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Nance-Horan syndrome (NHS) is an X-linked developmental disorder, characterized by bilateral congenital cataracts, dental anomalies, facial dysmorphism and mental retardation. Null mutations in a novel gene, NHS, cause the syndrome. The NHS gene appears to have multiple isoforms as a result of alternative transcription, but a cellular function for the NHS protein has yet to be defined. We describe NHS as a founder member of a new protein family (NHS, NHSL1 and NHSL2). Here, we demonstrate that NHS is a novel regulator of actin remodelling and cell morphology. NHS localizes to sites of cell-cell contact, the leading edge of lamellipodia and focal adhesions. The N-terminus of isoforms NHS-A and NHS-1A, implicated in the pathogenesis of NHS, have a functional WAVE homology domain that interacts with the Abi protein family, haematopoietic stem/progenitor cell protein 300 (HSPC300), Nap1 and Sra1. NHS knockdown resulted in the disruption of the actin cytoskeleton. We show that NHS controls cell morphology by maintaining the integrity of the circumferential actin ring and controlling lamellipod formation. NHS knockdown led to a striking increase in cell spreading. Conversely, ectopic overexpression of NHS inhibited lamellipod formation. Remodelling of the actin cytoskeleton and localized actin polymerization into branched actin filaments at the plasma membrane are essential for mediating changes in cell shape, migration and cell contact. Our data identify NHS as a new regulator of actin remodelling. We suggest that NHS orchestrates actin regulatory protein function in response to signalling events during development.

Heat shock factor-1 (HSF1) is the central regulator of heat-induced transcriptional responses leading to rapid expression of molecular chaperones that protect mammalian cells against proteotoxic stress. The main targets for HSF1 are specific promoter elements (HSE) located upstream of heat shock genes encoding a variety of heat shock proteins, including HSP70, HSP90, HSP27, and other proteins of the network. Herein we report that the zinc finger AN1-type domain-2a gene, also known as AIRAP, behaves as a canonical heat shock gene, whose expression is temperature-dependent and strictly controlled by HSF1. Transcription is triggered at temperatures above 40 degrees C in different types of human cancer and primary cells, including peripheral blood monocytes. As shown by ChIP analysis, HSF1 is recruited to the AIRAP promoter rapidly after heat treatment, with a kinetics that parallels HSP70 promoter HSF1-recruitment. In transfection experiments HSF1-silencing abolished heat-induced AIRAP promoter-driven transcription, which could be rescued by exogenous Flag-HSF1 expression. The HSF1 binding HSE sequence in the AIRAP promoter critical for heat-induced transcription was identified. Because its expression is induced at febrile temperatures in human cells, AIRAP may represent a new potential component of the protective response during fever in humans.

Several studies have shown that the motor system is involved in action perception, suggesting that action concepts are represented through sensory-motor processes. Such conclusions imply that motor system impairments should diminish action perception. To test this hypothesis, a group of 10 brain-damaged patients with hemiplegia (specifically, a lesion at the motor system that affected the contralesional arm) viewed point-light displays of arm gestures and attempted to name each gesture. To create the dynamic stimuli, patients individually performed simple gestures with their unaffected arm while being videotaped. The videotapes were converted into point-light animations. Each action was presented as it had been performed, that is, as having been produced by the observer's unaffected arm, and in its mirror reversed orientation, that is, as having been produced by the observer's hemiplegic arm. Action recognition accuracy by patients with hemiplegia was compared with that by 8 brain-damaged patients without any motor deficit and by 10 healthy controls. Overall, performance was better in control observers than in patients. Most importantly, performance by hemiplegic patients, but not by nonhemiplegic patients and controls, varied systematically as a function of the observed limb. Action recognition was best when hemiplegic patients viewed actions that appeared to have been performed by their unaffected arm. Action recognition performance dropped significantly when hemiplegic patients viewed actions that appeared to have been produced with their hemiplegic arm or the corresponding arm of another person. The results of a control study involving the recognition of point-light defined animals in motion indicate that a generic deficit to visual and cognitive functions cannot account for this laterality-specific deficit in action recognition. Taken together, these results suggest that motor cortex impairment decreases visual sensitivity to human action. Specifically, when a cortical lesion renders an observer incapable of performing an observed action, action perception is compromised, possibly by a failure to map the observed action onto the observer's contralesional hemisoma.

