Formation of extra digits in the interdigital spaces of the chick leg bud is not preceded by changes in the expression of the Msx and Hoxd genes

By in situ hybridization we studied the expression patterns of Msx and Hoxd genes during the late development of the chick leg autopodium (foot) and compared them to patterns during the experimental development of interdigital extra digits. Extra digits are induced in the third interdigital space after various experimental manipulations, such as transient isolation of the interdigit, or removal of the interdigital marginal ectoderm and mesoderm. Msx1 and Msx2 are normally expressed in the interdigital tissue programmed to die. Our experiment changes the fate of the interdigital tissue from cell death to chondrogenesis and provides a good model for studying Msx involvement in defining areas of programmed cell death. Among the proposed roles for Hoxd genes is their involvement in the specification of digit identity early in development. The induction of extra digits allows us to examine whether this new morphogenetic commitment of the interdigital tissue involves changes in the domains of expression of Hoxd genes. Our results show that extra digits develop without a previous modification of the normal pattern of expression of Msx or Hoxd genes. This observation does not support the correlation between the expression of Msx genes and programmed cell death and suggests a role for these genes in maintaining the interdigital tissue in an undifferentiated state. Our results show that an increased number of digits can be formed without modifications in the pattern of expression of the 5-located Hoxd genes and suggest the existence of latent or residual digit organization mechanisms past the time when digits are normally determined, independent of Hoxd gene expression.     

The structure of human S-phase chromosome fibres

Recentin situ hybridization studies suggested that within the range of 0.1–1.0 Mb, human interphase chromosomes follow a random walk model (i.e. they behave as flexible polymers without major constraints). However, chromosome structure may differ in the G1, S, and G2 phases, and phase-specific constraints may be masked if the chromosome analysis does not discriminate between the phases. Therefore, using confocal microscopy, we examined the structure of S-phase chromosomes labelled with 5-iododeoxyuridine after prolonged treatment with 5-fluorodeoxyuridine. In the S-phase, labelled 0.32 µ chromosome fibres mostly appear as semi-circles with an average diameter of 0.83 ±0.03 µ. These semi-circles are joined together to form different 3D structures, and two semicircles frequently adopt s- or-like conformations involving about 2.5 µ of the chromosome contour length (L). Morphometric analysis of the S-phase fibres suggests that our data fit both the random flexible polymer model and also a model in which two constrained semi-circles are attached to each other by a flexible joint, thus eliminating constraints at long distances (L more than 2 µ).     

“Nowadays it isn't easy to advise the young”: Grandmothers and granddaughters among Abaluyia of Kenya

Among Abaluyia of Kenya relative age (seniority) structures relationships hierarchically among co-wives, siblings and females of different generations. Ambiguous equality and affectionate informality in grandmother-granddaughter relationships mute the hierarchical implications of their different age and generation statuses. This facilitates grandmothers' educational roles although, as grandmothers say, Nowadays it isn't easy to advise the young. Increased physical, cognitive and experiential distances between these generations resulted from radical changes in the female lifecourse associated with modernization and delocalization. Nevertheless, reciprocal exchanges continue. Granddaughters are also intermediaries in exchanges between older women and their adult daughters. Some grandmothers are denied the companionship and assistance of granddaughters caught in the middle of mother-in-law/daughter-in-law conflicts. Many grandmothers assume parental responsibilities as they deal with the modern problem of daughters' premarital pregnancies. All in all, these intergenerational relationships reveal both cultural persistence and the effects of social change, and continue to have instrumental and expressive value for both grandmothers and granddaughters.     

Sex selection may be inadvertently performed in in-vitro fertilization--embryo transfer programmes

The present study aims to ascertain whether sex selection maybe inadvertently performed in human in-vitro fertilization (IVF)and embryo transfer (IVF-embryo transfer) programmes when selectingfor high quality embryos (those with the fastest cleaving ratesand/or the best morphology) at the fresh transfer cycle. Allpatients entering into the study were treated with gonadotrophinsafter pituitary suppression with gonadotrophin-releasing hormoneagonists (GnRHa) and had intrauterine embryo transfer on day2 post-insemination. These patients were retrospectively dividedinto three groups according to whether the difference in meannumber of cells between embryos transferred and all embryosavailable for transfer in a given cycle was less than (negativeselection), equal to (no selection) or greater (positive selection)than zero. In cycles resulting in singleton births, the sexratio of the resulting babies was significantly (P 0.005) shiftedtoward the female (88.8%) and to the male (90.0%) in the negativeand positive selection groups respectively. No shift in sexratio was observed in cycles resulting in multiple births. Maternalage was another independent factor affecting sex ratio at birth.Sex ratio was significantly (P 0.05) skewed in favour of males(62.7%) and females (71.4%) in women <35 and 235 years ofage respectively. Maternal age, number of embryos transferredand the event of selecting or not selecting the slowest cleavingembryos for transfer were entered automatically in a three-groupdiscriminant model for distinguishing cycles resulting in onlyboys, both boys and girls, and only girls. These data suggestthat (i) sex selection may be inadvertently performed in IVF-embryotransfer programmes when selecting for high quality embryosat the fresh transfer cycles; (ii) human endometria may be favourable,indifferent or hostile to either fast cleaving or slow cleavingembryos depending on maternal age; and (iii) ‘natural’sex selection may be performed for social, psychological ormedical reasons.     

