Comparison of different diluents based on liposomes and egg yolk for ram semen cooling and cryopreservation.

BACKGROUND: Sperm cryopreservation is an important tool for breed improvement, nonetheless, spermatozoids of rams are extremely sensitive to cryopreservation. AIMS: The present research was to compare a liposome-based (OptiXcell: OX) diluent, a commercial TRIS-egg yolk (Optidyl: OP) and a citrate egg yolk-based (CY) diluent on ovine semen quality through the cryopreservation process. METHODS: Semen was collected from four sexually mature Dorper rams during the natural breeding season. After collection, semen was evaluated and diluted in OX, OP or CY diluent and was cooled from 37°C to 4°C for 2 h (refrigerated semen, RS), after that semen-filled straws were placed in liquid nitrogen (LN) vapour for 10 min, then immersed into LN at -196°C (cryopreserved semen, CS) and stored until evaluation. RESULTS: For fresh semen (FS), similar values (P>0.05) were obtained from the 3 diluents [motility (4.2 ± 0.3), viability (75.4 ± 3.2), hypo-osmotic swelling test (HOST) (59.2 ± 2.1), and normality (84.7 ± 3.5)]. The motility values were higher for RS with OX and CY (4.0 ± 0.2 and 3.6 ± 0.3, respectively) compared to OP (3.0 ± 0.21; P<0.05). The viability was reduced after refrigeration and freezing (P<0.05). Refrigerated semen viability was similar for OX (65%), CY (63%) and OP diluents (60%; P>0.05), but for frozen semen, viability was lower in the CY diluent (P<0.05). Membrane integrity (HOST) in OX (53.6 ± 1.7) was similar to that in OP (50.7 ± 1.5; P>0.05) but higher than in CY (48.7 ± 1.5; P<0.05). CONCLUSION: No difference was found between the OX diluents and those made with egg yolk in terms of sperm parameters; however, the OX diluent was more efficient in protecting the integrity of membrane in freezing/thawing semen.

Validation of photo-identification as a mark-recapture method in the spotted eagle ray Aetobatus narinari.

The spotted eagle ray Aetobatus narinari is characterized by pigmentation patterns that are retained for up to 3·5 years. These pigmentations can be used to identify individuals through photo-identification. Only one study has validated this technique, but no study has estimated the percentage of correct identification of the rays using this technique. In order to carry out demographic research, a reliable photographic identification technique is needed. To achieve this validation for A. narinari, a double-mark system was established over 11 months and photographs of the dorsal surface of 191 rays were taken. Three body parts with distinctive natural patterns were analysed (dorsal surface of the cephalic region, dorsal surface of the pectoral fins and dorsal surface of the pelvic fins) in order to determine the body part that could be used to give the highest percentage of correct identification. The dorsal surface of the pectoral fins of A. narinari provides the most accurate photo-identification to distinguish individuals (88·2%).

Integrin-linked kinase mediates the hydrogen peroxide-dependent transforming growth factor-ß1 up-regulation.

Transforming growth factor type-ß1 (TGF-ß1) has been recognized as a central mediator in many pathological events related to extracellular matrix (ECM) proteins accumulation, where their locally increased expression has been implicated in the fibrosis process of numerous organs, including glomerular fibrosis in the kidney. We and others have reported the TGF-ß1 synthesis regulation by reactive oxygen species (ROS), and moreover we also described the implication of integrin-linked kinase (ILK) in the AP-1-dependent TGF-ß1 up-regulation. Thus, we propose here that hydrogen peroxide (H2O2)-dependent TGF-ß1 regulation may be mediated by ILK activation. First we confirmed the increase in TGF-ß1 expression in human mesangial cells (HMC) after treatment with H2O2 or with an alternative H2O2-generating system such as the glucose-oxidase enzyme (GOX). By using immunoblotting, immunofluorescence, and ELISA techniques, we demonstrate that extracellular H2O2 up-regulates TGF-ß1 transcription, as well as increases TGF-ß1 promoter activity. Furthermore, catalase-decreased intracellular H2O2 abolished TGF-ß1 up-regulation. The use of pharmacological inhibitors as well as knockdown of ILK with small interfering RNA (siRNA) demonstrated the implication of a PI3K/ILK/AKT/ERK MAPK signaling pathway axis in the H2O2-induced TGF-ß1 overexpression. Finally, we explored the physiological relevance of these findings by treating HMC with angiotensin II, a known stimuli of H2O2 synthesis. Our results confirm the relevance of previous findings after a more physiological stimulus. In summary, our results provide evidence that ILK activity changes may act as a mechanism in response to different stimuli such as H2O2 in the induced TGF-ß1 up-regulation in pathological or even physiological conditions.

Disseminated nocardiosis masking an atypical zygomycosis presentation in a kidney transplant recipient.

