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PubMed2006

Bioinorganic transformations of liver iron deposits observed by tissue magnetic characterisation in a rat model.

The magnetic properties and the ultrastructure, with special emphasis on the nanometric range, of liver tissues in an iron overload rat model have been investigated. The tissues of the animals, sacrificed at different times after a single iron dextran injection, have been characterised by magnetic AC susceptibility measurements together with transmission electron microscopy (TEM) and selected area electron diffraction (SAED) as helping techniques. It has been observed that few days after the iron administration the liver contains at least two iron species: (i) akaganéite nanoparticles, coming from iron dextran and (ii) ferrihydrite nanoparticles corresponding to ferritin. The magnetic susceptibility of the tissues depends not only on the elemental iron content but also on its distribution among chemical species, and varies in a remarkable regular manner as a function of the elapsed time since the iron administration. The results are of relevance with respect to non-invasive techniques for liver iron determination, directly or indirectly based on the magnetic susceptibility of the tissues, as biomagnetic liver susceptometry (BLS) and magnetic resonance (MRI) image treatment.

Animals↗
PubMed2004

Performing chest compressions in a confined space.

Standard cardiopulmonary resuscitation (CPR) may be difficult to perform in a confined space. This study set out to evaluate alternative techniques of chest compression, which may be easier to perform in such situations. Nineteen airline employees, trained in basic life support (BLS), were recruited to take part in the study. Following refresher training in standard one- and two-person CPR, they were taught two alternative techniques of chest compression: one-person over-the-head (OTH) and two-person straddle (STR). Their performances of chest compression during one-person standard CPR (St-1) and two-person standard CPR (St-2) were then compared with their performances during OTH and STR using a recording manikin. There were no statistically significant differences between the two-person methods of compression (St-2 and STR) for any of the parameters measured. There were no statistically significant differences between the one-person methods of chest compression (St-1 and OTH) for the average compression rate, the number of chest compressions achieved in a minute, or the average hands-off time per cycle. For OTH the average compression depth was significantly less than for St-1 (P = 0.0149) and there were more compressions of incorrect depth (P = 0.0400). The average duty cycle was significantly higher for OTH (P = 0.0045). 30.4% of compressions were incorrectly placed for OTH compared with 7.7% for St-1 (P = 0.0025). It was concluded that the quality of chest compression during two-person straddle CPR compares favorably with chest compression during standard two-person CPR, and may be useful in situations where space is limited. If only one rescuer is available to perform CPR, and limited space makes it impossible to carry out standard CPR, over-the-head CPR is an alternative method. However, in this study, hand placement during chest compression was poor, and additional training may be necessary before it can be considered a safe technique.

Adult↗
PubMed2004

Trials of teaching methods in basic life support (4): comparison of simulated CPR performance at unannounced home testing after conventional or staged training.

This study compares the retention of basic life support (BLS) skills after 6 and 12 months by lay persons trained either in a conventional manner, or using a staged approach. Three classes, each of 2h, were offered to volunteers over a period of 4 months. For the conventional group, the second and third classes consisted of review of skills. Those in the staged group were first taught chest compression alone; chest compression with ventilation in a ratio of 50:5 was introduced at the second class; full standard CPR was taught at the third class. A total of 495 volunteers entered the study, 262 being randomly allocated to conventional training, and 233 to staged training. More of those who received staged training attended a second (78 volunteers) and third class (41 volunteers), compared with those who received conventional training (36 and 17, respectively). The objective of this study, however, was to compare the strategies of the different training methods. A total of 291 volunteers (167 conventional and 124 staged training) were available for unannounced home testing of full conventional CPR 6 months after initial training, and 260 volunteers (135 conventional and 125 staged training) were tested at 12 months. At 6 months, those taught by the staged method were significantly better at time to first compression (P < 0.0001), compression rate (P = 0.024), and hand position (P = 0.0001). At 12 months, those taught by the staged method were significantly better at shouting for help (P = 0.005), time to first compression (P < 0.0001), and compression depth (P = 0.003). Those taught conventionally were significantly better at checking for a carotid pulse at both 6 and 12 months (P < 0.0001). These results suggest that training lay persons in basic life support skills using a staged approach leads to overall better skill retention at 6 and 12 months, and has other advantages including a greater willingness to re-attend follow-up classes.

Adult↗
PubMed2004

In-hospital cardiac arrest: survival depends mainly on the effectiveness of the emergency response.

