Antisense-mediated therapeutic pseudoexon skipping in TMEM165-CDG.

Deficiencies in glycosyltransferases, glycosidases or nucleotide-sugar transporters involved in protein glycosylation lead to congenital disorders of glycosylation (CDG), a group of genetic diseases mostly showing multisystem phenotype. Despite recent advances in the biochemical and molecular knowledge of these diseases, no effective therapy exists for most. Efforts are now being directed toward therapies based on identifying new targets, which would allow to treat specific patients in a personalized way. This work presents proof-of concept for the antisense RNA rescue of the Golgi-resident protein TMEM165, a gene involved in a new type of CDG with a characteristic skeletal phenotype. Using a functional in vitro splicing assay based on minigenes, it was found that the deep intronic change c.792+182G>A is responsible for the insertion of an aberrant exon, corresponding to an intronic sequence. Antisense morpholino oligonucleotide therapy targeted toward TMEM165 mRNA recovered normal protein levels in the Golgi apparatus of patient-derived fibroblasts. This work expands the application of antisense oligonucleotide-mediated pseudoexon skipping to the treatment of a Golgi-resident protein, and opens up a promising treatment option for this specific TMEM165-CDG.

Comparison of bacterial results from conventional cultures of the periprosthetic membrane and the synovial or pseudocapsule during hip revision arthroplasty.

INTRODUCTION: Bacterial identification is essential to diagnose and treat a revision for prosthetic loosening of an infected hip. The purpose of this study was to determine whether conventional cultures from the periprosthetic membrane are superior to synovial/pseudocapsule samples in the diagnosis of infection in hip revision arthroplasty. MATERIALS AND METHODS: We performed a prospective study including all hip revisions from October 2009 to October of 2011. Once the implants were removed and prior to the administration of the antibiotic prophylaxis, six periprosthetic samples from different sites were sent to the laboratory for culturing: two periprosthetic fluid samples, two solid material (synovial/pseudocapsule) samples and two swabs. Once the six samples were taken, antibiotic prophylaxis was administered and just as the implant was removed, two more solid samples of the periprosthetic membrane were obtained. RESULTS: Of a total of 86 hip revision surgeries, 22 were considered septic revisions postoperatively, of which 16 resulted in positive cultures. Of these 16 revisions, 14 obtained the same microbiologic diagnostic when considering either the synovial/pseudocapsule culture results or the solid membrane sample. CONCLUSIONS: We conclude that the membrane sample for a conventional culture is not superior to the synovial/pseudocapsule sample in detecting microorganisms.

[Perioperative management of non-deferrable oncologic surgeries during COVID-19 pandemic in Madrid, Spain. Is it safe?] / Manejo perioperatorio de cirugías oncológicas no diferibles durante la pandemia de covid-19 en Madrid, España ¿Es seguro?

INTRODUCTION: Surgical treatment during COVID-19 pandemic is controversial. Currently, most clinical guidelines advise to defer surgical patients during the COVID-19 pandemic, although the supporting data is sparse. We assumed that a COVID-19-free hospital, on the back of strong isolation measures and targeted screening, could reduce complications and enable us to continue treating high-risk patients. METHODS: Prospective study with retrospective analysis of 355 patients who had undergone nondeferrable oncological surgery between March 16th, 2020, and April 14th, 2020, at our institution. The aim of the study was to assess the hospital restructuring and surgical protocols to be able to safely handle non-deferrable surgeries during the first wave of the COVID-19 pandemic. We implemented structural changes and an updated surgical-anesthetic protocol in order to isolate COVID-19 patients from other surgical patients. Comprehensive targeted screening for COVID-19 patients was made. PCR tests were requested for suspected COVID-19 patients. We analyzed mortality and complications related to both surgery and COVID-19 during hospital admission and also 15 and 30 days after surgery. We compared it with a sample of similar patients in the pre-pandemic period. RESULTS: Of the 355 patients enrolled in our study, 21 were removed due to COVID-19 infection, leaving a total of 334 patients in our final analysis. Post-operative complications were found in 37 patients (11.07%). Two patients died after surgery (0.6%). At the end of the study, COVID-19-related adverse outcomes were detected in six patients (1.79%). When comparing the complications of our original sample with the complications that occurred in the pre-COVID era, we found no statistically significant differences. CONCLUSIONS: Our results show that the surgical treatment of oncologic patients during the COVID-19 pandemic is safe, as long as the hospital performs surgeries under strict isolation measures and a robust screening method. It is necessary to select COVID-19 free hospitals for this matter in this and future pandemics.

