Contraception during Coronavirus-Covid 19 pandemia. Recommendations of the Board of the Italian Society of Contraception.

Purpose: The Italian Society of Contraception identified as one of its priorities the need to give recommendations on management of contraception during Coronavirus-Covid 19 pandemiaMaterials and methods: A concise communication was produced which summarises in an easy-to-read format suitable for clinicians the management of the different contraceptives mostly used. Information how to manage contraception in different conditions is presented.Results: Women may, in general, continue to use either intrauterine and or hormonal contraceptives. The use of condom should be added to any hormonal contraceptive, when the contraceptive efficacy is reduced or when women stop the contraceptive method.Conclusion: At the present time, during the Coronavirus-Covid 19 pandemia, no data contraindicate the use of intrauterine or hormonal contraceptives. Conversely the use of an appropriate contraception is advocate to prevent unintended pregnancies.

Differential microRNA expression between decidual and peripheral blood natural killer cells in early pregnancy.

STUDY QUESTION: Have decidual natural killer (dNK) cells a different microRNA (miRNA or miR) expression pattern compared to NK cells circulating in the peripheral blood (pb) of healthy pregnant women in the first trimester of gestation? SUMMARY ANSWER: dNK cells have a unique miRNA profile, showing exclusive expression of a set of miRNAs and significant up- or down-regulation of most of the miRNAs shared with pbNK cells. WHAT IS KNOWN ALREADY: dNK cells differ from pbNK cells both phenotypically and functionally, and their origin is still debated. Many studies have indicated that miRNAs regulate several important aspects of NK cell biology, such as development, activation and effector functions. STUDY DESIGN, SIZE, DURATION: Decidua basalis and peripheral blood specimens were collected from women (n = 7) undergoing voluntary termination of gestation in the first trimester of pregnancy. dNK and pbNK cells were then highly purified by cell sorting. PARTICIPANTS/MATERIALS, SETTING, METHODS: miRNAs expression was analysed by quantitative RT-PCR (qRT-PCR)-based arrays using RNA purified from freshly isolated and highly purified pbNK and dNK cells. Results from arrays were validated by qRT-PCR assays. The bioinformatics tool ingenuity pathway analysis (IPA) was applied to determine the cellular network targeted by validated miRNAs and the correlated biological functions. MAIN RESULTS AND THE ROLE OF CHANCE: Herein, we identified the most differentially expressed miRNAs in NK cells isolated from peripheral blood and uterine decidua of pregnant women. We found that 36 miRNAs were expressed only in dNK cells and two miRNAs only in pbNK cells. Moreover, 48 miRNAs were commonly expressed by both NK cell preparations although at different levels: 28 were upregulated in dNK cells, while 15 were downregulated compared to pbNK cells. Validation of a selected set (n = 11) of these miRNAs confirmed the differential expression of nine miRNAs: miR-10b and miR-214 expressed only in dNK cells and miR-200a-3p expressed only in pbNK cells; miR-130b-3p, miR-125a-5p, miR-212-3p and miR-454 were upregulated while miR-210-3p and miR-132 were downregulated in dNK cells compared to pbNK cells. IPA network analysis identified a single network connecting all the miRNAs as well as their significant involvement in several classes of functions: 'Organismal injury, Reproductive system disease, Inflammatory disease' and 'Cellular development'. These miRNAs target molecules such as argonaute 2, tumour protein p53, insulin and other genes that belong to the same network and significantly influence cell differentiation and pregnancy. LIMITATIONS, REASONS FOR CAUTION: In the present study, the cellular network and biological functions modulated by miRNAs differentially expressed in dNK and pbNK cells were identified by IPA considering only molecules and relationships that were with confidence 'experimentally observed' in leucocytes. The decidual and pbNK cells that were analysed here are a heterogeneous population and further study will help to disentangle whether there are differences in miRNA production by the different subsets of NK cells. WIDER IMPLICATIONS OF THE FINDINGS: This is the first study describing a different miRNA expression profile in dNK cells compared to matched pbNK cells during the first trimester of pregnancy. Our findings improved the body of knowledge on dNK cell biology and strongly suggest further investigation into the roles of miRNAs that are differentially expressed in human dNK compared to pbNK cells. Our results suggest that specific miRNAs can modulate dNK cell origin and functions, highlighting a potential role of this miRNA signature in human development and diseases. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by grants from the Istituto Pasteur, Fondazione Cenci Bolognetti, the European NoE EMBIC within FP6 (Contract number LSHN-CT-2004-512040), Istituto Italiano di Tecnologia, and Ministero dell'Istruzione, dell'Università e della Ricerca (Ricerche Universitarie), and from Università Politecnica delle Marche. There are no conflicts of interest to declare.

