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News consumers today can access a wider variety of news sources than ever before, as trust in mainstream media is declining, particularly among conservatives, and distrust for people on opposite sides of the political aisle is also growing. Alternative news outlets maintain they are corrective to mainstream outlets, focusing on a particular political agenda, being more activist than their mainstream counterparts, and differentiating themselves from elite journalists. Through the lens of selective exposure, this study examined the extent to which consumers use mainstream news or selectively expose themselves to alternative news. It also explored how political ideology and trust related to alternative news use. Results showed that while partisans selectively exposed themselves to alternative sources that framed events in ways that confirmed their biases, the overwhelming majority of news consumers still turned to mainstream sources. However, conservatives are turning to mainstream sources less, and their use of conservative alternative news is driven partly by less trust in news and more interest in politics. Overall implications of this study contribute to understanding news consumer choices in that selective exposure occurs in the digital age.

Objective To evaluate pain improvement and recurrent stress incontinence (SUI) following painful synthetic midurethral sling (MUS) removal. Methods We conducted a retrospective review of patients who underwent synthetic MUS removal at our institution from 2009–2016 for the indication of pain. We recorded sling type (transobturator vs retropubic), complete vs partial removal, and presenting symptoms. Postoperative pain improvement was categorized as resolved (pain resolved, requiring no further therapy), improved (pain less bothersome, may require further therapy), or unresolved (no/minimal improvement, requiring further management). Recurrent incontinence and further reconstructive procedures were assessed. Results 87 patients (49 complete and 38 partial removal) with pain as the primary indication for removal were included. Median age at intervention was 54 years with median follow-up of 8 months. Overall, pain improved or resolved in 78.1% of cases. Complete removal was associated with significantly greater percentage of pain resolution (63.3%) compared to partial removal (26.3%) (P = 0.002) regardless of sling type. No significant differences in recurrent SUI were noted in complete vs partial removal. Additional reconstructive procedures were performed in 28 patients, most commonly sling placement, with no significant difference in complete (20.4%) vs partial (28.9%) removal groups (P = 0.36). The overall complication rate was low (5.7%), a majority of which were transfusions (4.6%). Conclusion Following MUS removal, most patients experienced resolution or improvement of pain. Complete sling removal was associated with significantly greater percentage of pain resolution compared to partial removal in both retropubic and transobturator slings. Rates of recurrent SUI and reintervention for SUI were not related to the extent of sling removal.

OBJECTIVE To assess the safety, feasibility and treatment outcomes of holmium laser enucleation of the prostate (HoLEP) as a same day surgery (SDS). METHODS HoLEPs performed from November 2013 to December 2018 at our institution were reviewed retrospectively. Inclusion criteria for same day surgery (SDS) included living in the local metropolitan area with access to local hospital and Eastern Cooperative Oncology Group (ECOG) 0-2, regardless of prostate size and anticoagulation status. Those patients who were discharged directly from the postoperative care unit were identified as SDS cases. Patients admitted overnight after HoLEP during the same period were used as a matched cohort. Patient demographics, disease characteristics and treatment outcomes were compared. RESULTS A total of 377 patients were identified, including 199 SDS and 178 non-SDS patients. No statistical difference was present between the 2 groups regarding the post-op complication and readmission rates. The non-SDS group had a significantly higher percentage of patients with history of urinary retention, lower pre-op Qmax, and larger prostate volume. The SDS group had shorter operative time, length of stay (LOS), and catherization time (all P CONCLUSION Same day outpatient surgery for HoLEP is safe in patients who live in close proximity and have ECOG status 0-2. Our readmission rate and complication rate are comparable to those reported in the literature with markedly decreased LOS. Long-term functional outcome is not compromised by SDS.