OBJECTIVES: To evaluate the factors affecting the outcome of in-hospital cardiac arrest. SETTING: A 1400-bed tertiary care teaching hospital with a dedicated cardiac arrest team (CAT). The CAT was immediately available in monitored areas (intensive care unit and emergency room). In the wards the staff had only BLS skills and automated external defibrillation was not available. METHODS: A 2-year prospective audit according to the Utstein style. RESULTS: A total of 114 cardiac arrests (37 with VF/VT and 77 with non-VF/VT) were included. Fifty-two cardiac arrests (46%) occurred in monitored areas, 62 (54%) occurred in non-monitored areas. The CAT arrival time in non-monitored areas was 3.98+/-1.73 min. Thirty-seven patients (32%) survived to hospital discharge. Cardiac arrests occurring in monitored areas had a significantly better outcome than those occurring in the wards. Patient survival in the wards was significantly higher when the CAT arrival time was less than 3 min. No patient whose CAT arrival time was longer than 6 min survived. CAT arrival time was significantly shorter (1.30+/-1.70) in survivors than in non-survivors (2.51+/-2.37; P<0.005). Sex, age and presence of bystanders were not significantly associated with survival. CONCLUSIONS: In our setting, where bystander defibrillation was not available, the survival of patients having cardiac arrest in non-monitored areas strongly depends on advanced life support provided by the CAT. A faster CAT response and early defibrillation from the ward staff are the most important improvements necessary to increase cardiac arrest survival in our setting.

Adult↗
PubMed2004

International EMS systems: France.

The EMS (Emergency Medical Service) system in France is a centrally based, two-tiered, physician-manned system. The first level is composed of BLS (Basic Life Support) fire department ambulances (called "VSAB") based at fire stations. The second level is composed of ALS (Advanced Life Support) physician staffed-ambulances. In France, there are two different levels of emergency department (ED). The first level is called "SAU" and has continuous coverage by surgeons, in Level 2 certain specialities may be available only on an "on-call" basis. Staffing patterns in the ED vary from one hospital to another. In general, EDs in university and major teaching hospitals are staffed by emergency physicians and residents from different specialties. In France medical schools are part of free public universities. The length of medical training varies from 8 to 11 years according to speciality. Emergency Medicine is not recognised as a stand alone specialty.

Emergency Medical Services↗
PubMed2005

A study comparing the usability of fully automatic versus semi-automatic defibrillation by untrained nursing students.

INTRODUCTION: Current international guidelines prefer the use of semi-automatic external defibrillators (SAEDs) over fully automatic external defibrillators (FAEDs). However, there is a lack of evidence supporting this recommendation. We conducted a study of usability with nursing students comparing the FAED version against the SAED version of the Lifepak CR Plus AED (Medtronic, Redmond, USA). We hypothesized that FAED use would limit the number of operator-device interactions, thereby increasing compliance by the rescuer, safety and speed. METHODS: Sixty-two untrained first year nursing students were randomized to use the FAED or the SAED in a simulated cardiac arrest scenario. During analysis and delivery of three shocks, the AED guided the user with six voice prompts per shock (18 voice prompts per student). Their performance with regard to efficacy and safety was assessed using video recording. RESULTS: All rescuers except for two were able to attach electrodes and deliver a series of three shocks. During rhythm analysis by the device, FAED users made 30/372 (8%) errors against 62/360 (17%) errors for SAED users (P < 0.001). During shock delivery, FAED users made 0/186 errors against 12/180 (7%) for SAED users (P < 0.001). FAED use eliminated long time intervals between the first to the third shock (range 47-49s for FAED versus 41-90s for SAED). CONCLUSION: Despite a lack of BLS skills and AED training, the majority of students demonstrated safe and effective use of the AED. The use of the FAED version of the CR Plus resulted in increased compliance with the protocol and reduced variability in time to deliver three shocks. Further research is needed to confirm these findings in other groups of first responders.

Adult↗
PubMed2005

Evaluation of a new approach to implement structured, evidence-based emergency medical care in undergraduate medical education in Germany.