Child neurology

Poster Session 1

Child neurology     

Reoperation Due to Complications after Gastric Restriction Operation

Background: In the past 4 years we performed operations on 90 patients who suffered from morbid obesity. Five different operative techniques were available, vertical banded gastroplasty (VBG), silastic ring vertical gastroplasty (SRVG), gastric banding operation, adjustable silicone gastric banding (ASGB) and biliopancreatic diversion (BPD). Methods: Two of these operations were mainly utilized. The ASGB was done routinely. The SRVG was used particularly for patients with hiatal hernia. Only one patient, who had a deformed pylorus, underwent the BPD. Results: Eleven patients had to be reoperated due to complications after the first operation of gastric restriction. They were divided into three groups depending on the type of complication: reoperation due to lack of compliance, due to technical failure, or due to other complications. In the last group we observed three patients with band perforation into the stomach without epigastric pain. This complication has, to our knowledge, only been described in very few cases. Conclusion: In some patients it remained difficult to reach adequate compliance, although we kept close contact with them after the operation. We do not yet know the reasons for the band perforation observed in three patients.     

Isolation perfusion in extracorporeal circulation with interleukin-2 and lymphokine-activated killer cells in the treatment of in-transit metastases from limb cutaneous melanoma

Background: Therapies of advanced melanoma patients with interleukin-2 (IL-2) and cytotoxic lymphocytes have produced interesting results, but a larger diffusion of these treatments is limited by the severe side effects due to IL-2 systemic infusion. A strictly regional administration of IL-2 and cells by an isolation perfusion (IP) in extracorporeal circulation (ECC) for the treatment of regional melanoma metastases could improve tolerability and efficacy of this specific modality of immunotherapy. Methods: Ten patients were submitted to adoptive immunotherapy with IL-2 and lymphokine-activated killer (LAK) cells by IP in ECC. The schedule of treatment included the first course of a 5-day systemic administration of IL-2 (Proleukin, EuroCetus 9–12 × 10⁶ IU/M²/day continuous infusion); autologous LAK cells were obtained via leukapheresis and after in vitro activation were given (range 8–28 × 10⁹) along with IL-2 (120-2,400 IU/ml of perfusion priming) to the affected limb by IP; IL-2 (9–12×10⁶ IU/m²/day) was also administered by systemic continuous infusion for 5 days starting on the day after IP. Results: All patients concluded the treatment without any major local or systemic toxicities. Clinical responses included one complete and six partial remissions; three patients had stable disease. All patients are alive. Follow-up after IP ranged from 12 to 35 months (median: 22). The analysis of circulating lymphocytes revealed the rapid disappearance of LAK cells, suggesting their extravasation and/or endothelial adhesion in perfused tissues. Conclusions: IP with IL-2 and LAK cells is a new approach for the treatment of in-transit metastases due to cutaneous melanoma. The treatment appears to be feasible and reliable. Further biological and immunological studies should permit amelioration of the present modality of treatment.     

Pregnancies in an 18-Year Follow-up after Biliopancreatic Diversion

Background: 239 pregnancies occurred in 1136 women who had undergone biliopancreatic diversion (BPD). Methods: There were 73 abortions, and 14 pregnancies are presently in their course. The 152 term pregnancies (six twins) occurred in 129 women 2-173 months (mean 42.7) after BPD. Mean age and current excess weight were 31.4 years (20-42) and 29.1% (-6.9-78.2), and mean excess weight loss was 72.9% (30.4-110.5). Results: Mean weight gain during pregnancy was 6.2 kg (-21-25). In 32 patients (21%), parenteral nutritional support was needed. In all the other patients (79%), the usual supplementations were given. Of the newborns, 122 were delivered at term (84.7%) with a mean weight of 2842.4 g (1760-4600 g) and a mean length of 48.5 cm (43-59 cm), while the 22 preterm babies (15.3%) weighed 2151.1 g (1400-3850 g) and had a length of 44.6 cm (33-56 cm). Forty infants (27.8%) were small for gestational age but 17 of them weighed more than 2500 g. Eleven twins (one abortion at 26th week) were also delivered, with a mean weight of 2088.6 g (1200-3100 g) and a mean length of 45.6 cm (35-50 cm). Delivery was spontaneous in 85 instances (56%), while vacuum extractor was used in one, and 66 cesarean sections were performed. There were two birth malformations, one infant died after surgery for meconium obstruction and two died from unknown causes. Of the 129 women, 35 had been infertile before BPD. Conclusions: Disappearance of infertility and decrease of pregnancy risk are to be considered among the beneficial effects of weight reduction following BPD.     

Responses to Median and Tibial Nerve Stimulation in Patients with Chronic Neuropathic Pain

Somatosensory evoked magnetic fields and electrical potentials were measured in eight patients with unilateral neuropathic pain. After median nerve stimulation on the painful side, the amplitudes of the evoked responses were enhanced 2 to 3 times at a latency of about 100 ms compared to the responses of the contralateral, unaffected side. After posterior tibial nerve stimulation an enhancement was found at latencies around 110 ms and 150 ms. The scalp distribution of the magnetic field at the latencies of abnormal responses was dipolar and the responses could be ascribed to a current dipole. Three (of the eight) patients underwent spinal cord stimulation (SCS) for their pain. The enhancement of the evoked responses to stimulation of the painful side decreased after spinal cord stimulation. After a long period of spinal cord stimulation only (e.g., a year) during which the patient reported to be pain free, these abnormal responses were no longer observed.