Objective To address renewed interest in nonmesh transvaginal Pelvic Organ Prolapse (POP) repair since the FDA reclassification of transvaginal mesh, our goal was to develop a transvaginal sacrospinous fixation for anterior and apical POP using only autologous fascia lata. We report our experience in 33 patients. Methods Autologous Anterior and Apical Pelvic Organ Prolapse (AAA-POP) repair utilizes a 4 × 14 cm piece of fascia lata harvested through a 3-4-inch upper thigh incision. The graft is reconfigured to provide apical fixation to the sacrospinous ligaments and distal fixation to the obturator fascia. Patients were followed by history, SEAPI scores, POP-Q scores, and Visual Analogue Pain Score. Treatment success was defined as absence of symptomatic anterior or apical POP. Results Thirty-three patients (mean age 63, mean follow-up 12 months) underwent AAA-POP. Treatment was successful in 31 patients (94%), and 2 failures were due to uterine prolapse. Mean harvest site Visual Analog Pain score was 0.27. Five and 7 patients developed nonbothersome thigh bulges and wound paresthesias, respectively. All 4 harvest-site seromas resolved, with 2 requiring simple aspiration. Eleven patients developed urinary retention, 10 (91%) of them after concurrent pubovaginal sling. All resolved after sling loosening (6 patients) or sling lysis (4 patients). Conclusion AAA-POP is an efficacious treatment for patients desiring nonmesh POP repair. Postoperative harvest site issues are minor and typically resolve with expectant management. Patients should be counseled about the potential risk of failure with a uterine-sparing approach and the risk of urinary retention with concurrent pubovaginal sling.

INTRODUCTION AND OBJECTIVE:Fascia lata is a versatile graft that has applications in various surgical fields. There is increased interest in the use of autologous fascia lata in pelvic floor recons...

INTRODUCTION AND OBJECTIVE:With the increasing prevalence of gender-affirming surgeries, management of the long-term complications of these surgeries is an important topic. One complication is prol...

OBJECTIVE To evaluate pain resolution and recurrent prolapse following removal of painful pelvic organ prolapse (POP) mesh. METHODS This was an IRB-approved retrospective review of patients who underwent POP mesh removal from 2009 to 2018 for the primary indication of pain and/or dyspareunia. Factors of interest included mesh type, complete versus partial removal, presenting symptom(s), and recurrent prolapse. Complete removal was defined as complete excision of all possible mesh, including arms, and partial removal was anything less. Postoperative symptom improvement was categorized as resolved, improved, or unresolved. RESULTS Ninety-two mesh removal patients were identified, 78 of whom underwent mesh removal for pain or dyspareunia, with mean follow-up of 17.25 months (range: 0.46-60.25). Overall, presenting symptoms resolved or improved in 85.9%. In the 45 complete removal patients, 46.7% resolved, 40.0% improved, and 11.1% unresolved compared to 51.5% resolved, 33.3% improved, and 12.1% unresolved in the 33 partial removal patients (P = .82). Recurrent symptomatic POP developed in 31.1% of patients after complete removal compared to 15.2% after partial removal (P = .12). Overall, 23 (30.3%) patients required additional reconstructive procedures. CONCLUSION Most patients experienced resolution or improvement in painful symptoms after any degree of mesh removal. Complete removal was not significantly associated with greater symptom improvement compared to partial removal. Complete removal was associated with a higher percentage of recurrent POP, but this association was not statistically significant. Less than one-third of patients required additional surgery.

The physical examination and pelvic imaging with MRI are often used in the pre-operative evaluation of pelvic organ prolapse. The objective of this study was to compare grading of prolapse on defecography phase of dynamic magnetic resonance imaging (dMRI) with physical examination (PE) grading using both the Pelvic Organ Prolapse Quantification (POP-Q) staging and Baden–Walker (BW) grading systems in the evaluation of pelvic organ prolapse (POP). We retrospectively reviewed the charts of 170 patients who underwent dMRI at our institution. BW grading and POP-Q staging were collected for anterior, apical, and posterior compartments, along with absolute dMRI values and overall grading of dMRI. For the overall grading/staging from dMRI, BW, and POP-Q, Spearman rho (ρ) was used to assess the correlation. The correlations between dMRI grading and POP-Q staging were compared to the correlations between dMRI grading and BW grading using Fisher’s Z transformation. A total of 54 patients were included. dMRI grading was not significantly correlated with BW grading for anterior, apical, and posterior compartment prolapse (p > 0.15). However, overall dMRI grading demonstrated a significant (p = 0.025) and positive correlation (ρ = 0.305) with the POP-Q staging system. dMRI grading for anterior compartment prolapse also demonstrated a positive correlation (p = 0.001, ρ = 0.436) with the POP-Q staging derived from measurement locations Aa and Ba. The overall dMRI grade is better correlated with POP-Q stage than with BW grade (p = 0.024). Overall and anterior compartment grading from dMRI demonstrated a significant and positive correlation with the overall POP-Q staging and anterior compartment POP-Q staging, respectively. The overall dMRI grade is better correlated with POP-Q staging than with BW grading.