Unmet needs and knowledge of postpartum contraception in Italian women.

PURPOSE OF INVESTIGATION: Clinic visits during pregnancy and puerperium provide a unique opportunity to counsel women on contraception practices. With the aim of evaluating postpartum contraceptive attitudes among urban women attending an antenatal care center and delivering in the same facility, a structured questionnaire was administered to assess desired and received information on contraception in the postpartum period. RESULTS: A total of 436 consecutive interviews were collected during the study period. Pregnancy was unplanned in 39% of the women interviewed. Overall, 269 women (61.7%) had decided to use a method of family planning during postpartum. Among the 112 women who stated they did not want to use a method during postpartum, almost 50% stated that they "did not think they needed it", due to a perceived lack of real risk. Of the 436 women interviewed, only 5.5 % women acknowledged that they had received information on contraceptive use. CONCLUSION: The present study indicates a need for ante- and postpartum counseling of women even in urban areas of Italy.

Mirena, an Italian experience.

AIM: The aim of this review was to evaluate contraceptive efficacy, compliance, effect on menstrual cycle of a levonorgestrel-releasing device (Mirena®) in an Italian setting. METHODS: During the 48 months of recruitment, 128 women accepted the insertion of the LNG-IUS satisfying all inclusion criteria; 71.6% of the women completing five years of use had heavy menstrual blood loss. General and pelvic examination, including an ultrasound scan were performed at recruitment and repeated at 1, 3, 6, months postinsertion. Each subject was given a menstrual diary to report both menstrual patterns and side effects. RESULTS: Over the 7 308 woman/months of observation, no pregnancy occurred. Menstrual blood flow decreased in all patients, both in terms of quantity and duration. In the vast majority of cases (93.7%), spotting disappeared within six months; within the first six cycles postinsertion, amenorrhoea occurred in 29.5% of all women. In five subjects the device was removed before the planned five years of observation: two women desired a pregnancy and three, in spite of a pre-existing condition of menorrhagia, requested removal within the first nine months, because they could not tolerate amenorrhoea. Three expulsions and one dislocation of the device were also detected, all within the first eight months. CONCLUSION: Although in Italy intrauterine contraception is poorly accepted, once started on Mirena® women found that the device represents a safe and effective contraceptive modality, particularly indicated in the presence of heavy or prolonged bleeding. The only important side effect reported was breast tenderness; in addition the transient appearance of ovarian cystic images was observed.

[Voluntary pregnancy termination over the first trimester. A five-year observation]. / Interruzione volontaria di gravidanza oltre il primo trimestre. Cinque anni di osservazione.

AIM: The law for legalising abortion was approved by the Italian Government in May 1978. In regulating legal abortion this law identifies two different scenarios: one where legal abortion is performed within 90 days of gestational age, and the second where it can be performed beyond this term but within 120 days: ''when pregnancy or delivery can cause a severe damage to the woman's life, in case of severe pathologies, as fetal relevant anomalies or malformations which can cause a severe damage to the woman's physical or psychological health''. Since during the last years an increase of requests for voluntary pregnancy termination (VPT) over 90 days of gestational age has been observed in Italy, it was decided to carry out a retrospective study on the reasons for requesting such an operation. METHODS: All interventions for VPT over 90 days of gestational age performed in the Department of Obstetrics and Gynecology in the University of Rome ''La Sapienza'' between January 2003 and December 2007 have been re-assessed, analysing age of women, obstetric anamnesis, reasons for VPT request, gestational age, mode of intervention, complications due to intervention and days of inpatient admission. RESULTS: During five years 255 women demanded to terminate a pregnancy over the first trimester. In all cases requested have been authorized following a psychological consult assessing a severe damage on psychological health by the Clinical Psychology Service of ''La Sapienza'' University, that in all cases was subsequent to a diagnosis of fetal anomalies, ascertained by a genetic test and/or ultrasound scan. Anomalies were genetic in 112 of cases (43.2%) and morphological, both single and multiple, in 143 of cases (56.8%). In most of the cases (65%) these anomalies have been assessed by ultrasound scan, while in 35% by cariotype analysis. CONCLUSION: After the legalisation in 1978, cases of abortion have constantly increased. More detailed data would be helpful to better understand and face this event.

