Leuprorelin Acetate Long-Lasting Effects on GnRH Receptors of Prostate Cancer Cells: An Atomic Force Microscopy Study of Agonist/Receptor Interaction

High cell-surface GnRH receptor (GnRH-R) levels have been shown to have a major influence on the extent of GnRH agonist-mediated tumor growth inhibition. The ability of the GnRH agonist leuprorelin acetate (LA) to induce a post-transcriptional upregulation of GnRH-R at the plasma membrane of androgen-sensitive (LNCaP) and -insensitive (PC-3) prostate cancer (PCa) cells has been previously demonstrated by Western blotting. Here we performed single molecule force spectroscopy by using Atomic Force Microscopy (AFM), which has proven to be a powerful tool allowing for investigation of living cell surface biological features, such as the so far unclear GnRH agonist/receptor interaction. Thus, in the hormone-insensitive PC-3 cells, we characterized the strength of the LA-receptor binding, and the amount and distribution of the functional receptor molecules on the cell surface. The effect of a long and continuous treatment (up to 30 days) with the agonist (10-11 and 10-6 M) on the same parameters was also investigated. A GnRH-R increase was observed, reaching the maximum (~80%) after 30 days of treatment with the highest dose of LA (10-6 M). The analogue-induced increase in GnRH-R was also demonstrated by Western blotting. In addition, two different receptor bound strengths were detected by AFM, which suggests the existence of two GnRH-R classes. A homogeneous distribution of the unbinding events has been found on untreated and treated PC-3 cell surfaces. The persistence of high receptor levels at the membrane of these living cells may warrant the maintenance of the response to LA also in androgen-unresponsive PCa. Moreover, the determination of ligand/receptor bond strength could shed light on the poorly understood event of LA/GnRH-R interaction and/or address structural/chemical agonist optimizations. \ua9 2013 Lama et al

Vision-related quality of life and symptom perception change over time in newly-diagnosed primary open angle glaucoma patients.

To evaluate the change over time of vision-related quality of life (QoL) and glaucoma symptoms in a population of newly-diagnosed primary open angle glaucoma (POAG) patients. Multicenter, prospective study. Consecutive newly-diagnosed POAG patients were enrolled and followed-up for one year. Follow-up visits were scheduled at 6 and 12 months from baseline. At each visit, vision-related QoL and glaucoma-related symptoms were assessed by the means of the 25-item National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) and the Glaucoma Symptom Scale (GSS), respectively. Trends over time for NEI-VFQ-25 and GSS scores were evaluated with longitudinal linear mixed models. One-hundred seventy-eight patients were included in the analysis. At baseline, early to moderate glaucoma stages were associated with higher scores for most GSS and NEI-VFQ-25 items, while lower best-corrected visual acuity was associated with lower scores for 4 of the 12 NEI-VFQ-25 items. During the follow-up, all the GSS scores, the NEI-VFQ-25 total score, and 7 of the 12 NEI-VFQ-25 scores significantly improved (p < 0.05). In multivariate model, higher increases of most GSS and NEI-VFQ-25 scores were modeled in patients with low scores at baseline. Vision-related QoL and glaucoma-related symptom perception significantly improved during the one-year follow-up in this population of newly diagnosed POAG patients

The mechanisms by which polyamines accelerate tumor spread

Increased polyamine concentrations in the blood and urine of cancer patients reflect the enhanced levels of polyamine synthesis in cancer tissues arising from increased activity of enzymes responsible for polyamine synthesis. In addition to their de novo polyamine synthesis, cells can take up polyamines from extracellular sources, such as cancer tissues, food, and intestinal microbiota. Because polyamines are indispensable for cell growth, increased polyamine availability enhances cell growth. However, the malignant potential of cancer is determined by its capability to invade to surrounding tissues and metastasize to distant organs. The mechanisms by which increased polyamine levels enhance the malignant potential of cancer cells and decrease anti-tumor immunity are reviewed. Cancer cells with a greater capability to synthesize polyamines are associated with increased production of proteinases, such as serine proteinase, matrix metalloproteinases, cathepsins, and plasminogen activator, which can degrade surrounding tissues. Although cancer tissues produce vascular growth factors, their deregulated growth induces hypoxia, which in turn enhances polyamine uptake by cancer cells to further augment cell migration and suppress CD44 expression. Increased polyamine uptake by immune cells also results in reduced cytokine production needed for anti-tumor activities and decreases expression of adhesion molecules involved in anti-tumor immunity, such as CD11a and CD56. Immune cells in an environment with increased polyamine levels lose anti-tumor immune functions, such as lymphokine activated killer activities. Recent investigations revealed that increased polyamine availability enhances the capability of cancer cells to invade and metastasize to new tissues while diminishing immune cells' anti-tumor immune functions