Background: Holmium laser enucleation of the prostate (HoLEP) is an attractive and well-studied alternative to transurethral resection of the prostate and open prostatectomy for the treatment of benign prostatic hyperplasia. There remains an established steep learning curve with relatively few complications described in the literature. A unique risk of HoLEP is injury of the bladder during morcellation of the adenoma and potential iatrogenic intraperitoneal bladder rupture. We present a rare complication of HoLEP demonstrated by two patients in which capsular perforation resulted in subsequent abdominal distention secondary to a large amount of irrigation fluid that leaked into the extraperitoneal space. Uniquely, these cases were managed differently, and serve as guidance to the HoLEP practitioner in postoperative management. Case Presentations: The first case involved a 74-year-old male who was found to have significant abdominal distention at the end of the procedure. Given an acute change in stability and concern for bladder injury during morcellation, a minilaparotomy was performed only to reveal extraperitoneal extravasation without intraperitoneal bladder injury or perforation. In the second case, a 78-year-old male undergoing HoLEP had a similar presentation of significant abdominal distention at the conclusion of morcellation. Given a low suspicion for any bladder injury, the patient was managed conservatively with diuretics. He was subsequently discharged on postoperative day 1. Conclusion: Capsular perforation is not a rare phenomenon that occurs during HoLEP. Rarely, perforations can lead to extravasation of irrigation fluid into the extraperitoneal space masquerading as a potential bladder injury related to morcellation because of the associated abdominal distention. This presentation can occur in large glands or early in a surgeon's learning curve when operative times are longer. When there is clear evidence to suggest there is no bladder injury, these cases can be managed conservatively and avoid the morbidity of an abdominal exploration.

Ten percent of patients undergoing prostatic urethral lift (PUL) require repeat surgical treatment within 3 years. We describe the feasibility and considerations of performing holmium laser enucleation of the prostate (HoLEP) as a salvage therapy after previous PUL.Men who had undergone HoLEP after PUL were retrospectively identified from three institutions with surgeons experienced in HoLEP. Subjects were characterized by age, time from PUL procedure (months) to HoLEP surgery, indication for retreatment, and pre-operative prostate volume by ultrasound. Outcomes of interest included enucleation time, morcellation time, morcellator type, weight of tissue resected, and Clavien complications. We also summarize findings related to the location of PUL device implants, and the effect of the implants on the enucleation and morcellation portions of the procedure.From December 15, 2015 to October 31, 2016, seven men aged 51-78 years underwent HoLEP at a median of 8.6 months (range 3-18) after PUL. The median prostate volume by transrectal ultrasound was 80 cmHoLEP can be performed safely and effectively post-PUL; however, device implants may be found in areas other than the intended location, and morcellation of the adenoma tissue is complicated by metallic implants of the PUL device.

INTRODUCTION AND OBJECTIVES:Since the reclassification of transvaginal mesh as a high-risk device, there is renewed interest in non-mesh pelvic organ prolapse (POP) repair. Our goal was to develop ...

Posterior compartment vaginal prolapse can be approached with multiple surgical techniques, including transvaginally, transperineally, and transanally, repaired with either native tissue or with the addition of an augment. Augment material for posterior compartment prolapse includes biologic graft (dermal, porcine submucosal), absorbable mesh (Vicryl polyglactin), or nonabsorbable synthetic mesh (polypropylene). Anatomic success rates for posterior compartment repair with augment has ranged from 54% to 92%. Augmented posterior compartment repair has not been shown to have superior outcome to native tissue repair. The focus of this article is on the transvaginal approach comparing native tissue repair with graft or mesh augmented repair.

PURPOSE: To evaluate the utility of a defecography phase (DP) sequence in dynamic pelvic floor MRI (DPMRI), in comparison to DPMRI utilizing only non-defecography Valsalva maneuvers (VM). MATERIALS AND METHODS: Inclusion criteria identified 237 female patients with symptoms and/or physical exam findings of pelvic floor prolapse. All DPMRI exams were obtained following insertion of ultrasound gel into the rectum and vagina. Steady-state free-precession sequences in sagittal plane were acquired in the resting state, followed by dynamic cine acquisitions during VM and DP. In all phases, two experienced radiologists performed blinded review using the H-line, M-line, Organ prolapse (HMO) system. The presence of a rectocele, enterocele and inferior descent of the anorectal junction, bladder base, and vaginal vault were recorded in all patients using the pubococcygeal line as a fixed landmark. RESULTS: DPMRI with DP detected significantly more number of patients than VM (p