Perioperative management of non-deferrable oncologic surgeries during COVID-19 pandemic in Madrid, Spain. Is it safe?

INTRODUCTION: Surgical treatment during Covid-19 pandemic is controversial. Currently, most clinical guidelines advise to defer surgical patients during the Covid-19 pandemic, although the supporting data is sparse. We assumed that a Covid-19-free hospital, on the back of strong isolation measures and targeted screening, could reduce complications and enable us to continue treating high-risk patients. METHODS: Prospective study with retrospective analysis of 355 patients who had undergone nondeferrable oncological surgery between March 16th, 2020, and April 14th, 2020, at our institution. The aim of the study was to assess the hospital restructuring and surgical protocols to be able to safely handle non-deferrable surgeries during the first wave of the Covid-19 pandemic. We implemented structural changes and an updated surgical-anesthetic protocol in order to isolate Covid-19 patients from other surgical patients. Comprehensive targeted screening for Covid-19 patients was made. PCR tests were requested for suspected Covid-19 patients. We analyzed mortality and complications related to both surgery and Covid-19 during hospital admission and also 15 and 30 days after surgery. We compared it with a sample of similar patients in the pre-pandemic period. RESULTS: Of the 355 patients enrolled in our study, 21 were removed due to Covid-19 infection, leaving a total of 334 patients in our final analysis. Post-operative complications were found in 37 patients (11.07%). Two patients died after surgery (0.6%). At the end of the study, Covid-19-related adverse outcomes were detected in six patients (1.79%). When comparing the complications of our original sample with the complications that occurred in the pre-covid era, we found no statistically significant differences. CONCLUSIONS: Our results show that the surgical treatment of oncologic patients during the Covid-19 pandemic is safe, as long as the hospital performs surgeries under strict isolation measures and a robust screening method. It is necessary to select Covid-19 free hospitals for this matter in this and future pandemics.

Perioperative management of non-deferrable oncologic surgeries during COVID-19 pandemic in Madrid, Spain. Is it safe? / Manejo perioperatorio de cirugías oncológicas no diferibles durante la pandemia de covid-19 en Madrid, España ¿Es seguro?

INTRODUCTION: Surgical treatment during COVID-19 pandemic is controversial. Currently, most clinical guidelines advise to defer surgical patients during the COVID-19 pandemic, although the supporting data is sparse. We assumed that a COVID-19-free hospital, on the back of strong isolation measures and targeted screening, could reduce complications and enable us to continue treating high-risk patients. METHODS: Prospective study with retrospective analysis of 355 patients who had undergone nondeferrable oncological surgery between March 16th, 2020, and April 14th, 2020, at our institution. The aim of the study was to assess the hospital restructuring and surgical protocols to be able to safely handle non-deferrable surgeries during the first wave of the COVID-19 pandemic. We implemented structural changes and an updated surgical-anesthetic protocol in order to isolate COVID-19 patients from other surgical patients. Comprehensive targeted screening for COVID-19 patients was made. PCR tests were requested for suspected COVID-19 patients. We analyzed mortality and complications related to both surgery and COVID-19 during hospital admission and also 15 and 30 days after surgery. We compared it with a sample of similar patients in the pre-pandemic period. RESULTS: Of the 355 patients enrolled in our study, 21 were removed due to COVID-19 infection, leaving a total of 334 patients in our final analysis. Post-operative complications were found in 37 patients (11.07%). Two patients died after surgery (0.6%). At the end of the study, COVID-19-related adverse outcomes were detected in six patients (1.79%). When comparing the complications of our original sample with the complications that occurred in the pre-COVID era, we found no statistically significant differences. CONCLUSIONS: Our results show that the surgical treatment of oncologic patients during the COVID-19 pandemic is safe, as long as the hospital performs surgeries under strict isolation measures and a robust screening method. It is necessary to select COVID-19 free hospitals for this matter in this and future pandemics.

