Search

Study question What is the transcriptomic response of decidualized endometrial stromal cells (DESCs) to the uptake of extracellular vesicles (EVs) secreted by human embryos? Summary answer Human embryo derived EVs can impact DESCs transcriptomic profile via a cargo that can induce a positive effect on the expression of some decidualization-related genes. What is known already The establishment of an appropriate embryo–endometrial communication is essential for the implantation process and a successful pregnancy. Embryo-released factors are able to elicit biological effects on the endometrium. Among these, EVs secreted by embryos carry molecules potentially able to modulate endometrial function and favour the implantation process. However, most results come from animal species as few studies were performed using EVs derived from human embryos, due to difficulty in sample collection or ethical reasons. Study design, size, duration EVs were isolated from one pool of IVF media where embryos were cultured between day 3 and 5 of development (blastocyst). Initially, DESCs(n = 4) were treated with different concentrations of EVs at different time points and RNA-sequencing was performed to define optimal conditions.Subsequently, EVs were isolated from one pool of IVF media where blastocysts diagnosed as euploid were cultured (EVe). To increase the reliability of the results, DESCs(n = 5) were treated with EVe using the optimal conditions. Participants/materials, setting, methods EVs were isolated by differential ultracentrifugation from spent media of in vitro cultured human blastocyst stage embryos and from an equal volume of control medium (not exposed to embryos). DESCs were in vitro decidualized (E2:10nM+ P4:1uM+cAMP:0.5mM) and treated with different concentrations of EVs (10µg,50µg,100µg) for 24hours and 48hours. RNA-sequencing analysis was then performed. Once established the conditions that allow to better detect differences in trascriptomic profile, RNA-sequencing analysis on DESCs treated specifically with EVe was performed. Main results and the role of chance The best response of DESCs to EVs uptake was induced by treatment with 10µg of EVs for 48hours. Using this condition, differential gene expression analysis allowed the identification of 166 upregulated and 124 downregulated genes in DESCs treated with EVe, when compared to DESCs treated with control. Based on Over-Representation analysis, DESCs treated with EVe were found to be enriched in transcripts of genes involved in signalling and cell communication. Moreover, based on Gene Set Enrichment Analysis results, EVe may regulate decidualization and the implantation process by targeting cholesterol biosynthesis and metabolism pathways, being among the most enriched pathways. The depletion of transcripts related to inflammatory and IL1R pathways in DESCs treated with EVe was also observed, suggesting a role of EVe in the fine regulation of inflammatory/immunomodulatory events occurring during implantation. At the intersection of genes whose expression was modified by EVe-treatment and those modified by the decidualization treatment, thirty-one transcripts exhibited the same trend, demonstrating that EVe exerted a positive effect on decidualization-related genes expression in DESCs. Specifically, expression of genes involved in the implantation process (PTGER2, IL15, WNT4) increased after decidualization and were further up-regulated after EVe-treatment, confirming the crucial role of EVs in the embryo-endometrium cross-talk. Limitations, reasons for caution Larger studies are needed to validate these findings. Wider implications of the findings EVs derived from human euploid embryos can evoke a specific transcriptional response in DESCs. Moreover, we speculate that these EVs may exert favourable effects on the endometrial tissue, increasing its receptivity for embryo implantation by modulating decidualization-related genes expression. Trial registration number not applicable