Objective To compare dynamic magnetic resonance imaging (dMRI) defecography phase findings with physical examination (PE) grading in the evaluation of pelvic organ prolapse (POP). Methods We retrospectively reviewed 274 consecutive patients who underwent dMRI with defecography. Baden-Walker grading of POP, absolute dMRI values, and grading by dMRI were collected for anterior, apical, and posterior compartments. Anatomically significant POP on PE was defined as Baden-Walker Grade ≥3 and on dMRI by dMRI Grade ≥2. A Spearman's Rank correlation was performed between absolute dMRI values and respective POP grades. Results A total of 178 female patients were included. Anatomically insignificant and significant cystoceles had a 26.4% (19/72) and 84.6% (66/78) agreement respectively. Anatomically insignificant and significant apical prolapse had a 2.0% (2/100) and 62.9% (17/27) agreement respectively. Anatomically insignificant and significant posterior prolapse had a 49.5% (51/103) and 78.7% (59/75) agreement respectively. PE detected only 30% (9/30) of total dMRI detected enteroceles and misdiagnosed 10% (3/30) of these patients with a rectocele. Conclusion The dMRI defecography phase correlated well for anatomically significant prolapse in anterior and posterior compartments. dMRI was superior to PE for enterocele detection and was better able to distinguish an enterocele from a rectocele. Thus, dMRI may have the greatest diagnostic value in cases where the presence of an enterocele is unclear in apical and/or posterior compartments.

Overactive bladder (OAB) is a urologic condition that negatively impacts quality of life. Sacral neuromodulation (SNM) is a safe and effective treatment option that has been studied extensively in patients with severe and refractory OAB. SNM significantly reduces urgency, frequency, and incontinence while also improving quality of life. Although SNM is not currently indicated for the treatment of OAB symptoms due to neurologic disease, recent literature suggests that these patients may also benefit from SNM. Other areas requiring further investigation include reduction of adverse events associated with SNM and the overall cost benefit ratio of SNM.

As transurethral resection of the prostate (TURP) continues to be the standard of care to alleviate voiding dysfunction in men with benign prostatic hyperplasia (BPH), novel techniques are being addressed to promote operative efficacy and long-term results for improved voiding. This review addresses recent literature over the past year on various transurethral technologies and procedures as well as the developing practice of prostatic arterial embolization (PAE) to improve lower urinary tract symptoms in the setting of BPH. The transurethral technologies include bipolarity, which has come to the forefront in the resection of large prostates as it reduces the risk of TUR syndrome, and plasmakinetic enucleation and diode laser enucleation which have both recently been demonstrated to improve tissue resection for larger prostate glands. The Oyster Procedure, described below, is a specific method of adenoma resection which has been demonstrated to be effective in the treatment of large obstructing prostates. HoLEP, which has been established as an effective tool for large prostates, has now been described to be useful in patients requiring retreatment of LUTS secondary to BPH. Prostatic artery embolization (PAE) has recently come to the forefront of a minimally invasive alternative to TURP with a reduction of symptoms and recovery time in patients who have had unsuccessful or refractory treatments for LUTS. While TURP aids in improving voiding dysfunction, it is known to affect sexual function. New implants, such as those explained in the UroLift procedure below, can improve LUTS while preserving sexual function. Finally, The GreenLight laser prostatectomy has now been demonstrated to be safe and effective in an office setting with conscious sedation, thereby reducing surgical risk with anesthesia for those with comorbidities. The studies discussed in this review focus on improving procedures for treating larger prostates, prostates with prior surgical interventions or refractory BPH, patients who wish to maintain sexual function, and assessing interventions for the elderly and those with comorbidities. As new technologies continue to expand, their use among both novice and experienced surgeons will be vital to advancing the treatments for BPH.

In order to obtain a global picture of how alveolar macrophages respond to influenza A virus (IAV) infection, we used a quantitative proteomics method to systematically examine protein expression in the IAV-infected primary human alveolar macrophages. Of the 1214 proteins identified, 43 were significantly up-regulated and 63 significantly down-regulated at > 95% confidence. The expression of an array of interferon (IFN)-induced proteins was significantly increased in the IAV-infected macrophages. The protein with the greatest expression increase was ISG15, an IFN-induced protein that has been shown to play an important role in antiviral defense. Concomitantly, quantitative real time PCR analysis revealed that the gene expression of type I IFNs increased substantially following virus infection. Our results are consistent with the notion that type I IFNs play a vital role in the response of human alveolar macrophages to IAV infection. In addition to the IFN-mediated responses, inflammatory response, apoptosis and redox state rebalancing appeared also to be major pathways that were affected by IAV infection. Furthermore, our data suggest that alveolar macrophages may play a crucial role in regenerating alveolar epithelium during IAV infection.

