Nandrolone Supplementation Promotes AMPK Activation and Divergent 18[FDG] PET Brain Connectivity in Adult and Aged Mice.

Decreased anabolic androgen levels are followed by impaired brain energy support and sensing with loss of neural connectivity during physiological aging, providing a neurobiological basis for hormone supplementation. Here, we investigated whether nandrolone decanoate (ND) administration mediates hypothalamic AMPK activation and glucose metabolism, thus affecting metabolic connectivity in brain areas of adult and aged mice. Metabolic interconnected brain areas of rodents can be detected by positron emission tomography using 18FDG-mPET. Albino CF1 mice at 3 and 18 months of age were separated into 4 groups that received daily subcutaneous injections of either ND (15 mg/kg) or vehicle for 15 days. At the in vivo baseline and on the 14th day, brain 18FDG-microPET scans were performed. Hypothalamic pAMPKT172/AMPK protein levels were assessed, and basal mitochondrial respiratory states were evaluated in synaptosomes. A metabolic connectivity network between brain areas was estimated based on 18FDG uptake. We found that ND increased the pAMPKT172/AMPK ratio in both adult and aged mice but increased 18FDG uptake and mitochondrial basal respiration only in adult mice. Furthermore, ND triggered rearrangement in the metabolic connectivity of adult mice and aged mice compared to age-matched controls. Altogether, our findings suggest that ND promotes hypothalamic AMPK activation, and distinct glucose metabolism and metabolic connectivity rearrangements in the brains of adult and aged mice.

Nasal Administration of Cationic Nanoemulsions as CD73-siRNA Delivery System for Glioblastoma Treatment: a New Therapeutical Approach.

Glioblastoma is the most devastating primary brain tumor. Effective therapies are not available, mainly due to high tumor heterogeneity, chemoresistance, and the difficulties imposed by blood-brain barrier. CD73, an enzyme responsible for adenosine (ADO) production, is overexpressed in cancer cells and emerges as a target for glioblastoma treatment. Indeed, ADO causes a variety of tumor-promoting actions, particularly by inducing tumor immune escape, whereas CD73 inhibition impairs tumor progression. Here, a cationic nanoemulsion to deliver CD73siRNA (NE-siRNA CD73R) via nasal route aiming glioblastoma treatment was developed. NE-siRNA CD73R was uptaken by glioma cells in culture, resulting in a parallel 60-80% decrease in AMPase activity and 30-50% in cell viability. Upon nasal delivery, NE-siRNA CD73R was detected in rat brain and serum. Notably, treatment with CD73siRNA complexes of glioma-bearing Wistar rats reduced tumor growth by 60%. Additionally, NE-siRNA CD73R treatment decreased 95% ADO levels in liquor and tumor CD73 expression, confirming in vivo CD73 silencing. Finally, no toxicity was observed in either primary astrocytes or rats with this cationic nanoemulsion. These results suggest that nasal administration of cationic NE as CD73 siRNA delivery system represents a novel potential treatment for glioblastoma. Graphical Abstract Glioblastoma is the most common and devastating form of primary brain tumor. CD73, a protein involved in cell-cell adhesion and migration processes and also responsible for extracellular adenosine (ADO) production, is overexpressed by glioma cells and emerges as an important target for glioma treatment. Indeed, ADO participates in tumor immune escape, cell proliferation, and angiogenesis, and CD73 inhibition impairs those processes. Here, a cationic nanoemulsion to deliver CD73 siRNA (NE-siRNA CD73R) via nasal route aiming glioblastoma treatment was developed. NE-siRNA CD73R knockdown in vitro and in vivo CD73. Upon nasal delivery of NE-siRNA CD73R, the treatment markedly reduced tumor volume by 60% in a rat preclinical glioblastoma model. The treatment was well tolerated, and did not induce kidney, liver, lung, olfactory, bone marrow, or behavior alterations. These results indicate that the nasal administration of NE as a CD73 siRNA delivery system offered an efficient means of gene knockdown and may represent a potential alternative for glioblastoma treatment.

