Junta de Andalucía

When expression of more than one gene is required in cells, bicistronic or multicistronic expression vectors have been used. Among various strategies employed to construct bicistronic or multicistronic vectors, an internal ribosomal entry site (IRES) has been widely used. Due to the large size and difference in expression levels between genes before and after IRES, however, a new strategy was required to replace IRES. A self-cleaving 2A peptide could be a good candidate to replace IRES because of its small size and high cleavage efficiency between genes upstream and downstream of the 2A peptide. Despite the advantages of the 2A peptides, its use is not widespread because (i) there are no publicly available cloning vectors harboring a 2A peptide gene and (ii) comprehensive comparison of cleavage efficiency among various 2A peptides reported to date has not been performed in different contexts. Here, we generated four expression plasmids each harboring different 2A peptides derived from the foot-and-mouth disease virus, equine rhinitis A virus, Thosea asigna virus and porcine teschovirus-1, respectively, and evaluated their cleavage efficiency in three commonly used human cell lines, zebrafish embryos and adult mice. Western blotting and confocal microscopic analyses revealed that among the four 2As, the one derived from porcine teschovirus-1 (P2A) has the highest cleavage efficiency in all the contexts examined. We anticipate that the 2A-harboring cloning vectors we generated and the highest efficiency of the P2A peptide we demonstrated would help biomedical researchers easily adopt the 2A technology when bicistronic or multicistronic expression is required.

Abstract The genetic make-up of an individual contributes to the susceptibility and response to viral infection. Although environmental, clinical and social factors have a role in the chance of exposure to SARS-CoV-2 and the severity of COVID-19 1,2 , host genetics may also be important. Identifying host-specific genetic factors may reveal biological mechanisms of therapeutic relevance and clarify causal relationships of modifiable environmental risk factors for SARS-CoV-2 infection and outcomes. We formed a global network of researchers to investigate the role of human genetics in SARS-CoV-2 infection and COVID-19 severity. Here we describe the results of three genome-wide association meta-analyses that consist of up to 49,562 patients with COVID-19 from 46 studies across 19 countries. We report 13 genome-wide significant loci that are associated with SARS-CoV-2 infection or severe manifestations of COVID-19. Several of these loci correspond to previously documented associations to lung or autoimmune and inflammatory diseases 3–7 . They also represent potentially actionable mechanisms in response to infection. Mendelian randomization analyses support a causal role for smoking and body-mass index for severe COVID-19 although not for type II diabetes. The identification of novel host genetic factors associated with COVID-19 was made possible by the community of human genetics researchers coming together to prioritize the sharing of data, results, resources and analytical frameworks. This working model of international collaboration underscores what is possible for future genetic discoveries in emerging pandemics, or indeed for any complex human disease.

In the cancer population, the term breakthrough pain typically refers to a transitory flare of pain in the setting of chronic pain managed with opioid drugs. The prevalence and characteristics of this phenomenon have not been defined, and its impact on patient care is unknown. We developed operational definitions for breakthrough pain and its major characteristics, and applied these in a prospective survey of patients with cancer pain. Data were collected during a 3 month period from consecutive patients who reported moderate pain or less for more than 12 h daily and stable opioid dosing for a minimum of 2 consecutive days. Of 63 patients surveyed, 41 (64%) reported breakthrough pain, transient flares of severe or excruciating pain. Fifty-one different pains were described (median 4 pains/day; range 1-3600). Pain characteristics were extremely varied. Twenty-two (43%) pains were paroxysmal in onset; the remainder were more gradual. The duration varied from seconds to hours (median/range: 30 min/1-240 min), and 21 (41%) were both paroxysmal and brief (lancinating pain). Fifteen (29%) of the pains were related to the fixed opioid dose, occurring solely at the end of the dosing interval. Twenty-eight (55%) of the pains were precipitated; of these, 22 were caused by an action of the patient (incident pain), and 6 were associated with a non-volitional precipitant, such as flatulence. The pathophysiology of the pain was believed to be somatic in 17 (33%), visceral in 10 (20%), neuropathic in 14 (27%), and mixed in 10 (20%). Pain was related to the tumor in 42 (82%), the effects of therapy in 7 (14%), and neither in 2 (4%). Diverse interventions were employed to manage these pains, with variable efficacy. These data clarify the spectrum of breakthrough pains and indicate their importance in cancer pain management.