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Background: Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. // Methods: We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung's disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. // Findings: We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung's disease) from 264 hospitals (89 in high-income countries, 166 in middle-income countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in low-income countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. // Interpretation: Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between low-income, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Hypotensive effect of pilocarpine after argon laser trabeculoplasty

The exact mechanism of action of argon laser trabeculoplasty (ALT) has only been presumptively proposed. There are two hypotheses of action of ALT: the first is the mechanical theory and the second is centered on the possibility of laser-induced modifications of trabecular cells. The goal of our investigation was to verify if pilocarpine determines a decrease of intraocular pressure mainly by a mechanical stretching of the meshwork; if pilocarpine could lower intraocular pressure after ALT; and thus if the mechanism of action of laser treatment could mainly be attributable to a mechanical strain of the trabecular meshwork. Twenty-one patients affected with primary open angle glaucoma had been recruited for the study. Twenty-one eyes underwent trabeculoplasty. All the patients were followed for 6 months. Statistical analysis of the data revealed that pilocarpine was not able to induce a further decrease of intraocular pressure after trabeculoplasty. Moreover, the maximum decrease of intraocular pressure induced by pilocarpine before laser treatment was not statistically different from the tensional values after ALT. The results of the present investigation suggest that the mechanism of action of trabeculoplasty is probably attributable to a mechanical stretching of the trabecular meshwork and the results show the inefficacy of pilocarpine after laser treatment in the management of glaucomatous patients

Comparative effects on intraocular pressure between systemic and topical carbonic anhydrase inhibitors: A clinical masked, cross-over study

The aims of the following study were to compare the efficacy of Dorzolamide 1% eye drops with systemic Acetazolamide on the ocular pressure diurnal curve in patients with maximal medical therapy. Three ocular pressure curve were performed in glaucomatous patients, already receiving maximal medical therapy. After a baseline curve, patients were pretreated either with Dorzolamide 1% eye drops or 250 mg Acetazolamide tablets in a double-blind cross-over study. Dorzolamide 1% eye drops and 250 mg Acetazolamide tablets significantly reduced intraocular pressure (IOP) at 2 and 4 h after pretreatment. Both treatments caused a significantly additional decrease of IOP despite maximal medical therapy. Dorzolamide 1% eye drops is as effective as Acetazolamide tablets in reducing the IOP curve. Topical carbonic anhydrase inhibitors may represent an additional and safer treatment for those patients with uncontrolled medical glaucoma. (C) 1997 The Italian Pharmacological Society

Hypotensive effect of pilocarpine after argon laser trabeculoplasty

The exact mechanism of action of argon laser trabeculoplasty (ALT) has only been presumptively proposed. There are two hypotheses of action of ALT: the first is the mechanical theory and the second is centered on the possibility of laser-induced modifications of trabecular cells. The goal of our investigation was to verify if pilocarpine determines a decrease of intraocular pressure mainly by a mechanical stretching of the meshwork; if pilocarpine could lower intraocular pressure after ALT; and thus if the mechanism of action of laser treatment could mainly be attributable to a mechanical strain of the trabecular meshwork. Twenty-one patients affected with primary open angle glaucoma had been recruited for the study. Twenty-one eyes underwent trabeculoplasty. All the patients were followed for 6 months. Statistical analysis of the data revealed that pilocarpine was not able to induce a further decrease of intraocular pressure after trabeculoplasty. Moreover, the maximum decrease of intraocular pressure induced by pilocarpine before laser treatment was not statistically different from the tensional values after ALT. The results of the present investigation suggest that the mechanism of action of trabeculoplasty is probably attributable to a mechanical stretching of the trabecular meshwork and the results show the inefficacy of pilocarpine after laser treatment in the management of glaucomatous patients

Comparative effects on intraocular pressure between systemic and topical carbonic anhydrase inhibitors: A clinical masked, cross-over study

The aims of the following study were to compare the efficacy of Dorzolamide 1% eye drops with systemic Acetazolamide on the ocular pressure diurnal curve in patients with maximal medical therapy. Three ocular pressure curve were performed in glaucomatous patients, already receiving maximal medical therapy. After a baseline curve, patients were pretreated either with Dorzolamide 1% eye drops or 250 mg Acetazolamide tablets in a double-blind cross-over study. Dorzolamide 1% eye drops and 250 mg Acetazolamide tablets significantly reduced intraocular pressure (IOP) at 2 and 4 h after pretreatment. Both treatments caused a significantly additional decrease of IOP despite maximal medical therapy. Dorzolamide 1% eye drops is as effective as Acetazolamide tablets in reducing the IOP curve. Topical carbonic anhydrase inhibitors may represent an additional and safer treatment for those patients with uncontrolled medical glaucoma. (C) 1997 The Italian Pharmacological Society