The expression of pluripotency genes and neuronal markers after neurodifferentiation in fibroblasts co-cultured with human umbilical cord blood mononuclear cells.

Human umbilical cord blood is an attractive source of stem cells; however, it has a heterogeneous cell population with few mesenchymal stem cells. Cell reprogramming induced by different methodologies can confer pluripotency to differentiated adult cells. The objective of this study was to evaluate the reprogramming of fibroblasts and their subsequent neural differentiation after co-culture with umbilical cord blood mononuclear cells. Cells were obtained from four human umbilical cords. The mononuclear cells were cultured for 7 d and subsequently co-cultured with mouse fibroblast NIH-3T3 cells for 6 d. The pluripotency of the cells was evaluated by RT-PCR using primers specific for pluripotency marker genes. The pluripotency was also confirmed by adipogenic and osteogenic differentiation. Neural differentiation of the reprogrammed cells was evaluated by immunofluorescence. All co-cultured cells showed adipogenic and osteogenic differentiation capacity. After co-cultivation, cells expressed the pluripotency gene KLF4. Statistically significant differences in cell area, diameter, optical density, and fractal dimension were observed by confocal microscopy in the neurally differentiated cells. Contact in the form of co-cultivation of fibroblasts with umbilical cord blood mononuclear fraction for 6 d promoted the reprogramming of these cells, allowing the later induction of neural differentiation.

Implementation of a nurse-delivered intravitreal injection service.

PURPOSE: The purpose of this study was to introduce nurse-delivered intravitreal injections to increase medical retina treatment capacity. METHODS: Indemnity, clinical governance, training, planning, and implementation issues were addressed. The outcome measures were patient safety, patient experience, and clinic capacity. RESULTS: No serious vision-threatening complications were recorded in a consecutive series of 4000 nurse-delivered intravitreal injections. A Mann-Whitney test showed a significant increase in intravitreal injections (P=0.003) in the medical retina service after introduction of nurse-delivered intravitreal injections. The majority of patients accepted and were satisfied with a nurse-delivered intravitreal injection. DISCUSSION: Nurse-delivered intravitreal injections appear safe, acceptable to patients, and are an effective means to increase intravitreal injection capacity in medical retina clinics.

Infantile glaucoma in Rubinstein-Taybi syndrome.

PURPOSE: Long-term follow-up of patients with Rubinstein-Taybi-associated infantile glaucoma. METHODS: Case series. RESULTS: Three cases of infantile glaucoma in association with Rubinstein-Taybi syndrome are presented. DISCUSSION: This report highlights the importance of measuring intraocular pressure in this condition, as glaucoma is one of the major preventable causes of blindness in childhood.

The dose-response effect of acute intravenous transplantation of human umbilical cord blood cells on brain damage and spatial memory deficits in neonatal hypoxia-ischemia.

Despite the beneficial effects of cell-based therapies on brain repair shown in most studies, there has not been a consensus regarding the optimal dose of human umbilical cord blood cells (HUCBC) for neonatal hypoxia-ischemia (HI). In this study, we compared the long-term effects of intravenous administration of HUCBC at three different doses on spatial memory and brain morphological changes after HI in newborn Wistar rats. In addition, we tested whether the transplanted HUCBC migrate to the injured brain after transplantation. Seven-day-old animals underwent right carotid artery occlusion and were exposed to 8% O(2) inhalation for 2 h. After 24 h, randomly selected animals were assigned to four different experimental groups: HI rats administered with vehicle (HI+vehicle), HI rats treated with 1×10(6) (HI+low-dose), 1×10(7) (HI+medium-dose), and 1×10(8) (HI+high-dose) HUCBC into the jugular vein. A control group (sham-operated) was also included in this study. After 8 weeks of transplantation, spatial memory performance was assessed using the Morris water maze (MWM), and subsequently, the animals were euthanized for brain morphological analysis using stereological methods. In addition, we performed immunofluorescence and polymerase chain reaction (PCR) analyses to identify HUCBC in the rat brain 7 days after transplantation. The MWM test showed a significant spatial memory recovery at the highest HUCBC dose compared with HI+vehicle rats (P<0.05). Furthermore, the brain atrophy was also significantly lower in the HI+medium- and high-dose groups compared with the HI+vehicle animals (P<0.01; 0.001, respectively). In addition, HUCBC were demonstrated to be localized in host brains by immunohistochemistry and PCR analyses 7 days after intravenous administration. These results revealed that HUCBC transplantation has the dose-dependent potential to promote robust tissue repair and stable cognitive improvement after HI brain injury.