Since June 2002, revised regulations in Germany allow medical faculties to implement new curricular concepts. The medical faculty of the Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen, Germany, decided to start a major reform experiment in winter 2003, focussing on an interdisciplinary integration of organs and organ systems such as the cardiovascular or respiratory system. Furthermore, students will have contact with patients at an early stage of their studies. Thus, re-organisation of course contents should lead to a chance to improve practical experience. With the public having the right to expect that physicians and all physicians in training possess a basic knowledge of emergency medical care and the necessary skills to manage acute problems, it was decided to start the first year of the Medical Reform Curriculum Aachen with 3 weeks interdisciplinary introduction into emergency medical care. The task consisted of defining interdisciplinary core objectives and the need to implement teaching and learning principles necessary for further education. Due to this, the content of this course should have practical relevance for the students concerning their practical experiences in the future. The result is an introductory course in emergency medical care in the first semester, coordinated with the lectures. Besides skill training on basics of emergency medical care (basic life support (BLS), early defibrillation), practical training in other lifesaving techniques (e.g., immobilisation skills) and basic principles of daily clinical care are included. In addition, personal safety and a standard algorithm for assessing the patient are covered by problem-based learning sessions. The course evaluation data clearly showed acceptance of the new approach and enhances possibilities of extending implementation of relevant topics concerning emergency medical care within the Medical Reform Curriculum Aachen.

Curriculum↗
PubMed2005

Two-rescuer CPR results in hyperventilation in the ventilating rescuer.

The "Guidelines 2000 for Cardiopulmonary Resuscitation and Emergency Cardiovascular Care--International Consensus on Science" recommend a tidal ventilation volume of 10 ml/kg body-weight without the use of supplemental oxygen during two-rescuer adult cardiopulmonary resuscitation (CPR). This relates to a ventilation volume of about 6.4 l/min. Additionally, the first aid provider ventilating the victim will breathe for him/herself during the external chest compression period adding another 3.2 l/min of ventilation. Finally, a deep breath is recommended before each ventilation to increase the end-expiratory oxygen concentration of the air exhaled. To investigate the effects of these recommendations, 20 healthy volunteers were asked to perform two-rescuer CPR in a lung model connected to a BLS-manikin. End-tidal carbon dioxide, oxygen saturation, and heart rate were recorded continuously. Capillary blood gas samples were collected and non-invasive blood pressure was recorded prior to the start of external chest compressions and immediately after the end of each measurement period. Furthermore, hyperventilation related symptoms reported by the volunteers were also recorded. The data reveal a significant decrease in capillary and end-tidal carbon dioxide pressure in the volunteers (P < 0.001). Additionally, in 75% of test persons multiple hyperventilation associated symptoms occurred. Ventilation during two-rescuer CPR performed according to the Guidelines 2000 may cause injury to the health of first aid providers. To minimize hyperventilation, both rescuers should exchange their positions at intervals of 3-5 min. These data challenge the recommendation to take a deep breath prior to each ventilation.

Adult↗
PubMed2005

A decade of in-hospital resuscitation: outcomes and prediction of survival?

OBJECTIVE: To provide survival rates and associated factors from a 10-year study of in-hospital cardiopulmonary resuscitation (CPR). DESIGN: Longitudinal prospective case register study of all adult in-hospital CPR attempts conducted from April 1993 to March 2003. SETTING: 1200-bed general hospital in Plymouth (UK). PATIENTS: 2121 adult in-hospital CPR attempts in Derriford Hospital, Plymouth during the period April 1993-March 2003. MAIN OUTCOME MEASURES: Immediate, 24 h, hospital discharge and 12 month survival rates. RESULTS: Following CPR the immediate survival rate (95% confidence interval (C.I.)) was 38.6% (36.5, 40.7), then 24.7% (22.8, 26.6) at 24 h, 15.9% (14.4, 17.6) at discharge and 11.3% (10.0, 12.7) at 12 months. The primary arrhythmia, age, duration of arrest and time of arrest were strongly related to survival at 24 h and discharge. There were very low survival rates for pulse-less electrical activity (PEA) and asystole compared to VT/VF arrests; survival rates were highest for those less than 60 years and decreased with increasing age. The longer the resuscitation the less the survival, and those who arrested at night were less likely to survive. The primary arrest, respiratory or cardiac, was also independently associated with survival at 24 h but not with hospital discharge. Sex and the commencement of basic life support (BLS) within 3 min was not an independent predictor of survival. CONCLUSION: The findings of this study show resuscitation survival rates from a 10-year study and indicate some of the key predictors of survival.

Adult↗
PubMed2006

Relationship between knowledge of cardiopulmonary resuscitation guidelines and performance.