Nance-Horan syndrome (NHS) is an X-linked developmental disorder characterized by congenital cataract, dental anomalies, facial dysmorphism and, in some cases, mental retardation. Protein truncation mutations in a novel gene (NHS) have been identified in patients with this syndrome. We previously mapped X-linked congenital cataract (CXN) in one family to an interval on chromosome Xp22.13 which encompasses the NHS locus; however, no mutations were identified in the NHS gene. In this study, we show that NHS and X-linked cataract are allelic diseases. Two CXN families, which were negative for mutations in the NHS gene, were further analysed using array comparative genomic hybridization. CXN was found to be caused by novel copy number variations: a complex duplication-triplication re-arrangement and an intragenic deletion, predicted to result in altered transcriptional regulation of the NHS gene. Furthermore, we also describe the clinical and molecular analysis of seven families diagnosed with NHS, identifying four novel protein truncation mutations and a novel large deletion encompassing the majority of the NHS gene, all leading to no functional protein. We therefore show that different mechanisms, aberrant transcription of the NHS gene or no functional NHS protein, lead to different diseases. Our data highlight the importance of copy number variation and non-recurrent re-arrangements leading to different severity of disease and describe the potential mechanisms involved.

The purpose of this paper is to analyse the nature of bureaucratization within public research bodies and its relationship to scientific performance, focusing on an Italian case-study. The main finding is that the bureaucratization of the research sector has two dimensions: public research labs have academic bureaucratization since researchers spend an increasing part of their time in administrative matters (i.e., preparing grant applications, managing grants/projects, and so on); whereas universities mainly have administrative bureaucratization generated by the increase over time of administrative staff in comparison with researchers and faculty. In addition, I show that research units with higher bureaucratization have lower scientific performance.

The regulatory function of invariant NKT (iNKT) cells for tolerance induction and prevention of autoimmunity is linked to a specific cytokine profile that comprises the secretion of type 2 cytokines like IL-4 and IL-10 (NKT2 cytokine profile). The mechanism responsible for iNKT cell differentiation toward a type 2 phenotype is unknown. Herein we show that costimulatory signals provided by the surface receptor signaling lymphocytic activation molecule (SLAM) on myeloid dendritic cells (mDC) to iNKT cells is crucial for NKT2 orientation. Additionally, we demonstrate that the impaired acquisition of an NKT2 cytokine phenotype in nonobese diabetic (NOD) mice that spontaneously develop autoimmune diabetes is due to defective SLAM-induced signals generated by NOD mDC. Mature mDC of C57BL/6 mice express SLAM and induce C57BL/6 or NOD iNKT cells to acquire a predominant NKT2 cytokine phenotype in response to antigenic stimulation with the iNKT cell-specific Ag, the alpha-galactosylceramide. In contrast, mature NOD mDC express significantly lower levels of SLAM and are unable to promote GATA-3 (the SLAM-induced intracellular signal) up-regulation and IL-4/IL-10 production in iNKT cells from NOD or C57BL/6 mice. NOD mice carry a genetic defect of the Slamf1 gene that is associated with reduced SLAM expression on double-positive thymocytes and altered iNKT cell development in the thymus. Our data suggest that the genetic Slamf1 defect in NOD mice also affects SLAM expression on other immune cells such as the mDC, thus critically impairing the peripheral differentiation of iNKT cells toward a regulatory NKT2 type.

Performance on tests of odour discrimination, naming, and matching was compared in patients with four distinct forms of neurodegenerative disease: Alzheimer's disease (AD), semantic dementia (SD), frontotemporal dementia (FTD), and corticobasal degeneration (CBD). The SD patients were found to have a severe impairment of identification from olfaction despite having normal discrimination, consistent with the multimodal semantic impairment characteristic of this patient group. The AD patients' poor odour discrimination suggests that a perceptual impairment is the root of their poor odour identification. Mild impairments in odour identification observed in FTD and CBD are consistent with their generalised executive dysfunction. The findings illustrate that breakdown in olfaction can occur at a perceptual or semantic level, analogous to the distinction between apperceptive and associative forms of deficit in the visual and auditory modalities. The findings add further insights into the nature of the semantic deficit in SD by exploring a hitherto neglected modality and may have relevance in explaining the altered eating habits commonly associated with SD.

