Evolution of a functionally intact but antigenically distinct DENV fusion loop.

A hallmark of dengue virus (DENV) pathogenesis is the potential for antibody-dependent enhancement, which is associated with deadly DENV secondary infection, complicates the identification of correlates of protection, and negatively impacts the safety and efficacy of DENV vaccines. Antibody-dependent enhancement is linked to antibodies targeting the fusion loop (FL) motif of the envelope protein, which is completely conserved in mosquito-borne flaviviruses and required for viral entry and fusion. In the current study, we utilized saturation mutagenesis and directed evolution to engineer a functional variant with a mutated FL (D2-FL), which is not neutralized by FL-targeting monoclonal antibodies. The FL mutations were combined with our previously evolved prM cleavage site to create a mature version of D2-FL (D2-FLM), which evades both prM- and FL-Abs but retains sensitivity to other type-specific and quaternary cross-reactive (CR) Abs. CR serum from heterotypic (DENV4)-infected non-human primates (NHP) showed lower neutralization titers against D2-FL and D2-FLM than isogenic wildtype DENV2 while similar neutralization titers were observed in serum from homotypic (DENV2)-infected NHP. We propose D2-FL and D2-FLM as valuable tools to delineate CR Ab subtypes in serum as well as an exciting platform for safer live-attenuated DENV vaccines suitable for naïve individuals and children.

Evolution of a Functionally Intact but Antigenically Distinct DENV Fusion Loop.

A hallmark of Dengue virus (DENV) pathogenesis is the potential for antibody-dependent enhancement, which is associated with deadly DENV secondary infection, complicates the identification of correlates of protection, and negatively impacts the safety and efficacy of DENV vaccines. ADE is linked to antibodies targeting the fusion loop (FL) motif of the envelope protein, which is completely conserved in mosquito-borne flaviviruses and required for viral entry and fusion. In the current study, we utilized saturation mutagenesis and directed evolution to engineer a functional variant with a mutated FL (D2-FL) which is not neutralized by FL-targeting monoclonal antibodies. The FL mutations were combined with our previously evolved prM cleavage site to create a mature version of D2-FL (D2-FLM), which evades both prM- and FL-Abs but retains sensitivity to other type-specific and quaternary cross-reactive (CR) Abs. CR serum from heterotypic (DENV4) infected non-human primates (NHP) showed lower neutralization titers against D2-FL and D2-FLM than isogenic wildtype DENV2 while similar neutralization titers were observed in serum from homotypic (DENV2) infected NHP. We propose D2-FL and D2-FLM as valuable tools to delineate CR Ab subtypes in serum as well as an exciting platform for safer live attenuated DENV vaccines suitable for naïve individuals and children.

Candida-associated denture stomatitis: clinical, epidemiological, and microbiological features.

OBJECTIVE: The identification of Candida spp. in denture stomatitis, the clinical manifestations, and the antifungal susceptibility profile lead to a correct and individualized therapeutic management of the patients. This study is aimed at investigating the clinical manifestations and epidemiological and microbiological characteristics of Candida-associated denture stomatitis. DESIGN: The samples were obtained by swabbing the oral mucosa of the subjects and then seeded onto Sabouraud Dextrose Agar and onto CHROMagar® Candida plates. The identification at the species level was confirmed by Matrix Assisted Laser Desorption Time of Flight Mass Spectrometry. Clinical classification was performed according to the criteria proposed by Newton (1962): (i) pinpoint hyperemia, (ii) diffuse hyperemia, and (iii) granular hyperemia. For carrying out the antifungal susceptibility testing, we adopted the CLSI M27-S4 protocol. RESULTS: C. albicans was the most prevalent species in our study. Regarding non-albicans Candida species, C. glabrata was the most common species isolated from the oral mucosa (n = 4, 14.8%), while in the prosthesis, it was C. tropicalis (n = 4, 14.8%). The most prevalent clinical manifestation was pinpoint hyperemia and diffuse hyperemia. Candida albicans, C. glabrata, and C. parapsilosis were susceptible to all the tested antifungals. Concerning fluconazole and micafungin, only two strains showed dose-dependent sensitivity (minimum inhibitory concentration (MIC), 1 µg/mL) and intermediate sensitivity (MIC, 0.25 µg/mL). One C. tropicalis strain was resistant to voriconazole (MIC, 8 µg/mL). CONCLUSIONS: C. albicans was the most common species found in oral mucosa and prosthesis. The tested antifungal drugs showed great activity against most isolates. The most prevalent clinical manifestations were Newton's type I and type II.