Study question How do fertility clinics’ websites of the UK, Belgium and Spain present the medical risks of egg donation and incentives? Summary answer Spanish and UK websites typically included more incentives to recruit egg donors compared to the Belgian websites. OHSS was overall the most discussed risk. What is known already People commonly turn to the internet for initial information. Primary presentations of information of a subject co-determine how an individual interprets the topic and thus influence later decisions. Considering the growing demand and clinics’ dependency on egg donors, some scholars have expressed concerns that clinics might (initially) misrepresent risks to recruit more egg donors. Offering appealing incentives may also encourage potential donors to dismiss possible risks and side-effects. Therefore, it is important to see how incentives (both monetary and non-monetary rewards) and risks are presented on the websites of fertility clinics, the first source of information for egg donors. Study design, size, duration This study is part of the EDNA-project, a multi-phased comparative study (2017-2021), that aims to explore the social, political, economic and moral configuration of egg donation in the United Kingdom, Belgium and Spain. In this study, we only focused on the medical risks of egg donation and incentives presented on the fertility clinics’ websites of the three countries. Participants/materials, setting, methods We analysed the websites of all Belgian fertility clinics (n = 18), and a maximum variation sample in the UK (n = 21) and Spain (n = 23). The sampling was based on the geographical location of the clinic, size/number of cycles performed each year, the clinic’s status (independent or part of a larger clinical group) and whether the clinic was public or privately funded in the UK. Frame analysis and content analysis were used for analysis in Nvivo12. Main results and the role of chance No misrepresentation of risks was found. There was an extensive variety in the representation of risks. There were differences between the websites of the three countries (e.g. the risks of having acne was only discussed on Spanish websites), but also remarkable differences within the websites of a particular country (of the 40 risks, 13 were one-off mentions, each found on a single website). A description of a risk was generally accompanied by a minimization or normalization of the risk, or a statement about the fertility clinic’s excellent care for their egg donors. These three approaches were often combined. Our analysis differentiated between incentives (i.e. external rewards) and emotional appeals. An appeal to emotions (e.g. empathy) can also motivate behavioural action (e.g. donation) but was not considered as an incentive since there is no external reward. While Belgian websites used almost no incentives, Spanish and UK websites used gratitude and a rewarding experience as incentives. However, only Spanish websites used free medical tests as incentives, while UK websites were the only ones that used discounts received with egg sharing as an incentive. All countries’ websites used emotional appeals by enticing feelings of empathy for the recipients. Limitations, reasons for caution Not all UK and Spanish fertility clinics’ websites were analysed. However, our international team of researchers applied a maximum variation sampling strategy. This generated samples of clinics per country that were as diversified as possible. Wider implications of the findings The study shows that incentives are more often used on the UK and Spanish websites than on the Belgian websites. All three countries’ websites used emotional appeals. It should be studied how effective these incentives and emotional appeals are, and if there is a correlation with the potential donors’ risk-perceptions. Trial registration number not applicable

Study question Could EVs secreted by aneuploid embryos a) serve for development of RNA biomarkers for PGT-A and b) elicit a relevant transcriptomic response in decidualised ESCs? Summary answer Aneuploid embryo EVs a) contain PPM1J, LINC00561, ANKRD34C and TMED10 in differential abundance from euploid EVs and b) induce up-regulation of MUC1 in decidualised ESCs. What is known already Embryo aneuploidy accounts for approximately 50% of all recurrent implantation failures in women >35 years old. PGT-A identifies euploid embryos to increase implantation probability but the technology is controversial as it requires an invasive embryo biopsy with an elusive long-term biosafety. The development of non-invasive methods to screen out aneuploid embryos is paramount. It is also critical to decode the embryo-endometrial dialog underlying implantation failure. We have previously reported that IVF embryos secrete EVs that can be internalised by ESCs, conceptualising that successful implantation to the endometrium is facilitated by EVs, which may additionally serve as biomarkers of ploidy status. Study design, size, duration Embryos destined for biopsy on days 5-7 for PGT-A were grown under standard conditions. Spent media (30μl) were collected from euploid (n = 175) and aneuploid embryos (n = 145) at both cleavage (days 1-3) and blastocyst (days 3-5) stage. Media samples from n = 35 cleavage embryos were pooled in order to obtain five euploid and four aneuploidy pools. Blastocyst media were pooled to create one euploid and one aneuploid pool. ESCs were obtained from five women undergoing diagnostic laparoscopy. Participants/materials, setting, methods The study was realised at a research hospital. EVs were isolated from euploid and aneuploid Day3 pools with differential ultracentrifugation and EV-RNA sequencing was performed following the SMARTer Stranded Total RNA-Seq approach. ESCs were decidualised (E2:10nM, P4:1uM, cAMP:0.5 mM twice every 48 hours) and treated for 24 hours with 50 ng/ml euploid or aneuploid EVs extracted from blastocyst media. RNA sequencing was performed on ESCs following the Truseq RNAseq protocol. Main results and the role of chance Aneuploid cleavage stage embryos (n = 4) secreted EVs that were less abundant in RNA fragments originating from the genes PPM1J (log2fc=-5.13, p = 0.011), LINC00561 (log2fc=-7.87, p = 0.010) and ANKRD34C (log2fc=-7.30, p = 0.017) and more abundant in TMED10 (log2fc=1.63 p = 0.025) compared to EVs (n = 5) from euploid embryos. Decidualisation per se induced downregulation of MUC1 (log2FC=-0.54, p = 0.0028) in ESCs as prerequisite for the establishment of receptive endometrium. The expression of MUC1 transcript in decidualised ESCs was significantly increased following treatment with aneuploid compared to euploid embryo-secreted EVs (log2FC=0.85, p = 0.0201). Limitations, reasons for caution The findings of the study may require validation utilising a second cohort of EVs samples. Wider implications of the findings This discovery that the RNA cargo of EVs secreted from aneuploid cleavage stage embryos is diverse from that of euploid embryos potentiates the development of non-invasive methodology for PGT-A. The upregulation of MUC1 in decidualised ESCs following aneuploid embryo EV treatment proposes a new mechanism underlying implantation failure. Trial registration number NA

