Combined treatment with myo-inositol, alpha-lipoic acid, folic acid and vitamins significantly improves sperm parameters of sub-fertile men: a multi-centric study.

OBJECTIVE: Reduction in motility and number of spermatozoa and change in their morphology are some of the most relevant causes of male infertility. Production of reactive oxygen species may affect motility, morphology and DNA stability of spermatozoa. This study aimed at evaluating the effect of combined treatment with myo-inositol, alpha-lipoic acid, folic acid, betaine and vitamins (namely, Sinopol®) on semen parameters of sub-fertile men. PATIENTS AND METHODS: We recruited 143 sub-fertile men, 26-53 years aged, no-smokers, without any testicular pathologies, with a normal endocrinological/metabolic profile, and no concomitant consumption of drugs. Out of them, 25 patients did not meet study inclusion criteria mainly due to the history of genital diseases that came to light after Sinopol® prescription. Among the 118 men that fulfilled inclusion criteria, 10 (8.4%) patients were lost at follow-up and in 8 (6.8%) cases the partner got pregnant spontaneously. Thus, 100 patients completed the study and semen analysis was performed before and after 90 days of treatment. RESULTS: Semen quality improved after 90 days of treatments, with a statistically significant increase of sperm concentration (p=0.0009), of number of spermatozoa (p=0.0017), of progressive motility (p=0.0047), of total motile sperm count (p=0.0010), and of normal sperm morphology (p<0.0001). CONCLUSIONS: For the first time we reported that a combination of nutraceuticals composed of myo-inositol, alpha-lipoic acid, folic acid, betaine and vitamins improves sperm parameters in sub-fertile men. We are aware that to clarify the clinical relevance of the data studies with larger sample sizes and longer durations are needed, as well as evaluation of myo-inositol and alpha-lipoic acid co-treatment effectiveness in improving the chances to obtain a pregnancy spontaneously or following assisted reproduction.

Seroprevalence of Trypanosoma cruzi in kidney transplant donors and recipients in Mexico City.

Infectious diseases are common causes of morbidity and mortality among kidney transplant recipients. Chagas disease (CD) has been recognized as an emerging infectious complication of transplantation caused by the parasite Trypanosoma cruzi. CD is prevalent in Mexico, particularly in the southern coastal region. The impact on Mexican kidney transplant programs has not been previously studied prospectively. From 2009 through 2010, serum samples from 59 kidney transplant donors and 405 renal transplant recipients were screened for antibodies against T. cruzi. Serum was initially screened using a locally developed ELISA test; positive results were confirmed by an indirect immunofluorescense test, in accordance with Panamerican Health Organization/World Health Organization guidelines. None of the donors were seropositive for T. cruzi, while 8 (1.97%) kidney transplant recipients were confirmed to be seropositive for T. cruzi. None of them have developed clinical manifestations of CD, although specific screening of recipients was not performed. A prospective study is planned to define the epidemiology and outcome of CD among kidney transplant donors and recipients in Mexico more thoroughly.

Immune response after rabies vaccine in a kidney transplant recipient.

A 48-year-old male kidney-transplant recipient was bitten by a rabid dog. His immunosuppressive treatment consisted of cyclosporine 60 mg b.i.d., mycophenolate mofetil (MMF) 250 mg t.i.d., and prednisone 5 mg. After wound care, he received 5 doses of purified vero cell rabies vaccine on days 0, 3, 7, 14, and 28, and human rabies immunoglobulin, according to international guidelines. Adequate levels of rabies virus neutralizing antibodies were observed after the administration of the third vaccine dose. However, a decrease of antibody titer was detected by day 28. Immunosuppressive medication was minimized, withdrawing MMF and reducing the dose of cyclosporine. Booster doses of the same vaccine were administered on days 38, 41, 45, 52, and 66. Adequate neutralizing antibody response was recovered during the ensuing 12 months, under reduced immunosuppression. Nineteen months after the incident, the patient remains with good graft function and is asymptomatic for rabies. It remains to be determined whether the attained immune response was either the result of the booster vaccinations or the reduction of immunosuppression alone. Nevertheless, such an outcome would have been possible only with the combined management strategy implemented.