Factores pronóstico a corto plazo en los ancianos atendidos en urgencias por síndrome coronario agudo con elevación del segmento ST / Short-term prognostic factors in elderly patients treated in emergency departments

Objetivo: Investigar los factores asociados a la mortalidad a corto plazo en los pacientes ancianos que acuden a urgencias por síndrome coronario agudo con elevación del segmento ST (SCAEST).Método: Estudio multicéntrico, longitudinal, observacional, analítico-prospectivo y sin intervención. Se incluyó a pacientes de 70 o más años atendidos en 42 hospitales españoles. Se analizaron 17 variables independientes que pudieran influir en la mortalidad a 30 días. Los datos se obtuvieron a partir de un registro creado para este estudio, de la historia clínica o de la entrevista con el paciente o sus familiares. Se realizó un estudio multivariable mediante regresión logística. Resultados: Se incluyó a 1.137 pacientes, 340 (29,9%) fallecieron a los 30 días de la consulta en urgencias. Cuatro variables se asociaron de forma significativa con la mortalidad: la edad (odds ratio [OR] = 2,71; intervalo de confianza [IC] del 95%, 2,02-3,64), la no realización de angioplastia primaria (AP) (OR = 3; IC del 95%, 1,32-6,81)la clasificación de Killip avanzada (OR = 10,19; IC del 95%, 6,99 -14,85) y la localización anterior del infarto (OR = 1,39; IC del 95%, 1,03-1,86).Conclusiones: Encontramos diversos factores disponibles tras la valoración en urgencias, como la edad, que determinan un mal pronóstico a corto plazo del paciente anciano que consulta por un SCAEST. Ni la clase de Killip, ni la localización anterior del infarto agudo de miocardio ni la edad son susceptibles de modificación, no así la realización de una AP que, a diferencia del tratamiento fibrinolítico, es un factor independiente de no mortalidad a los 30 días (AU)

Objective: To identify factors associated with short-term mortality in patients of advanced age who come to the emergency department with acute coronary syndrome with ST segment elevation.Methods: Prospective longitudinal observational multicenter analytic study without interventions. Patients aged 70 yearsor older who were treated at 42 Spanish hospitals were included. Seventeen independent variables that might influence30-day mortality were analyzed. The information was extracted from the medical records or obtained during interviews with the patient or a family member; it was then recorded in a database developed for this study.Results: A total of 1137 patients were included; 340 (29.9%) died within 30 days of the emergency department visit.Four variables conferred significant risk of mortality. These were age (odds ratio [OR], 2.71; 95% confidence interval [CI],2.02-3.64); lack of primary angioplasty (OR, 3; 95% CI, 1.32-6.81); advanced Killip class (OR, 10.19; 95% CI, 6.99-14.85); and anterior location of the lesion (OR, 1.39; 95% CI, 1.03-1.86).Conclusions: We identified several factors, such as age, that are recorded during emergency department assessment andthat predict poor short-term outcome in the elderly patient treated for acute coronary syndrome with ST segment elevation. Although Killip class, location of the acute myocardial infarction, and age cannot be modified, we did identify a factor (performance of primary angioplasty) that, unlike fibrinolytic treatment, is independently associated with a better outcome in terms of 30-day mortality (AU)

A pilot study on the combination treatment of reduced-fluence photodynamic therapy, intravitreal ranibizumab, intravitreal dexamethasone and oral minocycline for neovascular age-related macular degeneration.