Immunosuppressive agents increase the vulnerability of solid organ transplant patients to opportunistic infections. An atypical clinical presentation of a bacterial and fungal co-infection makes diagnosis and treatment even more challenging in this population. A 54-year-old hypertensive woman underwent a cadaveric kidney transplant after years on hemodialysis. Her treatment included mycophenolate, tacrolimus, and prednisone. By post-transplant week 8, she had pneumonia followed by progressive visual changes and seizures. Diagnostic work-up, consisting of magnetic resonance imaging of the brain and chest x-ray, showed several cerebral ring-enhancing lesions, and a pulmonary cavitary lesion. Disseminated nocardiosis was suspected and therapy was started. Skin biopsy was taken from a nodular lesion and culture confirmed Nocardia species infection. During hospitalization, neurological deficit persisted with worsening of brain lesions. She underwent excision of a brain abscess and the final pathologic report showed mucormycosis, revealing the patient's co-infection by 2 different pathogens. After therapy with liposomal amphotericin B and posaconazole, she has remained stable for more than 1 year. Disseminated nocardiosis masked and delayed the diagnosis and treatment of a more aggressive and worrisome organism. Mucormycosis, as a non-fatal isolated brain abscess without rhinal involvement, is an atypical presentation, and only a few cases have been reported.

Factors and outcome in BK virus nephropathy in a Hispanic kidney transplant population.

UNLABELLED: BK virus nephropathy (BKVN) is an increasingly recognized cause of kidney allograft loss and is thought to be related to the newer, more potent immunosuppressive agents. Conflicting information has been reported on risk factors for BK infection. PURPOSE: To determine incidence, associated factors, and outcome of BKVN in our kidney transplant population in order to improve identification and management. METHODS: Kidney transplants from January 2000 to December 2005 were retrospectively reviewed. Data were collected for patients with biopsy-proven BKVN including age, sex, body mass index (BMI), etiology of renal failure, other medical diseases, donor type, surgical complications, rejection and infection, time to diagnosis, induction, immunosuppressive and antiviral therapy, and clinical outcome. A control group of patients matched for sex, age, type of graft, etiology of kidney disease, and BMI, was established for comparison. STUDY GROUP: During this period, 20 (4%) of 497 transplanted patients were diagnosed with BKVN. Thirteen (65%) were males, 8 (40%) were young adults (ages 21-40), and 18 (90%) received grafts from cadaveric donors (P=0.05). Twelve (60%) had hypertensive renal disease, 2 (10%) also had diabetes, and 16 (80%) had a BMI >25 (P=0.01). Lymphoceles occurred in 5 patients (25%). Mean creatinine level at diagnosis was 2.7 mg/dL and mean time to diagnosis was 23 months. Ten patients (50%) had leukopenia at or within a year before biopsy (P=0.001). Viruses other than BK occurred in 9 patients: varicella zoster virus in 3, cytomegalovirus in 2, herpes simplex virus in 1, molluscum contagiosum in 1, Epstein-Barr virus in 1, and human papillomavirus in 1. Eighteen patients (90%) had related rejection (P= 0.001) and 4 (20%) suffered allograft loss (P= 0.001). Basiliximab (living donors) and anti-thymocyte globulin (cadaver donors) were given for induction. All patients were on triple therapy; 15 on prednisone and sirolimus, with either tacrolimus in 8, cyclosporine in 4, mycophenolate in 1, or mycophenolate and tacrolimus in 2. The other 5 received prednisone with tacrolimus and mycophenolate. Graft loss occurred in 2 patients on tacrolimus and mycophenolate, 1 patient on tacrolimus and sirolimus, and 1 patient on cyclosporine and sirolimus. Immunosuppression was decreased in all patients. Two were given cidofovir for 6 months and had stable creatinine levels at the end of the study. Records were reviewed until April 2007. There were no deaths in this cohort. CONTROL GROUP: The number of rejections experienced by patients with BKV was much higher (P<0.0001), but the rate of graft loss was similar between the 2 groups (P=0.19). Viral co-infection was more frequent in patients with BKV (P=0.04). No episodes of leukopenia were reported for any of the patients in the control group (P=0.001). Immunosuppression with tacrolimus and sirolimus was more frequent in the BKV group, but this was not statistically significant (P=0.18, 0.28, respectively). The number of lymphoceles was larger in patients with BKV, but the difference was not statistically significant (P=0.35). CONCLUSION: BKVN is present in our transplant population and results in a high rate of allograft rejection with varying rates of graft loss. Associated factors were deceased donor and immunosuppression with potent agents, particularly tacrolimus and sirolimus. We also found a higher frequency of obesity, viral co-infection, and leukopenia. Routine screening and timely biopsy could prove cost-effective and significantly reduce morbidity.

Cutaneous aspergillosis: a report of six cases.