[Intrauterine death: experience at a tertiary centre of Italy]. / La morte intrauterina: esperienza in un centro terziario.

AIM: In order to analyse causes of stillbirths, we collected all the cases observed from January 1993 to December 2006 at the Department of Gynecological Sciences, Perinatology and Child Care, University ''La Sapienza'', Rome, Italy. METHODS: For each case, age of the patient, parity, country of origin, gestational age at the moment of stillbirth, clinical condition before pregnancy, pathologies occurred during pregnancy, possible therapies and autopsy of the fetus, have been collected. To evaluate and classify the obtained data, both the NICE (Neonatal and Intrauterine Death Classification according to Etiology) and the ReCoDe (Relevant Condition at Death) classifications have been utilised; the first one being more suitable than the second for our case series. RESULTS: Results showed that among 25892 labours, 186 were intrauterine deaths (7.2%). In 1999 we noticed a decrease in the number of labours of approx. 30%, due to a reduction in the number of inpatients available spaces. The number of stillbirths presented a slithering line until 2001, while after then a marked decrease has been observed. CONCLUSION: A high percentage of stillbirths had to be classified as ''unknown causes'' (26.9%). Additional prospective research, in order to achieve a better classification, is needed. All the new cases, should be classified using the most appropriate parameter, drawing attention to all the possible issues, and centralizing the data acquired.

Hormonal contraception: state of the art and future perspectives.

Attempts at controlling female fertility through the utilisation of hormonal extracts date back to the beginning of the 20th century, however, practical applications in the human species were only possible when Gregory Pincus had the excellent idea of mimicking the effect of progesterone in blocking ovulation and, through this mechanism, inhibit fertility. It is important to notice that, over its 50 years of history, the original method for hormonal contraception (HC), universally known as ''the pill'', developed into a variety of modalities that today utilise a number of new routes of administration. HCs of today are exclusively made up of either oestrogen-progestin combinations or progestins alone. This does not mean that no attempt has been made to use other classes of steroids for contraception. The best known compounds tested are the so-called ''antiprogestins'', more precisely called selective progesterone receptor modulators (SPRM). Hormonal contraceptives, originally administered in the form of daily pills, can be utilised today through seven different routes: intramuscularly, intranasally, intrauterus, intravaginally, orally, subcutaneously, and transdermally. New strategies in the field of oral contraception include further dose reduction, the synthesis of new active molecules and new administration schedules. Newly marketed contraceptive rings to be inserted in the vagina, offer a novel approach by providing a sustained release of steroids and low failure rates. The transdermal route is now well established, as a contraceptive patch, a spray or a gel. The intramuscular route has also seen new products with the marketing of improved monthly injectable preparations, containing an oestrogen and a progestin. A new minipill (progestin-only preparation) containing desogestrel has been recently marketed in a number of countries and is capable of inhibiting ovulation in most women. After the first device capable of delivering progesterone directly into the uterus was withdrawn, a new system releasing locally 20 microg over 5 years, is today widely marketed with excellent contraceptive and therapeutic performance. Finally, several subcutaneously implanted systems have been also developed in the form of contraceptive ''rods'', where the polymeric matrix is mixed with the steroid and ''capsules'', made of a hollow polymer tube filled with free steroid crystals. Last, but not least, work is in progress to provide effective emergency contraception after an unprotected intercourse. Very promising in this area is the use of SPRM.

