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Pregnant ewes were implanted with 1 g testosterone between days 30–80, 50–100, 70–120 or 90–140 of gestation. Ewes treated between days 30–80 and 50–100 showed increased aggressive behaviour and clitoral enlargement, whereas this was not seen in the day 70–120 or 90–140 groups. Although the implants released similar amounts of testosterone at all stages of gestation, plasma testosterone concentrations were lower in the day 70–120 and 90–140 groups. The increased testosterone clearance in late gestation could be a result of increased placental aromatization. Masculinization of the genital tubercle was complete in female lambs in the day 30–80 group, partial in the day 50–100 and 70–120 groups, and absent in the day 90–140 group. These results therefore suggest that the ‘critical period’ for masculinization of the external genitalia is between days 40 and 50 of foetal life. In contrast to this anatomical masculinization, one aspect of behavioural masculinization had a later critical period, since female lambs of 30–80, 50–100 and 70–120 day groups adopted male-like urination postures, whereas the day 90–140 offspring showed the normal female pattern.

Active immunization of sheep against androstenedione results in an increase in ovulation rate that is associated with increased plasma levels of LH and progesterone, but not FSH. Although immunized ewes have more activated follicles the secretion rate of oestradiol is not increased. An experiment was conducted to examine the effect of androstenedione immunity on the ovarian secretion and peripheral plasma concentrations of inhibin.Merino ewes in which the left ovary had been autotransplanted to a site in the neck were divided into control (n= 5) and androstenedione-immune (n= 6) groups. Ovarian and jugular venous blood was collected every 10 min at two stages of the follicular phase, 21–27 h and 38–42 h after a luteolytic dose of an analogue of prostaglandin F2α(PG), and every 15 min for 6 h on day 10 of the subsequent luteal phase. The ewes were monitored regularly for luteal function by measurement of the concentration of progesterone and preovulatory LH surges. The concentration of inhibin in jugular and ovarian venous plasma was determined by radioimmunoassay and ovarian secretion rates and peripheral concentrations are expressed as pg of 1–26 peptide fragment of the α chain.The ovarian secretion rate of inhibin tended to be greater in androstenedione-immune ewes at all stages of the oestrous cycle measured, with this difference being statistically significant (P<0·05) during the luteal phase (100±40 and 260±80 (s.e.m.) pg/min for control and immune groups respectively). The pattern of ovarian inhibin secretion exhibited pulsatile-like fluctuations which were not associated with LH pulses. Peripheral concentrations of inhibin were generally higher in immunized than in control ewes with this difference being significant (P< 0·01) from day 4 to 14 of the luteal phase (59±5 and 110±7 ng/1 for control and immune respectively). The ovarian secretion rate of immunoactive inhibin was greater (P<0·01) during the follicular phase than during the luteal phase in both groups of ewes, and peripheral concentrations of inhibin increased (P< 0·001) following injection of PG in ewes from both treatment groups.We concluded that androstenedione immunity results in an increase in ovarian inhibin secretion, an effect that can probably be attributed to the greater number of large oestrogenic follicles present in the ovaries of these ewes. Furthermore, this increase in the concentration of inhibin may override any decrease in the negative feedback effects of ovarian steroid produced by immunization and, hence, explain the paradoxical findings of normal concentrations of FSH and raised concentrations of LH in ewes which are immunized against androstenedione.Journal of Endocrinology(1990) 127,285–296