Time of occurrence of kidney acute antibody-mediated allograft rejection/acute cellular rejection and cell senescence: implications for function outcome.

BACKGROUND: Late versus early acute antibody-mediated rejection (AAMR) or acute cellular rejection (ACR) episodes are associated with poorer kidney function and graft survival. We explored whether cell senescence upon detection of AAMR ± ACR contributes to these results. METHODS: We reviewed the renal transplant database of 2 Institutions. Biopsies performed for acute graft dysfunction from January 2000 to March 2007 were analyzed for morphological criteria of AAMR with or without ACR (n = 17 from 17 patients). Immunoperoxidase staining for p16(INK4B) was performed on the remaining paraffin-embedded tissue in 9 of 17 cases. The average number of positive cells/high power field (HPF) was calculated in every case. Cases with rejection were grouped according to the time of presentation: early (<3 months n = 8) versus late (>3 months; n = 9). Graft function was obtained using the Modification of Diet in Renal Disease (mDRD) glomerular filtration rate estimate (eGFR) before, during rejection, and at the last visit, to calculate ΔeGFR. RESULTS: Nuclear expression of p16(INK4B) was 12.2 ± 11.3 cells/HPF in 4 of 8 biopsies performed at a median of 23 (range = 4-80) days (early AAMR ± ACR), and 59.8 ± 51.3 cells/HPF in 5 of 9 biopsies performed at a median of 1171 (range = 279-3210) days (late AAMR ± ACR). eGFR before rejection was 48.5 ± 7.6 mL/min, and 43.7 ± 4.3 mL/min for early and late rejection episodes, respectively (P = not significant [NS]). ΔeGFR of 12.5 ± 25.9 mL/min (early rejection), and -13.7 ± -12.3 mL/min (late rejection), versus last follow-up visit (P = .02) occurred at a median of 143.9 ± 94.1 and 69.6 ± 35.1 weeks after the rejection episodes, respectively. CONCLUSIONS: Even though the number of biopsies analyzed for p16(INK4a) was small, it was evident that the number of cells expressing this marker of senescence was higher among biopsy specimens obtained with late rejection episodes. This finding suggests the presence of injuries prior to the rejection episode. The significantly lower eGFR at last follow-up in the late rejection group may translate to a reduced capacity of the repair process to sustain nephron function.

Role of major histocompatibility complex and ethnicity in acute renal graft rejection.

BACKGROUND: The major histocompatibility complex (MHC) plays a main role in antigen presentation. Class I, II, and III genes form defined "blocks" of conserved DNA sequences (conserved extended haplotypes) that are useful to follow the ancestry of a population. Each variant encodes a specific peptide that determines a particular individual's immune response. In addition, differential expression of HLA antigens in certain physiological and pathological conditions may participate in the pathogenesis of allograft rejection versus tolerance. OBJECTIVES: The aim of this study was to determine whether the specific HLA ancestry was associated with acute renal graft rejection among the Mexican mestizo population. MATERIALS AND METHODS: We studied 544 Mexican mestizo renal donors and their respective recipients for their serologically determined HLA and based on antigens haplotype assignments. The acute rejection group was compared with the nonrejection group among donors and recipients, correspondingly. RESULTS: Frequent Mexican alleles were observed in this study. Moreover, HLA-B*61/-DR*04, HLA-A*35/-DR*06 (Amerindian ancestry), HLA-A*68/-DR*01, HLA-A*28/-B*65/-DR*01 (African ancestry), and HLA-A*33/-B*65 (Caucasian ancestry) in donors were associated with acute renal graft rejection episodes. CONCLUSION: Knowing the ancestry of a donor's HLA molecules may help to individualize immunosuppressive therapy for posttransplant surveillance, because they may be more membrane-exposed in parenchymal cells, making them more susceptible of being recognized by the recipient's immune system.

Gastrointestinal stromal tumours: experience with multislice CT.