Skin invasion by Aspergillus is infrequent. We here describe six immunocompromised patients with skin manifestations caused by Aspergillus. A heart transplant recipient developed a primary cutaneous aspergillosis; two patients (one with chronic granulomatous disease and another treated with a high dose of corticosteroids) presented with nodular lesions secondary to haematogenous dissemination; and three patients with acute myelogenous leukaemia had skin dissemination by contiguity from orbit and sinus invasion. A. flavus was isolated in the three cases of leukaemia; the infection was due to A. fumigatus in the transplant recipient; A. fumigatus and A. versicolor were isolated in the patients with the secondary aspergillosis. In most cases, amphotericin B was useful, with clinical and mycological remission in four patients. A patient with leukaemia died without undergoing treatment, and a child carrier of chronic granulomatous disease died after only 12 days of treatment.

Mutations in TWIST, a basic helix-loop-helix transcription factor, in Saethre-Chotzen syndrome.

Saethre-Chotzen syndrome is one of the most common autosomal dominant disorders of craniosynostosis in humans and is characterized by craniofacial and limb anomalies. The locus for Saethre-Chotzen syndrome maps to chromosome 7p21-p22. We have evaluated TWIST, a basic helix-loop-helix transcription factor, as a candidate gene for this condition because its expression pattern and mutant phenotypes in Drosophila and mouse are consistent with the Saethre-Chotzen phenotype. We mapped TWIST to human chromosome 7p21-p22 and mutational analysis reveals nonsense, missense, insertion and deletion mutations in patients. These mutations occur within the basic DNA binding, helix I and loop domains, or result in premature termination of the protein. Studies in Drosophila indicate that twist may affect the transcription of fibroblast growth factor receptors (FGFRs), another gene family implicated in human craniosynostosis. The emerging cascade of molecular components involved in craniofacial and limb development now includes TWIST, which may function as an upstream regulator of FGFRs.

Cutaneous ulcers due to Candida albicans in an immunocompromised patient--response to therapy with itraconazole.

The patient, a 68-year-old female, with chronic anaemia and a previous history of carcinoma of the breast treated surgically, had been receiving therapy with methyl prednisone, in addition to gold, for pemphigus vulgaris. She developed deep cutaneous ulcers on the lower leg. All had well-defined edges and were covered with purulent and serosanguinous exudates. On histopathology the ulcers were deeply infiltrated with yeasts and mycelium and Candida albicans was isolated on culture. There was no evidence of systemic candidosis. Complete healing was obtained using itraconazole in a dose of 200 mg daily for 45 days: treatment with prednisone was continued throughout. The response to antifungal therapy alone suggests that Candida was largely, if not wholly responsible, for this unusual clinical condition.

[Norwegian scabies caused by Sarcoptes scabiei variety hominis]. / Escabies noruega por Sarcoptes scabiei variedad hominis.

A rare case of Norwegian scabies in a 16 years old female patient is reported. There also was Silver Roussel's syndrome and cellular immunologic deficiency. The case was confirmed at the biopsy, direct examination and immunologic examinations. Clinically, by erythema and adherent, abundant, whitish desquamation. The treatment resulted in a full recovery.

[Goldenhar's syndrome]. / Síndrome de Goldenhar

We are presenting five cases of Goldenhar's syndrome studied at the Clinical Genetics Unit of the Hospital Infantil of México. We do not know exactly the type of inheritance of this congenital malforamtion; in one of our cases, it seems to be related to advanced paternal age. We can consider the possibility of autosomal dominant neomutation.

A severe infantile micromelic chondrodysplasia which resembles Kniest disease.

This paper describes 3 infants with a severe, generalized chondrodysplasia with short limbs, shortness of stature, relative micrognathia and neonatal respiratory distress in all cases, cleft palate in two and dislocation of lenses in one. They died at 7 and 10 weeks and at 17 months respectively. No autopsy was performed on any of the 3 patients. Roentgenological manifestations include short, broad tubular bones with metaphyseal widening, bowing of leg, thigh and forearm bones, neonatal accelerated carpal bone maturation, short, broad pelvis with wide, flared iliac wings, many gross vertebral abnormalities including most prominently coronal clefts of almost any vertebral body, and short ribs with flared anterior ends. Roentgenographically the condition has some similarities with Kniest disease, or more correctly, the Kniest chondrodysplasia. However, we think that the clinical and roentgenological manifestations are sufficiently unique to permit delineation of the condition of our 3 patients as a "new" entity different from the Kniest chondrodysplasia. Similar cases have been described by Rolland et al. from France and by Dinno et al. from the U.S.A. The later case and our 3 patients were sporadic cases, but the former had a probably affected sibling, suggesting that this disorder is an autosomal recessive trait.

[The pediatrician and the intersexuality problems. I. Classification]. / El pediatra ante los problemas de intersexualidad. I. Clasificación

The author discusses the classification of human intersexuality and considers chromosomic sex, genetic sex, type of sex apparent in the newborn, legal sex, social sex and psychological sex.

[ADAM syndrome]. / Síndrome ADAM

We present six cases of ADAM syndrome, that we have been studying at the Unit of Clinical Genetics of the Hospital Infantil de México. We wish to stress how it is possible using medical photography to appreciate the different characteristics of ADAM syndrome: A-Amniotic, D-Deformation, A-Adhesions, M-Mutilation.