraction wave in the free wall of the guinea pig left ventricle

We studied the variation from the simultaneous contraction of the normal left ventricle (LV). The pattern of the contraction along the LV long axes was assessed on the LV free wall on seven guinea pig hearts in situ with ultra fast video system and epicardial markers by means of the latitude and the size of the areas defined by markers. We found that the contraction occurs as a continuous contraction wave from the apex towards the base, which might yield functional adaptation of these two regions to diastolic and systolic function, respectively.     

Recommendations for the classification of group A rotaviruses using all 11 genomic RNA segments

Recently, a classification system was proposed for rotaviruses in which all the 11 genomic RNA segments are used (Matthijnssens et al. in J Virol 82:3204-3219, 2008). Based on nucleotide identity cut-off percentages, different genotypes were defined for each genome segment. A nomenclature for the comparison of complete rotavirus genomes was considered in which the notations Gx-P[x]-Ix-Rx-Cx-Mx-Ax-Nx-Tx-Ex-Hx are used for the VP7-VP4-VP6-VP1-VP2-VP3-NSP1-NSP2-NSP3-NSP4-NSP5/6 encoding genes, respectively. This classification system is an extension of the previously applied genotype-based system which made use of the rotavirus gene segments encoding VP4, VP7, VP6, and NSP4. In order to assign rotavirus strains to one of the established genotypes or a new genotype, a standard procedure is proposed in this report. As more human and animal rotavirus genomes will be completely sequenced, new genotypes for each of the 11 gene segments may be identified. A Rotavirus Classification Working Group (RCWG) including specialists in molecular virology, infectious diseases, epidemiology, and public health was formed, which can assist in the appropriate delineation of new genotypes, thus avoiding duplications and helping minimize errors. Scientists discovering a potentially new rotavirus genotype for any of the 11 gene segments are invited to send the novel sequence to the RCWG, where the sequence will be analyzed, and a new nomenclature will be advised as appropriate. The RCWG will update the list of classified strains regularly and make this accessible on a website. Close collaboration with the Study Group Reoviridae of the International Committee on the Taxonomy of Viruses will be maintained.     

Differential effects of osmotic and SSR149415 challenges in maternally separated and control rats: The role of vasopressin on spatial learning

Maternal separation (MS) has been demonstrated to up-regulate the hypothalamic vasopressin (VP) system. Intracerebrally released VP has been demonstrated to affect several types of animal behaviour, such as active/passive avoidance, social recognition, and learning and memory. However, the role of VP in spatial learning remains unclear. In the present study, we investigated the effects of an osmotic challenge and a V1b receptor-specific (V1bR) antagonist, SSR149415, on spatial learning of maternally separated and animal facility reared (AFR) adult male Wistar rats. The osmotic challenge was applied by injecting a hypertonic saline solution, 1 h before the Morris water maze test (MWM). V1bR antagonist SSR149415 (5 mg/kg) was injected i.p. twice (1 h and 30 min) previous to the MWM. A combined treatment with both osmotic challenge and the SSR149415 was applied to the third group whereas rats for basal condition were injected with isotonic saline. Under basal condition no differences between AFR and MS groups were observed. MS rats showed severe impairment during the MWM after the osmotic challenge, but not after the administration of SSR149415. For AFR rats, the opposite phenomenon was observed. The joint application of SSR149415 and osmotic challenge restored the spatial learning ability for both groups. The differential impairment produced by osmotic stress-induced up-regulation and SSR149415 induced V1bR blockage in MS and control rats suggested that VP involvement in spatial learning depends on the individual intrinsic ligand-receptor functional state.     

Immunogenicity of human mesenchymal stem cells in HLA-class I-restricted T-cell responses against viral or tumor-associated antigens

Human mesenchymal stem cells (MSC) are immunosuppressive and poorly immunogenic but may act as antigen-presenting cells (APC) for CD4(+) T-cell responses; here we have investigated their ability to serve as APC for in vitro CD8(+) T-cell responses. MSC pulsed with peptides from viral antigens evoked interferon (IFN)-gamma and Granzyme B secretion in specific cytotoxic T lymphocytes (CTL) and were lysed, although with low efficiency. MSC transfected with tumor mRNA or infected with a viral vector carrying the Hepatitis C virus NS3Ag gene induced cytokine release but were not killed by specific CTL, even following pretreatment with IFN-gamma. To investigate the mechanisms involved in MSC resistance to CTL-mediated lysis, we analyzed expression of human leukocyte antigen (HLA) class I-related antigen-processing machinery (APM) components and of immunosuppressive HLA-G molecules in MSC. The LMP7, LMP10, and ERp57 components were not expressed and the MB-1 and zeta molecules were downregulated in MSC either unmanipulated or pretreated with IFN-gamma. Surface HLA-G was constitutively expressed on MSC but was not involved in their protection from CTL-mediated lysis. MSC supernatants containing soluble HLA-G (sHLA-G) inhibited CTL-mediated lysis, whereas those lacking sHLA-G did not. The role of sHLA-G in such inhibition was unambiguously demonstrated by partial restoration of lysis following sHLA-G depletion from MSC supernatants. In conclusion, human MSC can process and present HLA class I-restricted viral or tumor antigens to specific CTL with a limited efficiency, likely because of some defects in APM components. However, they are protected from CTL-mediated lysis through a mechanism that is partly sHLA-G-dependent.     

Electromyogram changes during sustained contraction after resistance training in women in their 3rd and 8th decades

The present study aimed at investigating the neuromuscular adaptations to 6 weeks of resistance training in women in their third (6 experimental, 8 controls) and eighth decades (8 experimental, 8 controls). The surface electromyogram (sEMG) was measured from the biceps brachii muscle during constant-force isometric contractions lasting 12 s at 80% of maximal voluntary contraction (MVC). All the signals were analysed adopting in the time domain the root mean square (RMS) as a measure of amplitude and in the frequency domain the median frequency (MDF) of the power spectrum. Quantitative analysis was performed from the 3rd to the 6th second, to describe the early phase of the contraction (“Early”), starting from point at which 80% of the MVC was reached, and from the 9th to the 12th second, to describe the last part of the constant-force sustained contraction (“Late”). After training, the MVC increased by 22.4% in the young (P < 0.0001) and by 13.4% in the older (P < 0.05) women. The “Early” RMS increased by 60.4% with respect to the pre-training condition in the young (P < 0.01) but not in the older women. In contrast, the “Late” RMS increased by 46.7% in the older (P < 0.05) but not in the young women. The MDF remained unchanged in both groups. These results indicate that young and older women showed different training-induced adaptation of the motor unit (MU) activation pattern, in order to keep a constant level of force during a sustained isometric contraction at 80% of MVC. Accepted: 11 March 2000