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1.
Sci Rep ; 8(1): 1174, 2018 01 19.
Article in English | MEDLINE | ID: mdl-29352119

ABSTRACT

The Old World non-human primates (NHP) - baboons (Papio spp.) share similarities with humans regarding fetal and placental development and some pregnancy-related complications. Information about the mechanism of birth and complications arising during parturition in these species is relatively sparse. In this manuscript, we add information from a series of pathological and observational cases to highlight insights and selected complications of birth in Papio spp, based on video-recording of the delivery process, X-ray, MRI, and ultrasound evaluations in pregnant baboons. Additionally, we abstracted pathology records obtained from perinatal loss in a large baboon colony during a 17 year period. The presented cases provide important information for the management of pregnancy and delivery in Papio spp.


Subject(s)
Papio/physiology , Parturition/physiology , Animals , Female , Gestational Age , Labor, Obstetric , Male , Papio/anatomy & histology , Pregnancy , Pregnancy Complications
3.
AJP Rep ; 5(1): e60-6, 2015 Apr.
Article in English | MEDLINE | ID: mdl-26199802

ABSTRACT

Background Improvements in congenital heart disease (CHD) screening are needed based on the lack of sensitivity of current screening methods and the understanding that the early detection of certain CHDs may improve outcomes. Fetal venous circulation has caught medical attention, and two studies demonstrated that it is feasible to register pulmonary vein flow velocity waveforms (FVWs) during early gestation. Meanwhile, the latter study proposed pulmonary vein A-wave reversal as a marker of cardiac anomaly. Methods We report a series of six consecutive fetuses with confirmed cardiac anomalies that underwent first-trimester screening, including pulmonary vein FVWs, at our center during 2013. CHD was confirmed by late pregnancy echocardiography, and in three cases fetal autopsies were performed. Result/Discussion The ductus venosus (DV) and nuchal translucency (NT) predicted 50% of CHD cases, whereas the combination of markers identified 66.6% of CHD cases. When adding pulmonary vein assessment, the rate of detection rose to 83.3%. Total five of six cases of CHD had reversal of pulmonary vein A-wave during early pregnancy. The sixth case with CHD and nonreversal of A-wave was described as right ventricle hypoplasia with type 1 tricuspid atresia and persistent ductus arteriosus. Conclusion This is the first series reporting pulmonary vein end-diastolic reversal as a CHD screening add-on during early pregnancy. The addition of pulmonary vein FVW assessment to the current CHD screening bundle could increase the rate detection of cardiac anomalies. This pilot study suggests that pulmonary vein end-diastolic flow reversal favors detection of left-sided CHD over the right-sided ones.

4.
Am. j. obstet. gynecol ; 212(6)Jun. 2015. tab
Article in English | BIGG - GRADE guidelines | ID: biblio-965276

ABSTRACT

OBJECTIVE: We sought to provide evidence-based guidelines for the diagnosis and management of fetal anemia. METHODS: A systematic literature review was performed using MEDLINE, PubMed, EMBASE, and the Cochrane Library. The search was restricted to English-language articles published from 1966 through May 2014. Priority was given to articles reporting original research, in particular randomized controlled trials, although review articles and commentaries were consulted. Abstracts of research presented at symposia and scientific conferences were not considered adequate for inclusion. Evidence reports and published guidelines were also reviewed, and additional studies were located by reviewing bibliographies of identified articles. GRADE (Grading of Recommendations Assessment, Development, and Evaluation) methodology was used for defining the strength of recommendations and rating the quality of evidence. Consistent with US Preventive Task Force guidelines, references were evaluated for quality based on the highest level of evidence. RESULTS AND RECOMMENDATIONS: We recommend the following: (1) middle cerebral artery peak systolic velocity (MCA-PSV) measured by ultrasound Doppler interrogation be used as the primary technique to detect fetal anemia; (2) amniotic fluid delta OD450 not be used to diagnosis fetal anemia; (3) MCA-PSV assessment be reserved for those patients who are at risk of having an anemic fetus (proper technique for MCA-PSV evaluation includes assessment of the middle cerebral artery close to its origin, ideally at a zero degree angle without angle correction); (4) if a fetus is deemed at significant risk for severe fetal anemia (MCA greater than 1.5 multiples of the median or hydropic), fetal blood sampling be performed with preparation for an intrauterine transfusion, unless the pregnancy is at a gestational age when the risks associated with delivery are considered to be less than those associated with the procedure; (5) if a fetus is deemed at significant risk for severe fetal anemia, the patient be referred to a center with expertise in invasive fetal therapy; (6) MCA-PSV be considered to determine the timing of a second transfusion in fetuses with anemia, and, alternatively, a predicted decline in fetal hemoglobin may be used for timing the second procedure; and (7) pregnancies with a fetus at significant risk for fetal anemia be delivered at 37-38 weeks of gestation unless indications develop prior to this time.(AU)


