Combined hyperosmolarity and inflammatory conditions in stressed human corneal epithelial cells and macrophages to evaluate osmoprotective agents as potential DED treatments.

PURPOSE: To develop an easy-to-perform combined model in human corneal epithelial cells (HCECs) and Balb/c mice macrophages J774.A1 (MP) for preliminary screening of potential ophthalmic therapeutic substances. METHODS: HCECs were exposed to different osmolarities (350-500 mOsm/L) and MTT assay was employed for cell survival and flow cytometry to assess apoptosis-necrosis and relative cell size (RCS) distribution. Effectiveness of Betaine, L-Carnitine, Taurine at different concentrations (ranging from 20 mM to 200 mM) was studied. Also, mucoadhesive polymers such as Hyaluronic acid (HA) and Hydroxypropylmethylcellulose (HPMC) (0.4 and 0.8%) were evaluated. Cells were pre-incubated with the compounds (8h) and then exposed to hyperosmotic stress (470 mOsm/L) for 16h. Moreover, anti-inflammatory activity was performed in LPS-stimulated MP. RESULTS: Exposure to hyperosmotic solutions between 450 and 500 mOsm/L promoted the highest cell death after 16h exposures (p < 0.0001) with a drop in viability to 34.96% ± 11.77 for 470 mOsm/L. Pre-incubation with Betaine at 150 mM and 200 mM provided the highest cell survival against hyperosmolarity (66.01% ± 3.65 and 65.90% ± 0.78 respectively) while HA 0.4% was the most effective polymer in preventing cell death (42.2% ± 3.60). Flow cytometry showed that Betaine and Taurine at concentrations between 150-200 mM and 20-80 mM respectively presented the highest anti-apoptotic activity. Also, HA and HPMC polymers reduced apoptotic-induced cell death. All osmoprotectants modified RCS, and polymers increased their value over 100%. L-Carnitine 50 mM, Taurine 40 mM and HA 0.4% presented the highest TNF-α inhibition activity (60%) albeit all of them showed anti-inflammatory inhibition percentages higher than 20% CONCLUSIONS: HCECs hyperosmolar model combined with inflammatory conditions in macrophages allows the screening of osmoprotectants by simulating chronic hyperosmolarity (16h) and inflammation (24h).

Transplant renal artery stenosis: Study of incidence using doppler ultrasound, risk factors and analysis is effect in graft outcomes.

INTRODUCTION AND OBJECTIVES: Transplant renal artery stenosis (TRAS) is a vascular complication after kidney transplantation which estimated incidence is 13%. It could cause refractory arterial hypertension, kidney dysfunction and premature death in transplant recipients. METHODS: We carried out a retrospective study including every patient who underwent renal transplantation between 2014 and 2020. They were evaluated with a systematic post-transplant renal Doppler ultrasound. To identify independent risk factors for transplant renal artery stenosis we performed a multivariate analysis. RESULTS: Seven hundred twenty-four kidney transplants were included, 12% ​​were living donors and 88% were deceased donors. The mean age was 54.8 in recipients and 53 in donors. Transplant renal artery stenosis was diagnosed in 70 (10%) recipients, the majority in the first 6 months after surgery. 51% of patients with transplant renal artery stenosis were managed conservatively. The multivariate analysis showed diabetes mellitus, graft rejection, arterial resuture and donor body mass index as independent risk factors for transplant renal artery stenosis. Survival of the grafts with transplant renal artery stenosis was 98% at 6 months and 95% at two years. CONCLUSIONS: The systematic performance of Doppler ultrasound in the immediate post-transplant period diagnosed 10% of transplant renal artery stenosis in our cohort. Despite the above risk factors, an adequate monitoring and treatment could avoid the increased risk of graft loss in patients with transplant renal artery stenosis.

New trends towards glaucoma treatment: Topical osmoprotective microemulsions loaded with latanoprost.

The chronic use of hypotensive agents eventually leads to ocular surface damage and poor patient compliance during glaucoma management. Thus, new sustained drug delivery systems are needed. This work aimed to develop osmoprotective latanoprost-loaded microemulsion formulations as new potential glaucoma treatments with ocular surface protective properties. The microemulsions were characterized and latanoprost encapsulation efficacy determined. In-vitro tolerance, osmoprotective efficacy, cell internalization as well as cell-microemulsion interactions and distribution were performed. In vivo hypotensive activity was conducted in rabbits to assess intraocular pressure reduction and relative ocular bioavailability. Physicochemical characterization showed nanodroplet sizes within 20-30 nm, being in vitro tolerance within 80 and 100% viability in corneal and conjunctival cells. Besides, microemulsions exhibited higher protection under hypertonic conditions than untreated cells. Cell fluorescence lasted for 11 days after short exposure to coumarin-loaded microemulsions (5 min) showing extensive internalization in different cell compartments by electronic microscopy. In vivo studies exhibited that a single instillation of latanoprost-loaded microemulsions reduced the intraocular pressure for several days (4-6 days without polymer and 9-13 days with polymers). Relative ocular bioavailability was 4.5 and 19 times higher than the marketed formulation. These findings suggest the use of these microemulsions as potential combined strategies for extended surface protection and glaucoma treatment.