Griffiths phase and spontaneous exchange bias in La1.5Sr0.5CoMn0.5Fe0.5O6.

La1.5Sr0.5CoMn0.5Fe0.5O6 (LSCMFO) compound was prepared by solid state reaction and its structural, electronic and magnetic properties were investigated. The material forms in rhombohedral [Formula: see text] structure, and the presence of distinct magnetic interactions leads to the formation of a Griffiths phase above its FM transition temperature (150 K), possibly related to the nucleation of small short-ranged ferromagnetic clusters. At low temperatures, a spin glass-like phase emerges and the system exhibits both the conventional and the spontaneous exchange bias (EB) effects. These results resemble those reported for La1.5Sr0.5CoMnO6 but are discrepant to those found when Fe partially substitutes Co in La1.5Sr0.5(Co1-x Fe x )MnO6, for which the EB effect is observed in a much broader temperature range. The unidirectional anisotropy observed for LSCMFO is discussed and compared with those of resembling double-perovskite compounds, being plausibly explained in terms of its structural and electronic properties.

Mansonella ozzardi infections in Indians of the Southwestern part of the state of Bolivar, Venezuela.

Mansonella ozzardi infections were found in 80/139 (58%) of Indians living in the forest of the upper Caura River Basin located in the southwestern part of Bolivar State, Venezuela. In four (3%) of the blood samples a second type of microfilaria which could not be identified was also found.

Lagochilascaris minor infection in Venezuela. Report of a case.

A Lagochilascaris minor infection in a 6-year-old Venezuelan girl with a 4-year history of nasal obstruction is reported. Treatment with levamisole resulted in clinical and radiological cure. Eggs of L. minor were observed in the patient's feces; these were cultured to second stage larvae.

Thoracoscopic resection of ectopic parathyroid glands.

BACKGROUND: The vast majority of parathyroid glands in hyperparathyroidism can be resected through a cervical approach. In approximately 2% of the cases, the ectopic gland is in the mediastinum in a location that requires a thoracic approach. METHODS: We report 7 such cases that were resected using video-assisted thoracic surgery to avoid the need for an open surgical procedure. RESULTS: All glands were successfully identified preoperatively and subsequently resected. Hospital stay averaged less than 3 days with only one minor complication. CONCLUSIONS: Ectopic mediastinal parathyroid glands may be safely and accurately resected using video-assisted thoracic surgery to avoid open approaches.

Effects of acrylamide and 2,5-hexanedione on brain mitochondrial respiration.

The effects of acrylamide (ACR) and 2,5-hexanedione (2,5-HD) on brain mitochondrial respiration were assessed. Mitochondria were isolated from whole brains or brain regions of control and neurotoxicant-treated rats. Direct in vitro exposure of isolated brain mitochondria to ACR (1 mM final concentration) had no effect on respiration, whereas direct exposure to 2,5-HD (1 mM final concentration) inhibited state 3 respiration. Chronic treatment of rats with ACR (50 mg/kg/day x 10 days) did not affect respiration of mitochondria isolated from cortex or brainstem. However, in mitochondria from cerebellum of ACR treated rats, pyruvate + oxaloacetic acid (pyr/oaa) supported oxygen consumption was decreased significantly in both states 3 and 4. In addition, the ADP/O ratio was reduced in this brain structure. In all brain regions of 2,5-HD (400 mg/kg/day x 24 days) intoxicated rats, pyr/oaa supported state 3 respiration was reduced. Glutamate + malate (glu/mal) supported respiration was diminished only in mitochondria isolated from brain stem of 2,5-HD treated rats. In contrast, the non-neurotoxic analogs, 1,6-hexanediol and N,N'-methylene-bis-acrylamide did not alter mitochondrial respiration in parallel experiments. Thus, both ACR and 2,5-HD produce a substrate-dependent, toxicologically specific inhibition of brain mitochondrial respiration. This inhibition of mitochondrial energy production might play a role in the neurotoxic mechanisms of action for these chemicals.

Lead-induced alterations in retinal cGMP phosphodiesterase trigger calcium overload, mitochondrial dysfunction and rod photoreceptor apoptosis.