AIM: To assess the safety and efficacy of the combined treatment of reduced-fluence verteporfin photodynamic therapy (PDT), intravitreal ranibizumab, intravitreal dexamethasone and oral minocycline for choroidal neovascularisa- tion (CNV) secondary to age-related macular degeneration (AMD). METHODS: Nineteen patients with subfoveal CNV secondary to AMD were recruited into the trial. All study eyes (n = 19) received a single cycle of reduced-fluence (25 mJ/cm(2)) PDT with verteporfin followed by an intravitreal injection of ranibizumab 0.3 mg/0.05 ml and dexamethasone 200 µg at baseline. Oral minocycline 100 mg daily was started the following day and continued for 3 months. Patients were followed up monthly for 12 months. Repeat intravitreal ranibizumab was given if best-corrected visual acuity (BCVA) deteriorated by >5 letters on the Early Treatment Diabetic Retinopathy Study (ETDRS) chart or central retinal thickness (CRT) on ocular coherence tomography increased >100 µm. RESULTS: Eighteen patients completed the 12-month study. Stable vision (loss of ≤15 ETDRS letters) was maintained in 89% eyes (16/18). The mean change in BCVA was -5.0 ± 10.5 ETDRS letters. The mean number of ranibizumab injections was 3.4 (range 2-6). The mean reduction in the CRT was 66.3 µm (±75). CONCLUSION: This open-label clinical trial has demonstrated the safety in terms of adverse effects and maintenance of stable vision of combination treatment with verteporfin, ranibizumab, dexamethasone and minocycline in exudative AMD. However, the outcomes with reduced-fluence PDT combination therapy does not differ significantly with outcomes of clinical trials on combination treatment with standard dose PDT and intravitreal ranibizumab in neovascular AMD.

Anti-vascular endothelial growth factor agents for diabetic maculopathy.

The management of diabetic macular oedema is changing. The therapeutic armamentarium for diabetic macular oedema (DMO) includes a new group of drugs that inhibit vascular endothelial growth factor (VEGF). These anti-VEGF agents are already being used widely in DMO in clinical practice despite that several phase III trials on these drugs are still underway. There are no established protocols on the use of these agents in DMO, but short-term results are appealing. This review provides an update on the use of anti-VEGF agents in DMO. Although intravitreal delivery of anti-VEGF agents is a relatively safe procedure, the long-term local and systemic effects of these agents in the diabetic population remain unknown. In this regard, this review also highlights the need for close surveillance of the use of these drugs in this high-risk population.

Enzyme replacement therapy in mucopolysaccharidosis type II (Hunter syndrome): a preliminary report.

Mucopolysaccharidosis type II (MPS II; Hunter syndrome) is an X-linked disease caused by a deficiency of the enzyme iduronate-2-sulphatase (IDS), which results in the lysosomal accumulation of glycosaminoglycans (GAG). This paper describes a knockout mouse model of MPS II which has been used to assess the effect of enzyme replacement therapy. Therapy with IDS results in a marked decrease in urinary GAGs, as well as reduced GAG accumulation in several tissues. These studies have been used to support the first clinical trial of recombinant IDS in patients with Hunter syndrome.

Esophageal candidoma in a patient with acquired immunodeficiency syndrome.

Oral thrush and esophagitis caused by Candida are common in patients infected with the human immunodeficiency virus. We present the case of a 33-year-old man with acquired immunodeficiency syndrome who developed dysphagia during a hospitalization for pneumonia. Signs and symptoms were consistent with Candida esophagitis. Despite therapy with fluconazole, the patient's symptoms persisted. At upper endoscopy, a 1-cm, polypoid esophageal mass at 30 cm from the incisors and several other nodular lesions were observed; white plaques were noted throughout the esophagus. Biopsy specimens of the mass contained hyphal forms consistent with Candida species. Therapy with amphotericin B improved the patient's symptoms, and resolution of the mass was confirmed by repeat upper endoscopy. We believe this is the first case in the medical literature of a Candida mass (candidoma) causing dysphagia in a patient with acquired immunodeficiency syndrome. Candidoma should be considered in the differential diagnosis of dysphagia in patients with human immunodeficiency virus infection or immunosuppression due to other causes.