Subject(s)
Humans , Fetal Nutrition Disorders/diagnosis , Fetal Nutrition Disorders/therapy , Blood Transfusion, Intrauterine , Hydrops Fetalis , Cordocentesis , Amniocentesis
5.
Placenta ; 34(11): 983-9, 2013 Nov.
Article in English | MEDLINE | ID: mdl-24008071

ABSTRACT

INTRODUCTION: Maternal obesity (MO) remains a serious obstetric problem with acute and chronic morbidities for both mothers and offspring. The mechanisms underlying these adverse consequences of MO remain unknown. Endocannabinoids (ECB) are neuromodulatory lipids released from adipocytes and other tissues. Metabolic crosstalk between placenta and adipocytes may mediate sequelae of MO. The goal of this study was to elucidate placental and systemic ECB in MO. MATERIAL AND METHODS: Placentas, sera, and subcutaneous fat were collected at Cesarean sections performed near term (0.9 G) in four non-obese (nOB) and four obese (OB) baboons (Papio spp.). Concentrations of anandamide (AEA) and 2-arachidonoylglycerol (2-AG) were measured by liquid chromatography coupled to tandem mass spectrometry. AEA and 2-AG pathways were characterized in placentas by Q-RT-PCR, Western blot and immunohistochemistry. RESULTS: Placental 2-AG levels were lower and maternal fat AEA levels were higher in OB (1254.1 ± 401.3 nmol/kg and 17.3 ± 4 nmol/kg) vs. nOB (3124.2 ± 557.3 nmol/kg and 3.1 ± 0.6 nmol/kg) animals. Concentrations of 2-AG correlated positively between maternal fat and placenta (r = 0.82, p = 0.013), but correlated negatively with maternal leptin concentrations (r = -0.72, p = 0.04 and r = -0.83, p = 0.01, respectively). CONCLUSION: This is the first study to demonstrate differential ECB pathway regulation in maternal fat and placenta in MO. Differential regulation and function exist for AEA and 2-AG as the major ECB pathways in placenta.


Subject(s)
Disease Models, Animal , Endocannabinoids/metabolism , Obesity/metabolism , Placenta/metabolism , Pregnancy Complications/metabolism , Receptors, Cannabinoid/metabolism , Subcutaneous Fat, Abdominal/metabolism , Animals , Arachidonic Acids/blood , Arachidonic Acids/metabolism , Biological Transport , Chromatography, High Pressure Liquid , Endocannabinoids/blood , Female , Gene Expression Regulation, Developmental , Glycerides/blood , Glycerides/metabolism , Leptin/blood , Obesity/blood , Obesity/pathology , Papio , Placenta/pathology , Polyunsaturated Alkamides/blood , Polyunsaturated Alkamides/metabolism , Pregnancy , Pregnancy Complications/blood , Pregnancy Complications/pathology , Receptor, Cannabinoid, CB1/genetics , Receptor, Cannabinoid, CB1/metabolism , Receptor, Cannabinoid, CB2/biosynthesis , Receptor, Cannabinoid, CB2/genetics , Receptor, Cannabinoid, CB2/metabolism , Receptors, Cannabinoid/biosynthesis , Receptors, Cannabinoid/genetics , Subcutaneous Fat, Abdominal/pathology , Tandem Mass Spectrometry
6.
J Med Primatol ; 42(4): 204-10, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23621893

ABSTRACT

BACKGROUND: Abruptio placentae is a serious problem with a high rate of maternal and fetal mortality and documented sexual dimorphism in reoccurrence. Macaca fascicularis is a well-described reproductive model; however, there are no data available regarding sexual dimorphism in abruptio placentae in these species. METHODS: A retrospective study of pathology and medical records in a large colony of M. fascicularis was performed. Placental specimens were analyzed. RESULTS: The incidence of placenta abruptio in the colony was 15.7/1000 births. In the abruptio placentae group, male fetuses had lower placental disk length and increased femur length compared with female fetuses. The feto-pacental ratio and fetal weight were lower in the male fetuses in the abruption group compared with those in the stillbirth group without abruption placentae. CONCLUSION: This is the first documentation of male bias in placental and fetal development in abruptio placentae in non-human primates.