[Infertility: a global perspective]. / Infertilitá: una visione globale.

Infertility has always existed; however, today it has a different meaning because the possibility exists, not only to treat it in some three quarters of all cases, but also, in a number of instances, to prevent it. At the same time, this improved scenario created a number of important new issues concerning public health policies and the social impact of infertility; these issues can be summarized with two words: equity and ethics. Indeed, there are intolerable differences in access to infertility care, depending on the social-economic status, as well as the country in which a couple lives. It is generally believed that, overall, in the general population a persistent form of infertility affects some 7-8% of all couples; in Europe the prevalence of infertility has been estimated at around 14%. There are important regional differences in the incidence and causes of sterility. Whereas the best known factor in western countries is an increase in the age in which women attempt to conceive, in Africa, the most important cause is represented by sexually transmitted infections. Over the last fifty years there have been major break-throws in the diagnosis and treatment of infertility, although it must be stressed that the importance and validity of therapy must be in all cases evaluated against expectant management. This is because in couples without clear signs of functional or organic pathology, conception will occur spontaneously in between 80 and 90% of the cases. At the same time, if attempts to conceive fail over a period of three years, even in women 30 years or younger, the probability of pregnancy decreases to about 40%. Finally, it must be stressed that advances in assisted reproductive technology have created not only important successes, but also significant ethical problems: on the one hand, the debate is open on the limits of artificial interventions in the field of reproduction; on the other the possibility exists that a partner may ''impose'' an infertility on the other.

Effects on cycle control and bodyweight of the combined contraceptive ring, NuvaRing, versus an oral contraceptive containing 30 microg ethinyl estradiol and 3 mg drospirenone.

BACKGROUND: The objective of this study was to compare cycle control, cycle-related characteristics and bodyweight effects of NuvaRing with those of a combined oral contraceptive (COC) containing 30 microg of ethinyl estradiol and 3 mg of drospirenone. METHODS: A randomized, multicentre, open-label trial in which 983 women were treated (intent-to-treat population) with NuvaRing or the COC for 13 cycles. RESULTS: Breakthrough bleeding or spotting during cycles 2-13 was in general less frequent with NuvaRing than that with the COC (4.7-10.4%) and showed a statistically significant odds ratio of 0.61 (95% confidence interval: 0.46, 0.80) with longitudinal analysis. Intended bleeding was significantly better for all cycles with NuvaRing (55.2-68.5%) than that with the COC (35.6-56.6%) (P < 0.01). Changes from baseline in mean bodyweight and body composition parameters were relatively small for both groups with no notable between-group differences. CONCLUSION: NuvaRing was associated with better cycle control than the COC, and there was no clinically relevant difference between the two groups in bodyweight.

[Emergency contraception]. / La contraccezione d'emergenza.

A specific formulation has been approved for use in Italy for emergency contraception (EC) in 2000. As expected, marketing of this levonorgestrel (LNG) only formulation has been accompanied by an increased interest and, often, controversies leading to even strong opposition on the part of those ethically opposed to the use of any method that may act after fertilization. At present, several trials on the exact mechanism of action and safety have been conducted, giving good reason for simplifying access, providing it free or over the counter, in several European countries. EC, also known as ''the morning after pill'' or postcoital contraception, is a modality of preventing the establishment of an unwanted pregnancy after unprotected intercourse and thus, probably, of reducing the number of voluntary pregnancy terminations. Two different forms are available: the hormonal and the intrauterine. Hormonal estrogen only EC was first proposed in the 60s and in 1974 Yuzpe following his studies proposed for the first time his combined regimen, that showed better efficacy and lower side effects. More recently, a new regimen, consisting of LNG, administered alone at the dose of 1.5 mg, was introduced and found in clinical trials to be more effective than the Yuzpe regimen, if taken as early as possible, within 72 h, thereby replacing the latter in common use. Mechanism of action of both hormonal preparations used for EC is inhibiting or delaying ovulation, therefore a prefertilization action. No effect has been reported on the process of implantation nor on an ongoing pregnancy. The WHO have developed a third regimen based on the use of the selective progesterone receptor modulator (antiprogestin) Mifepristone and conducted trials with different dosages, reporting similar efficacy and safety compared to LNG. Intrauterine EC was first proposed by Lippes in 1976. It has the advantage of being effective if inserted within 5 days after unprotected intercourse and the disadvantage of a greater complexity. In addition, this modality is truly interceptive, acting by preventing implantation. Pregnancy rates reported following EC using a device >300 mm2 of copper are consistently low (0.1-0.2%).