A thermo-responsive adsorbent-heater-thermometer nanomaterial for controlled drug release: (ZIF-8,EuxTby)@AuNP core-shell.

An adsorbent-heater-thermometer nanomaterial, (ZIF-8,EuxTby)@AuNP, based on ZIF-8 (adsorbent), containing Eu3+ and/or Tb3+ ions (thermometer) and gold nanoparticles (AuNPs, heater) was designed, synthetized, characterized, and applied to controlled drug release. These composite materials were characterized as core-shell nanocrystals with the AuNPs being the core, around which the crystalline ZIF-8 has grown (shell) and onto which the lanthanide ions have been incorporated or chemosorbed. This shell of ZIF-8 acts as adsorbent of the drugs, the AuNPs act as heaters, while the luminescence intensities of the ligand and the lanthanide ions are used for temperature monitoring. This thermo-responsive material can be activated by visible irradiation to release small molecules in a controlled manner as established for the model pharmaceutical compounds 5-fluorouracil and caffeine. Computer simulations and transition state theory calculations shown that the diffusion of small molecules between neighboring pores in ZIF-8 is severely restricted and involves high-energy barriers. These findings imply that these molecules are uploaded onto and released from the ZIF-8 surface instead of being inside the cavities. This is the first report of ZIF-8 nanocrystals (adsorbents) containing simultaneously lanthanide ions as sensitive nanothermometers and AuNPs as heaters for controlled drug release in a physiological temperature range. These results provide a proof-of-concept that can be applied to other classes of materials, and offer a novel perspective on the design of self-assembly multifunctional thermo-responsive adsorbing materials that are easily prepared and promptly controllable.

Dartos flap interposition in the surgical repair of rectourethral fistulas.

BACKGROUND: Multilayer closures are important techniques in urethral fistula repair. We report our experience with dartos flap interposition in the repair of rectourethral fistula. METHODS/RESULTS: A tip of a V-shaped scrotal skin flap is mobilized and de-epithelialized to develop a dartos flap. Two patients with a rectourethral fistula underwent fistula closure covered with this dartos flap. The surgical repair was successful in both cases and neither patient showed any late complication. CONCLUSIONS: A scrotal dartos flap is well vascularized and easy to mobilize. We believe this technique is helpful in preventing postoperative complications following complex posterior urethral surgery.

Sonographic characteristics of the urinary tract in healthy neonates.

PURPOSE: In infancy it has been reported that intravesical pressure is generally higher in males than in females. We investigated whether there are significant differences in the sonographic characteristics of the urinary tract in male and female neonates. MATERIALS AND METHODS: The study comprised 280 consecutive full-term newborns, including 146 males and 134 females. Kidneys were assessed for hydronephrosis and graded according to Society for Fetal Urology guidelines. Kidney measurements included maximum longitudinal length and largest longitudinal area. Bladder measurements included bladder dimensions and wall thickness. Bladder wall thickness was measured only in neonates with an estimated bladder volume of greater than 10 ml. RESULTS: Of the 280 neonates 114 had mild renal pelvic dilatation in at least 1 kidney and the incidence in males was significantly higher than in females (53% versus 27%). Kidney longitudinal length and largest area were significantly greater in males on each side. Mean bladder volume was essentially equal in the 169 male and female neonates with an estimated bladder volume of greater than 10 ml. but mean bladder wall thickness was significantly greater in males than in females (1.63 versus 1.38 mm.). CONCLUSIONS: Our study shows that there are marked differences in sonographic findings in male and female neonates in regard to renal pelvic dilatation, renal size and bladder wall thickness. These differences should be considered when sonography is done for screening for urinary tract anomalies in newborns.

Common sheath reimplantation with ureteral plication: a useful technique for the management of ectopic ureterocele.

BACKGROUND: When salvaging the upper pole kidney in duplex ectopic ureterocele, primary bladder level surgery with common sheath ureteral reimplantation has the definite advantage of allowing the reconstruction of the entire collecting system through a single lower abdominal incision. However, there are several complications associated with a common sheath reimplantation in a child with a very dilated upper pole ureter, such as vesicoureteral reflux or ureterovesical stenosis. METHODS/RESULTS: To avoid these complications, ureteral plication over the common ureteral sheath in two children with duplex ectopic ureterocele was used. Postoperatively, neither child showed reflux or recurrent urinary tract infection and both showed a marked improvement of the upper pole collecting system. CONCLUSION: This technique allows for a simple and definitive reconstruction in cases of duplex ectopic ureterocele, particularly with dilated upper pole ureter.