One of the most effective techniques in the rehabilitation of visual field defects is based on implementation of oculomotor strategies to compensate for visual field loss. In the present study we develop a new rehabilitation approach based on the audio-visual stimulation of the visual field. Since it has been demonstrated that audio-visual interaction in multisensory neurons can improve temporally visual perception in patients with hemianopia, the aim of the present study was to verify whether a systematic audio-visual stimulation might induce a long-lasting amelioration of visual field disorders. Eight patients with chronic visual field defects were trained to detect the presence of visual targets. During the training, the visual stimulus could be presented alone, i.e. unimodal condition, or together with an acoustic stimulus, i.e. crossmodal conditions. In the crossmodal conditions, the spatial disparity between the visual and the acoustic stimuli were systematically varied (0, 16 and 32 degrees of disparity). Furthermore, the temporal interval between the acoustic stimulus and the visual target in the crossmodal conditions was gradually reduced from 500 to 0 ms. Patients underwent the treatment for 4 h daily, over a period of nearly 2 weeks. The results showed a progressive improvement of visual detections during the training and an improvement of visual oculomotor exploration that allowed patients to efficiently compensate for the loss of vision. More interesting, there was a transfer of treatment gains to functional measures assessing visual field exploration and to daily-life activities, which was found stable at the 1 month follow-up control session. These findings are very promising with respect to the possibility of taking advantage of human multisensory capabilities to recover from unimodal sensory impairments.

The correct approach for endometriosis management is still unclear. This review explores recent data concerning diagnosis and treatment of endometriosis, trying to define guidelines for the most appropriate diagnostic approach and therapeutic regimen. At present, laparoscopy is still considered the gold standard in endometriosis diagnosis. The risks and the diagnostic limitations of laparoscopy and the inaccuracy of clinical examination justify the considerable efforts made to improve the diagnosis with imaging techniques. The therapeutic approach is still far from being defined as causal and focuses on management of clinical symptoms of the disease rather than on the disease itself. A first-line medical therapy should be tried in patients with pelvic pain not asking for a pregnancy. Surgical treatment is considered the best treatment for women with pain and or pelvic mass who wish to become pregnant in a short time. For infertile patients, medical therapy has a limited role. The 2 treatment options include surgery or in vitro fertilization (IVF). According to our results, it seems that correct management of infertile women with endometriosis is a combination of surgery and IVF in women who did not obtain post-surgery pregnancy spontaneously.

Purpose of Investigation: To evaluate how many women required the so-called "emergency contraception" at our outpatient service and what the actual role is of this kind of pharmacological administration in interfering with ovulation and pregnancy, paying particular attention to the ethical and medico-legal aspects of this subject., Methods: During the period from 1 December 1998 to 30 November 2003, emergency contraception was prescribed to a total of 1,160 women. With regard to the contraceptives used, in most cases (1,132, 97.6%) a combined oral estrogen-progestogen pill (ethinyloestradiol 0.05 mg plus levonorgestrel 0.25 mg) was prescribed; in some cases (20 patients, 1.8%) danazol (400 mg), in four women (0.3%) a progestin-only pill (levonorgestrel 0.75 mg), and in four other women (0.3%) an intrauterine device., Results: It does not come out that there were any pregnancies in our study patients since none of them, who were told to come back for follow-up, were seen at our termination of pregnancy service or delivery room., Conclusion: The "Yuzpe regimen" of a combined oral estrogen-progestogen pill has been the most commonly used method for emergency contraception. A new method recently proposed, a progestin-only pill with levonorgestrel 0.75 mg, is having better results than the previous one, with a lower incidence of side-effects and higher efficacy. Moreover, the treatment with this method does not interfere in case of a pregnancy already being carried and cannot interrupt it.

Purpose of Investigation: The management of fetal ovarian cysts is still controversial despite the improvement in prenatal diagnosis with ultrasonography. Some studies suggest an aggressive management, while others opt for a conservative one. The prognosis of the majority of congenital ovarian cysts is good since they have a benign origin. Sometimes, however, complications such as torsion or rupture can occur which often require surgical intervention after delivery. In this paper we report our experience and a brief review of the literature., Methods: The authors report on 32 pregnant women in whom ultrasonography revealed the presence of an echo-rare or echo-free area in the fetal abdomen suggestive of an ovarian cyst. All women were followed-up during pregnancy with serial ultrasound examinations. Postnatal ultrasound controls confirmed the prenatal diagnosis in all cases. The diameters of the cysts ranged from 2.7 to 7.5 cm., Results: In the 16 cases (50%) in which the cyst diameter was below 4 cm, periodic ultrasound examinations revealed a tendency towards spontaneous regression of the cysts. In the other 16 cases (50%) in which the cyst diameter exceeded 4 cm, cystectomy was necessary due to subsequent complications (torsion in 6 cases, 37.5%, and intracystic hemorrhage in the other 10, 62.5%)., Conclusion: The most appropriate clinical approach in the management of benign feto-neonatal ovarian cysts is to adopt a wait-and-see policy, assessing the course of the condition by means of periodic ultrasound monitoring. Only when tumefactions measure more than 4 cm in diameter with attendant complications is surgical therapy indicated. Without complications, however, aspiration of the cystic contents is possible even in ovarian cysts exceeding 4 cm in diameter.