Reasons for requesting emergency contraception: a survey of 506 Italian women.

OBJECTIVE: To evaluate the reason for requesting emergency contraception (EC), previous use of contraceptive methods and provision route in a Family Planning Clinic in Italy. METHODS: Women requesting EC were interviewed, through a questionnaire containing questions on demographic characteristics, about their reasons for requesting EC, their prior contraceptive use, their reasons for not using an effective contraceptive method (or possible reasons for its failure) and specifically about the so-called 'provision route' (i.e. whether and where they had previously requested EC receiving a negative response). RESULTS: Almost 70% of all women requesting EC were aged between 18 and 25 years. Some 80% of all women were in a stable relationship with their partner, with fewer than 20% having had an occasional intercourse. The vast majority of women (83%) reported prior use of a modern contraceptive method, i.e. 64% with a condom, 27% for combined oral contraceptives and 1.1% for the intrauterine device (IUD). In addition, 15% of the women had used more than one method (oral pills and condoms). Concerning the reasons for requesting EC, condom breakage or slipping was the most frequently cited (64%), followed by totally unprotected intercourse (28%), failed withdrawal (5%) and forgetting one or more pill (only 1.1%). CONCLUSIONS: More than one-third of the women interviewed had previously used an emergency contraceptive modality; although no one did so more than four times. Therefore, it can be inferred that-at least in the present series-EC had not been used as a routine contraceptive method. Finally, it seems clear that in Italy, even in large cities, information about the availability, proper usage and mechanism of action is lacking. This seems due to information being spread by word of mouth between peers and friends, with more formal communication channels lagging behind.

[Preterm premature rupture of membranes in the 19th week of pregnancy. A clinical case]. / Rottura pretermine prematura delle membrane (pPROM) alla 19a settimana di gravidanza. Caso clinico.

A case of premature rupture of membranes occurred at 19 weeks of pregnancy is reported. The patient underwent a genetic amniocentesis at 16 weeks of gestational age. During the hospitalization period, she presented amniotic fluid leakage responsible of olygohidramnios. She was treated with antibiotic, spasmolytic, tocolytic and cortisone therapy. All subsequent parameters were evaluated: blood cell count, heart rate frequency, body temperature. Ultrasound examinations were performed every two weeks. Cervical smears to detect infections were normal. Labour started at 28 weeks and the patient delivered spontaneously a 1010 g male baby, Apgar 4/8. The heaviest complication was a cerebral hemorrhage and the subsequent frontoparietal hematoma which progressively reduced. To date the neurological prognosis is good.

[Use of intrauterine device by nulliparous women. Prospective study and preliminary data]. / Uso del dispositivo intrauterino nelle nullipare. Studio prospettico e dati preliminari.

BACKGROUND: The aim of the study, that is still on the way, was to evaluate the negative effects of an intrauterine device, used for more than 2 years, on future fertility of young women who due to medical or personal reasons couldn't use hormonal contraceptives. METHODS: A prospective study was started in 1987 in a group of 515 nulliparous healthy women (age 20-30) using an intrauterine device (Nova T, Schering, Multiload Cu 275 Organon) as contraceptive method. Two hundred and twentyfour women wished to become pregnant following the IUD extraction. RESULTS: Within 12 months, 221 (98.7%) of them became pregnant demonstrating the harmlessness of the contraceptive method on future fertility. All pregnancies had a normal course, except one abortion at the 8th week of gestational age. There hadn't been pregnancies during the observation period, 25 (4.8%) women requested the IUD be removed before the end of the study because of side effects. DISCUSSION: IUD did not cause a reduction in fertility or an increase in ectopic pregnancy in our group of patient.