OBJECTIVE: Despite widespread training with CPR guidelines, CPR is often poorly performed. We explore relationships between knowledge of CPR guidelines and performance (compression rate, compression depth, compression to ventilation ratio, and ventilation volume). METHODS: Sixty professional EMTs were sampled at 26 randomly ordered EMS response stations from an urban system of 31 stations. A recording manikin and video model were used to assess performance in a standardized scenario, and a survey was used to assess guideline knowledge. Survey and performance outcomes were categorized prospectively as correct or incorrect based on the International CPR Guidelines from 2000. Relationships were modeled with logistic regression. Covariates included years of work experience, frequency of CPR performance, and ALS versus BLS EMT level. RESULTS: Compression rate was between 80 and 120 min(-1) in 56% (33/59) of trials. Compression depth was 1.5-2 in. in 39% (23/59), compression to ventilation ratio approximated to 15:2 in 42% (25/59), and ventilation volume was 800-1,200 cm(3) in 13% (8/60). Accurate knowledge of the CPR guidelines was associated with better performance of chest compression rate and compression to ventilation ratio. Adjusted OR (95% CI) were 4.6 (1.2-18.1) for compression rate, 1.7 (0.4-6.5) for compression depth, 4.5 (1.1-18.5) for compression to ventilation ratio, and 9.0 (0.2-351) for ventilation volume. CONCLUSIONS: Although accurate knowledge of guidelines is associated with increased odds of correct performance of some aspects of CPR, overall performance remains poor.

Adult↗
PubMed2006

The 2005 compression-ventilation ratio in practice: cycles or time?

AIM: The purpose of this study was to determine how long it takes rescuers to complete five cycles of cardiopulmonary resuscitation (CPR) using a compression-ventilation (CV) ratio of 30:2. MATERIALS AND METHODS: Twenty subjects, who were all members of the medical service at a motor racing circuit and trained in basic life support (BLS), were instructed to provide five cycles of CPR with a CV ratio of 30:2 using a manikin (Little Anne Adult CPR Manikin, Laerdal, Stavanger, Norway). The time taken to deliver the first two breaths and to complete all five cycles was recorded. RESULTS: The median time to deliver the first two breaths was 7.3 s (IQR 6.5-9.6 s) and the median time to complete five cycles with a CV ratio of 30:2 was 105.0 s (IQR 92.0-112 s). Many of the subjects found it difficult to count five cycles when using this CV ratio. CONCLUSIONS: Five cycles of CPR using a CV ratio of 30:2 takes approximately 1 min 45 s to complete. Using this CV ratio, trained individuals find it difficult to count out five cycles of CPR. It may be simpler to train individuals to give CPR for a specified time (2 min) instead of a specific number of cycles.

Allied Health Personnel↗
PubMed2006

Emergency medical service systems in Japan: past, present, and future.

Emergency medical services are provided by the fire defence headquarters of the local government in Japan. There is a one-tiered EMS system. Ambulances are staffed by three crew members trained in rescue, stabilisation, transport, and advanced care of traumatic and medical emergencies. There are three levels of care provided by ambulance personnel including a basic-level ambulance crew (First Aid Class One, FAC-1), a second level (Standard First Aid Class, SFAC), and the highest level (Emergency Life Saving Technician, ELST). ELSTs are trained in all aspects of BLS and some ALS procedures relevant to pre-hospital emergency care. Further development of an effective medical control system is imperative as the activities of ambulance crews become more sophisticated. A marked recent increase in the volume of emergency calls is another issue of concern. Currently, private services for transportation of non-acute or minor injury/illness have been introduced in some areas, and dispatch protocols to triage 119 calls are being developed.

Advanced Cardiac Life Support↗
PubMed2006

The 'ABC for life' programme - teaching basic life support in schools.

BACKGROUND: Less than 1% of the general public know how to assess or manage someone who has collapsed. It has been estimated that if 15-20% of the population were capable of performing cardiopulmonary resuscitation (CPR), mortality of out of hospital cardiac arrest could be decreased significantly. Training basic life support (BLS) skills to school children would be the most cost effective way of achieving this goal and ensuring that a large proportion of the population acquire basic life saving skills. AIMS: To assess retention of knowledge of basic life support 6 months after a single course of instruction in cardiopulmonary resuscitation designed specifically for school children. SETTING: School pupils in a rural location in one region of the United Kingdom. METHODS: A course of instruction in cardiopulmonary resuscitation - the 'ABC for life' programme - specifically designed to teach 10-12-year-old school children basic life support skills. The training session was given to school pupils in a rural location in Northern Ireland. A 22 point questionnaire was used to assess acquisition and retention of basic life support knowledge. RESULTS: Children instructed in cardiopulmonary resuscitation showed a highly significant increase in level of knowledge following the training session. While their level of knowledge decreased over a period of 6 months it remained significantly higher than that of a comparable group of children who had never been trained. CONCLUSION: A training programme designed and taught as part of the school curriculum would have a significant impact on public health.