Classic neurological accounts and some contemporary theories of semantic memory assume that concepts are acquired through a learning process that draws together information experienced in each of our verbal and nonverbal modalities. These accounts embody three critical assumptions: semantic representations are amodal; the mapping between surface form and meaning varies for different modalities; and the representations are dynamic. The influence of these three factors was revealed in data collected over a 4-year longitudinal period in two patients with semantic dementia. Semantic assessment revealed a parallel decline in verbal and nonverbal aspects of conceptual knowledge, reflecting a gradual degradation of a single amodal semantic system. As expected, when the patients' semantic impairment was mild, they presented with profound anomia but relatively preserved object use. Over time, performance on all semantic tasks including object use declined. High item-by-item consistency across these tasks was observed in all testing sessions. The impact of dynamic semantic representations was revealed by a striking clinical finding. Although unable to name many of the objects in isolation, their performance was significantly facilitated if they were asked to name while they demonstrated the use of each object. These results are discussed in the context of contemporary models of semantic memory.

Between January 1996 and December 2001, at the Department of Gynecology, Perinatology and Human Reproduction of the University of Florence, 49 ectopic pregnancies were submitted to medical treatment. The treatment schedule consisted of the administration of 100 mg of intravenous methotrexate (MTX). The patients included in this study fulfilled the following requisites: gestational period <8 weeks; diameter of the ectopic gestational sac <4 cm; serum level of human chorionic beta-gonadotropin (beta-hCG) <5000 IU/ml; absence of clinical and ultrasound signs of tube rupture with initial hemoperitoneum; hematochemical tests compatible with chemotherapic treatment. All patients were followed with a dosage of serum beta-hCG repeated every 2-3 days after chemotherapy and with an ultrasound every 3-4 days. In case of documented success of treatment the patient was hospitalized for no more than 3 days after administration of the drug. In 1 case therapy took place in a day-hospital regimen. Medical treatment was effective in 35 patients out of 49 (71.4%) and led to negative beta-hCG in a median time of 11 days, with a range between 2 and 48 days. In the 14 non-responsive cases (28.6%), after a mean time of 6 days we proceeded to a traditional surgical approach or laparoscopy. In none of the cases did we find significant pharmacological toxicity, while in 9 patients (18.3%), severe painful symptoms appeared immediately after treatment, but resolved within 24 hours. Our results are interesting and in agreement with other experiences found in the literature. In our opinion, the advisability of a second administration in case of slow response, the comparison with an analogous intramuscular treatment, a more precise definition of the eligibility criteria, long-term follow-up of the patients, especially in case of subsequent pregnancies should all be further considered.

The dual-route model of writing assumes two basic procedures involved in writing: lexical and non-lexical. The lexical route is fundamental in opaque orthographies such as English; from its impairment surface dysgraphia arises. Evidence for the role of a lexical route in transparent languages such as Italian, which have a regular orthography, has been more limited. We report a case study of ES, an Italian patient suffering from degenerative brain disease, who presented with a selective disorder of writing. He showed the unusual phenomenon of inserting an apostrophe inappropriately in the spellings of words. Neuropsychological evaluation provided evidence of loss of orthographic meaning of the apostrophe and a pattern of writing performance consistent with surface dysgraphia. There was also evidence of an accompanying surface dyslexia. We conclude that examination of apostrophe use provides a valuable means of detecting surface dysgraphia in the Italian language. The findings point to the need for cognitive models of writing to account for "dumb symbols" of language such as the apostrophe. This unique case provides a further example of the variety of clinical presentations of focal cerebral degeneration.