PURPOSE: The aim of this study was to evaluate the possibilities of multislice computed tomography (MSCT) in the identification and characterisation of gastrointestinal stromal tumours (GISTs). MATERIALS AND METHODS: MSCT images of 27 patients affected by GIST were analysed. MSCT scans were performed before and after contrast medium injection, and bowel distension was obtained with the administration of water, air or a diluted Gastrografin solution. Images were evaluated for presence and site of the tumour, origin and growth pattern relative to the bowel wall, density, relationship with adjacent structures and evidence of lymph nodes and metastases. RESULTS: GISTs were located in the stomach in 18/27 patients, in the small bowel in seven, in the oesophagus in one and in the rectum in one. Tumour size ranged from 1.5 cm to 21 cm. An extramucosal origin was definitely established in 23/27 cases. In 15/27 cases, the lesions exhibited extramural growth, and in 17/27 cases, they were homogeneous after contrast medium injection. The borders were regular in 17 cases. Hepatic metastases were detected in 5/27 cases, and lymphadenomegaly was found in one case only. CONCLUSIONS: Nowadays, MSCT can be considered an essential tool for the diagnosis of GISTs, as it enables one to detect the disease, define its relationships and search for possible metastases.

The Von Restorff effect in ageing and Alzheimer's disease.

The purpose of the study was to evaluate the Von Restorff effect in normal ageing and in Alzheimer's disease (AD). A shortened paradigm was administered to three groups of subjects: young volunteers, elderly volunteers and patients with early-stage AD (MMSE>20). Each subject was presented with 25 lists of 10 words each, the target word appearing in double font size. A free recall phase followed the presentation of each list; after completion of the battery, a size recognition test was administered and subjects were inquired regarding the strategy employed and perception of target words. The total number of recalled words differed significantly among the three groups (young volunteers 144.4+/-38.6, elderly volunteers 86.5+/-17.6, patients 44.2+/-14.6). A significant difference in percentage of recall was found between target and non-target words in young (60.0+/-13.8% vs. 45.7%+/-15.0%, p<0.001) and in elderly (31.2+/-11.4% vs. 20.2+/-6.9%, p<0.001) volunteers, but not in patients (10.7+/-6.9% vs. 11.8+/-7.3%). The present study highlights that the Von Restorff effect can be detected in healthy elderly subjects, and that it is significantly reduced in patients in the early stage of AD. On the basis of the findings of the present study it is not possible to disentangle the contribution of visual-perceptual and encoding impairment, both of them potentially contributing to the observed reduction.

Prevalence of HLA antibodies and its impact on graft function in a group of kidney transplant recipients: a cross-sectional study.

UNLABELLED: HLA alloantibodies (Abs) are associated with chronic rejection and poorer graft survival. The current study was designed to document the prevalence of HLA Abs in a group of kidney transplant recipients (KTR) and its impact on graft function. PATIENTS AND METHODS: 283 KTR transplanted between January 1990 and December 2003 who had a functional graft were invited to participate. 198 KTR were enrolled. HLA class I and II Abs were measured by Luminex-One Lambda. Graft function was assessed by DeltaCr and GFR calculated by the Levey formula. RESULTS: Median post-kidney transplant (post-KT) follow-up was 51.4 (4.3 to 176.3) months. Forty-four (22.2%) KTR were found to have class I and/or class II Abs. Eleven had both class I and II Abs, ten were positive only for class I, and 23 for class II. Overall, no significant difference was seen in renal function. The DeltaCr for Ab positive and Ab negative were -0.24+/-0.84 and -0.17+/-0.60 mg/dL (P=0.54), respectively. The GFR for Ab positive and Ab negative were 64.4+/-26 and 60.2+/-20 mL/min (P=0.25), respectively. No statistically significant difference was found between HLA Abs and number of HLA mismatches, gender, blood transfusions, pre-KT pregnancies, DGF, history of acute rejection, and chronic allograft nephropathy. Adjusting analysis by transplant year showed no significant difference. CONCLUSION: The prevalence of HLA antibodies was similar to previous reports. In this cross-sectional study, the presence of HLA antibodies was not related to a negative impact on renal function.

Effect of rapamycin on cytokine profile in kidney transplant recipients treated with triple drug therapy.