The secretion rates of bioactive inhibin, oestradiol and progesterone were measured during the mid-luteal phase and at various times during the follicular phase of the cycle by a sensitive bioassay using sheep pituitary cells in culture in 12 Booroola ewes with and without copies of the Fecundity (F) gene in which the left ovary had been auto-transplanted to the neck.Inhibin secretion was high during the luteal phase and fell in the early follicular phase in all genotypes (P < 0.01). In Booroola ewes with a F/- genotype, inhibin secretion then increased again, towards luteal rates, in the mid and late follicular phases. In Booroola ewes without a copy of the F gene (+/+) inhibin secretion remained low at all three sampling times in the follicular phase. The secretion rate of inhibin at 36 h (P < 0.1) and 48 h (P < 0.01) were significantly lower in ewes from the +/+ (no copy of the gene) ewes than in F/(one copy of the gene) ewes. Oestradiol secretion was low during the luteal phase and increased steadily during the early (24 h) to a plateau in the mid (36 h; P < 0.01) and late (48 h; P < 0.05) follicular phase. Progesterone secretion was high during the luteal phase, and decreased to a very low rate by 24 h after prostaglandin (PG) treatment (P < 0.001) and remained low. At 24 h after PG the concentration of FSH was significantly lower (P < 0.01) than that during the luteal phase and remained suppressed until the onset of the LH surge. There were no significant differences in LH concentrations. We conclude that (1) the secretion of inhibin by the ovary is highest in the luteal phase and (2) inhibin secretion is significantly raised during the mid to late follicular phase in Booroola ewes with a copy of the Fecundity gene compared with those without.

Summary.Cytogenetic examination was made of 103 13–14-day-old blastocysts and 116 24–32-day-old fetuses from untreated and androstenedione-7–HSA-immunized Merino ewes. There were no differences in the chromosome composition of blastocysts or fetuses from treated or untreated ewes and so the data were combined. At Days 13–14 a 1N/2N mosaic and a 2N – 1/2N/4N mosaic embryo were observed. In addition, 52 of the blastocysts were 2N/4N mosaics, with 8 of these also containing 8N cells, and one blastocyst was a 2N/8N mosaic. No aneuploid fetuses were observed, but 80 of the 116 fetuses contained polyploid cells, including 4N, 6N and 8N cells. The polyploid cells observed in the blastocysts and fetuses should not be considered as abnormal cells as they appear to be a normal part of the developmental processes leading to trophoblast formation and fetal differentiation.

Six ewes were immunized against a prostaglandin F-2α-protein conjugate. Between 24 and 82 days after immunization the regular cyclic occurrence of oestrus was abolished in all six ewes. Further investigations of the immunized animals revealed that the blockade of oestrus was due to a persistence of the CL and the constantly elevated (>2 ng/ml) blood levels of progesterone. Surgical enucleation of the persistent CL was promptly followed by a fall in progesterone concentrations (<0·5 ng/ml), normal oestrus and a subsequent return to a state of constantly elevated blood progesterone levels.These results show that neutralization of the biological activity of PGF by active immunization against PGF-2α results in a failure of luteal regression and provide evidence that endogenous PGF is involved in normal luteal regression in this species.

A.R.C. Animal Breeding Research Organisation, King's Buildings, West Mains Road, Edinburgh EH9 3JQ, and †MRC Unit of Reproductive Biology, 39 Chalmers Street, Edinburgh EH2 9ER(Received 10th April 1975)The new CL is formed from the granulosa cells of the preovulatory follicle. This process involves well-characterized changes in the morphology of the granulosa cells that are usually accompanied by an increase in progesterone secretion (functional luteinization). In many animals, morphological luteinization of the granulosa cells is apparent some hours before the rupture of the follicle (Baird et al., 1975). Functional luteinization at this time can be inferred from an increase in the concentration of progesterone in the peripheral plasma in women (Yussman & Taymor, 1970). These changes are coincident with, and dependent on, the preovulatory surge of LH. Although the morphology of the granulosa cells in the ewe changes after the onset of oestrus and immediately before the rupture of

M.R.C. Unit of Reproductive Biology, Department of Obstetrics and Gynaecology, University of Edinburgh, 39 Chalmers Street, Edinburgh EH3 9ER(Received 14th June 1974)The biological activity of a steroid can be readily neutralized by high-affinity antibodies directed against the steroid, thus providing a useful method of studying its effects. Biological neutralization with antibodies to low molecular weight hormones can be produced either by active immunization against an hormonal hapten (Scaramuzzi, Caldwell, Tillson & Thorneycroft, 1970), or by the passive transfer of antibodies to the hormone (Vande Wiele, Ferin, Raziano, Dyrenfurth & Mikhail, 1970). The widespread use of these methods has not been possible in the past because of difficulties in generating antisera to steroid haptens and the relatively small quantities of antisera produced. The availability of reliable quick methods for the production of antisera to hormones of low molecular weight (Vaitukaitis, Robbins, Nieschlag & Ross, 1971) has been reflected by