[Risk factors for ectopic pregnancy. Case-control study]. / Fattori di rischio per la gravidanza ectopica. Studio caso-controllo.

BACKGROUND: To analyze risk factors for ectopic pregnancy in an Italian population. METHODS: A retrospective case-control study has been carried out on 213 cases of ectopic pregnancy and 213 controls. The control subjects were women who delivered spontaneously at the same time and in the same hospital of the cases considered. RESULTS: Smoking more than 10 cigarettes/die doubles the risk of ectopic pregnancy. IUD causes an increased risk in women using this contraceptive method for less than two years. A history of abdominal surgery was associated with about a double risk of ectopic pregnancy. The risk was higher in cases of cesarean section and pelvic surgery. CONCLUSIONS: Abdominal surgery (particularly pelvic surgery and cesarean sections), use of IUD for less than two years and cigarette smoking are the risk factors found to be a cause for ectopic pregnancy in the population studied. It was not possible to evaluate the history of pelvic inflammatory disease because there were not enough data. For this reason it is suggested it will be possible in the future to compile a register data, collecting all the cases of ectopic pregnancy as a part of a prevention campaign.

Comparative clinical evaluation of the effect on carbohydrate and lipid metabolism of two norethisterone-containing hormonal contraceptives: Mesigyna and TriNovum.

The effect on carbohydrate and lipid metabolism of two hormonal contraceptive preparations containing norethisterone (commercially known as Mesigyna and TriNovum) was studied in a total of 60 women, before and after 6 months of treatment. Carbohydrate metabolism was evaluated by means of a euglycemic glucose clamp test; lipid metabolism was monitored by measuring total cholesterol, HDL-cholesterol, LDL-cholesterol, VLDL-cholesterol, and triglycerides. The two groups were properly matched with the exception of pretreatment levels of cholesterol and LDL-cholesterol. At the end of treatment, no difference was found within or between groups in fasting glucose and insulin levels and in glucose rate of disappearance. A significant increase in total cholesterol, HDL-cholesterol, and VLDL-cholesterol was found in both groups at the end of the treatment period; in addition, TriNovum caused a significant increase also in triglycerides. In conclusion, the safety of both preparations with regard to carbohydrate metabolism was confirmed using the most accurate method available; furthermore, changes in lipid metabolism were such as to have little clinical significance.

[Causes for transfer of obstetric patients to an intensive care unit (1990-19960]. / Cause di trasferimento di pazienti ostetriche ad un centro di rianimazione (anni 1990-1996).

BACKGROUND: In order to have an updated review of the causes for transferring patients from our Institute to Intensive Care Unit (ICU), a study has been conducted on patients admitted from 1990 to 1996. METHODS: Of the 28 women transferred, one underwent dilatation & curettage of uterine cavity after abortion, 3 had a normal delivery and 24 underwent a cesarean section. RESULTS AND CONCLUSIONS: The complications were as follows: respiratory failure (5 cases); suspected or confirmed lung embolism (1); cerebral ischemia (1); septic complications (1); cardiocirculatory complications (19 cases; 7 of them were due to DIC; 6 due to cardiocirculatory failure; 6 due to haemorrhagic shock). These patients were divided into 3 groups, those with pre-existing pathologies which worsened (2 cases); those due to deterioration of pathology arose during pregnancy (17) and, finally, complications during delivery (9). In total the cases of mortality were 7, all of them following cardiocirculatory complications.

[Contraception and voluntary termination of pregnancy. Survey of a sample of 500 women]. / Contraccezione e interruzione volontaria della gravidanza. Indagine conoscitiva su di un campione di 500 donne.