BACKGROUND AND PURPOSE: We aimed to establish the prevalence, characteristics, and outcomes of intracranial atherosclerosis (ICAS) in China by a large, prospective, multicenter study. METHODS: We evaluated 2864 consecutive patients who experienced an acute cerebral ischemia<7 days after symptom onset in 22 Chinese hospitals. All patients underwent magnetic resonance angiography, with measurement of diameter of the main intracranial arteries. ICAS was defined as ≥50% diameter reduction on magnetic resonance angiography. RESULTS: The prevalence of ICAS was 46.6% (1335 patients, including 261 patients with coexisting extracranial carotid stenosis). Patients with ICAS had more severe stroke at admission and stayed longer in hospitals compared with those without intracranial stenosis (median National Institutes of Health Stroke Scale score, 3 versus 5; median length of stay, 14 versus 16 days; both P<0.0001). After 12 months, recurrent stroke occurred in 3.27% of patients with no stenosis, in 3.82% for those with 50% to 69% stenosis, in 5.16% for those with 70% to 99% stenosis, and in 7.27% for those with total occlusion. Cox proportional hazards regression analyses showed that the degree of arterial stenosis, age, family history of stroke, history of cerebral ischemia or heart disease, complete circle of Willis, and National Institutes of Health Stroke Scale score at admission were independent predictors for recurrent stroke at 1 year. The highest rate of recurrence was observed in patients with occlusion with the presence of ≥3 additional risk factors. CONCLUSIONS: ICAS is the most common vascular lesion in patients with cerebrovascular disease in China. Recurrent stroke rate in our study was lower compared with those of previous clinical trials but remains unacceptably high in a subgroup of patients with severe stenosis.

STUDY DESIGN: Retrospective study. OBJECTIVES: To report a technique of vertebral column resection through a single posterior approach and its preliminary results in the treatment of moderate to severe spinal deformities with limited flexibility. SUMMARY OF BACKGROUND DATA: Vertebral column resection is a formidable operation reserved for moderate to severe deformities with limited flexibility. The authors devised a technique of vertebral column resection through a single posterior approach that offers significant advantages over the anterior-posterior vertebral column resection. METHODS: Seventy spinal deformity patients treated by posterior vertebral column resection were reviewed. Minimum follow-up was 2 years (range 2-3.3 years). There were 34 males and 36 females with a mean age of 27.4 years at the time of the operation. Etiologic diagnoses were adult scoliosis in 7, congenital kyphoscoliosis in 38, and postinfectious kyphosis in 25. The surgery consisted of temporary stabilization of the vertebral column with segmental pedicle screw fixation, resection of the vertebral column at the apex of the deformity via the posterior route, followed by gradual deformity correction and global fusion. RESULTS: The total number of resected vertebrae was 143: 76 in thoracic and 67 in lumbar. Mean operation time was 4 hours, 31 minutes with average blood loss of 2333 mL. The deformity correction was 61.9% in the coronal plane and 45.2 degrees in the sagittal plane. Complications were encountered in 24 patients: 2 complete cord injuries in severe adult scoliosis and thoracic kyphosis patient who had significant preoperative cord compromise, 6 hematomas, 4 root injuries (all incomplete), 5 fixation failures, 2 infections, and 5 hemopneumothoraxes. CONCLUSIONS: Posterior vertebral column resection is an effective alternative for moderate to severe deformities with limited flexibility. However, it is a technically demanding and exhausting procedure with possible risks for major complications.

Approximately 30% of postmenopausal white women in the United States have osteoporosis, and 16% have osteoporosis of the lumbar spine in particular. Bone density of the spine is positively associated with greater height and weight, older age at menopause, a history of arthritis, more physical activity, moderate use of alcoholic beverages, diuretic treatment, and current estrogen replacement therapy, whereas later age at menarche and a maternal history of fracture are associated with lower levels of density. Low bone density leads to an increased risk of osteoporotic fractures. Fracture risk also increase with age. Vertebral fractures affect approximately 25% of postmenopausal women, although the exact figure depends on the definition used. Recent data show that vertebral fracture rates are as great in men as in women but, because women live longer, the lifetime risk of a vertebral fracture from age 50 onward is 16% in white women and only 5% in white men. Fracture rates are less in most nonwhite populations, but vertebral fractures are as common in Asian women as in those of European heritage. Other risk factors for vertebral fractures are less clear but include hypogonadism and secondary osteoporosis; obesity is protective of fractures as it is of bone loss. Compared with hip fractures, vertebral fractures are less disabling and less expensive, costing approximately $746 million in the United States in 1995. However, they have a substantial negative impact on the patient's function and quality of life. The adverse effects of osteoporotic fractures are likely to increase in the future with the growing number of elderly people.