Aestivation experiments on URGINEA MARITIMA Bak. — Bulbs of Urginea maritima Bak. have been subjected to artificial aestivation in different months (March, April, May, June) in order to find out the most appropriate period for forced blossoming. The bulbs, once taken out of the soil, have been kept for a whole month at 28° C, with atmospheric humidity not strictly controlled but never falling below 48%.Aestivation in March and April, when the plant is in its full vegetative activity, caused suppression of the sexual phase. Aestivation in May, when the vegetative activity of the plant is approaching its end, caused total flowering anticipated of about 20 days. Aestivation in June, when the plant is at the beginning of its resting period, had no effect on flowering.These results seem to show that the leaves create in the course of their activity the necessary conditions for flowering and that this critical stage is reached, in the plants under examination, between April and May.In the plants treated in May that is after they had reached the critical stage, the flowering was not suppressed; on the contrary it occurred earlier than in the plants left in their natural habitat. The advancement of the time of flowering corresponded more or less to the anticipation of the artificial as compared with the naturally occurring aestivation.

The author has examined the cambial activity and the formation of the annual ring in Quercus aegilops from February 1954 to February 1955. Material was collected at Tricase (Lecce) the only Italian locality where this oriental species grows although it is doubtful whether it is spontaneous.Cambial activity starts in the middle of April both in the main stem and in the branches following flowering and bud development. It goes on for about five months until the second half of August forming a complete growth ring. This annual ring shows the normal sequence of early wood with wide vessels formed during the second half of April and the whole of May, intermediate wood with medium and small size vessels, parenchima, and fibres formed in June-July, and late wood with few small size vessels, parenchima, and fibres formed from the end of July to middle August.There is no reawakening of cambial activity in autumn even if the climatic conditions are favourable. Tylosis are very easily formed in the wood vessels and by this process the plant can withstand the drought of the Apulian summer.

The flora of a Quercus pubescens wood, near Ruvo di Puglia, has been studied.The presence of Pirus amygdaliformis, and of various weeds, together with the disappearence of some species like Bivonaea albiflora, indicate a certain amount of degradation due to grazing and cutting. The biological spectrum shows a noticeable abundance of therophytes (45%).The mixture of typical mediterranean «maquis» plants and of more mesophilous species suggests that at Ruvo the basal Quercus Ilex horizon finds its ecological boundary, Q. pubescens substituting green oak and decidous plants outbalancing evergreens.

Attainment of puberty in ewe lambs is a function of season of birth, breed, age and weight (Hulet, 1968). Foote, Sefidbakht and Madsen (1970) found that the first ovulation in ewe lambs was very seldom accompanied by estrus, however, the hormonal changes responsible for the first ovulation and first estrus are not well documented. A higher gonadotropic potency of the pituitary before puberty with a decrease at puberty has been demonstrated by Mansour (1959).Estrogen has been implicated in the ovulatory release of luteinizing hormone (LH) in mature ewes (Goding et al., 1969; Piper and Foote, 1968), a response which was prevented by concomitant injection of progesterone or by immunization against estradiol-17β in ovariectomized ewes (Scaramuzzi et al., 1971; Caldwell et al., 1970). Pretreatment with estradiol-17β enhanced the occurrence of estrus and ovulation in ewe lambs in response to progestin and pregnant mares serum gonadotropin (PMS) treatments (Bennett and Foote, 1968; Foote and Matthews, 1969; Burfening and Van Horn, 1970).