Alveolar type II epithelial cells (ATIIs) are one of the primary targets for influenza A pneumonia. The lack of a culture system for maintaining differentiated ATIIs hinders our understanding of pulmonary innate immunity during viral infection. We studied influenza A virus (IAV)-induced innate immune responses in differentiated primary human ATIIs and alveolar macrophages (AMs). Our results indicate that ATIIs, but not AMs, support productive IAV infection. Viral infection elicited strong inflammatory chemokine and cytokine responses in ATIIs, including secretion of IL-8, IL-6, MCP-1, RANTES, and MIP-1β, but not TNF-α, whereas AMs secreted TNF-α as well as other cytokines in response to infection. Wild-type virus A/PR/8/34 induced a greater cytokine response than reassortant PR/8 virus, A/Phil/82, despite similar levels of replication. IAV infection increased mRNA expression of IFN genes IFN-β, IL-29 (IFN-λ1), and IL-28A (IFN-λ2). The major IFN protein secreted by type II cells was IL-29 and ATIIs appear to be a major resource for production of IL-29. Administration of IL-29 and IFN-β before infection significantly reduced the release of infectious viral particles and CXC and CC chemokines. IL-29 treatment of type II cells induced mRNA expression of antiviral genes MX1, OAS, and ISG56 but not IFN-β. IL-29 induced a dose-dependent decrease of viral nucleoprotein and an increase of antiviral genes but not IFN-β. These results suggest that IL-29 exerts IFN-β-independent protection in type II cells through direct activation of antiviral genes during IAV infection.

Contains fulltext : 80484.pdf (Publisher’s version ) (Closed access) The processing of pro-interleukin-1beta depends on activation of caspase-1. Controversy has arisen whether Toll-like receptor (TLR) ligands alone can activate caspase-1 for release of interleukin-1beta (IL-1beta). Here we demonstrate that human blood monocytes release processed IL-1beta after a one-time stimulation with either TLR2 or TLR4 ligands, resulting from constitutively activated caspase-1 and release of endogenous adenosine triphosphate. The constitutive activation of caspase-1 depends on the inflammasome components, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), and NALP3, but in monocytes caspase-1 activation is uncoupled from pathogen-associated molecular pattern recognition. In contrast, macrophages are unable to process and release IL-1beta solely by TLR ligands and require a second adenosine triphosphate stimulation. We conclude that IL-1beta production is differentially regulated in monocytes and macrophages, and this reflects their separate functions in host defense and inflammation.

OBJECTIVES To investigate whether angiotensin II (AII) receptor antagonism decreases the inflammation and oedema in acute murine experimental autoimmune cystitis (EAC), as interstitial cystitis (IC) might have an autoimmune component and AII has been implicated in autoimmune-mediated vascular congestion, oedema and scarring. MATERIALS AND METHODS Female Balb/cAN mice were divided into three treatment groups (eight in each group) that were autoimmunized with bladder homogenate to induce EAC. One group received an AII type 1 receptor (AT1) antagonist, one group an AII type 2 receptor (AT2) antagonist, and one group remained untreated (EAC). A control and sham-injected group were also included. After 10 weeks, bladders were removed, sectioned, and stained with haematoxylin and eosin. RESULTS Grossly, there was no thickening or adhesions in the bladders of the control or sham-injected mice. In five of seven surviving EAC bladders, there were dense adhesions to surrounding peritoneal structures. There were also adhesions and bladder thickening in all of the AT2 antagonist-treated mice (though in a milder form) but in only two of seven surviving AT1 antagonist-treated mice. There was no inflammation or oedema in the sham and control groups. All the EAC bladders were inflamed, with submucosal oedema and urothelial detachment from the lamina propria. In the AT1 antagonist-treated mice there was no inflammation or oedema. By contrast, all AT2 antagonist-treated mice had moderate inflammation and minor detachment of the urothelium from the lamina propria. CONCLUSIONS AT1 receptor blockade ameliorated the inflammatory infiltration, submucosal oedema, and urothelial detachment associated with EAC in mice. This was achieved to a lesser extent by AT2 receptor blockade. If some patients with IC have a pathophysiology similar to that of EAC mice, there might be potential benefit from AII receptor blockade.