Lead exposure results in the selective apoptotic loss of rods and bipolar cells. During and following developmental lead exposure rod/retinal cGMP phosphodiesterase expression and activity are delayed in onset and decreased, [Ca2+] is elevated, and mitochondrial ATP synthesis is decreased. In vitro studies, using retinas incubated in Ca2+ and/or Pb2+, demonstrate that rods selectively die by apoptosis, retinal mitochondrial ATP synthesis is decreased, mitochondrial cytochrome c is released and caspase activity is increased. These results suggest that lead-induced rod and bipolar cell apoptosis is triggered by Ca2+ and Pb2+ overload due to altered cGMP phosphodiesterase activity and that mitochondrial alterations play a central role in this process.

Pharmacological strategies to block rod photoreceptor apoptosis caused by calcium overload: a mechanistic target-site approach to neuroprotection.

PURPOSE: Photoreceptor apoptosis and resultant visual deficits occur in humans and animals with inherited, and disease-, injury- and chemical-induced retinal degeneration. Our aims were three-fold: 1) to determine the kinetics of rod apoptosis and Ca2+ overload in Pde6b9rd1) mice and developmentally lead-exposed rats, 2) to establish a pathophysiologically-relevant model of Ca2+ overload/rod-selective apoptosis in isolated rat retina and 3) to examine different mechanistic based neuroprotective strategies that would abrogate or mollify rod Ca2+ overload/apoptosis. METHODS: Retinal morphometry and elemental calcium content ([Ca]) determined the kinetics of rod apoptosis and Ca2+ overload. A multiparametric analysis of apoptosis including rod [Ca], a live/dead assay, rod oxygen consumption, cytochrome c immunoblots and caspase assays was combined with pharmacological studies of an isolated rat retinal model of rod-selective Ca2+ overload/apoptosis. RESULTS: Ca2+ overload preceded rod apoptosis in mice and rats, although the extent and kinetics in each differed significantly. The isolated rat model of rod Ca2+ overload/apoptosis showed that blockade of Ca2+ entry through rod cGMP-activated channels with L-cis diltiazem was partially neuroprotective, whereas blockade of Ca2+ entry into rods through L-type Ca2+ channels with D-cis diltiazem or verapamil provided no protection. Inhibition of the mitochondrial Na+/Ca2+ exchanger with D-cis diltiazem provided no protection. CsA and NIM811, mitochondrial permeability transition pore (mPTP) inhibitors, blocked all Ca(2+)-induced apoptosis, whereas the caspase-3 inhibitor DEVD-fmk only blocked the downstream cytochrome c-induced apoptosis. CONCLUSIONS: The successful pharmacological neuroprotective strategies for rod Ca2+ overload/apoptosis targeted the rod cGMP-activated channels or mPTP, but not the rod L-type Ca2+ channels.

[Natural infection of Speothos venaticus (Carnivora: Canidae) by adult stages of Lagochilascaris sp]. / Infección natural de Speothos venaticus (Carnivora: Canidae) por estadios adultos de Lagochilascaris sp.

An adult male Speothos venaticus Lund (bush dog) was found killed on a rural road in the Northeast of Bolívar State, in a locality where a patient with lagochilascariasis has lived for the past 16 years. The animal was frozen for 1 month, and after removal of the head and skin, was autopsied. Two adult gravid females of Lagochilascaris sp. were found in the trachea. Certain morphological characteristics of the soft parts differed from the description given for Lagochilascaris minor Leiper, 1909; however, stable characters, such as the form of the interlabials, the location of the vulva, and particularly the size and number of depressions of the egg shell appear to indicate that the worms are of the above mentioned species. Unfortunately, no males were found for study of the spicules and ejaculatory duct. In histological sections of the larynx and the trachea from the deep tissues, parts of degenerated worms were found, with characteristics attributable to Lagochilascaris sp.

[Infection induced in wild rodents Dasyprocta leporina (Rodentia: Dasyproctidae), with larval eggs of Lagochilascaris minor (Nematoda: Ascarididae)]. / Infección inducida en el roedor selvático Dasyprocta leporina (Rodentia: Dasyproctidae), con huevos larvados de Lagochilascaris minor (Nematoda: Ascarididae).