The auth. describes the vegetation of the «Murgia S. Elio» mountain (500 m. s. l.), near Altamuta (Bari); the vegetation is open as the cretaceous limestone is often bare, and is composed mainly of therophytic, and suffruticose or chamaephytic plants.

The concentrations of progesterone, oestrogen and luteinizing hormone (LH) in ovarian venous plasma, and of progesterone, follicle-stimulating hormone, LH and prolactin in peripheral plasma were measured in 47 Welsh Mountain sheep at accurately timed stages around oestrus. The histology and ultrastructure of the pre-ovulatory and newly ruptured follicles in these animals were also examined.The highest ovarian vein levels of oestrogen were invariably obtained from the ovary containing the largest non-atretic follicle. A significant amount of oestrogen was already being secreted by day 13, when both the peripheral and ovarian venous blood progesterone levels were still high.On day 15 the corpus luteum stopped secreting progesterone; coincident with this decline there was a maximal increase in oestrogen secretion from the largest non-atretic follicle. The thecal cells of this follicle enlarged concomitantly with the rise in oestrogen, and maximal development of the smooth endoplasmic reticulum was observed at or after the oestrogen peak.By 6 h after the onset of oestrus, progesterone concentration was still very low, and oestrogen values had fallen in most animals, while LH showed a pronounced increase. The maximal level of prolactin was also found at this time.By 15 h after the onset of oestrus, both progesterone and oestrogen values were low and LH had returned to baseline levels except in a few animals. Ovulation had occurred in 11 out of 12 animals by 30 h after the onset of oestrus and the recently ruptured follicle was just beginning to secrete a little progesterone. Luteinizing hormone and oestrogen values were very low.

The effects of repeated injections of oestradiol benzoate (ODB) on the behavioural oestrous responses of ovariectomized ewes were studied using a pool of 144 animals. Oestradiol benzoate (15·6–421·2 μg) injected at intervals of 4 or 6 days produced a gradual decrease in the number of ewes showing induced oestrous behaviour. The response eventually fell below 10% at which time the ewes were defined to be refractory to oestrogenic stimulation of behavioural oestrus. When a continuous variable was used as a measure of response an immediate effect was observed; after the second injection of ODB the duration of oestrous behaviour decreased while the time to onset of oestrous behaviour showed a corresponding increase when compared with the first injection. However the proportion of ewes responding remained unaltered, indicating that continuous variables are a more sensitive measure of refractoriness. Treatment of refractory ewes with the progestagens progesterone, SC-9880 or Nilevar restored normal oestrous behaviour. These results are interpreted to mean that oestrogens (15–400 μg) act to increase thresholds for subsequent oestrogen stimulation of behavioural oestrus, while progestagens (1–10 mg) act to decrease this threshold.

A radioimmunoassay for ovine LH is described in which the plasma values are lower than those reported in most other radioimmunoassay studies. Simultaneous measurements of estrogen and LH in the ovarian venous effluent of sheep were performed by radioimmunoassay. Levels of both hormones were elevated during the follicular phase. The first rise in estrogen levels was detected 1 day before the LH rise. The measurement of LH in the peripheral and ovarian venous blood collected from the same sheep at the same time (± 5 min) indicated no significant lowering of LH in the ovarian venous effluent when compared to jugular levels.

Fourteen synthetic steroids and androstenedione were examined in ovariectomized ewes for oestrogenic activity when administered alone and with oestradiol benzoate by intramuscular injection. None of the compounds investigated was active when administered alone, as assessed by the vaginal smear assay, and only androstenedione produced a behavioural response. Androstenedione had a MED of 8·8 mg. but was less active when administered intravenously. Several steroids acted as anti-oestrogens when injected with oestradiol benzoate. Eight steroids inhibited the behavioural response and four the vaginal response. An additive response was found with androstenedione for behavioural response and with 17β-ethyl-17-hydroxy-19-nor-4-androsten-3-one (SC-5914) for vaginal response.Vaginal and behavioural responses were not necessarily related, and responses obtained in the ewe to particular steroids were not identical with those obtained in laboratory animals by other workers using similar tests.