Development of an osmoprotective microemulsion as a therapeutic platform for ocular surface protection.

Self-emulsified osmoprotective ophthalmic microemulsions (O/A) were prepared by combining betaine/leucine, clusterin/oleanolic acid, and hyaluronic acid or Dextran. The microemulsions contained an internal oily phase (1.2%), an external aqueous phase (96.3%), cosolvents (1%), and surfactants (1.5%). Physicochemical characterization and in vivo and in vitro tolerance were analyzed. The formulations' osmoprotective in vitro activity was assayed in a hyperosmolar model in human corneal cells. Average internal phase sizes were 16-26 nm for the microemulsions including Dextran. Addition of hyaluronic acid increased the size range (25-39 nm). Addition of osmoprotectants did not change nanodroplet size. The formulations were isotonic (280-290 mOsm/L) with neutral pH (≈7) and zeta potential (-10 to 0 mV), low surface tension (≈35-40mN·m-1), and low viscosity (≈1 mPa·s), except for the microemulsions containing hyaluronic acid (≈4-5 mPa·s). SEM and cryo-TEM showed that all formulations exhibited sphere-shaped morphology with good cell tolerance (≈100%) and were stable at 8 °C for 9 months. Osmoprotective formulations were well tolerated in vitro and in vivo, protecting cells from hypertonic stress. We therefore developed stable microemulsions compatible with the ocular surface that could constitute a novel tool for treatment of ophthalmic diseases.

[Endoscopic modified technique of ureteral resection during nephroureterectomy]. / Modificación técnica de la desinserción ureteral endoscópica en la nefroureterectomía.

We show a technical modification of the ureteral endoscopic resection with which we try to avoid comunication between urine and surgical bed in order to prevent tumor local spread of upper urotelial tumor.

[Ileal neobladder with double chimney. Ureterointestinal anastomosis]. / Neovejiga ileal con doble chimenea. Anastomosis ureterointestinal.

Radical cystoprostatectomy is accepted as the standard treatment for muscle-invasive bladder cancer. During last years the indications for orthotopic neobladders have increased due to their advantages over other kind of diversions. Hautmann neobladder is one of the most commonly used. Several modifications have been later described. For example, after perform the W-shape pouch ureters can be anastomosed to a not-detubularized bowel segment (chimney modification). Here is described a modification of the Hautmann neobladder with two chimneys. Each ureter is spatulated in a golf club manner and anastomosed to the open end of each bowel loop. This kind of anastomosis provides several advantages. It is possible to use shorter ureteral segments by increasing the length of bowel used. It allows an anastomosis without tension, and less ischemia so the risk of stenosis and fistula is decreased. It is not necessary to perform additional enterothomies and in case of reintervention it is easier to access each anastomosis without damaging the other one.

[Bilateral brucellosic psoas abscess: one case is related and literature review]. / Absceso de psoas brucelósico bilateral: presentación de un caso y revisión de la literatura.

The Brucellosis is a zoonotic and unfrequent infection but it is endemic in Spain. It is a well documented cause of fever of unknown origin with varied and nonspecific symptoms. The onset of symptoms of brucellosis may be abrupt or insidious, developing over several days to weeks. Virtually any organ system can be involved with brucellosis and localization of the process may cause focal symptoms. The most frequent focal presentation is the osteoarthicular. Some times it can complicates with para-perivertebral abscess. The unilateral psoas abscess(PA) is uncommon and the bilateral afection is exceptional. The manifestations of PA usually are insidious, the classic tiad of: feber, lumbar pain and functional impotence is rare. For the diagnosis the serology c tests and cultures are necesary. The imaging techniques like: Ultrasonography and Tomography have improved the diagnosis and treatment of this pathologies' complications. The recommended therapy is the use of doxycicline and streptomicine. Some times the use of percutaneous drainage or open surgery is necessary.

Uncontrolled non-heartbeating donors (types i-ii) with normothermic recirculation vs. heartbeating donors: evaluation of functional results and survival.