The main aim of the study is to assess clinical and functional outcomes of arthroscopic outside-in repair of isolated radial tears of the midbody of lateral meniscus in professional athletes and to evaluate the return to the sport activity after surgery.A retrospective data collection on professional athletes with isolated complete lesion of the midbody of lateral meniscus, treated with arthroscopic outside-in repair was carried out. Outcome measures included functional assessment, Limb Symmetry Index (LSI) and Hamstring Quadriceps Ratio (HQR) and Lysholm score collected before surgery and at 4-month follow-up. Data on return to sport practice and re-injury were also retrieved.Fourteen patients satisfied the selection criteria. Full return to professional sport activity (Tegner 10) was registered in the 86% of the cohort at 4 months after the surgery. Functional testing of the athletes showed a return of the LSI and HQR to the pre-surgical condition, demonstrating a full recovery of the functional ability and muscle strength. Similarly, clinical evaluation through Lysholm score showed an improvement, reaching an average of 97.7 points at 4 months follow-up.A good functional recovery and a high rate of return to play has been observed in a population of professional athletes, at 4 months after outside-in repair.

Systemic anaplastic large cell lymphoma (sALCL) is a rare histological entity expressing the CD30 antigen that comprises around 11% of peripheral T-cell lymphoma. We analysed the outcome of patients with relapsed/refractory sALCL treated with autologous stem cell transplantation (auto-HCT). We included 65 adult patients (42 males; median age, 44 years); 24 patients had an ALK-ve sALCL. Fifty-one patients had chemosensitive disease at the time of transplant. Ten patients (15%) were treated with brentuximab vedotin (BV) before auto-HCT (median number of doses: 5). The median follow-up for surviving patients was 35 months (3-71). Three-year cumulative incidence of nonrelapse mortality and of relapse were 1.7% and 34%, respectively. Three-year progression-free survival and overall survival were 64% and 73%, respectively. No prognostic factors for any of the outcomes analysed were found in univariate analysis. There were no significant differences in any of the outcomes between patients who had received BV and the remainder. This is the largest analysis presented so far analysing the role of auto-HCT in patients with relapsed/refractory sALCL, showing a promising PFS and OS in this high-risk population. The potential impact of the administration of BV as salvage strategy before the procedure needs to be further elucidated.

Enzyme-Based electrodes represent an important class of biosensors that allow to electrochemically measure the products derived from a target analyze transformation.

Codon optimized DNA and aminoacidic sequence of human pro-relaxin H2. (DOCX 11 kb)

Osteoporosis is the most common bone disease, affecting millions of people and causing a high risk of fractures and a loss of quality of life. It is characterized by low bone mass and microarchitectural deterioration of bone tissue, with a consequent increase in bone fragility and susceptibility to fracture. A primary method of prevention, in order to reduce the risk of fractures, is represented by an appropriate lifestyle and a correct diet. There are potentially numerous nutrients and dietary components that can influence bone health, and these range from macronutrients to micronutrients as well as bioactive food ingredients. The purpose of this review is to overview osteoporosis, including its definition, etiology, and incidence, and then provide some information on possible dietary strategies for optimizing bone health and preventing osteoporosis. A correct diet to prevent osteoporosis should contain adequate amounts of calcium, vitamins D and K, protein, and fatty acids. The effects of these elements are briefly discussed, reporting on their correlation with bone benefits.

Bioconjugation is a rapidly expanding field because of the numerous potential applications of bioconjugate materials. We explored the usefulness of branched porphyrins as rigid scaffolds, bearing multiple sites for bioconjugation. To this end, we first selected the tetrakis(p-[aminomethyl] phenyl) porphyrin (TAMPP) macrocycle and developed a straightforward synthetic protocol, able to provide the desired tetraphenylporphyrin, carrying four functional amino groups. The partially protection of the amino groups by tert-butoxy-carbonyl allowed the selective and specific decoration of the porphyrin with different peptide sequences. To explore the utility of the macrocycle as molecular scaffold for bioconjugation, we selected peptide sequences able to function as thrombin inhibitors. In particular, two peptide sequences, named CS3 and ES7, able to interact, respectively, with the thrombin catalytic site and the fibrinogen recognition exosite, were joined onto the porphyrin macrocycle, providing the multisite-directed inhibitor CS3-TAMPP-ES7. This multisite inhibitor and its MnIII complex are able to inhibit ?-thrombin-catalyzed hydrolysis of Tos-Gly-Pro-Arg-nitroanilide with inhibition constants in the micromolar range, as well as the hydrolysis of the natural substrate fibrinogen. The inhibitor is resistant against enzymatic degradation by thrombin and is highly selective. The MnIII complex is capable of interacting with clot-bound thrombin and partially inhibits clot growth in the presence of fibrinogen. The results herein reported are very promising, suggesting the potential of the newly developed conjugate as new imaging agents for clot detection.