Reliability of 99mtechnetium dimercapto-succinic acid uptake 2 hours after injection in hydronephrosis.

PURPOSE: The accumulation of radioactivity in the dilated collecting system potentially influences the calculation of differential renal function on the radionuclide test. We focused on this reservoir effect in unilateral hydronephrosis and assessed the reliability of calculating differential renal function by dimercapto-succinic acid (DMSA) uptake 2 hours after injection. MATERIALS AND METHODS: Unilateral partial ureteral obstruction was created in 8-week-old rats. Four weeks after surgery the animals were sacrificed 2 and 24 hours after the injection of tracer. The DMSA uptake rates of the renal parenchyma and collecting system were measured separately by autowell gamma counter. Differential function was calculated according to renal parenchymal and whole kidney (parenchyma and collecting system) uptake. RESULTS: There was a higher accumulation of DMSA in the dilated renal pelvis at 2 than at 24 hours. However, DMSA uptake in the collecting system was extremely small in comparison to that in the parenchyma. As a result; differential renal function calculated using parenchymal uptake was similar to that calculated using whole kidney uptake measured 2 and 24 hours after injection. CONCLUSIONS: Our data suggest that the pure reservoir effect of DMSA uptake at early measurement is much smaller than reported in previous experimental animal studies. Further clinical studies are needed to reexamine this reservoir effect in children with unilateral hydronephrosis.

[Pregnancy after augmentation cystoplasty. A case report].

A 31-year-old woman with a history of spina bifida occulta became pregnant after ileocystoplasty for her neurogenic bladder. During the pregnancy she had frequent episodes of febrile urinary tract infections, and progressive hydronephrosis appeared in the second trimester. At 25 weeks gestation she was complicated by severe pyelonephritis requiring the intervention with the placement of double pigtail ureteral stent. However, long term efficacy of ureteral stent was questionable and this indwelling catheter caused bacteriuria which was not eradicated by intravenous antibiotics. Classical cesarean section was performed at 32 weeks of gestation due to the fear of fetal distress. Neobladder and mesenteric blood supply were adherent to the anterior surface of the uterus. Urinary tract infection is extremely common during pregnancy after enterocystoplasty. The most important point is prophylactic antibiotics throughout the pregnancy. At the time of cesarean section, a reconstructive urological surgeon should be part of the operative team and take great care to avoid injury to the blood supply of cystoplasty.

[A case of urachal carcinoma associated with elevated levels of CA19-9].

A 53-year-old man was admitted with the chief complaint of gross hematuria. Cystoscopic examination revealed a non-papillary tumor at the dome of the bladder. The plasma level of CA19-9 was markedly elevated before treatment. Total cystectomy with en bloc removal of the tumor and radical lymphadenectomy were performed under the diagnosis of urachal carcinoma. The plasma CA19-9 level decreased to the normal level after operation and was well correlated with postoperative course. The monitoring of CA19-9 may be useful for follow up in patients showing a high elevation of CA19-9.

[Adenovirus-induced kidney graft pyelonephritis following renal transplantation].

A 16-year-old female received a kidney transplantation from her mother 13 months before she suddenly noticed gross hematuria and painful micturition, and developed high fever with chills. The serum creatinine (S-Cr) level rose from 1.5 to 2.6 mg/dl, but there was no clinical sign of acute rejection. Despite the treatment with antibiotics and gamma-globulin, the the high fever and hematuria did not improve. The adenovirus antibody titer elevated from x8 to x1,024, while adenovirus was not isolated from the urine. On the 15th day of the disease, hematuria disappeared spontaneously and on the 19th day she became afebrile. The S-Cr level also was normalized spontaneously. Histological examination of the graft biopsy on the 14th day, showed severe tubulointerstitial nephritis localized in the renal medulla and full type intranuclear inclusions were revealed in tubular epithelial cells. From these findings, we diagnosed this case as adenovirus-induced kidney graft pyelonephritis associated with acute hemorrhagic cystitis.