Cardiopulmonary Resuscitation↗
PubMed2000

Genetic alterations of the retinoblastoma-related gene RB2/p130 identify different pathogenetic mechanisms in and among Burkitt's lymphoma subtypes.

Alterations of cell cycle-associated genes probably contribute to the pathogenesis of Burkitt's Lymphoma (BL), in addition to c-myc translocation. Mutations disrupting the nuclear localization signal of the retinoblastoma-related gene RB2/p130 have been documented recently in BL cell lines and primary tumors. Given the importance of the RB2/p130 gene in controlling cell growth, mutations of this gene may result in uncontrolled cell proliferation. We tested the expression and genomic organization of the RB2/p130 gene in relation to the proliferative features of a series of BL samples collected from the endemic and sporadic regions, regardless of whether the samples were acquired immune deficiency syndrome (AIDS)-related. The expression of the Rb2/p130, p107, and cell proliferation-related proteins (cyclin A and B) was determined by immunohistochemistry. The structures of exons 19 through 22 of the RB2/p130 gene, encoding for the B domain and C terminus, were analyzed by polymerase chain reaction (PCR) analysis and single-strand conformation polymorphism (SSCP) technique. The direct PCR products were sequenced to identify the actual mutations. Our results suggest that BL is composed of a mixture of molecular types with distinct genetic and phenotypic patterns, probably resulting from different pathogenetic mechanisms. In endemic BL, the RB2/p130 gene is mutated in most of the cases, and the protein is restricted to the cytoplasm. In AIDS-related BL, high levels of nuclear expression of the wild-type pRb2/p130, p107, and cell proliferation-related proteins were detected. This finding is in line with the molecular mechanisms observed in virus-linked oncogenesis. Sporadic BLs were mainly characterized by the low nuclear values of the wild-type pRb2/p130 and, conversely, the high values of p107. The increased cell proliferation due to different alterations of cell growth control by Rb-related proteins may be the first step in lymphomagenesis, during which additional genetic changes, including missense mutations of c-myc, may subsequently occur.

Adolescent↗
PubMed2002

Comparison of two fatal occupational injury surveillance systems in the United States.

INTRODUCTION: Using different methods, two national systems compile fatal occupational injury data in the United States: the National Institute for Occupational Safety and Health (NIOSH) National Traumatic Occupational Fatalities (NTOF) surveillance system, and the Bureau of Labor Statistics (BLS) Census of Fatal Occupational Injuries (CFOI). The NTOF uses only death certificates, while CFOI uses multiple sources for case ascertainment. METHODS: Through overall and case-by-case comparisons, this study compares these systems and evaluates counts for the nation and by state for worker and case characteristics. RESULTS: From 1992 through 1994, NTOF reported an average of 84% of the number of traumatic occupational fatalities reported in CFOI. This percentage changed somewhat when a case-by-case comparison was conducted--88% of the NTOF cases were matched directly to the CFOI cases. Although CFOI captured a larger number of fatalities annually, the additional fatalities did not follow a discernable pattern. IMPACT ON INDUSTRY: By understanding the distribution of fatalities, targeted efforts to reduce them will benefit all industries.

Accidents, Occupational↗
PubMed1999

Emergency medical services for children: a general pediatrician's perspective.