Subject(s)
Abruptio Placentae/veterinary , Macaca fascicularis , Monkey Diseases/epidemiology , Abruptio Placentae/epidemiology , Abruptio Placentae/physiopathology , Animals , Female , Fetal Development , Fetal Weight , Fetus/pathology , Male , Monkey Diseases/pathology , Monkey Diseases/physiopathology , Placenta/pathology , Pregnancy , Retrospective Studies , Sex Factors , Stillbirth/epidemiology , Stillbirth/veterinary
7.
Urol Int ; 90(3): 249-52, 2013.
Article in English | MEDLINE | ID: mdl-23221307

ABSTRACT

INTRODUCTION: Among the medical and surgical options which have been proposed in the last years for the management of male stress urinary incontinence (SUI), stem cell therapy represents a new frontier treatment. The aim of this paper is to update the current status of stem cell therapy in animal and human studies for the management of iatrogenic male SUI. MATERIAL AND METHODS: A PubMed review of the literature on stem cell therapy for the treatment of male SUI was performed. RESULTS: Regarding animal studies, bone marrow-, muscle- and adipose-derived stem cells have been widely studied, showing regeneration of the urethral sphincter and recovery of the damaged pelvic nerves. With regard to human studies, only four papers are available in the literature using muscle- and adipose-derived stem cells which reported a significant improvement in sphincteric function and incontinence with no severe side effects. CONCLUSIONS: In spite of these promising results, further studies are needed with longer follow-ups and larger numbers of patients in order to clarify the potential role of stem cell therapy for the treatment of male SUI.


Subject(s)
Iatrogenic Disease , Regeneration , Stem Cell Transplantation/methods , Urethra/surgery , Urinary Incontinence, Stress/surgery , Animals , Cell Self Renewal , Humans , Male , Recovery of Function , Risk Factors , Stem Cell Transplantation/adverse effects , Treatment Outcome , Urethra/physiopathology , Urinary Incontinence, Stress/diagnosis , Urinary Incontinence, Stress/etiology , Urinary Incontinence, Stress/physiopathology
8.
Placenta ; 33(4): 278-84, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22265925

ABSTRACT

INTRODUCTION: Placental abruption is a serious condition that increases perinatal morbidity and mortality. Clinical prevention and treatment options are limited, especially in human preterm deliveries. Knowledge of the mechanisms that keep the placenta in place during pregnancy is critical for developing strategies for the prevention of abruption. Failure of physiological transformation of spiral arteries has been described as a major contributing factor of the placental abruption development. Baboons (Papio spp.) share striking similarities with humans in regard to placental structure, utero-placental blood flow, and fetal development; however, the mode of trophoblast invasion is shallow in baboons. This fact prompted the hypothesis that the incidence of placental abruption will be increased in baboons compared to humans. MATERIAL AND METHODS: Baboon placentas were collected between 2002 and 2008. Two independent veterinary pathologists evaluated the slides. A certified physician pathologist performed additional histology. RESULTS: Placental abruption was diagnosed in 22 baboons among 2423 live births during the study period (0.9% prevalence). The most common clinical presentations were fetal demise and vaginal bleeding. The most common pathological findings were intraplacental hemorrhages with or without hematoma formation (86.4%). Other findings consisted of neutrophil infiltration (50%), decidual necrosis (22.7%), decidual vascular congestion and inflammation, villous congestion and retroplacental hemorrhage/hematoma (each 18.2%). These pathologic findings were the same for term and preterm deliveries. CONCLUSION: This is the first systematic study of placental abruption in non-human primates, analyzing a large colony of baboons. Despite differences in trophoblast invasion, the clinical features observed in placental abruption affecting baboons resembled those reported in humans. The cluster of placental pathological findings in baboons also agreed with clinical reports, but the prevalence of these findings differed between baboons and humans. We discuss a mechanism of anti-abruption forces that offset shallow trophoblast invasion observed in baboons.