Objective: To evaluate the efficacy and safety of ultrasound (US) guidance as compared to laparoscopic monitoring during operative hysteroscopy., Study Design: Prospective, open study including 81 patients undergoing operative hysteroscopy under US guidance for uterine septum and submucous myoma. Clinical and surgical outcomes were compared with those in an historical control group of 45 patients undergoing the same operation under laparoscopic guidance., Results: US guidance proved satisfactory in all patients, and there were no complications due to insufficient visualization of the pelvic structures; in no case was conversion to laparoscopic guidance required. US scanning was most useful in determining the outer limit of the intramural component of submucous partial intramural myoma, allowing complete resection. During metroplasty, US guidance allowed extension of the resection beyond the normal limit conventionally defined by hysteroscopy; none required reintervention. By comparison, in the control group, a second attempt was required because the operation was insufficiently radical in four patients., Conclusion: US guidance was used successfully as the only visual aid for hysteroscopic surgery, comparing favorably with laparoscopy in terms of efficacy and safety.

T-cell activation requires co-stimulation through receptors such as CD28 and antigen-specific signalling through the T-cell antigen receptor. Here we describe a new murine costimulatory receptor-ligand pair. The receptor, which is related to CD28 and is the homologue of the human protein ICOS, is expressed on activated T cells and resting memory T cells. The ligand, which has homology to B7 molecules and is called B7-related protein-1 (B7RP-1), is expressed on B cells and macrophages. ICOS and B7RP-I do not interact with proteins in the CD28-B7 pathway, and B7RP-1 co-stimulates T cells in vitro independently of CD28. Transgenic mice expressing a B7RP-1-Fc fusion protein show lymphoid hyperplasia in the spleen, lymph nodes and Peyer's patches. Presensitized mice treated with B7RP-1-Fc during antigen challenge show enhanced hypersensitivity. Therefore, B7RP-1 exhibits co-stimulatory activities in vitro and in vivo. ICOS and B7RP-1 define a new and distinct receptor-ligand pair that is structurally related to CD28-B7 and is involved in the adaptive immune response.

Preantral ovarian follicles isolated from prepubertal sheep ovaries were individually cultured for 6 days in the presence of increasing doses of FSH (ranging from 0.01 to 1 microg/ml) and under two different oxygen concentrations, 20% and 5% O2. Follicle development was evaluated on the basis of antral cavity formation as well as the presence of healthy cumulus oocyte complexes. Follicle growth was enhanced by FSH addition to culture medium, while the use of a low oxygen concentration slightly stimulated this process. However, when follicles were cultured in the presence of high doses of FSH (1 microgram/ml) and under low oxygen concentration, a high proportion of them showed the presence of an antral cavity and of a healthy cumulus-oocyte complex. In addition, under this specific culture condition sheep preantral follicles released higher levels of estradiol as compared to those secreted at lower FSH concentrations or under 20% O2. When the meiotic competence of oocytes derived from follicles cultured at 1 microgram/ml FSH was assessed, no significant difference was recorded between the two oxygen groups. These results show that the culture conditions here identified are beneficial to in vitro growth and differentiation of sheep preantral follicles.

We report here that immunization of human PBMC reconstituted SCID mice (hu-PBL-SCID mice) with in vitro cultured autologous dendritic cells (DC) pulsed with prostate specific antigen (PSA) complexed to a PSA-specific mouse IgG2a (PSA-IgG2a) consistently and reproducibly stimulates PSA-specific human IgG production. On day 0, female PBMC were used to reconstitute SCID mice and to generate DC in vitro. DC cultures were pulsed with PSA or PSA-IgG2a on day 6. The previously reconstituted hu-PBL-SCID mice were immunized with either PSA-pulsed DC and PSA, PSA-IgG2a-pulsed DC and PSA-IgG2a, or additional PBMC and PSA-IgG2a on day 7. Mice immunized with PSA-IgG2a-pulsed DC had, on the average, up to 31.5 times greater PSA-specific IgG serum concentrations than control mice. Competition ELISA confirmed the PSA specificity of serum IgG. Immunoblot analysis suggested that sera IgG preferentially recognized conformational epitopes on PSA. Therefore, our results represent a major step toward cloning human tumor-associated Ag-specific human mAbs from hu-PBL-SCID mice. In addition, flow cytometry showed that PSA-pulsed DC express significantly more B7.1, B7.2, CD40, and MHC class II surface molecules than mock-treated DC, but PSA-IgG2a-pulsed DC only had significantly enhanced B7.2 surface expression. Interestingly, PSA-specific IgG responses were reproducibly stimulated by DC expressing more B7.2, a molecule associated with Th2-type immune deviation, but not by those expressing more B7.1 and CD40, molecules associated with Th1-type immune deviation. Thus, our results show that stimulation with either Ag or Ag complexed to mAb yields DC with different phenotypes and APC effector functions.