A novel class of synthetic, multisite-directed thrombin inhibitors, known as hirunorms, has been described recently. These compounds were designed to mimic the binding mode of hirudin, and they have been proven to be very strong and selective thrombin inhibitors. Here we report the crystal structure of the complex formed by human alpha-thrombin and hirunorm V, a 26-residue polypeptide containing non-natural amino acids, determined at 2.1 A resolution and refined to an R-factor of 0.176. The structure reveals that the inhibitor binding mode is distinctive of a true hirudin mimetic, and it highlights the molecular basis of the high inhibitory potency (Ki is in the picomolar range) and the strong selectivity of hirunorm V. Hirunorm V interacts through the N-terminal tetrapeptide with the thrombin active site in a nonsubstrate mode; at the same time, this inhibitor specifically binds through the C-terminal segment to the fibrinogen recognition exosite. The backbone of the N-terminal tetrapeptide Chg1"-Val2"-2-Nal3"-Thr4" (Chg, cyclohexyl-glycine; 2-Nal, beta-(2-naphthyl)-alanine) forms a short beta-strand parallel to thrombin main-chain residues Ser214-Gly219. The Chg1" side chain fills the S2 subsite, Val2" is located at the entrance of S1, whereas 2-Nal3" side chain occupies the aryl-binding site. Such backbone orientation is very close to that observed for the N-terminal residues of hirudin, and it is similar to that of the synthetic retro-binding peptide BMS-183507, but it is opposite to the proposed binding mode of fibrinogen and of small synthetic substrates. Hirunorm V C-terminal segment binds to the fibrinogen recognition exosite, similarly to what observed for hirudin C-termninal tail and related compounds. The linker polypeptide segment connecting hirunorm V N-and C-terminal regions is not observable in the electron density maps. The crystallographic analysis proves the correctness of the design and it provides a compelling proof on the interaction mechanism for this novel class of high potency multisite-directed synthetic thrombin inhibitors.

The aim of this study was to evaluate the impact of reduced peripheral insulin sensitivity, beta cell hypersecretion and reduced hepatic insulin clearance in the hyper-insulinaemia of lean and obese PCOS patients. A total of 35 women with polycystic ovary syndrome (PCOS) and 10 lean normo-ovulatory controls underwent an oral glucose tolerance test and an euglycaemic-hyper-insulinaemic clamp study. PCOS patients were classified into four groups according to their BMI and insulin secretion (normo-lean; normo-obese; hyper-lean; hyper-obese), and results were compared between groups and with the controls. All the PCOS groups showed significantly higher insulin secretion than controls; there were no differences in insulin response to glucose load between lean and obese normo- and hyper-insulinaemic patients. Secretion of c-peptide was greater in PCOS groups than controls. All the hyper-insulinaemic PCOS patients had lower values of hepatic insulin clearance, independent of BMI, when compared either with controls (P < 0.001) or with PCOS normo-insulinaemic women (P < 0.01). Normo- and hyper-insulinaemic obese patients had similar total body glucose utilization (M value), which was lower than in lean PCOS subjects and controls. Our results suggest that evaluation of insulin resistance alone does not fully characterize the PCOS population; differences in liver metabolism of insulin are present in obese insulin resistant subjects and in lean patients with normal insulin sensitivity when divided into normo- and hyper-insulinaemic subgroups. Insulin resistance and hyper-insulinaemia may represent two distinct features of the insulin disorder in PCOS: the former appear to reflect the presence of obesity, while the latter may be a primary feature of PCOS.