In order to better evaluate the link between contraceptive attitudes and induced abortion, 500 women who underwent voluntary termination of pregnancy (VTP) at the First Institute of Obstetrics and Gynaecology of the University of Rome "La Sapienza" were interviewed. They were young (median age 28 years), medium to high educated (61%), non married (52.4%), nulliparous (59.2%) women. Forty-two percent of them had the first sexual intercourse before 18 years of age and 72.6% at least five years before. Ninety-five women (19%) admitted a previous voluntary termination of pregnancy. Withdrawal (305 women), oral contraceptives (246 women) and condom (223 women) were the most widely used methods of fertility regulation, but the pill had been taken only for short periods of time, never exceeding one year. Withdrawal resulted to be the most utilized method during the cycle in which conception occurred (49.4%); no method had been employed by 34.8% of the women. The diffusion of modern methods of fertility regulation greatly influences the number of induced abortions in Italy, as clearly indicated by the progressive reduction of VIP starting from 1982 paralleled by a constant increase in OCs diffusion. Our data, although limited in number, confirm that induced abortion is the consequence of an insufficient use of modern methods of fertility regulation.

Diagnosis and classification of the acute leukemias: recent advances and controversial issues.

Although morphology and cytochemistry continue to be the mainstay of the diagnosis of acute leukemia (AL), new developments in immunophenotyping, cytogenetics, molecular biology, and in vitro assays have dramatically improved our understanding of this disease and enabled the identification of entities with distinct clinico-biologic features. Immunophenotyping is essential for diagnosing and subclassifying acute lymphoblastic leukemia (ALL) and is also very helpful in certain types of acute myeloid leukemias (AML), such as AML with minimal differentiation or acute megakaryoblastic leukemia. Cytogenetic findings are clinically relevant for diagnosis and prognosis. Nonrandom chromosomal abnormalities such as t(15;17)(q22;q12) or t(1;19)(q23;p13) have been so closely associated with distinct types of acute leukemias that their recognition can allow diagnosis independent of the other criteria. Molecular analysis is a powerful method in the assessment of the malignant potential, clonality, and classification of the ALs. It has become clear that in some leukemias a proportion of patients exhibit the biologically relevant molecular defect in the absence of a karyotypic equivalent. On the other hand apparently uniform chromosomal abnormalities such as the t(1;19), t(9;22), t(8;14), or t(15;17) may differ at the molecular level. In vitro assays can evaluate the growth pattern and cell-cycle kinetics of leukemic cells, as well as their sensitivity to therapeutic agents. All these data are relevant to the management of AL. Because the French-American-British (FAB) classification does not fully correlate with much of this new information, alternative classifications have been proposed. In this review we concentrate on recent diagnostic contributions resulting from advances in biotechnology and discuss some of the points that arouse controversy in the single classifications.

Contribution of immunophenotypic and genotypic analyses to the diagnosis of acute leukemia.

Diagnostic accuracy in acute leukemia (AL) can be improved if traditional morphology and cytochemistry are supplemented with immunophenotypic and genotypic analyses. This multiparameter approach is of crucial importance for the management of patients, as it enables the identification of leukemic syndromes with distinct biological features and response to treatment. Immunophenotyping using monoclonal antibodies has been universally accepted as a useful adjunct to morphological criteria. This technique is particularly valuable in diagnosing and subclassifying acute lymphoblastic leukemia and is also essential in certain types of acute myeloid leukemia (AML), such as AML with minimal differentiation or acute megakaryoblastic leukemia. Cytogenetic findings can be quite helpful in establishing the correct diagnosis and can add information of prognostic significance. A number of specific chromosomal abnormalities have been recognized that are very closely, and sometimes uniquely, associated with morphologically and clinically distinct subsets of leukemia. An even more basic understanding of normal and malignant hematopoietic cells has begun to evolve as molecular biology begins to unravel gene misprogramming by Southern and Northern blot analysis, the polymerase chain reaction, and fluorescence in situ hybridization. With the extensive use of these techniques it has become apparent that a proportion of leukemias exhibit the biologically relevant molecular defect in the absence of a karyotypic equivalent. On the other hand, apparently uniform chromosomal abnormalities such as the t(1;19) (q23;p13), t(9;22) (q33;q11), t(8;14) (q24;q32), or t(15;17) (q21;q21) may differ at the molecular level. Data collected from these modern technologies have introduced a greater complexity, which needs to be taken into consideration to improve both the diagnostic precision and the reproducibility of current classifications.