Cultures of differentiating fetal human type II cells have been available for many years. However, studies with differentiated adult human type II cells are limited. We used a published method for type II cell isolation and developed primary culture systems for maintenance of differentiated adult human alveolar epithelial cells for in vitro studies. Human type II cells cultured on Matrigel (basolateral access) or a mixture of Matrigel and rat tail collagen (apical access) in the presence of keratinocyte growth factor, isobutylmethylxanthine, 8-bromo-cyclicAMP, and dexamethasone (KIAD) expressed the differentiated type II cell phenotype as measured by the expression of surfactant protein (SP)-A, SP-B, SP-C, and fatty acid synthase and their morphologic appearance. These cells contain lamellar inclusion bodies and have apical microvilli. In both systems the cells appear well differentiated. In the apical access system, type II cell differentiation markers initially decreased and then recovered over 6 d in culture. Lipid synthesis was also increased by the addition of KIAD. In contrast, type II cells cultured on rat tail collagen (or tissue culture plastic) slowly lose their lamellar inclusions and expression of the surfactant proteins and increase the expression of type I cell markers. The expression of the phenotypes is regulated by the culture conditions and is, in part, reversible in vitro.

Introduction: For many years, transurethral resection of the prostate (TURP) has been accepted as the gold standard to surgically alleviate obstructive voiding dysfunction in men with benign prostatic hyperplasia (BPH). This historical standard has been challenged repeatedly over the last decade by consistent data demonstrating the superiority of Holmium enucleation of the prostate (HoLEP). This review summarizes the literature comparing HoLEP and traditional therapies for BPH that are widely used and have long term efficacy data, primarily TURP, open prostatectomy (OP), and alternative laser therapies (PVP, ThuLEP, etc). Results: Patients undergoing HoLEP have greater improvements in post-operative Qmax, greater reduction in post-operative subjective symptom scores, and lower rates of repeat endoscopic procedures for recurrent symptoms at 5-10 year follow up compared with TURP, OP, and other laser therapies. Furthermore, patients undergoing HoLEP benefit from significantly shortened catheterization times, decreased length of hospital stay (LOS), and fewer serious post-operative complications. In particular, randomized controlled trials (RCT) have demonstrated that HoLEP can be used to resect adenomas greater than 100 grams with equivalent efficacy to open prostatectomy, but with radically decreased morbidity. Conclusion: Numerous large, RCTs demonstrate HoLEP to be objectively superior to other surgical therapies for BPH. The urologic community should embrace HoLEP as the new gold standard for surgical BPH therapy, especially in men with large prostates who would otherwise be considered for an OP or staged TURP. The only obstacle to widespread implementation of HoLEP remains its difficult learning curve when compared with traditional transurethral resection. Further allocation of resources towards appropriate mentoring and teaching of HoLEP is warranted, particularly in residency training programs.

Ischemia/reperfusion injury is a leading cause of renal damage and antisense gene therapy has been shown to ameliorate its effects. However, this approach has been limited by current delivery methods that require high concentrations of intravenous nucleic acids lacking specificity for targeting tissues. To overcome these limitations we developed a novel murine partial nephrectomy model to evaluate polyethylene-glycol (PEG) hydrogel tissue sealant as a topical oligonucleotide delivery system.A total of 18 male C57BL/6 mice underwent left partial nephrectomy with vascular occlusion. Hydrogel primer and then sealant were applied to the cut surface and photopolymerized. Using this method 16 additional mice received hydrogel primer mixed with Cy5 labeled fluorescent oligonucleotide (10 to 100 microg). Kidneys were harvested at various time points and assessed for oligonucleotide penetration using fluorescence microscopy.A survival rate of 100% (34 subjects) was obtained using this mouse model of partial nephrectomy. PEG hydrogel provided adequate protection against renal hematoma and intraperitoneal blood. Fluorescent images revealed that 50 microg was the minimum dose resulting in complete progressive cellular penetration with time. In addition to direct diffusion from the application site, movement of oligonucleotide through the subcapsular space into the cortex was an observed mechanism of distribution.A murine partial nephrectomy model was successfully created using PEG hydrogel. In addition to achieving hemostasis, hydrogel served as a successful depot for delivering oligonucleotides throughout the kidney.

