The oxidizing agent tertiary butyl hydroperoxide induces disturbances in spindle organization, c-meiosis, and aneuploidy in mouse oocytes

It has been recently proposed that a concomitant generationof oxidative stress of oocytes with increasing maternal agemay be a major factor responsible for the age-related increasein aneuploid conceptions. As a preliminary step in the testingof this hypothesis, we need to confirm that oxidative stressin itself can induce errors in chromosome segregation. In orderto achieve this goal, germinal vesicle (GV)-stage mouse oocytesfrom unstimulated ICR and (C57BLxCBA) F₁ hybrid female micewere matured in vitro for 9 h for metaphase I (MI) oocytes or16 h for metaphase II (MII) oocytes in the presence of varyingconcentrations of the oxidizing agent tertiary-butyl hydroperoxide(tBH). MII oocytes from (C57BLxCBA) F₁ hybrid mice were fixedand C-banded for karyotyping analysis. MI and MII oocytes fromICR mice were fixed and stained with the DNAfluorescent probe4',6-diamidino-2-phenylindole (DAPI) to detect abnormalitiesin chromosomal distribution. Meiosis I and meiosis II spindlesfrom ICR mice were visualized by confocal immunofluorescencemicroscopy. Data from these experiments demonstrate that in-vitroexposure of mouse oocytes to tBH during meiosis I reduces thelength (pole-to-pole distance) and width (diameter at the equatorof the spindle) of meiosis I and meiosis II spindles. This reductionis associated with an increase in the percentage of oocytesshowing chromosome scattering and clumping on the MII plate,and of aneuploidy (hyperhaploidy) in MII oocytes. However, tBHat the concentrations used in the present study has only a minimalnegative effect on the frequency of meiotic maturation. Theseresults suggest that oxidative stress during meiotic maturationin vitro may induce chromosomal errors that are undetectablein the living oocyte and whose developmental consequences maybecome manifest after fertilization. aneuploidy/meiosis/mouse oxidative stress/spindle/tertiary butyl hydroperoxide     

Carrier detection in Duchenne and Becker muscular dystrophy Argentine families

In order to offer carrier detection, genetic counseling, and prenatal diagnosis to families with Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) in our country, segregation analysis of highly polymorphic short tandem repeats (STR) (dC-dA)n: (dG-dT)n loci was utilized. The risks to females of 15 DMD/BMD families (9 familial and 6 sporadic) were evaluated on STR, pedigree and serum creatine kinase (SCK) data. From the 36 females at risk of being carriers (not including 8 obligate carriers), results of STR analysis were compatible with carrier status in 7 and not compatible in 20. In 9 females, no information regarding carriership was derived from the STR analysis. Prenatal diagnosis is now possible on the carrier females. Previously identified deletions in the central part of the gene were confirmed by STR analysis in 3 families. Five new alleles were identified in Argentine individuals; allele frequencies differed from those of North American people. Results derived from this study are useful for carrier detection and genetic counseling in DMD/BMD. One case of probable mosaicism in an unaffected father was detected on a pedigree basis in a family with DMD patients.     

Redescription of <Emphasis Type="Italic">Bacciger microacetabularis</Emphasis> (Martorelli et Suriano, 1983) nov. comb. parasitizing <Emphasis Type="Italic">Paralichthys orbignyanus</Emphasis> (Pisces,Paralichthyidae) from Argentina

In this paper Steringotrema microacetabularis (Suriano et Martorelli, 1983) is redescribed and transferred to Bacciger Nicoll, 1924 in the Faustulidae Poche, 1926 based on newly collected material from the type-host, Paralichthys orbignyanus Valenciennes, 1839 and the type-locality, Mar Chiquita coastal lagoon, Buenos Aires Province, Argentina. A careful re-examination of the new specimens shows that some anatomical characteristics were ignored or incompletely described by the previous authors and they are included herein. The species is tentatively transferred to Bacciger with which it appears to have closest affinity. Despite the anatomical differences detailed in this paper, confirmation of this proposal must await further work, including molecular studies.     

Use of UroLume as an alternative to the treatment of recurrent stenosis of bulbar urethra and obstructive prostatic hyperplasia in patients with high surgical risk

This device consists of a woven in a form of a tubular mesh, made up for the use in the urethra. We report the clinical and uroflujometric results in 10 patients with urethral stricture and 4 with obstructive prostatic hyperplasia. The study includes a 7 year period since October of 1993 up until June 2000. All patients were evaluated pre and post stent insertion with periodic follow ups to assess the prostatic symptoms score, quality of life assessment, peak urinary flow rate, mean flow rate and post-void residual urine volume. The results were the following: For the group with urethral stricture the I-PSS decreased from 26.8 to 5.4 points, the peak flow rate increased from 8.24 to 16.12 mL/sec and the post-void residual urine volume decreased from 42.7 to 31.6 mL. By 12-month follow-up most endoprostheses were 90% covered with urothelium, only one of these patients required stent extraction with no sphincter lesion. For the prosthatic hyperplasia group the I-PSS decreased from 20.2 to 8 points. The peak flow rate increased from 6.95 to 14.5 mL/sec and the post-void residual urine volume decreased from 49 to 18.3 mL. By 12-month follow-up two patients were 95% covered with epithelium, and the other two were 70%. There have not been significant problems related to infections, migration, incontinence or erectile disfunction. An 80% of patients have shown some irritative symptoms (urgency, frequency or dysuria) at least during the first month after stent insertion. These 7 year results suggest that Urolume urethral endoprostheses can be a long-term effective treatment alternative to these patients.     