The relationship between insulinaemia and obesity and glucose tolerance and the impact of pregnancy as risk factor for carbohydrate abnormalities were investigated in 91 consecutive patients with polycystic ovary syndrome (PCOS) aged 26-32 years. Fifteen normoglycaemic patients became pregnant within 6 months of the pregestational study using pharmacological induction of ovulation. Plasma concentrations of insulin and glucose after an oral glucose tolerance test (OGTT) were determined by immunoradiometric assay and glucose oxidase technique respectively. OGTT patients were classified according to their response as normoinsulinaemic (n = 46) or hyperinsulinaemic (n = 45). Impairment of glucose metabolism occurred in 12.1% (n = 11, 10 obese and one lean) of all PCOS subjects. Based on insulin secretion, 6.5% of normoinsulinaemic and 13.3% of hyperinsulinaemic patients had an impaired glucose tolerance and 2.3 and 2.2% respectively a non-insulin-dependent diabetes mellitus. Obese patients had higher values for area under the curve for insulin response to OGTT (I-AUC values) than lean patients, and the percentage above ideal body weight was greater in hyperinsulinaemic than in normoinsulinaemic patients. All hyperinsulinaemic (7/15) subjects who became pregnant developed an impairment of glucose metabolism during pregnancy. It is concluded that the PCOS population was at higher risk of developing carbohydrate abnormalities than the normal population of a similar reproductive age. Furthermore, those with abnormal insulin secretion at the pregestational stage may, during pregnancy, develop an impaired gestational glucose tolerance or gestational diabetes.

Summary DESIGN There are conflicting data on hypothalamic-pituitary-adrenal (HPA) axis function in women with polycystic ovary syndrome (PCOS). We have evaluated the HPA axis responses to naloxone in patients with PCOS compared to control subjects. PATIENTS Twenty PCOS patients and 10 control women participated in the study. MEASUREMENTS On days 5–6 Of a spontaneous or progestin induced cycle each patient received an intravenous bolus (5 mg) of naloxone (time 0 min), followed by a 2-mg naloxone infusion In 100 ml of 0.9% saline over one hour. Samples were collected at −30, 0, 15, 30, 60, 90 and 120 minutes. ACTH and cortisol levels were measured in all plasma samples. RESULTS PCOS patients showed significantly greater response than controls to naloxone of ACTH (peak value 261 vs 172% of basal value) and cortisol (peak value 237 vs 165% of basal value); also, ACTH and cortisol incremental areas were higher In PCOS patients (P < 0.05 and P < 0.04 respectively). The cortisol/ACTH ratio of AUCs percentage increase was found to be near unity for all patients without significant difference between PCOS and control groups, suggesting a direct correspondence between ACTH circulating levels and adrenal cortisol production. CONCLUSIONS Polycystic ovary syndrome patients showed a hypothalamic-pituitary-adrenal axis hyper-responsiveness to naloxone infusion compared with control subjects. These data support the hypothesis that this disturbance could be central in origin.

Sepsis is a major cause of significant morbidity and mortality in neutropenic patients. Blood culture remains the gold standard in the microbiological diagnosis of bacterial or fungal bloodstream infections, but it has clear limits of rapidity and sensitivity. The objective of the study was to compare the real-time polymerase chain reaction (RT-PCR) with automated blood cultures (BC) method in detection in whole blood of pathogens in febrile neutropenic patients with hematological malignancies.A total of 166 consecutive febrile neutropenic patients were enrolled. Blood samples for cultures and SeptiFast testing were obtained at the onset of fever, before the implementation of empirical antibiotic therapy.Forty (24.1%) samples out of the 166 blood samples tested, were positive by at least one method. Twenty-three (13.9%) samples were positive by blood culture and 38 (22.9%) by multiplex real-time PCR. The analysis of concordance evidenced a low correlation between the two methods (n = 21; 52.5%), mainly due to samples found negative by culture but positive with the Septi-Fast assay. Sensitivity, specificity, and positive and negative predictive values of RT-PCR were 91.3%, 88.1%, 55.3%, and 98.4%, respectively, compared with BC.Multiplex real-time PCR assay improved detection of the most bacteria associated with febrile neutropenia episodes. Further studies are needed to assess the real advantages and clinical benefits that molecular biology tests can add in diagnosis of sepsis.

This study was designed to investigate the growth hormone (GH) secretory response to the growth hormone-releasing hormone (GHRH) test in women with polycystic ovary syndrome (PCOS). A total of 25 patients with PCOS (13 obese and 12 non-obese) and 15 normal ovulatory women (seven obese and eight non-obese) were included in this study. In the follicular phase patients were subjected to an oral glucose tolerance test (OGTT); 2 days later they underwent a GHRH test. Basal plasma concentrations of gonadotrophins, steroids and sex hormone-binding globulin were measured. Insulin and GH were assayed under the OGTT and GHRH test respectively. Based on the insulin response to OGTT in the PCOS group, nine patients were classified as normoinsulinaemic and 16 as hyperinsulinaemic; none of the patients of the control group had a hyperinsulinaemic response to OGTT. Obese patients showed a trend towards a lower GH response to GHRH. Moreover, hyperinsulinaemic patients showed a significantly (P < 0.05) lower area under the curve for secretion of GH (AUC-GH) as compared to normo-insulinaemic patients. All PCOS patients exhibited a markedly decreased response of GH to GHRH compared with the control population. Obese and hyperinsulinaemic PCOS patients were both found to have a lower response of GH to GHRH than all other groups. Despite the fact that obesity and hyperinsulinaemia have an additive influence on the impairment of GH secretion, our results suggest that other factors may also negatively affect GH secretion in PCOS.