Subject(s)
Abruptio Placentae/pathology , Abruptio Placentae/physiopathology , Disease Models, Animal , Monkey Diseases/pathology , Monkey Diseases/physiopathology , Papio , Abruptio Placentae/epidemiology , Abruptio Placentae/immunology , Animals , Animals, Laboratory , Female , Fetal Death/etiology , Hematoma/etiology , Hemorrhage/etiology , Monkey Diseases/epidemiology , Monkey Diseases/immunology , Neutrophil Infiltration , Placenta/blood supply , Placenta/immunology , Placenta/pathology , Placentation , Pregnancy , Prevalence , Risk Factors , Texas , Uterine Hemorrhage/etiology
9.
Biomed Environ Sci ; 23(4): 300-4, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20934118

ABSTRACT

OBJECTIVE: To present the protocol and the early results of a urothelial carcinoma (UC) screening analysis performed in a risk population of coke workers. METHODS: Between June 2006 and October 2008, 171 male workers (mean age 43 years), employed in a Ligurian coke plant (Italiana Coke S.r.l) and exposed to polycyclic aromatic hydrocarbons (PAHs) for a median period of 16 years, underwent screening for UC. Urological evaluation included medical history, physical examination, routine laboratory tests, urine analysis, urinary cytology and uCyt+ assay. In the event of signs and symptoms suggestive of UC or positive urinary tests, the workers were also subjected to urinary ultrasonography and cystoscopy with biopsy of any suspicious lesions. RESULTS: Regarding the laboratory tests, 19/171 (11%) uCyt+ samples were considered inadequate and were excluded from the outcomes assessment. Overall, urine analysis, cytology and uCyt+ were positive in 18/152 (12%) subjects who showed no evidence of UC at the scheduled check-ups. No significant association was identified between marker positivity and occupational activity. CONCLUSIONS: Our results fail to show an increased risk of UC among the coke workers evaluated. However, they will need to be confirmed in the future by a larger enrollment and a longer follow-up in order to assess the definitive risk for UC after exposure to coke.


Subject(s)
Coke , Early Detection of Cancer , Occupational Diseases/diagnosis , Occupational Diseases/epidemiology , Urinary Bladder Neoplasms/diagnosis , Urinary Bladder Neoplasms/epidemiology , Adult , Air Pollutants, Occupational/toxicity , Biomarkers, Tumor/analysis , Early Detection of Cancer/methods , False Positive Reactions , Humans , Italy , Male , Middle Aged , Occupational Diseases/chemically induced , Risk , Urinary Bladder Neoplasms/chemically induced , Young Adult
10.
Indian J Urol ; 26(2): 279-83, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20877609

ABSTRACT

OBJECTIVES: The increasing number of prostatectomies entails an increasing number of patients suffering from iatrogenic incontinence despite improved surgical techniques. The severity of this problem often requires invasive treatments such as periurethral injection of bulking agents, artificial urinary sphincter (AUS) implantation, and sub-urethral sling positioning. The artificial urethral sphincter has represented, until today, the gold standard but, in the recent years, sling systems have been investigated as minimally invasive alternative options. Today, three different sling procedures are commonly performed: bone-anchored, readjustable, and trans-obturator slings systems. The aim of this review is to critically report the current status of sling systems in the treatment of iatrogenic male incontinence. MATERIALS AND METHODS: MEDLINE and PubMed databases were searched and all articles between 1974 and 2009 were evaluated. RESULTS: With regard to bone-anchored, readjustable, and trans-obturator slings systems, cure rates ranged between 58.0% and 86.0%, 55.5% and 73.0%, and 40.0% and 63.0%, respectively, while major complication rates ranged between 0 and 14.5%, 10.0 and 22.2%, and 0 and 10.0%, respectively. CONCLUSIONS: Suburethral slings are the only alternative techniques which can be favorably compared with the AUS, showing more advantages with respect to AUS implantations which are mainly represented by a quick and less invasive approach, low morbidity, and low costs. In spite of the difficulty in identifying the most effective sling procedure, overall, sling systems can be recommended for patients with persistent mild or moderate incontinence. However, the indication can also be extended to patients with severe incontinence, after appropriate counseling, allowing AUS implantation in the event of sling failure.