UNLABELLED: The aim of this study was to explore differences in the cytokine profile among de novo kidney transplant recipients treated with either Rapamycin (Rapa) + cyclosporine (CsA) + prednisone (P) or CsA + azathioprine (Aza) + P. PATIENTS AND METHODS: Among the 13 adult kidney transplant recipients studied, seven received Rapa + CsA + P while the remaining six received CsA + Aza + P with their living donors serving as controls (n = 13). Spontaneous production of IL-2, IFNgamma, IL-10, and TGF-beta were measured by ELISA in supernatants from 24-hour cultured unstimulated peripheral blood mononuclear cell (PBMC) at time zero (the day before the transplant), and at 3 and 6 months posttransplant. Cytokines were also measured 1 month after CsA withdrawal in the Rapa + CsA + P group. RESULTS: From time zero to the end of the study, IL-2, IFNgamma, and IL-10 were present at low or undetectable levels in all three groups. TGF-beta tended to increase in supernatants from patients under Rapa + CsA + P at 6 months posttransplant and at 1 month after CsA withdrawal without correlation to Rapa blood levels. TGF-beta remained stable throughout the study period for patients included in the CsA + Aza + P group. There was no difference in this cytokine level between these study groups at any given time. CONCLUSIONS: This study showed no differences in the spontaneous cytokine profiles evaluated in patients treated with both therapeutic schemes.

Psychological and social aspects in management of Alzheimer's patients: an inquiry among caregivers.

A survey in the cities of Genoa and Savona (Italy) was performed to examine stress levels in caregivers of patients with Alzheimer's disease in the context of a project of the Italian Ministry of Health named Cronos. It offered free anticholinesterase inhibitor therapy to patients who addressed dedicated Neurological Units; in this occasion caregivers could be invited to express the main difficulties encountered in managing demented people during an interview conducted by health personnel of the Neurophysiology Service. Caregivers were mainly women, daughters or spouses, with a medium educational level, retired, housekeepers, employees or teachers; they claimed a lowering of economic standard of living of the family owing to extra expenses for assistance. Satisfaction was expressed towards specialists, while support by general practitioners and other sanitary services was usually lacking and money contribution from the government or territorial services was considered inadequate. From the emotional point of view, caregivers claim loss of free time, friendships and hobbies, and feel isolated in the social context; sometimes the patient's death is thought of as a solution. A strong need for information and support is clearly emerging and any further interventions should take these requirements into consideration.

Identification of two homologous antigenic peptides derived from L1 HPV-16 and 18 proteins specific for the HLA-B*3901 allele.

In this work we present evidence that the homologous peptides IHSMNSTIL and IHSMNSSIL derived from L1 HPV-16 and 18 proteins respectively, and with high specificity for the allele HLA-B*3901, according with an algorithm prediction program, induced T cell stimulation in patients with advanced cervical cancer positive for HPV-16 or 18 infection and for the HLA-B*3901 allele. Interestingly, T lymphocytes derived from a patient with HPV-18 infection and stimulated with the peptide IHSMNSTIL were capable to kill a cervical cancer cell line named Rova, derived from the tumor of the same patient. In addition, the cytotoxic activity was strongly increased when this cell line was previously treated with hrIFN-gamma. These results suggest that the CTL immune response to L1 HPV-16 and 18 protein derived epitopes is maintained in patients with advanced cervical cancer within specific alleles, and opens the possibility that homologous epitopes may be used in the generation of prophylactic vaccines for cervical tumors bearing different HPV-types.

[Tinea faciei in a newborn]. / Tinea faciei in etá neonatale.

Dermatophytosis in newborns and infants has been regarded as very unusual; a case recently observed of a 2-month-old child affected by tinea faciei is described. Twenty days before, localized erythematous infiltrated patches at the upper left eyelid and eyebrows with papules and pustules, were observed. Rarefaction of the eyebrows and partially loss of the eyelashes associated with scattered areas of erythematopapulous lesions on the face, were seen. Mycological examination was positive and trichophyton rubrum colonies grew. Tinea capitis is more common than tinea faciei in newborn and infants. Differential diagnosis includes: seborrheic dermatitis, atopic dermatitis with associated impetigo, candidiasis, bacterial folliculitis. The interhuman transmission by relatives is the most probable modality of trans-mission, as previously reported in the literature. A widespread of the dermatoses was due to previous application of topical corticosteroids. The lesions resolved completely after treatment with miconazole cream, applied twice daily for three weeks.