In order to test the possible negative effect of corticotropin-releasing factor (CRF) on the pituitary-gonadal axis, two gonadotropin-releasing hormone (GnRH) tests, on two consecutive days, were performed in five normally cycling women in the midluteal phase of the menstrual cycle. The first GnRH test was performed after 2 h during which time basal blood samples were collected every 10 min. On the second day, the GnRH test was repeated after a 2-h CRF infusion which was continued throughout the GnRH test. Our results demonstrate that while CRF increases cortisol levels it affects neither the basal nor the GnRH-stimulated gonadotropin levels. Similarly, both basal and luteinizing hormone (LH)-stimulated progesterone concentrations were not modified by the CRF infusion thereby also excluding a possible direct effect of the peptide on corpus luteum steroidogenesis.

Patients and methods 62 patients affected by oligoarticular JIA that referred to our Departments in the period between December 2004 and June 2008, were included in the study. 34 cases (54%) had persistent oligoarthritis and the only affected joints were knee (45%) and ankle (9%). The remaining patients presented extended oligoarthritis and the involved joints, in addition to knee and ankle, were elbow (3%), wrist (3%), interphalangeal (6%). Triamcinolone hexacetonide and Methylprednisolone acetate were used. Patients were followed up for three years. MTX was started only if NSAIDs and IAC failed. Clinical pattern and serologic data, such as ESR and ANAs, were evaluated in order to correlate them with treatment response to predict the outcome of IAC procedure.

The urokinase plasminogen activator receptor (u-PAR) focuses the proteolytic activity of the urokinase plasminogen activator (u-PA) on the endothelial cell surface, thus promoting angiogenesis in a protease-dependent manner. The u-PAR may exist in a glycophosphatidylinositol-anchored and in a soluble form (soluble u-PAR [Su-PAR]), both including the chemotactic Ser⁸⁸ -Arg-Ser-Arg-Tyr⁹² internal sequence.To investigate whether Su-PAR may trigger endothelial cell signaling leading to new vessel formation through its chemotactic Ser⁸⁸ -Arg-Ser-Arg-Tyr⁹² sequence.In this study, the formation of vascular-like structures by human umbilical vein endothelial cells was assessed by using a matrigel basement membrane preparation. First, we found that Su-PAR protein promotes the formation of cord-like structures, and that this ability is retained by the isolated Ser(88) -Arg-Ser-Arg-Tyr⁹² chemotactic sequence, the maximal effect being reached at 10 nmol L⁻¹ SRSRY peptide (SRSRY). This effect is mediated by the α(v) β₃ vitronectin receptor, is independent of u-PA proteolytic activity, and involves the internalization of the G-protein-coupled formyl-peptide receptor in endothelial cells. Furthermore, exposure of human saphenous vein rings to Su-PAR or SRSRY leads to a remarkable degree of sprouting. Finally, we show that Su-PAR and SRSRY promote a marked response in angioreactors implanted into the dorsal flank of nude mice, retaining 91% and 66%, respectively, of the angiogenic response generated by a mixture of vascular endothelial growth factor and fibroblast growth factor type 2.Our results show a new protease-independent activity of Su-PAR that stimulates in vivo angiogenesis through its Ser⁸⁸ -Arg-Ser-Arg-Tyr⁹² chemotactic sequence.

Lymphangiomatosis is a well-recognized congenital benign tumour, frequently seen in infancy and childhood, characterized by the presence of multiple lymphangiomas. Diffuse lymphangiomatosis also involving bony tissue is called Gorham's disease. This condition generally affects somatic soft tissue, where lymphatics are normally found. A predilection of this affection for bone, thoracic and neck involvement is well known, while involvement of abdominal viscera is very unusual. In bone this non malignant proliferation of lymphatic channels results in destruction and resorption of the osseous matrix. We report on a child in whom lymphangiomatosis involved both the bone and the spleen. A review of 166 cases is also reported.

Total hip replacement surgery carries the risk of thromboembolic complications, which could be fatal. Over the last three decades however, the risk has decreased considerably thanks to progress made in the understanding of the physiopathogenetic mechanism of thromboembolic disease and perioperative prophylaxis. It is the purpose of this study to discuss the main medical and surgical preventive measures that must be carried out before, during and immediately after surgery. The old concept of deciding thromboembolic prophylaxis after surgery is now obsolete.