11.
World J Urol ; 27(5): 607-12, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19455340

ABSTRACT

OBJECTIVES: To compare the oncological and functional outcomes reported after radical retropubic prostatectomy (RRP) versus brachytherapy (BT) in the treatment of low-risk prostatic cancer (CaP). METHODS: Between May 1999 and October 2002, 200 patients (mean age 65.3 +/- 8.7) were enrolled and randomized into two groups of 100 patients each to undergo RRP (group 1) or BT (group 2). Prior to and following treatment, all patients were evaluated by physical examination, PSA assay and compilation of IPSS, IIEF-5 and EORTC-QLQ-C30/PR25 questionnaires. Oncological results were reported at 5 years, while functional outcomes were reported at 6 months, and 1 and 5 years mean follow-up. RESULTS: Of the 200 patients studied, 174 completed the 5-year follow-up assessment. With regards to oncological outcomes, similar 5-year biochemical disease-free survival rates were reported for RRP (91.0%) or BT (91.7%). At 6 months and 1 year, both techniques produced a significant decrease in quality of life aspects, while group 2 patients reported a significantly higher and longer lasting rate of urinary irritative disorders and better erective function than group 1. No differences in functional outcomes were encountered after 5 years in either group. CONCLUSIONS: RRP and BT are two different options for the treatment of low-risk CaP, which produce different short-term sequelae in terms of urinary disorders and erective functions, but similar biochemical disease-free survival. Further studies with a higher number of patients and a longer follow-up are needed to evaluate their comparative effectiveness on overall disease-specific survival and metastatic disease.


Subject(s)
Brachytherapy , Prostatectomy , Prostatic Neoplasms/therapy , Aged , Humans , Male , Middle Aged , Prospective Studies , Risk Factors , Surveys and Questionnaires , Treatment Outcome
12.
Urologia ; 76(2): 87-9, 2009.
Article in Italian | MEDLINE | ID: mdl-21086303

ABSTRACT

The RALP is the most modern technology available for the treatment of intracapsular prostate cancer (CaP), which can produce a shorter learning curve and better results than the traditional techniques. METHODS. Between March 2005 and March 2008, 90 patients (64.3 ys, range 52-71) with intracapsular CaP underwent RALP at our institute. Before surgery the patients underwent routine examinations and filled in IIEF, IPSS and EORTC-QLQC30/PR 25 questionnaires. Patients were followed up with PSA assay, physical examination and compilation of the questionnaires. Median follow-up was 12.5 months (range 1-35 months). RESULTS. Mean operative time was 230 min. Discharge and catheter removal were at day 7.4 and 8.2, respectively, after surgery. Pathological staging reported pT2 and pT3 in 57 (63%) and 33 patients (37%), respectively. Positive surgical margins were assessed in 30 patients (33%), particularly 8.7% in pT2 tumors. The one-year biochemical disease-free survival rate was 90%. Regarding the functional results, 81 patients (90%) were perfectly continent while a mild and a moderate incontinence were reported in 7 (8%) and 2 (2%) patients, respectively. Mean IPSS score decreased from 8 to 4; among the patients who underwent bilateral nerve sparing RALP and no adjuvant therapy, 31 (70.4%) reported satisfactory sexual intercourses. Concerning postoperative quality of life, mean EORTC-QLQC30/PR 25 questionnaires scores were very similar before and after RALP. CONCLUSIONS. After 90 cases of RALP the oncological and functional results are definitely promising. However, a wider number of patients and a greater follow-up are needed to confirm these data particularly as regards the functional results.

13.
Urologia ; 76(2): 104-6, 2009.
Article in Italian | MEDLINE | ID: mdl-21086308

ABSTRACT

The anti-incontinence methods "tension free" may be insufficient in the treatment of stress urinary incontinence (IUS) due to intrinsic sphincteric dysfunction (ISD). We report our findings on the use of the suburetral sling with adjustable tension "Remeex" sistem in the treatment of 24 patients. METHODS. Between May 2002 and February 2008, 24 patients with IUS of type III, were subjected to suburetral sling "Reemex." Positioning. The intervention provides a vaginal access to the positioning of suburetral sling and an access to the positioning of a varitensor which the wires are connected at the sling seats, recovered by the passage of a Stamey needle carrier of. The average operative time was approximately 70 minutes, the resignation was in I-II day. The tension of the sling was adjusted the day following intervention by turning the screw connected to the varitensor. Patients were followed with physical examination and completed the Korman's questionnaire about the quality of life. RESULTS. At a follow-up average 30 months, 21 patients (87.5%) were perfectly continent with improvement of quality of life. Among the complications, wound infection occurred in 2 patients (8%); 1 (4%) with mild recurrence IUS; 1 (4%) reported "de novo" urgency, 1 (4%) reported urinary retention. CONCLUSIONS. Our data show that the use of the suburetral sling "ReMeEx" is a effective option in the treatment of IUS due to ISD which is a condition often secondary to urogynecologic surgery and refractory to common techniques antincontinence.