Thirty-four episodes of pneumococcal bacteremia were identified in 15 patients over 5 years in 10 hospitals in Franklin County, Ohio. Twelve patients each had 2 episodes of pneumococcal bacteremia, 2 had 3, and 1 had 4. All patients had predisposing conditions, with lymphoma, multiple myeloma, and chronic obstructive pulmonary disease being the most frequent. The mean interval between the first and second episode was 268 days. Serotyping and genotyping were performed on 29 isolates. The same serotypic and genotypic patterns were found for sequential isolates from four patients; three of these patients had a recurrence between 22 and 90 days after a previous episode. Seven (24%) of the 29 isolates were serotype 23F; four isolates (14%) were not susceptible to penicillin. All of our patients received appropriate antimicrobial therapy and appeared to be clinically cured of their initial infection. For patients with recurrent pneumococcal disease, alternate preventive measures such as immunization with conjugate pneumococcal vaccine and/or prophylactic antibiotic therapy should be considered.

The SCIDhu PBL model of human Ig production was modified by using human interleukin-6 (hIL-6) secreting tumors for continuous hIL-6 production, in vivo. On day one, SCID mice were injected subcutaneously with 200 microliters PBS (control mice), 10(4) SP2/0-Ag14 cells (IL-6+ mice) or 10(4) hIL-6 secreting SP2/0-hIL6.17 cells (IL-6- mice). The mice were reconstituted with human PBMC on day two and immunized with 100 micrograms of tetanus toxoid (TT) on days two and fifteen. Serum hIL-6 concentrations in IL-6+ mice ranged between 2.9 and 38.1 ng/ml by days 26-33. IL-6+ mice had enlarged spleens and lymph nodes (LN). Flow cytometry and histology showed that SCIDhu PBL mouse spleen, LN and peritoneal exudate cells (PEC) contained mostly murine myeloid lineage cells. In addition, many more human B cells, T cells and IL-2R(+)-activated lymphocytes were present in spleen, LN and PEC of IL-6+ mice. Despite enhanced lymphocyte engraftment and activation, by day 14 IL-6+ mice produced up to 6-fold less TT-specific IgG relative to total IgG than either control group. TT-specific and total Ig sera concentrations were equivalent in all three groups on days 26-33. Our results suggest that sustained circulating hIL-6 enhanced human delayed-type hypersensitivity (DTH)-like inflammatory responses with consequential inhibition of TT-specific IgG production in SCIDhu PBL mice.

Traditional hybridoma fusion technology requires complete medium with serum supplements to support the growth of hybridoma cells. Serum is also required for subcloning of hybridoma cells to support low density cell growth. IL-6 has been shown to enhance the growth of hybridomas and stimulate antibody production by B cells. We found that the serum requirement in media used for generation of hybridomas can be totally eliminated by substituting with 300 units/ml of IL-6. Stable hybridoma cell lines were generated to peptide and protein antigens using serum-free adapted P3.653 myelomas as the fusion partner and medium containing IL-6. Our results indicate that, in general, the fusion efficiencies of serum-free IL-6 supplemented fusions are lower than the fusions employing serum containing media (40%-60% vs. 80%-100%). However, in spite of the lower fusion efficiency, the number of antigen-specific clones generated using IL-6 was equal to or greater than fusions using serum supplements. The use of IL-6 instead of serum in the generation of monoclonal antibodies (MAbs) has several advantages. We are able to eliminate the costly need for serum in media by using IL-6 that is prepared in house. In addition, we eliminate the need for time-consuming serum-free adaptation of hybridoma cell lines prior to transfer to hollow fiber bioreactors.