A crystal-state structural analysis of t-Boc-L-Ala-β-Ala-NHMe, t-Boc-Aib-β-Ala-NHMe, and t-Boc-Aib-Aib-β-Ala-NHMe has been performed by X-ray diffraction. While the conformation adopted by t-Boc-L-Ala-β-Ala- NHMe and t-Boc-Aib-β-Ala-NHMe is essentially extended, f-Boc-Aib-Aib- β-Ala-NHMe is folded into two consecutive intramolecularly hydrogen-bonded structures of the i + 3 → i type (β-bends), with Aib(1)-Aib(2) and Aib(2)-β-Ala(3), respectively, as corner residues. Owing to the presence of the β-aminoacid, the latter β-bend is characterized by an unusual C11 hydrogen-bonded ring. These results indicate that: (i) a β-aminoacid may be incorporated into a β-bend without a major perturbation of the overall geometry of this folded conformation, and (ii) the propensity of the β-Ala residue for β-bend formation is rather low, unless other conformational constraints (e.g. a preceding β-bend) are present in the linear peptide molecule.

Die Rontgenstereometrieanalyse (RSA) ist eine etablierte Methode in der Chirurgie des Bewegungsapparates. Das Ziel dieser experimentellen Studie war es, die Anwendung der digitalen Radiographie in der RSA in Hinblick auf Reduktion des Flachendosisproduktes und Verbesserung der Bildqualitat zu analysieren. Material und Methoden: Eine Knieprothese wurde in ein Leichenknie implantiert. In das Polyathyleninlay wurden funf, in das proximale Tibiaplateau 17 Tantalkugeln eingesetzt. Fur die Rontgenuntersuchung stand eine Obertischrohre (General Electrics) und eine Mobilette (Siemens) zur Verfugung. Fur die konventionellen Rontgenaufnahmen wurde ein Film- Folien System (Folie: SE- Folie grun {Gadolinium Oxisufid}; Agfa Gevaert verwendet. Die digitalen Aufnahmen wurden mit dem Agfa Diagnostik Center (ADC) (Folie: Phosphorzusammensetzung BaSrFBr: Eu; Speicherfolien-EK: 100) angefertigt. Das Auflosungsvermogen betragt beim Kassettenformat 24*30 und einer EK von 200, 2,5 LP/mm. Die Dokumentation erfolgte auf Laserfilmen im Format 8*10 inch. Mit einer Ionisationskammer (Diamentor M4, PTW- Freiburg) erfolgte die Dokumentation des Flachendosisproduktes. Die Bildqualitat wurde durch drei erfahrene Untersucher anhand einer subjektiven Qualitatsskala bewertet. Ergebnisse: Es wurden 30 digitale und 20 konventionelle Bildpaare ausgewertet. Bei einer Rohrenspannung von 109 kV und 3,2 mAs betrug das Flachen-Dosisprodukt fur digitale und konventionelle Aufnahmen a.p. 7,2 cGy/cm2 und seitlich 8,5 cGy/cm2. Die Bildgute war bei beiden Verfahren zufriedenstellend. Die Kriterien fur eine RSA-Analyse waren erfullt. Bei 66 kVund 3,2 mAs ergab sich ein Flachen- Dosisprodukt von a.p. 2,7 cGy/cm2 und seitlich 2,5 cGy/cm2. Mit digitaler Technik war die Bildgute befriedigend und die Kriterien fur eine RSA-Analyse waren erfullt. Die konventionellen Rontgenaufnahmen liesen eine RSA-Analyse nicht zu. Eine weitere Dosisreduktion auf ein Flachen- Dosisprodukt von 1,3 cGy/cm2 im seitlichen und im a.p. Strahlengang lies sich nur in digitaler Technik erreichen (66 kV; 1,6 mAs). Die Kriterien fur eine RSA-Analyse waren hier bei einem guten bis zufriedenstellenden Bildeindruck erfullt. Zusammenfassungg: Diese Untersuchung zeigte erstmalig, das eine Reduktion des Flachen-Dosis- Produktes um circa 82% im anterioren und um circa 85% im lateralen Strahlengang bei gutem bis zufriedenstellendem Bildeindruck und erfolgreicher RSA-Analyse moglich ist. Die digitale Radiographie bietet zusatzlich die Moglichkeit der Nachverarbeitung sowie der Einbindung in digitale Kommunikations- und Speichersysteme (PACS).