Endosperm embryo interrelationships during germination of PINUS PINEA L. (2nd Note). — Experiments have been performed with unripe fresh seeds paralleled with unripe stored seeds (50 days of storage).The effect of storage is very similar to the effect of further permanence on the tree.It is confirmed the importance of the endosperm in the germination process already put in evidence in the 1rst note.The relationships existing between endosperm and embryo during the germination of the seed become more and more strict as the seed ripens, so that in a ripe seed the embryo develops in a normal seedling only in connection of its own endosperm.In an unripe seed the embryo is uncompletely controlled by its endosperm.Some remarks have been made about the degree of maturity of the different parts of the embryo.

Regeneration and vegetative multiplication in ENTEROMORPHA COMPRESSA (L.) Grev. — Fragments of young filaments of Enteromorpha compressa were cultured in order to determine its regenerative and multiplicative power and time limit.The original fragments cultured in vitro produced a new strain of aposporical plants, by means of buds. These aposporical individuals, when grown up, bear rhizoids with propagula from which a new strain of free algs take place. The latter generation is supplied with a fixation apparatus and show regular shape and clear polarity.

With the aim of a quantitative check of the Landau-Pomerančuk picture of the movement of foreign atoms in liquid helium II, an experiment has been performed in which some elementary charged particles could be dragged perpendicular to their motion in an electric field by a stream of excitations in non-turbulent flow. Using known values for the mobility of charged particles in He II, the drift velocity of the charges parallel to the stream of excitations has been found to be equal to the normal fluid velocity in the temperature range above 1.18 °K. Working under similar conditions, the apparatus has then been used to obtain mobilities at temperatures near 1 °K. The mobility is found to go exponentially with decreasing temperature, and this behaviour is discussed in terms of elastic scattering of the charges by the rotons.

An experiment was performed in 20 Merino ewes in which ovarian venous blood was collected by venepuncture at surgery and at two stages of the oestrous cycle. The ovarian venous concentrations of inhibin, oestradiol-17ß and progesterone were determined. The results demonstrate that during the luteal phase of the oestrous cycle the ovarian venous blood draining an ovary containing luteal tissue contains significantly more inhibin bioactivity than ovarian venous blood from an ovary not containing luteal tissue. During the follicular phase the concentration of inhibin bioactivity in ovarian venous blood was reduced compared with the luteal phase. From this data we conclude that the sheep corpus luteum secretes inhibin bioactivity into the ovarian venous blood.

We have investigated the possible relation between serum levels of CA 15.3 and disease status in 110 patients after radical mastectomy for breast cancer, with metastatic diffusion. Its persistant elevation was usually related to a very poor prognosis.In patients who died within 18 months the marker was always elevated. In case of progression of the disease, the marker level appeared to be consistently correlated with the general clinical condition. In healthy patients with stable disease the marker remained near the normal range.

M.R.C. Unit of Reproductive Biology, 2 Forrest Road, Edinburgh, EHI 2QW(Received31 October1977)Exposure to testosterone during development masculinizes the genitalia and behaviour of ewes (Clarke, Scaramuzzi & Short, 1976a; Clarke, 1977) and causes ovulatory failure (Clarke, Scaramuzzi & Short, 1977). Androgenized ewes do not release luteinizing hormone (LH) after oestrogen treatment during anoestrus (Clarke, Scaramuzzi & Short, 1976b). These experiments were performed to determine the site of action (hypothalamus or anterior pituitary gland) of prenatally administered androgens in blocking the preovulatory release of LH.Finnish Landrace × Dorset Horn ewes were used in their second breeding season after exposure to testosterone between days 30 and 80 (D30–80, n= 6), 50 and 100 (D50–100, n= 1), 70 and 120 (D70–120, n= 6) or 90 and 140 (D90–140, n= 5) of prenatal life (Clarke et al.1976a). Eight normal ewes served as controls. To facilitate