OBJECTIVE: Non-heartbeating donors (NHBD) are an alternative to heartbeating donors (HBD). Our objective was to compare functional results and kidney survival from NHBDs and HBDs. MATERIAL AND METHODS: A retrospective study comparing the results of 236 normothermically preserved kidneys from type i and ii type NHBDs with the results of 250 from HBDs that were transplanted in our center between 2005 and 2012. Homogeneity between groups was tested and we evaluated the presence of delayed graft function (DGF) associated with pretransplant variables of the donor and recipient. RESULTS: Both groups show homogeneity in pretransplant characteristics in terms of: age, HLA incompatibilities, and recipient hemodialysis time. Average follow-up time was 33 months (range 0-87) for NHBDs and 38 months (range 0-90) for HBDs. 5.5% of NHBDs showed primary non-function (PNF) vs. 4% of HBDs (P=.42) and 80.9% of DGF vs. 46.8% of HBDs (P<.001). At the end of the follow-up, there were no statistically significant differences in the survival of grafts (92.8% for NHBD vs. 93.6% for HBD, P=.71) and recipients (99.1% NHBD vs. 98.6% HBD, P=.28). CONCLUSIONS: Although the DGF percentage was greater for NHBDs, final creatinine as well as graft and recipient survival were similar for both groups. Therefore, in our experience, kidneys from NHBDs have similar results to those from HBDs and are an excellent source of organs for transplantation.

[Obturator nerve block in transurethral surgery]. / Bloqueo del nervio obturador en la cirugía transuretral.

The obturator nerve passes in close proximity to the bladder as it courses through the pelvis. During transurethral operations, resection may result in stimulation of the obturator nerve, causing violent adductor contraction. Bladder perforation and incomplete tumor resection are the most important complications. All techniques proposed since transurethral surgery began, until nowadays are reviewed: neuromuscular blockade, electric circuit modifications, transparietal endoscopic blockade, periprostatic and subvesical infiltration, obturator nerve blockade and the "3 in 1 block" described by Winnie. Practical advices are proposed finally.

[Melanoma in urology: our experience and review of the literature]. / El melanoma en urología: nuestra experiencia y revisión de la literatura.

Melanomas make up 2% of all body tumours, but the figure is increasing mainly in countries with longer sun hours daily. Genitourinary location is even more infrequent. That explains our clinical report when we discovered the three cases seen in our unit, one in a female urethra, one suprarenal and the third one in the bladder.

[Leukemoid reaction as a paraneoplastic manifestation of bladder carcinoma]. / Reacción leucemoide como manifestación paraneoplásica del carcinoma vesical.

Sometimes vesical malignant tumours present themselves with highly characteristic paraneoplastic signs like hypercalcemia, or more rarely like a leukaemoid reaction within the symptomatic setting typical of vesical neoplasia. We present a case of vesical urothelioma where the leukaemoid reaction was present in the diagnosis and later in the tumour relapse after the appropriate treatment.

[Intradiverticular bladder tumors]. / Tumores vesicales intradiverticulares.

Retrospective study conducted on 58 patients with vesical diverticulum seen in our unit between 1975 and 1992, of which a total of 11 (18.9%) patients had vesical tumours. Of these, 6 (10.3%) were intradiverticular and 5 (8.6%) extradiverticular. Sixty-six percent of patients with intradiverticular tumours also had prior or concurrent history of extradiverticular vesical tumours. The most frequent clinical presentation was gross haematuria. The abdomino-pelvic CT is the most sensitive morphological examination although in 33% cases it resulted in overstaging. Curative treatment was only possible in the 4 patients with urothelial tumours, in whom 3 partial cystectomies with pelvian lymphadenectomy (2 pT1 G2 and 1 pT3 G3) and 1 TUR (T1 G2) were performed. The 2 (33%) remaining patients had advanced locoregional epidermoid carcinoma (T4 N+). All patients with urothelial carcinoma are alive with follow-up ranging between 6 months for the one infiltrant case and 136 months for a surface tumour undergoing partial cystectomy. Prognosis for both epidermoid carcinoma was ominous with mean survival time of 9 months. Review of the literature and discussion of epidemiological, clinical, diagnostic, therapeutic and prognostic issues.

[Tumor pathology in horseshoe kidney]. / Patología tumoral en el riñón en herradura.

Within renal aberrations, the most frequent one is the horseshoe kidney. Adenocarcinoma is the most common primary renal neoplasia. The association of tumour and horseshoe kidney is uncommon, with only 135 cases described in the literature. Presentation of 4 patients with tumours in horseshoe kidney: 3 adenocarcinoma and one urothelial tumour of renal pelvis. As a result of our experience we consider that arteriography is a key imaging technique for pre-operative study of these patients, medial laparotomy being the choice access for approach, and prognosis depending on tumour staging, with no influence from the malformation.

[Technique of bladder catheterization after transurethral prostatic resection]. / Técnica de cateterización vesical tras la resección transuretral de próstata.