Objective: The purpose of this contribution is to investigate the pathophysiology of the abnormal endometrial development in hyperstimulated IVF cycles., Study Design: In 12 IVF-patients who did not have embryo transfer because of failure of oocyte fertilization, serum values of 17 beta-estradiol, progesterone, FSH, LH, total and free testosterone, and androstenedione were measured on the pick-up day and were evaluated with respect to the values normally expressed in the day of ovulation; in the endometrial specimens collected 2 days later, at the time of embryo replacement, estrogen and progesterone receptors were immunohistochemically determined and dating by the Noyes method was performed., Results: 17 beta-Estradiol values are constantly higher, and progesterone levels are, only in four cases, higher than expected for the day of ovulation in a natural cycle. These hormonal patterns can only partially explain the pattern of steroid receptors: progesterone receptors are expressed sparsely both in glands and stroma, while estrogen receptors are abundant in the glands and absent in the stroma. In 11 of 12 patients an abnormal endometrial development with stromal advancement was observed: this morphological picture of the endometrium could partially be explained only in the four cases presenting high progesterone levels by serum values and endometrial receptor content of estrogen and progesterone., Conclusions: The abnormal endometrial development in hyperstimulated IVF cycles could only in part be explained by estrogen and progesterone, and other factors have to be considered.

Purpose: This multicenter study was carried out to compare the efficacy of intrauterine insemination (IUI) and intraperitoneal insemination (IPI) associated with multiple follicular development as treatment for unexplained infertility., Method: A total of 205 couples completed the trial. Sixty-seven couples underwent treatment with IPI (group A) and 138 couples underwent treatment with IUI (group B)., Results: Clinical pregnancy was obtained in 23 couples in group A (pregnancy rate: 34.3%) and in 36 couples in group B (pregnancy rate: 26.1%). No significant difference was observed between group A and group B. As for the evolution of pregnancies and the incidence of twin pregnancies, no significant difference was observed between the two groups., Conclusions: Because IUI and IPI allow us to obtain same results and IPI is more invasive than IUI, the latter technique can be considered the method of choice and IPI should be used when IUI is difficult to perform, as in the presence of a tight cervical canal.

Background: Bilateral vulvar edema associated with severe ovarian hyperstimulation syndrome is described for the first time., Case: A 28-year-old woman underwent gonadotropin-releasing hormone analogue and gonadotropin treatment for in vitro fertilization and embryo transfer. On day 18 of the cycle, the patient reported mild abdominal discomfort that became severe during the following four days. One the 22nd day of the cycle, bilateral ovarian enlargement and ascites were present. The vulva showed massive edema and fissures. Ovarian hyperstimulation syndrome (OHSS) therapy consisted of human albumin, lactated Ringer's solution and heparin. The vulvar edema was treated with topical hydrocortisone ointment, ice packs and topical gentamycin twice a day. After one week of treatment, the vulva was normal., Conclusion: We think that the vulvar edema in this case was the result of decreased oncotic pressure and increased hydrostatic pressure, as occur during OHSS.

Gene amplification is an important mechanism of increased gene expression in a number of human solid tumors. We have recently identified and cloned sequences from a novel DNA amplification unit in malignant fibrous histiocytoma. The amplified sequences are derived from chromosome 12q13-14 and encode a gene designated SAS (sarcoma amplified sequence). In the present study, a series of soft tissue sarcomas was studied to characterize further the phenomenon of SAS amplification. Seven of 22 (32%) malignant fibrous histiocytomas and three liposarcomas contained SAS amplification. Strikingly, all of the tumors with SAS amplification occurred in central sites (i.e., in the abdominal or inguinal regions) rather than in the extremities (i.e., in the arms of legs). These observations demonstrate that SAS amplification occurs with a significant frequency in mesenchymal tumors and is particularly associated with abdominal disease.

Amplification of cellular oncogenes occurs frequently in several human cancers and is an important mechanism of increased gene expression. Identification of amplified genes in tumor cells has proved to be a useful approach for understanding genetic alterations in cancer. Previous procedures for isolating probes from amplified DNA sequences have relied on tissue culture cells, limiting the range of tumors that can be studied and raising questions of in vitro artifact. We have circumvented these problems by combining in gel renaturation of amplified sequences with the polymerase chain reaction. Using this approach, we have identified and partially cloned a DNA amplification unit from biopsies of human malignant fibrous histiocytoma. This amplification unit is derived from chromosome 12q13-14, a site commonly involved in rearrangements in soft tissue tumors, and contains at least one transcribed region (designated SAS, for sarcoma amplified sequence).

A 15-year-old female with acquired von Willebrand disease associated with hypothyroidism is presented and discussed. The patient was initially seen for right lower quadrant abdominal pain and menorrhagia. Once hypothyroidism was diagnosed and treated her coagulation abnormality resolved.