14.
Urologia ; 76(1): 41-4, 2009.
Article in Italian | MEDLINE | ID: mdl-21086328

ABSTRACT

UNLABELLED: INTRODUCTION. Brachytherapy with real-time technique allows the ultrasound transperineal implant of needles and the release of I125 seeds in the prostate, with intraoperative dosimetry carried out by a special software. After performing this seed-releasing technique on 160 patients, we present 15 cases treated with strand seeds using the QuickLink®system. MATERIALS AND METHODS. INCLUSION CRITERIA: CaP clinical stage T1-T2, PSA ≤10 ng/ml, prostate volume ≤ 50g, Gleason score ≤3+3=6, Q maximum >13-15 ml/sec., and I-PSS score <10. METHODICAL. In ultrasound-guided transrectal technique the needles are inserted near the prostatic capsule. A 5mm section plan is performed; the computer allows identifying the contours of prostate, rectum, urethra, and the position of needles to be inserted. The total activity is defined, as well as the number and location of the sources to be included; isodoses are calculated, the correct position of seeds and spacers is printed. The strand seeds are prepared on the basis of this scheme, using the QuickLink® system, and placed into the prostate tissue. RESULTS. The QuickLink® system allows reducing the operative time (110 min, range 95-125). The pelvic CT assessing the post-planning has shown a more regular distribution of seeds. The cut-off of D90≥140 Gy was reached in all patients, with doses at urethra and rectum within the prescribed limits. CONCLUSIONS. The brachytherapy real-time with strand seeds QuickLink® combines the flexibility of planning intraoperative real-time with a faster process, and less tendency for the relocation of the seeds.

15.
Biochemistry ; 40(48): 14686-95, 2001 Dec 04.
Article in English | MEDLINE | ID: mdl-11724583

ABSTRACT

Plasmin (Pm), the main fibrinolytic protease in the plasma, is derived from its zymogen plasminogen (Plg) by cleavage of a peptide bond at Arg(561)-Val(562). Streptokinase (SK), a widely used thrombolytic agent, is an efficient activator of human Plg. Both are multiple-domain proteins that form a tight 1:1 complex. The Plg moiety gains catalytic activity, without peptide bond cleavage, allowing the complex to activate other Plg molecules to Pm by conventional proteolysis. We report here studies on the interactions between individual domains of the two proteins and their roles in Plg activation. Individually, all three SK domains activated native Plg. While the SK alpha domain was the most active, its activity was uniquely dependent on the presence of Pm. The SK gamma domain also induced the formation of an active site in Plg(R561A), a mutant that resists proteolytic activation. The alpha and gamma domains together yielded synergistic activity, both in Plg activation and in Plg(R561A) active site formation. However, the synergistic activity of the latter was dependent on the correct N-terminal isoleucine in the alpha domain. Binding studies using surface plasmon resonance indicated that all three domains of SK interact with the Plg catalytic domain and that the beta domain additionally interacts with Plg kringle 5. These results suggest mechanistic steps in SK-mediated Plg activation. In the case of free Plg, complex formation is initiated by the rapid and obligatory interaction between the SK beta domain and Plg kringle 5. After binding of all SK domains to the catalytic domain of Plg, the SK alpha and gamma domains cooperatively induce the formation of an active site within the Plg moiety of the activator complex. Substrate Plg is then recognized by the activator complex through interactions predominately mediated by the SK alpha domain.