The obstacle that represents the bladder neck posterior lip after a prostate transuretral resection is, sometimes, the cause of sugtrigonal perforations when a bladder catheterization is performed after the surgery. We explain an easy procedure that allows a safe catheterization, reducing the risk of complications added to those of the surgery.

[Abdominal paragangliomas. The experience of the Hospital 12 de Octubre]. / Paragangliomas abdominales. Experiencia del Hospital 12 de Octubre.

This is a revision of our experience between 1975 and 1992 over a total of 35 patients who underwent surgery in 40 occasions for pheochromocytoma. If we dismiss one case of pheochromocytoma with early metastatic malignant presentation and which was not removed, a total of 6 patients with abdominal paraganglioma (17%) have undergone surgery in 10 occasions. Three of them (50%) were multiple and recurrent, an one (16%) become malignant after multiple recurrences. Average age was 34.6 years with balanced sex distribution. No patient had neurofibromatosis, Von Hippel Lindau disease, MEN syndromes or Carney's triad and only one case, which corresponded to the malignant pheochromocytoma, showed direct familial background. All patients were hypertensive. Four cases (66%) presented with catecholamine crisis, one case was an incidentaloma that at the 24-hour monitoring presented hypertensive crisis during the sleep, and the last one was diagnosed while investigating a case of sustained HTA in a young female. All patients had high urinary catecholamine excretion. To establish location of the tumour MIBG scanning (90% sensitivity) was used in first place followed by guided CT (80% sensitivity) since both techniques have a good correlation in order to design the surgical approach. Angiography was reserved for cases where the other two techniques were inconclusive or when it was necessary to obtain a better profile of the surgical anatomy. A total of 15 paraganglioma were removed, the most frequent location being renal parahilar (26%) followed by preaortic in Zuckerkandl's organ (20%) and vesical (20%). The most relevant intraoperative complications were HTA crisis related to anaesthetics manoeuvres and tumoral handling (90%), and hypotension following tumour exercises (10%). During the postoperative period, three patients required blood transfusion and one of them had to be re-intervened for bleeding caused by unnoticed damage to the right renal vein. Currently, three patients (50%) are disease free, one with residual mild HTA controlled with diuretics. Two patients (33%) are awaiting for re-intervention due to recurrence (one with multiple extra-abdominal recurrence) after one and two prior interventions respectively. The last one (16%) is the malignant pheochromocytoma, operated four times for recurrence which currently shows lung metastasis with adequate drug control of clinical manifestations, after polychemotherapy failure and 27-months follow-up since metastasis has been detected. Revision of existing literature and discussion of issues related to signs and symptoms, diagnosis, surgical preparation and approach, as well as prognostic implications related to paraganglioma as compared with adrenal-located pheochromocytoma.

[Genitourinary malacoplakia]. / Malacoplaquia genitourinaria.

Malacoplakia is a chronic inflammatory disease the etiology of which remains obscure. It has a very low incidence and affects primarily the genitourinary tract, although it has been described in some other organs. This paper presents a historic insight of the clinical cases diagnosed in this centre, and includes a review and update of several issues related to this entity such as pathogenesis, pathological anatomy and treatment. Also, the peculiarities related to the involvement of each separate organ with regard to diagnosis, prognosis and treatment are described.

Dual kidney transplantation involving organs from expanded criteria donors: a review of our series and an update on current indications.

BACKGROUND: Our purpose was to review our kidney transplantation program based on the use of expanded criteria donors, and to determine current indications for dual kidney transplantation (DKT). In 1996, a program was initiated to transplant kidneys from donors of over 60 years performing single or dual transplantation. METHODS: In 1996, a program was initiated to transplant kidneys from donors of over 60 years performing single or dual transplantation. DKT were performed with donors >75 and donors between 60 and 74 years of age and glomerulosclerosis of >15%. The kidneys of donors between 60 and 74 years of age and with glomerulosclerosis of <15% were used for single kidney transplantation (SKT). In 2005, we started to perform SKT despite glomerulosclerosis being >15%, taking into account donor and recipient characteristics. RESULTS: From 1996 to 2004, 222 SKTs and 88 DKTs were performed. Graft survival after 1 and 4 years was, respectively, 91% and 78% for SKT and 95% and 79% for DKT. In 2005, we started to perform SKT despite glomerulosclerosis being >15%, taking into account donor and recipient characteristics. From 2005 to 2011, 328 SKT and 32 DKT were performed. During this period most kidneys used for DKT were from female donors >75 years old, weighing <65 kg, with a creatinine of >1 mg/dL and glomerulosclerosis of >15%. The recipients for DKT were mostly male, <70 years old and whose weight was >75 kg. CONCLUSION: DKT from expanded criteria donors shows good outcomes. However, in many cases SKT may fulfill the need of the recipient. The archetype for DKT is an older female weighing <65 kg and the most common recipient is an overweight male who is <70 years old.