A.R.C. Animal Breeding Research Organisation, The King's Buildings, West Mains Road, Edinburgh, EH9 3JQ and †M.R.C. Unit of Reproductive Biology, Department of Obstetrics and Gynaecology, University of Edinburgh, Edinburgh, EHI 2QW(Received12 May1977)It has been suggested that the sensitivity of ewes to the negative and positive feedback effects of oestrogen can be related to the rate of ovulation of the breed (Land, 1976; Land, Wheeler & Carr, 1976), and may be associated with different patterns of oestrogen secretion as inferred by Land, Thompson & Baird (1972). However, no apparent differences between the concentrations of oestrogens in jugular venous plasma from ewes with differing rates of ovulation have been reported (Bindon, Blanc, Pelletier, Terqui & Thimonier, 1975; Scaramuzzi & Land, 1976). The pulsatile nature of the ovarian secretion of oestradiol (Baird, Swanston & Scaramuzzi, 1976) and the fact that the concentrations of oestrogen in peripheral plasma are so

Summary.In ewes immunized against androstenedione, ovulation rate was increased (P< 0·001) although the number of live lambs/nursing ewe (1·37), but not the sex ratio (10♀♀:1♂), was similar to that of controls (1·33).

M.R.C. Unit of Reproductive Biology, 2 Forrest Road, Edinburgh, EH1 2QW(Received11 June1976)Testosterone treatment of female mammals during a critical period of foetal or neonatal life affects their postpubertal endocrine and behavioural activity. For example, it prevents the occurrence of regular ovulatory cycles in adult rats (Barraclough & Gorski, 1961), guinea-pigs (Brown-Grant & Sherwood, 1971), hamsters (Swanson & Brayshaw, 1973) and sheep (Short, 1974), which is apparently due to a failure of oestrogen to facilitate the release of luteinizing hormone (positive feedback) (Brown-Grant, 1974; Short, 1974). Positive feedback is a sexually dimorphic character in sheep and is only shown by ewes (Short, 1974; Karsch & Foster, 1975). Female sheep foetuses exposed to testosterone from days 20 or 60 of gestation until birth not only failed to show positive feedback (Short, 1974), but were incapable of displaying behavioural oestrus, even when given 800 μg oestradiol benzoate (OB) (I.

ALTHOUGH low rates of ovulation constitute a major limitation to increased productivity of sheep and cattle little is known about the mechanisms that control ovulation either within or among species. Androstenedionem a week androgen secreted by ovaries1and adrenals1, is quantitatively the major steroid secreted by the ovary before ovulation in several species2and as such it might have a role in determining ovulation rate; it is also an important precursor for the biosynthesis of oestrogens, by the ovary3. Although androstenedione can act as a prehormone by virtue of its peripheral metabolism to oestrogens4a direct hormonal action has only been demonstrated in the female rhesus monkeys5where it seems to be important for sexual behaviour. We report here that by actively immunizing sheep against androstenedione a slight but significant increase in ovulation rate can be induced.

Most investigators studying oestrous behaviour in sheep use the incidence of such behaviour as a quantal response, an animal being scored as showing or as not showing behavioural oestrus. This procedure has been useful in studying the endocrinology of oestrus in this species but has not clarified the effects of the ovarian hormones on certain quantitative aspects of oestrus such as its duration. In only one study (Lindsay, 1966) have quantitative measurements of the duration of oestrous behaviour in the ovariectomized ewe been recorded and these results suggest that the length of the induced oestrus might be related to the dose of oestradiol benzoate.Fifty Border Leicester × Merino ewes, ovariectomized 2 years previously, were randomly divided into five groups. The ewes were then injected with 20 mg progesterone every second day for 10 days, followed by a final injection of 10 mg progesterone 2 days later. Forty-eight hours after