Subject(s)
Plasminogen/metabolism , Streptokinase/metabolism , Binding Sites/physiology , Electrophoresis, Polyacrylamide Gel , Enzyme Activation , Humans , Kinetics , Models, Molecular , Protein Binding/physiology , Protein Folding , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Surface Plasmon Resonance
16.
J Mol Biol ; 308(4): 705-19, 2001 May 11.
Article in English | MEDLINE | ID: mdl-11350170

ABSTRACT

The X-ray crystal structure of a complex of a modified recombinant kringle-2 domain of human plasminogen, K2Pg[C4G/E56D/L72Y] (mK2Pg), containing an upregulated lysine-binding site, bound to a functional 30 residue internal peptide (VEK-30) from an M-type protein of a group A Streptococcus surface protein, has been determined by molecular replacement methods using K4Pg as a model, and refined at 2.7 A resolution to a R-factor of 19.5 %. The X-ray crystal structure shows that VEK-30 exists as a nearly end-to-end alpha-helix in the complex with mK2Pg. The final structure also revealed that Arg17 and His18 of VEK-30 served as cationic loci for Asp54 and Asp56 of the consensus lysine-binding site of mK2Pg, while Glu20 of VEK-30 coordinates with Arg69 of the cationic binding site of mK2Pg. The hydrophobic ligand-binding pocket in mK2Pg, consisting primarily of Trp60 and Trp70, situated between the positive and negative centers of the lysine-binding site, is utilized in a novel manner in stabilizing the interaction with VEK-30 by forming a cation-pi-electron-mediated association with the positive side-chain of Arg17 of this peptide. Additional lysine-binding sites, as well as exosite electrostatic and hydrogen bonding interactions involving Glu9 and Lys14 of VEK-30, were observed in the structural model. The importance of these interactions were tested in solution by investigating the binding constants of synthetic variants of VEK-30 to mK2Pg, and it was found that, Lys14, Arg17, His18, and Glu20 of VEK-30 were the most critical amino acid binding determinants. With regard to the solution studies, circular dichroism analysis of the titration of VEK-30 with mK2Pg demonstrated that the peptidic alpha-helical structure increased substantially when bound to the kringle module, in agreement with the X-ray results. This investigation is the first to delineate structurally the mode of interaction of the lysine-binding site of a kringle with an internal pseudo-lysine residue of a peptide or protein that functionally interacts with a kringle module, and serves as a paradigm for this important class of interactions.


Subject(s)
Antigens, Bacterial/chemistry , Antigens, Bacterial/metabolism , Kringles , Peptide Fragments/metabolism , Plasminogen/chemistry , Plasminogen/metabolism , Streptococcus/chemistry , Amino Acid Sequence , Amino Acid Substitution/genetics , Antigens, Bacterial/genetics , Binding Sites , Calorimetry , Circular Dichroism , Crystallography, X-Ray , Humans , Hydrogen Bonding , Ligands , Lysine/metabolism , Models, Molecular , Molecular Sequence Data , Peptide Fragments/chemistry , Peptide Fragments/genetics , Protein Binding , Protein Structure, Secondary , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Static Electricity , Streptococcus/genetics , Thermodynamics
17.
Arch Ital Urol Androl ; 72(4): 228-34, 2000 Dec.
Article in Italian | MEDLINE | ID: mdl-11221043

ABSTRACT

The target of this work is to evaluate the translabial ultrasonography (US) reliability as valid alternative to chain cystography in the pre and post operative assessment of patients with stress urinary incontinence (SUI). From June 1996 to May 1999, we studied 448 patients ranging in age from 35 to 90 years old with SUI from defect of anatomic support. Patients underwent translabial ultrasonography. The translabial US was performed with the patient in lithotomic position using a linear 7.5 MHz probe. The bladder was slightly filled, and the probe positioned longitudinally at the introitus to evaluate downwards and posterior rotation of bladder neck and urethra in basal conditions as well as during the abdominal strain. The evaluation of the anterior urethral angle of the 448 patients who underwent translabial US showed that, during the abdominal strain, all the patients with SUI had a very significative rotation of the urethral axis compared to continent women. Translabial US is a quick, simple, reliable non-invasive procedure. It may be used routinely for the pre and post-operative evaluation of anti-incontinence surgery.