Nasopancreatic drainage: a novel approach for treating internal pancreatic fistulas and pseudocysts.

Internal pancreatic fistulas are rare but debilitating complications of chronic pancreatitis. Fistulous tracts from the pancreatic duct to the peritoneal or pleural cavities have been treated by medical therapy and surgical management, with success rates of 41% and 89%, respectively. Endoscopic stent placement for internal and external pancreatic fistulas has also been shown effective. We report on three patients with histories of chronic alcohol abuse and pancreatitis. Two patients presented with dyspnea and pleuritic chest pain. Imaging studies revealed pleural effusions, and endoscopic retrograde cholangiopancreatography (ERCP) demonstrated a patent fistulous tract from the pancreatic duct to the pleural cavity in each patient. Chemical analysis of the pleural fluid indicated pancreatic origin. The third patient, who had left-upper-quadrant abdominal pain and a small pleural effusion, had a large noncommunicating pseudocyst adjacent to the stomach. Nasopancreatic drains, along with chest tube drainage, were placed in the patients with pancreatic pleural fistulas. The patient with the pseudocyst received nasocystic drainage via the stomach. Drainage was measured until closure of the fistulas or cyst. Additionally, simply by injecting contrast medium, we were able to monitor the closure of fistulas without ERCP. The fistulas closed within 7 days, and the pseudocyst resolved within 14 days. Following discharge, all three patients were pain free, without evidence of recurrent fistulas or pseudocyst. In conclusion, the use of nasopancreatic/cyst drainage is an effective and convenient way to treat internal, communicating collections and pseudocysts of pancreatic origin. Furthermore, this method provides a simple means of assessing closure of fistulas and pseudocysts.

Heat stress induces in leaves an increase of the minimum level of chlorophyll fluorescence, Fo: A time-resolved analysis.

A time-resolved study of the effects of heat stress (23 to 50°C) on Fo level of chlorophyll fluorescence of leaves having different antenna content has been performed in order to elucidate the causes of heat induced increase of Fo in vivo. The multi-exponential deconvolution of the decays after a picosecond flash at Fo have shown that the best fit in both wild-type and the mutant chlorina F2 of barley leaves is obtained with three components in the temperature range utilized (100, 400 and 1200 ps at 23°C). In intermittent light greened pea leaves, a fourth long lifetime component (4 ns at 23°C) is needed. The comparison of the three types of leaves at 23°C shows that the content of the LHCII b complex does not affect the lifetimes of the two main components (100 and 400 ps) and affects their preexponential factors. This result suggests that in the PS II unit the exciton transfer from LHC IIb to the rest of the antenna is irreversible. The effects of heat stress on individual lifetime components, Ti, included several changes. Utilizing for PS II unit an extended 'Reversible Radical Pair' model, having three compartments, to interpret the variations of Ti and Ai induced by temperature increases, it can be inferred that heat determines: (i) an irreversible disconnection of a monor antenna complex which is not the LHC IIb complex, this effect is induced by temperatures higher than 40°C; (ii) a decrease of the quantum efficiency of Photosystem II photochemistry which is due to several effects: a decrease of the rate of charge separation, an increase of P(+)I(-) recombination rate constant and a decrease of the stabilization of charges. These effects on Photosystem II photochemistry start to occur above 30°C and are partially reversible.

Concentrations of morphologically normal, motile spermatozoa: Mg, Ca and Zn in the semen of infertile men.

Semen from infertile men (n = 23) has been compared with that of control subjects (n = 25). Whereas the concentrations of morphologically normal, motile sperms, Mg, Ca and Zn fell within the acceptable limits for all the control subjects, only two infertile men qualified by all five parameters. Of the patient group, seven were abnormal on all counts; sperm motility, Mg and Zn were low in 16, Ca in 19 and abnormal morphology was encountered in 12. Since there was no linear correlation between any two parameters, it is possible that each factor may singly or jointly influence the physiological integrity of the spermatozoa. The results are discussed from a consideration of pathological manifestations known to occur in deficiency of these trace elements à propos their role in determining the fertility index of the semen.