Certain chemokines act as natural antagonists of human immunodeficiency virus (HIV) by blocking key viral coreceptors, such as CCR5 and CXCR4, on the surface of susceptible cells. Elucidating the structural determinants of the receptor-binding and HIV-inhibitory functions of these chemokines is essential for the rational design of derivative molecules of therapeutic value. Here, we identify the structural determinants of CCR5 recognition and antiviral activity of the CC chemokine RANTES, showing that critical residues form a solvent-exposed hydrophobic patch on the surface of the molecule. Moreover, we demonstrate that the biological function is critically dependent on dimerization, resulting in the exposure of a large ( approximately 180 A2), continuous hydrophobic surface. Relevant to the development of novel therapeutic approaches, we designed a retroinverted RANTES peptide mimetic that maintained both HIV- and chemotaxis-antagonistic functions.

We report on a 63-year-old patient with Gaucher disease type I who developed severe bone involvement with destructive lesions and huge soft tissue extension in both humeri that appeared to evolve slowly. The clinical course and histopathological findings in our patient suggested a progressive extraosseous extension of the storage cells into the soft tissue, accompanied by a striking increase of fibrotic tissue and resulting in an impressive deformity. The extraordinary bone involvement in this patient expands our knowledge on the most severe skeletal complications of untreated Gaucher disease.

Recent studies on the conformational preferences of the Dphig (C(alpha,alpha)-diphenylglycine) residue showed that this C(alpha,alpha)-disubstituted glycine has a structural versatility. In fact, depending on the nature of the following or preceding residue, Dphig can assume either folded or extended conformations. We have carried out the analysis of the conformational preferences of the Dphig residue in tripeptides containing consecutive Dphig residues. The crystal structures of Z-Dphig-Dphig -Dphig-OMe (a; Z = benzyloxycarbonyl; OMe = methyl ester), Z-Aib-Dphig-Dphig-OMe (b; Aib = alpha-aminoisobutyric acid), and Z-Ac(3)c-Dphig-Dphig-OMe (c; Ac(3)c = alpha-amino-cyclopropan carboxylic acid), are here reported. The Dphig residues adopt the fully extended conformation in the three tripeptides examined. Together with our previous findings on Dphig containing peptides, the structures of the peptides here examined, indicate that the presence of adjacent Dphig residue in the sequence further stabilizes the extended conformation.

We have investigated the conformational preferences of a newly synthesized C(alpha,alpha) symmetrically disubstituted glycine, namely alpha,alpha-dicyclopropylglycine (Dcp). We report here the crystal structure of a fully protected dipeptide containing Dcp, namely Z-Dcp(1)-Dcp(2)-OCH(3). Both Dcp residues are in a folded conformation. The overall peptide structural organization corresponds to an alpha-pleated sheet conformation, similar to that observed in linear peptides made up of alternating D- and L-residues and in Z-Aib-Aib-OCH(3) (Aib: alpha,alpha-dimethylglycine). These preliminary data suggest that the Dcp could represent an alternative as molecular tool to stabilize folded conformations.

Objective: To investigate the involvement of opioid tone, obesity, and hyperinsulinemia in GH secretion in women with polycystic ovary syndrome (PCOS). Design: Controlled clinical study. Setting: Catholic University of Sacred Heart School of Medicine in Rome, Italy. Patient(s): Twenty-two patients with PCOS and 14 healthy, normally ovulating volunteers, matched for age and body mass index. Intervention(s): Patients underwent a GH-releasing hormone (GHRH) test and an oral glucose tolerance test before and after 4–5 weeks of treatment with 50 mg/d of naltrexone. Main Outcome Measure(s): Serum concentrations of GH, insulin, glucose, steroids, and gonadotropins, as well as the GH area under the curve (AUC-GH) and the insulin area under the curve (AUC-I), were measured before and after naltrexone treatment. Result(s): In patients with PCOS, the administration of naltrexone increased the GH response to the GHRH test without interfering with the insulin response to the oral glucose tolerance test. However, the GH response to the GHRH test was improved significantly only in lean patients with PCOS, whereas obese patients with PCOS did not show any improvement in GH secretion. In obese control subjects, the treatment reduced plasma basal insulin concentrations and increased the AUC-GH, whereas in lean control subjects, the treatment reduced the GHRH-induced response. In normoinsulinemic patients with PCOS, the GH response to the GHRH test increased significantly after treatment, whereas the AUC-I was not affected. In hyperinsulinemic patients with PCOS, treatment with naltrexone significantly reduced the AUC-I, whereas the AUC-GH increased only in lean hyperinsulinemic patients with PCOS. Conclusion(s): Naltrexone treatment improves GHRH-induced GH secretion in patients with PCOS. However, this GH response is heterogeneously represented in relation to both obesity and hyperinsulinism.