Entrainment by a palatable meal induces food-anticipatory activity and c-Fos expression in reward-related areas of the brain

Rats maintained under restricted feeding schedules (RFS) develop food-anticipatory activity and entrainment of physiological parameters. Food entrainment is independent of the suprachiasmatic nucleus and depends on food-entrainable oscillators (FEO). Restricted feeding schedules lead animals toward a catabolic state and to increase their food driven motivation, suggesting that in this process metabolic- and reward-related mechanisms are implicated. This study explored if motivation driven by a palatable meal is sufficient to produce food-entrainment. To address this question, we evaluated whether daily fixed access to a highly palatable meal entrained (PME) locomotor activity, serum glucose and free fatty acids concentrations in rats maintained without food deprivation. The entrained response of PME rats was compared with rats entrained to RFS. In a second experiment, we used c-Fos-IR to identify structures in the central nervous system involved with PME. Rats showed anticipatory activity to a daily palatable meal, with a lower intensity than rats entrained to RFS. Anticipatory activity persisted at least for four cycles after interrupting palatable meal, suggesting that this persistence depends on an endogenous oscillator. Glucose and free fatty acids were not entrained in PME rats. c-Fos expression in limbic system nuclei was in phase with PME time, but not in the hypothalamus. Results suggest 1) that food deprivation, i.e. a catabolic state is not necessary for the expression of anticipatory activity; 2) that an increase in the motivational state due to taste and/or nutritional contents of palatable meal is sufficient to entrain behavior; and 3) that structures in the limbic system are involved in this entrainment process. The present study indicates that metabolic and motivational mechanisms are involved in food entrainment, and suggests that the FEO may be a multi-oscillatory system distributed over different regulatory systems in the brain.     

Nonmyeloablative stem cell transplantation is an effective therapy for refractory or relapsed hodgkin lymphoma: results of a spanish prospective cooperative protocol.

We report the results of reduced-intensity conditioning allogeneic stem cell transplantation (allo-RIC) in patients with advanced Hodgkin lymphoma (HL). Forty patients with relapsed or refractory HL were homogeneously treated with an RIC protocol (fludarabine 150 mg/m(2) intravenously plus melphalan 140 mg/m(2) intravenously) and cyclosporin A and methotrexate as graft-versus-host disease (GVHD) prophylaxis. Twenty-one patients (53%) had received >2 lines of chemotherapy, 23 patients (58%) had received radiotherapy, and 29 patients (73%) had experienced treatment failure with a previous autologous stem cell transplantation. Twenty patients (50%) were allografted in resistant relapse, and 38 patients received hematopoietic cells from an HLA-identical sibling. Five patients (12%) died from early transplant-related mortality (before day +100 after allo-RIC). One-year transplant-related mortality was 25%. Acute GVHD developed in 18 patients (45%). Chronic GVHD developed in 17 (45%) of the 31 evaluable patients. The response rate 3 months after the allo-RIC was 67% (21 [52%] complete remissions and 6 [15%] partial remissions). Eleven patients received donor lymphocyte infusions (DLIs) for disease relapse. The response rate after DLI was 54% (3 complete remissions and 3 partial remissions). Overall survival (OS) and progression-free survival (PFS) were 48% +/- 10% and 32% +/- 10% at 2 years, respectively. Refractoriness to chemotherapy was the only adverse prognostic factor for both OS (63% +/- 12% versus 35% +/- 13%; P = .05) and PFS (55% +/- 16% versus 10% +/- 9%; P = .006). For patients with failure of a prior autologous hematopoietic stem cell transplantation, results were especially good for those who experienced late relapses (>/=12 months: 2-year OS and PFS were 75% +/- 16% and 70% +/- 18%, respectively). These data suggest that allo-RIC is feasible in heavily pretreated HL patients and has an acceptable early transplant-related mortality. Results are better in patients allografted in sensitive disease. Both responses observed after the development of GVHD and DLI may suggest a graft-versus-HL effect. Allo-RIC has to be considered an effective therapeutic approach for patients who have had treatment failure with a previous autologous hematopoietic stem cell transplantation.     

Atrial automaticity and atrioventricular conduction in athletes: contribution of autonomic regulation

Little is known about the sinoatrial automatism and atrioventricular conduction of trained individuals who present a normal resting electrocardiogram. We used transesophageal atrial stimulation, a minimally invasive technique, to evaluate aerobically trained athletes (n=10) and sedentary individuals (n=10) with normal resting electrocardiograms, to test the hypothesis that parasympathetic tone, as detected by heart rate variability, could be associated with changes in sinoatrial automatism and atrioventricular conduction. Corrected sinus node recovery time tended to be longer in athletes than in sedentary individuals, but this difference did not reach statistical significance. The Wenckebach point occurred at a lower rate in athletes than in the controls. Over a 24-h period of measurement, the mean RR interval was longer in the athletes than in the sedentary individuals. The mean square root of successive differences (rMSSD) tended to be higher in athletes than in controls, but this difference did not reach statistical significance. There was a moderate correlation (r=0.48, P < 0.05) between the index of atrioventricular conduction, the rate at the Wenckebach point, and the logarithmically transformed rMSSD. Thus, as a corollary to its effects on the sinus node, where increased parasympathetic tone, decreased sympathetic tone, and non-autonomic components may contribute to sinus bradycardia, it is possible that athletic training may also induce intrinsic adaptations in the conduction system, which could contribute to the higher prevalence of atrioventricular conduction abnormalities observed in athletes. Accepted: 2 January 2000     

Checkpoint Blockade Immunotherapy Induces Dynamic Changes in PD-1−CD8+ Tumor-Infiltrating T Cells

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