INTRODUCTION AND OBJECTIVES: Dynamic Pelvic MRI (DPMRI) remains an emerging diagnostic modality in the evaluation of Pelvic Organ Prolapse (POP), but study quality remains variable, and is typically dependent upon performance of a proper valsalva maneuver during the dynamic phase of the procedure. We sought to determine whether the addition of a defecography phase (DP) to routine DPMRI would increase its clinical utility over standard valsalva maneuver (VM) in the evaluation of POP. METHODS: Fifty-six female patients undergoing preoperative evaluation for surgical correction of symptomatic POP were evaluated by DPMRI utilizing a protocol incorporating both VM and DP. Ultrasound gel was injected into the vagina and rectum followed by (1) steady state free precession images in the sagittal plane, (2) dynamic cine acquisitions during VM, and (3) dynamic cine acquisitions during DP. All images were separately reviewed in a blinded fashion by two radiologists experienced in DPMRI. The HMO system was used for quantification of pelvic floor descent. Inferior descent of the ano-rectal junction, bladder base and vaginal vault was recorded in all patients, utilizing the pubococcygeal line (PCL) as a fixed landmark. IRB approval was obtained for the study. RESULTS: DP produced a greater degree of pelvic organ descent compared to VM for all recorded measures in all 56 patients. 100%(56/56) demonstrated rectocele, 71% (40/56) cystocele and 71% (40/56) vaginal vault descent on DP, compared with 86% (48/56), 14% (8/56) and 43% (24/56) on VM, respectively. The average descent of the bladder base was 3.4 cm with DP, compared to 0.4 cm with VM (p

We retrospectively compared our initial experience with the hand-assisted and retroperitoneal laparoscopic nephrectomy techniques to determine if there are important differences between these approaches.Twenty-four laparoscopic cases consisting of 12 hand-assisted and 12 retroperitoneal nephrectomies were compared. All cases but one were radical nephrectomies with intact specimen extraction performed for suspected stage T1 neoplasms. Data were collected from medical records and a postoperative questionnaire. To determine if significant learning curves existed, the first six nephrectomies in each group were compared with the second six nephrectomies on the basis of operative criteria. The two groups did not differ significantly in age, body mass index, ASA rating, or number of previous abdominal operations.Although the mean tumor volume was greater in the hand-assisted group than the retroperitoneal group, the difference did not quite reach statistical significance (91.19 v 24.7 cc3; P = 0.06). The mean operative time, estimated blood loss, narcotic use (milligrams of intravenous morphine equivalent), hours to oral intake, hospital stay, and estimated percent activity at 2 weeks for the hand-assisted group (238.33 min, 293.75 mL, 35.7 mg, 17.56 hours, 4.4 days, 74.75%, respectively) were not significantly different from the values in the retroperitoneal group (255.83 min, 141.67 mL, 24.5 mg, 22.36 hours, 3.6 days, 76.91%). We found no significant difference in the mean operative times for the first and second six cases in either group.In the initial experience and comparison of hand-assisted and retroperitoneal laparoscopic nephrectomy, we found no significant differences in operative time, estimated blood loss, narcotic usage, hours to oral intake, hospital stay, or activity level at 2 weeks postoperatively. A randomized trial is under way at our institution.

Human coronavirus strain 229E (HCoV-229E) commonly causes upper respiratory tract infections. However, lower respiratory tract infections can occur in some individuals, indicating that cells in the distal lung are susceptible to HCoV-229E. This study determined the virus susceptibility of primary cultures of human alveolar epithelial cells and alveolar macrophages (AMs). Fluorescent antibody staining indicated that HCoV-229E could readily infect AMs, but no evidence was found for infection in differentiated alveolar epithelial type II cells and only a very low level of infection in type II cells transitioning to the type I-like cell phenotype. However, a human bronchial epithelial cell line (16HBE) was readily infected. The innate immune response of AMs to HCoV-229E infection was evaluated for cytokine production and interferon (IFN) gene expression. AMs secreted significant amounts of tumour necrosis factor alpha (TNF-α), regulated on activation normal T-cell expressed and secreted (RANTES/CCL5) and macrophage inflammatory protein 1β (MIP-1β/CCL4) in response to HCoV-229E infection, but these cells exhibited no detectable increase in IFN-β or interleukin-29 in mRNA levels. AMs from smokers had reduced secretion of TNF-α compared with non-smokers in response to HCoV-229E infection. Surfactant protein A (SP-A) and SP-D are part of the innate immune system in the distal lung. Both surfactant proteins bound to HCoV-229E, and pre-treatment of HCoV-229E with SP-A or SP-D inhibited infection of 16HBE cells. In contrast, there was a modest reduction in infection in AMs by SP-A, but not by SP-D. In summary, AMs are an important target for HCoV-229E, and they can mount a pro-inflammatory innate immune response to infection.