Specimens of Dasyprocta leporina (Linnaeus, 1758) were raised out their natural environment. At three to four months of age they were orally inoculated with Lagochilascaris minor (Leiper, 1909) eggs obtained from a native patient. The eggs were incubated for more than 80 days so that it was possible to obtain, by mechanical compression, larvae that could be maintained alive in liquid medium for 48 hours or more. The animals were sacrificed 14-46 days after infection and tangled larva in inflammatory nodules were found in skeletal muscle without foreign body reaction, abscess formation or calcification. The development of the nodules did not seem to affect the hosts. The larvae obtained were similar to those described by SPRENT as the third stage of these helminths. When white mice were inoculated with similar material it was not possible to recuperate larva from their tissues nor were nodules found. Based on these results it is postulated that the helminth does not present a pulmonary cycle and that its development requires an intermediary host.

Evaluation in white mice of the infectivity of eggs of Lagochilascaris minor (Nematoda: Ascarididae), incubated by decortication with sodium hypochlorite (NaOCl).

White mice were used to study the infectivity of the eggs of Lagochilascaris minor Leiper, 1909 after incubation in liquid media, with or without preservative substances. Potassium bichromate (K2Cr2O7) at 1% restrict hatching, while 1% formalin gave a greater larval yield. Incubation of eggs in distilled water, in Roux or Falcon flasks gave a good yield, whether the eggs were obtained from human feces or from experimentally infected cats. Treatment of eggs with Sodium hypochlorite (NaOCl) at 5.25% for 2 min prior to inoculation, produced a notable increment of the larval yield in the infections.

[Mansonelliasis in the southeast Venezuelan Orinoquia region]. / Mansonelosis en el area Sur-oriental de la Orinoquia Venezolana.

Physical exam and skin biopsy were performed and peripheral blood samples drawn from 10% of the inhabitants of 26 jungle villages of the State of Bolívar, Venezuela. One hundred and fifty three Indians and an inhabitant of mixed blood from 13 different communities were found to be infected with mansonelliasis representing a global index of 36.40% in the endemic area. The parasitosis was found to be concentrated in three areas. The largest, located in the southwest of the country and forming part of an extent infected area embarking neighboring parts of Venezuela and Brazil, contains villages with a parasite index of 80.76% and 94.44%. A second focus is located in the southeast, its carriers, as those of the first, are infected with Mansonella ozzardi. In a third, central eastern focus infections with Mansonella pertans were found. The patients infected with M. ozzardi were apparently asymptomatic and their physical exam was normal, even though individual parasite densities up to 30,000 microfilariae/ml blood and 22,000/gr skin were found.

Value of tonometry in postoperative high risk patients with digestive surgery.

Intramucosal pH (pHi) in splanchnic organs is a reliable index of local tissular perfusion, and can be measured by tonometry. At the Surgical Intensive Care Unit we used tonometry to determine tissular perfusion in patients who underwent major digestive surgery. We report a prospective study of 20 patients with elective and emergency surgery. All of them underwent gastric tonometry and 10 of them, who had colonic disease, also underwent sigmoid tonometry. The values below pHi = 7.30 were associated with increased morbidity and mortality.

[Hemodynamic changes during laparoscopic surgery. Preliminary study]. / Alteraciones hemodinámicas durante la cirugía laparoscópica. Estudio preliminar.

The haemodynamic changes produced by laparoscopic surgery in children have been evaluated. A transesophageal echocardiographic study on 13 patients (7 males and 6 females, 10.8 +/- 2.7 years old) has been performed before, during and after peritoneal CO2 gas insufflation. A change on the Doppler waves pattern of the infradiaphragmatic veins along with an increase in blood flow velocity were observed. Pulmonary veins suffered minimal changes showing an increase on either the systolic pressure and on atrial contraction. Diastolic retrograde flow in aorta reflected an increase in peripheral vascular resistance. Cardiac output increased minimally. All haemodynamic changes returned to basal after gas peritoneal desufflation. These data show that laparoscopic surgery could produce important cardiac derangements in children with ventricular dysfunction or in patients presenting left ventricular outlet obstruction. In these patients laparoscopic surgery might be precluded.