The EMSC movement is still in its infancy, and there is much that remains to be done. The primary care pediatrician plays a major role in the EMSC system and should continue to advocate for efficient, high-quality pediatric emergency care. In summary, there are several ways that the office-based pediatrician can and should become involved with EMSC: 1. Pediatricians should emphasize safe and injury prevention at each health maintenance visit throughout a child's life. 2. Pediatricians should encourage all parents to become certified in BLS/CPR. Ideally, training in CPR should be provided during prenatal and childbirth classes. 3. Pediatricians should advocate for injury prevention and safety campaigns in their communities. They can also become involved with efforts to develop legislation dealing with issues in injury prevention and safety. 4. Pediatricians should ensure that all children receive the appropriate immunizations. 5. Pediatricians need to maintain office emergency preparedness. All office personnel should maintain certification in BLS as a minimum and ideally, PALS. Equipment used for pediatric resuscitation should be available and functional. Monthly mock codes should be scheduled to ensure that all personnel clearly know their roles and responsibilities in the event of an emergency. 6. Pediatricians should maintain their skills in emergency pediatrics. In addition, they should maintain certification in PALS. Continuing medical education (CME) workshops and conferences in emergency pediatrics are available throughout the year. Also, pediatricians can maintain their airway management skills by practicing endotracheal intubation in the operating room setting. 7. Pediatricians must become familiar with the prehospital care providers, EDs, and transport services in their communities. Association with a pediatric intensive care unit at a tertiary care center would also be beneficial. 8. Pediatricians must be available for consultation to local EDs. They must realize that, in many instances, they may represent the physician who is most experienced with caring for the critically ill or injured child. 9. Pediatricians can serve as medical advisors to the EMS systems in their communities. 10. Pediatricians should stay well informed on issues pertaining to EMSC.

Child↗
PubMed2003

Host cell-dependent expression of latent Epstein-Barr virus genomes: regulation by DNA methylation.

Epstein-Barr virus (EBV) is a ubiquitous human gammaherpesvirus associated with a wide spectrum of malignant neoplasms. Expression of latent (growth transformation-associated) EBV genes is host cell specific. Transcripts for EBV-encoded nuclear antigens (EBNAs) are initiated at one of the alternative promoters: Wp, Cp (for EBNA1-6), or Qp (for EBNA1 only). Wp is active shortly after EBV infection of human B cells in vitro but is progressively methylated and silenced in established lymphoblastoid cell lines (LCLs). In parallel Cp, an unmethylated, lymphoid-specific promoter is switched on. In contrast, Cp is methylated and silent in Burkitt's lymphoma (BL) cell lines, which keep the phenotype of BL biopsy cells (group I BL lines). These cells use Qp for the initiation of EBNA1 messages. Qp is unmethylated both in group I BLs (Qp on) and in LCLs (Qp off). Thus, DNA methylation does not play a role in silencing Qp. In LCLs and nasopharyngeal carcinoma (NPC) cells, transcripts for latent membrane protein 1 (LMP1) are initiated from LMP1p, a promoter regulated by CpG methylation. LMPlp is silent in group I BL lines but can be activated by demethylating agents. Promoter silencing by CpG methylation involves both direct interference with transcription factor binding (Wp, Cp) and indirect mechanisms involving the recruitment of histone deacetylases (LMPlp). A dyad symmetry sequence(DS) within oriP (the latent origin of EBV replication) and intragenic RNA polymerase III control regions of EBER 1 and 2 transcription units are invariably unmethylated in EBV-carrying cells.

Animals↗
PubMed2003

Glutathione synthesis during in vitro maturation of buffalo (Bubalus bubalis) oocytes: effects of cysteamine on embryo development.

It was demonstrated that cysteamine supplementation during in vitro maturation (IVM) improves embryo development by increasing glutathione (GSH) synthesis in several species. An improved developmental competence of oocytes matured in the presence of cysteamine was also recorded in buffalo species. The purpose of this work was to investigate (1) if glutathione is de novo synthesized during in vitro maturation of buffalo oocytes, (2) if cysteamine improves buffalo embryo development via an increase in GSH synthesis, and (3) if the inhibition of glutathione synthesis by buthionine sulfoximide (BSO), in the presence or absence of cysteamine, affects subsequent embryo development and GSH synthesis.Cumulus-oocytes complexes (COCs), recovered from slaughtered animals, were matured in vitro in TCM199+10% fetal calf serum (FCS), 0.5 microg/ml FSH, 5 microg/ml LH and 1 microg/ml 17-beta-estradiol in the absence or presence of cysteamine (50 microM), with or without 5mM BSO. Glutathione content was measured by high-performance liquid chromatography (HPLC) and fluorimetric analysis in immature oocytes and in oocytes matured in the different experimental conditions. In a second experiment, the mature oocytes were in vitro fertilized and cultured for 7 days in order to assess development to blastocysts (BLs). It was demonstrated that buffalo oocytes synthesize glutathione during in vitro maturation and that cysteamine increases glutathione synthesis. Furthermore, the promoting effects of cysteamine on embryo development and GSH synthesis were neutralized by buthionine sulfoximide. These results indicate that glutathione plays a critical role on buffalo embryo development.

Animals↗