The histopathology of Entomophaga grylli pathotype 2-infected tissues was investigated. Fat body, male reproductive system, gastrointestinal tract, central nervous system, and skeletal muscle of infected and uninfected grasshoppers were compared to determine the pathological events during the infection. The pathotype 2 protoplasts adhered to fat body and penetrated neural tissue early in the infection, whereas the skeletal muscle was penetrated after host death. Ultrastructural examination of infected fat body suggested one contributing cause of death was nutrient depletion.

The whitefly Bemisia tabaci contains a primary prokaryotic endosymbiont housed within specialized cells in the body cavity. Two DNA fragments from the endosymbiont, totaling 33.3 kilobases, were cloned and sequenced. In total, 37 genes were detected and included the ribosomal RNA operon and genes for ribosomal RNA proteins. The guanine plus cytosine of the DNA was 30.2 mol%, different from that of endosymbionts of other plant sap-sucking insects.

Kinesins are a group of related molecular motor proteins that have great potential as targets for antimitotic drug development. We have developed two novel assays, one end-point and one kinetic, that are useful for the discovery and optimization of kinesin modulators. Both assays measure inorganic phosphate (Pi) generated by microtubule-activated kinesin adenosine triphosphatase activity. The assays were validated using the mitotic Eg5 kinesin-specific inhibitor, monastrol. A panel of nine kinesin motor domain proteins, representing 8 of the 14 classes of kinesins, was screened. The coefficient of variation for both assays was determined to be 4-14% depending on the panel member. Using the Eg5 kinetic assay with monastrol the IC50 value was 12 microM, which agrees well with previously published results. Two other closely related mitotic kinesins (AnBimC and MKLP1) were found to have IC50 values in the millimolar range. The other panel members (kinesin heavy chain, chromokinesin KIF4A, KIF3C, CENP-E, MCAK, and KIFC3) were not significantly inhibited by millimolar levels of monastrol. It is anticipated that screening of the nine-member panel of kinesins in these assays will serve as a platform for the discovery and development of specific kinesin modulators.

The genome of the Lymantria dispar multinucleocapsid nucleopolyhedrovirus (LdMNPV) was sequenced and analyzed. It is composed of 161,046 bases with a G + C content of 57.5% and contains 163 putative open reading frames (ORFs) of >/=150 nucleotides. Homologs were found to 95 of the 155 genes predicted for the Autographa californica MNPV (AcMNPV) genome. More than 9% of the LdMNPV genome was occupied by 16 repeated genes related to AcMNPV ORF2. Readily identifiable homologs of several genes that have been reported to play important roles in the AcMNPV life cycle are not present; these include ie-2, a transcriptional transactivator, and gp64, a major envelope glycoprotein of the nonoccluded form of the virus. A number of genes lacking in AcMNPV but present in other baculoviruses were identified; these include two viral enhancing factor homologs, a second copy of a conotoxin-like gene, and a dutpase homolog. Although a single gene predicted to encode a large subunit of ribonucleotide reductase was found, two different copies of the small subunit gene were present. In addition, homologs of genes not previously reported for baculoviruses were identified, including a predicted protein with homology to DNA ligases and another that has motifs most closely related to a yeast mitochondrial helicase. Thirteen homologous regions (hrs) containing 54 repeated sequences that include 30-bp imperfect palindromes were identified. The imperfect palindromes are related to those from other baculoviruses.

Viral infections in the lung are an important cause of morbidity and mortality in humans worldwide. In order to design strategies for vaccination and therapy, we need to understand the molecular mechanisms that initiate and regulate effective antiviral immune responses in the lung. This requires animal models in which in vitro mechanistic studies can be correlated with infections in vivo. We study rat coronavirus (RCoV) infection in rats and cultures of primary differentiated rat alveolar epithelial cells as a model for the early events in respiratory coronavirus pathogenesis. Intratracheal inoculation of adult rats with RCoV resulted in infection of the lung, primarily in alveolar type I cells. Infection caused transient weight loss and focal lesions and recruitment of neutrophils in the lung, followed by clearance of virus and resolution of inflammation. Thus, adult rats mounted an effective innate immune response to RCoV infection in the lung. In agreement, in primary cultures of differentiated rat alvoelar type I cells, RCoV replicated and induced expression of neutrophil chemotactic CXC chemokines. This in vitro model will elucidate the molecular interactions between RCoV and alveolar epithelial cells that initiate and regulate the innate antiviral immune response in the lung. This work was supported by NIH grants AI-059576 and HL-029891, and NCRR/NIH grant P20 RR015587.