Insheep, the seasonal patterns of reproductive activity are driven primarily by the annual photoperiodic cycle, but can also respond to other environmental factors, such as nutrition, yet little is known about the mechanisms underlying this interaction. This study was designed to define the interaction between photoperiodic and nutritional cues on seasonal patterns of ovarian activity, and to determine if there is a central interaction between these cues. Groups of Ile-de-France ewes were maintained in two nutritional states (restricted and well fed) under a simulated annual photoperiod of 8–16 h of light per day over two breeding seasons. At the end of the first breeding season, half of the animals of each group were ovariectomized (OVX) and fitted subcutaneously with estradiol implants. Low nutritional status shortened the season of ovarian activity, determined from the pattern of progesterone concentrations, by modifying the timing of seasonal transitions between periods of ovarian activity and anestrus. The same results were observed for the seasonal rhythm of neuroendocrine activity, assessed in the OVX ewes, from the pattern of luteinizing hormone concentrations. These results were then confirmed for neuroendocrine activity induced by a photoperiodic treatment. We conclude that nutrition centrally modulates the interpretation of photoperiod to affect seasonal reproductive transitions. The mechanisms of this interaction are discussed in the paper.

Background: Despite the popularity of splenectomy has decreased dramatically in the past few years, the surgical approach remains the best therapy for patients with refractory Immune Thrombocytopenic Purpura (ITP) in terms of high and durable rate of response (Vesely et al, Ann Intern Med2004; 140: 112). The recent introduction of anti-CD20 antibodies and thrombopoietins of second generation such as AMG 531 and Eltrombopag may have a relevant role (Kuter et al, Lancet2008; 371: 362) but their long-term safety and efficacy have not been still established. In parallel with new drugs, there has been an evolution in the surgery of splenectomy as well (Dolan et al, Am J Hematol2008; 83: 93). Actually, the laparoscopic surgery is considered the standard approach and the ITP represents the most common indication in 50–80% of all the laparoscopic splenectomies. Methods: The aim of this study is to evaluate the long-term complete and partial haematological response (CR + PR), as well as the short and long-term complications, of 40 patients (30 females and 10 males; median age: 38 years - range 6–71) with unresponsive ITP after one or more medical approaches and underwent laparoscopic splenectomy at our Institution from 1999 through 2006. The 40 patients accounted for 22.2% of 181 patients diagnosed in those years. An abdominal CT scan to evaluate the presence of accessory spleens was performed in all cases. All patients received meningococcal, pneumococcal and haemophilus influenzae vaccine one week before splenectomy. For 4 or 5 days before splenectomy the patients were treated with high doses of intravenous Immunoglobulins. Anti-thrombotic prophylaxis was performed with low molecular weight heparin (LMWH) for 10 days and afterwards with cardioaspirin (ASA) if the platelet count exceeded 500x10E9/L. Results: No cases required conversion to laparotomic splenectomy. An accessory spleen was found in 2 patients (5%). Immediate haematological response rate was of 100%. At date, after a median follow-up of 78 months (range 28–112 months), 36 patients (90%) remain in CR or PR with a platelet count more than 50x10E9/L and 2 patients are taking ASA. Four patients (10%) relapsed; out of which, 2 patients have a platelet count less than 10x10E9/L. Short and long-term mortality rate was 0%. Immediate postoperative complications rate was 5%: we observed 2 cases of hemoperitoneum related to a trocar’s tube and to an active bleeding, respectively, both resolved with new laparoscopic approach. The mean postoperative hospital stay was 4,5 days (range 4–8). Neither cases of bacterial sepsis in the postoperative or during the follow-up time, nor cases of splenic-portal vein thrombosis (SPVT) and no cases of neoplasms occurred. Conclusions: Our experience suggests that laparoscopic splenectomy is an excellent approach to patients with refractory ITP in terms of safety, efficacy and costs. With respect to laparotomic splenectomy, the use of laparoscopy is likely to make the splenectomy even safer and therefore suitable for a larger number of patients. Undoubtedly there is a great expectation for the new drugs (Rodeghiero et al, Am J Hematol2008; 83: 91) and we agree that only controlled comparative clinical trials (Vianelli et al, Haematologica2005; 90: 72) will be able or not to say a final word and to challenge the role of splenectomy.