The molecular characteristics of two human doxorubicin-resistant cell lines were examined specifically for MDR1 gene amplification by Southern analysis and for overexpression of its messenger RNA. The 285-fold doxorubicin-resistant colon adenocarcinoma subline, LoVo/DR5, was found to overexpress the mRNA for P-glycoprotein without the concomitant requirement of MDR1 gene amplification, suggesting that relatively high levels of P-glycoprotein mediated multiple drug resistance may occur by transcriptional activation of the gene. Despite a similar in vitro selection strategy and in contrast to LoVo/DR cells, the 220-fold doxorubicin-resistant fibrosarcoma subline, HT1080/DR4, did not overexpress P-glycoprotein mRNA nor was the MDR1 gene amplified. In-gel renaturation studies were performed to determine the nature of a putative HSR-bearing chromosome 7 found in HT1080/DR4 cells; however, at a level of sensitivity nearing 20 copies of an amplified DNA fragment per haploid genome, no amplified sequences could be detected. These results suggest that doxorubicin resistance is multifactorial and alternative mechanisms of multiple drug resistance remain to be determined. LoVo/DR5 cells should prove to be a useful model for investigating transcriptional activation of the MDR1 gene; HT1080/DR4 cells should be an excellent model for the study of non-P-glycoprotein mediated multiple drug resistance.

This study analyzed a group of morbidly obese patients who underwent jejunoileal bypass over a 10 year period. Patients underwent either a Payne or Scott procedure. Weight loss was reported in terms of percent of excessive weight loss. Results showed that patients who had a Payne bypass had an average excess weight loss of 75 percent, whereas those with the Scott bypass had an average excess weight loss of 61 percent. All patients lost weight. Complications in this series were lower than what has been traditionally reported. The problem that occurred most often was kidney stones (8 percent of the patients). There has been only one death to date. The low incidence of complications can be attributed to the fact that the patients were evaluated, operated on, supervised, and followed by one doctor and his associated nurses. Thus, it is proposed that jejunoileal bypass, compounded with good patient care and careful patient selection, can produce positive results with a minimum number of problems.

A randomized polycentric study was programmed to establish the effects of daily administration of ferritin iron from early pregnancy to puerperium. 254 women with normal iron balance at the beginning of their pregnancy were randomized receiving no supplements or 40 mg iron daily. At the end of pregnancy iron balance was still normal only in one third of the pregnant women of the first group versus two third of the second group. 204 women who were iron-deficient received daily 40 or 120 mg of iron; in this group anemia developed less frequently (13% versus 29%) and iron balance normalized in one subject on four; the great majority of these women remained iron-deficient. Unwanted effects of minimal or mild relevance, and almost always sporadic were observed in 6.5% of cases and with the reduction or withdraw of the treatment in only 1.4% of cases. These results showed that daily administration of ferritin iron during pregnancy is effective and well tolerated; furthermore they suggest that the treatment must be done with at least 60 mg daily in women with normal iron balance and protracted also after the puerperium in iron deficient subjects.

The diagnosis of thoracic outlet syndrome rests on clinical findings. A study of 21 patients with the syndrome is reported. Six of the patients underwent surgery and the remainder were treated conservatively. Surgery completely eliminated the symptoms in five patients; one patient had minor symptoms after surgery. In the patients managed conservatively, half had partial relief of symptoms and the other half did not respond to therapy. Physical therapy is initially recommended in most patients.

With increasing awareness of cost-containment measures, early discharge after parathyroidectomy for primary hyperparathyroidism is to be evaluated. This report concerns the Cleveland Clinic experience with the last 70 consecutive patients treated for primary hyperparathyroidism from June 1981 to June 1983. The mean postoperative hospital stay was 1.4 days. Most patients were discharged on the morning of the second postoperative day receiving oral calcium supplements for 3 weeks. Three patients were discharged on the morning of the first postoperative day. The overall mortality rate was 0%. Morbidity included two patients (2.9%) who developed symptoms of mild tetany after discharge that responded to an increased dose of oral calcium. The usual practice of most surgeons of delaying hospital discharge after neck exploration for primary hyperparathyroidism for 5 to 7 days in fear of symptomatic hypocalcemia is unnecessary. Considerable cost containment can be achieved by sparing patients from 3 to 5 extra days of hospitalization.