Subject(s)
Urinary Incontinence, Stress/diagnostic imaging , Adult , Aged , Aged, 80 and over , Female , Humans , Middle Aged , Reproducibility of Results , Ultrasonography/methods , Vulva
18.
J Pept Res ; 56(6): 438-45, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11152303

ABSTRACT

The determinants of binding of a peptide lacking C-termini-exposed lysine residues to a kringle domain were investigated using an up-regulated lysine binding kringle (K2Pg[C4G/E56D/K72Y]) of plasminogen and a peptide (a1-PAM) with a sequence derived from a surface-exposed M-like streptococcal protein. Significant kringle-induced chemical shifts in a His side-chain of a1-PAM were revealed by two-dimensional NMR. Further studies using isothermal titration calorimetry (ITC) provided support for the involvement of His12 in the peptide/ protein complex. In an effort to screen a1-PAM-derived truncation peptides, a combinatorial mixture, a1deltaa2-PAM[H12X] (where X=Pro, Arg, His, Trp, Lys, Ala, Phe, Asp and Gly), was analyzed using the surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI) platform. The major peptide that remained bound to the surface of the K2Pg[C4G/ E56D/K72Y]-containing chip was that containing His12, corresponding to the wild-type sequence. Minor peaks, representing binding, were obtained for Lys12-, Arg12- and Trp12-containing peptides. Individual peptides containing these amino acids were then examined using ITC and the binding constants obtained correlated with the relative strengths of binding estimated from the SELDI-based screen.


Subject(s)
Bacterial Proteins , Carrier Proteins/chemistry , Histidine/chemistry , Plasminogen/chemistry , Calorimetry , Humans , Kinetics , Kringles , Ligands , Magnetic Resonance Spectroscopy , Peptide Biosynthesis , Peptides/chemistry , Protein Binding , Protein Conformation , Protein Structure, Tertiary , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Thermodynamics , Time Factors
19.
Prog Urol ; 9(3): 562-6, 1999 Jun.
Article in French | MEDLINE | ID: mdl-10434337

ABSTRACT

OBJECTIVES: Anterolateral transabdominal incisions provide good exposure for supramesocolonic and inframescolonic surgery. However, these incisions section and denervate the rectus abdominis, oblique and transversus abdominis muscles with marked loss of active muscle control in a large number of patients. In 1974, Giuliani described an anterolateral transbdominal approach for renal tumours, which provides good visualization and good access to the renal pedicle, as well as good exposure caudally as far as the aortic bifurcation and cranially as far as the diaphragm. The authors report a new anatomical technique using this incision, which splits the muscles and preserves the nerves thereby avoiding the abdominal muscle hypotonia. MATERIAL AND METHODS: From March 1996 to March 1998, Giuliani's surgical incision was performed in 35 patients undergoing radical nephrectomy for renal cancer (24 on the left side and 11 on the right side). The mean age of the patients was 63.2 years (range: 42 to 80 years) and the mean follow-up was 11.6 months. RESULTS: Tone and active control of muscles of the abdominal wall were completely preserved in all of these 35 patients. However, all patients presented a slight sensory loss in the low portion of the transverse skin incision close to the umbilicus, which improved with time and resolved completely in about 50% of cases. CONCLUSION: The mahor advantage of this anatomical incision compared to the conventional technique is to eliminate permanent functional deficits and hypotonia of the abdominal wall. This anatomical approach also allows easy and perfectly safe wound closure in layers, by reconstructing the anterior abdominal wall.


Subject(s)
Abdominal Muscles/innervation , Abdominal Muscles/surgery , Kidney Neoplasms/surgery , Nephrectomy/methods , Adult , Aged , Dermatologic Surgical Procedures , Humans , Middle Aged , Muscle Tonus
20.
Ophthalmologica ; 212 Suppl 1: 25-6, 1998.
Article in English | MEDLINE | ID: mdl-9730742

ABSTRACT

Intraocular lens (IOL) power calculation in 46 pseudophakic eyes (extracapsular cataract extraction with IOL in posterior chamber), utilizing a Javal keratometer, a Sonomed A 2000 echobiometer (probe 10 MHz, velocity=1,548 m/s) and the SRK2 formula, although there was a statistically significant reduction of the axial length, both in normal and hyperopic eyes, demonstrated no statistically significant differences of IOL power, when compared to the power previously calculated in the phakic eye.


Subject(s)
Eye/anatomy & histology , Lenses, Intraocular , Pseudophakia/diagnostic imaging , Aged , Aged, 80 and over , Anterior Chamber/anatomy & histology , Anterior Chamber/diagnostic imaging , Cataract Extraction , Cornea/anatomy & histology , Cornea/diagnostic imaging , Eye/diagnostic imaging , Female , Follow-Up Studies , Humans , Lens Implantation, Intraocular , Male , Middle Aged , Polymethyl Methacrylate , Refraction, Ocular , Ultrasonography
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