Identification of peptides presented by HLA class I molecules on cervical cancer cells with HPV-18 infection.

In this work we eluted peptides from purified class I MHC molecules, isolated from a novel human cervical carcinoma cell line (INBL), generated in our laboratory and positive for HPV-18 infection. A fraction of these peptides was capable of stimulating T lymphocytes obtained from a donor matched for HLA-Cw4 and who was also HPV-18+. Direct N-terminal Edman degradation of these peptides, revealed the sequence (XQFPIFLQF) that matched 85% with the sequence NVFPIFLQM localized in between the 54 and 62 residues of the HPV-18 L1 protein. After stimulation with the synthetic peptide NVFPIFLQM, T lymphocytes from the donor were capable to lyse INBL cells. Our results provide evidence of the existence of naturally occurring viral epitopes presented on cervical cancer cells by the HLA-Cw4 allele, that could be useful for immunotherapy on this type of patient.

HLA class I and class II alleles and haplotypes in Mexican mestizos established from serological typing of 50 families.

We describe new information on the frequency and association of class II antigens (HLA-DR and HLA-DQ) of the major histocompatibility complex (MHC) in Mexicans. The study includes HLA-B typing and its association with the HLA-DR antigens determined in 50 families, which included 100 individuals. This family study allowed the establishment of the precise composition of the 200 HLA haplotypes, which cannot be obtained from unrelated individuals. The predominant antigens in decreasing order of frequency were B35, B39, and B61 at the B locus; DR4, DR5, and DR8 at the DR locus; and DQ3 at the DQ locus. The most common HLA-B,HLA-DR haplotype (considering broad specificities) was B16,DR4, with a frequency of 8.0%. Five HLA-B,HLA-DR haplotypes showed significant delta values (observed vs. expected frequencies) after correcting for the number of comparisons. On the other hand, the most common HLA-DR,HLA-DQ haplotypes were DR4,DQ3 and DR5,DQ3 with a frequency higher than 10%. Ten of the 17 HLA-DR,HLA-DQ haplotypes had significant postcorrection delta values.

Interaction of Candida albicans, macrophages and fluconazole: in vitro and ex vivo observations.

In recent years, medical interest in evaluating the interaction among mycetes, phagocytes, and antimycotic drugs has increased notably due to higher incidences of fungal infections in immunocompromised subjects and to the long-term therapy they require. In this study the in vitro and ex vivo interaction of fluconazole, at plasma concentrations, with mouse macrophages was evaluated in the presence of different inocula of Candida albicans. The results showed that fluconazole did not interfere negatively with phagocyte functions; conversely, according to different experimental conditions, it was able to increase both phagocytosis and intracellular killing of candida, probably exerting its action on the yeast rather than on the phagocyte. A higher enhancement of macrophage functions was observed in vitro when the drug was present in the medium with macrophages and candida in a 1:1 ratio.

Effect of rufloxacin upon non-specific immune defences: in-vitro, ex-vivo and in-vivo results.

This study investigated in-vitro, ex-vivo and in-vivo the immunomodulatory effects of rufloxacin. 0.5 MIC of rufloxacin significantly enhanced human macrophage phagocytosis and increased intracellular killing of Klebsiella pneumoniae in vitro. Pre-incubation of K. pneumoniae with rufloxacin made the bacteria more susceptible to both phagocytosis and intracellular killing by human macrophages than control organisms. Following pre-exposure of macrophages to 0.5 MIC of rufloxacin, there was a significant increase in the intracellular killing of K. pneumoniae compared with the controls, indicating the ability of rufloxacin to cross biological membranes and to remain active within phagocytes. Ex-vivo experiments show that iv administration of rufloxacin in mice lead to an increase in both phagocytic and microbicidal intracellular activity by phagocytes. In-vivo models of experimental infections showed that prophylactic administration of rufloxacin increased the survival of mice after challenge with Candida albicans.