Wine is one of the most consumed beverages around the world. It is composed of alcohols, sugars, acids, minerals, proteins and other compounds, such as organic acids and volatile and phenolic compounds (also called polyphenols). Polyphenols have been shown to be highly related to both (i) wine quality (color, flavor, and taste) and (ii) health-promoting properties (antioxidant and cardioprotective among others). Polyphenols can be grouped into two big families: (i) Flavonoids, including anthocyanidins, flavonols, flavanols, hydrolysable and condensed tannins, flavanones, flavones and chalcones; and (ii) Non-flavonoids, including hydroxycinnamic acids, hydroxybenzoic acids, stilbenes, tyrosol and hydroxytyrosol. Each group affects in some way the different properties of wine to a greater or a lesser extent. For that reason, the phenolic composition can be managed to obtain singular wines with specific, desirable characteristics. The current review presents a summary of the ways in which the phenolic composition of wine can be modulated, including (a) invariable factors such as variety, field management or climatic conditions; (b) pre-fermentative strategies such as maceration, thermovinification and pulsed electric field; (c) fermentative strategies such as the use of different yeasts and bacteria; and (d) post-fermentative strategies such as maceration, fining agents and aging. Finally, the different extraction methods and analytical techniques used for polyphenol detection and quantification have been also reviewed.

AIMS AND OBJECTIVES: To evaluate at what age parous and nonparous women were diagnosed with breast cancer. Factors taken into account for parous women were whether they had breastfed their children, and if so, the length of the lactation period. Other factors considered for both groups were obesity, family histories of cancer, smoking habits and alcohol consumption. BACKGROUND: Breast cancer is the most common form of cancer in younger women in Western countries. Its growing incidence as well as the increasingly early age of diagnosis led us to carefully analyse its possible causes and the preventive measures to be taken. This is a particularly important goal in epidemiological research. DESIGN: A retrospective study of the clinical histories of patients diagnosed with breast cancer at the San Cecilio University Hospital in Granada (Spain). METHODS: In this study, we analysed 504 medical records of female patients, 19-91 years of age, who had been diagnosed and treated for breast cancer from 2004-2009 at the San Cecilio University Hospital in Granada (Spain). Relevant data (age of diagnosis, period of lactation, family history of cancer, obesity, alcohol consumption and smoking habits) were collected from the clinical histories of each patient and analysed. A conditional inference tree was used to relate the age of diagnosis to smoking habits and the length of the lactation period. RESULTS: The conditional inference tree identified significant differences between the age of the patients at breast cancer diagnosis, smoking habits (p < 0·001) and lactation period if the subjects had breastfed their children for more than six months (p = 0·006), regardless of whether they had a family history of cancer. CONCLUSIONS: Our study concluded that breastfeeding for over six months not only provides children with numerous health benefits, but also protects mothers from breast cancer when the mothers are nonsmokers. RELEVANCE TO CLINICAL PRACTICE: Nurses play a crucial role in encouraging new mothers to breastfeed their children, and this helps to prevent breast cancer.

We investigated short-term effects (up to 24 h) of air exposure and confinement, and long-term effects (up to 11 days) of confinement, to elucidate signalling pathways in the stress response of gilthead sea bream Sparus aurata L. Plasma glucose and lactate were taken as indicators of sympathetic activation, and alpha-melanocyte stimulating hormone (alpha-MSH), adrenocorticotrophic hormone (ACTH) and cortisol as indicators of activation of the brain-pituitary-interrenal (BPI) axis. Air exposure for 3 min resulted, within 30 min, in an increase in plasma concentrations of cortisol, alpha-MSH, glucose, lactate, osmolality and plasma Na, Cl and Mg. Plasma ACTH and beta-endorphin and plasma K, Ca and P did not change. We conclude that air exposure mainly activates the brain-sympathetic-chromaffin cell (BSC) axis. In fish confined at a density of 70 kg/m(3) (compared with 4 kg/m(3) in controls), cortisol, ACTH and alpha-MSH increased within 1 h, indicating activation of the BPI axis. Plasma glucose, Na, Cl and Mg increased with an 8 h delay compared with the response to air exposure. No changes in plasma lactate, osmolality, K, Ca and P were observed. Long-term confinement induced a biphasic cortisol response with peaks at 1 h and at 2 and 3 days. A gradual increase in plasma beta-endorphin concentrations peaked at 7 days; the concentration of alpha-MSH increased rapidly within 1 h and then declined to control values 4 days after the onset of confinement. No changes in ACTH were detected. Our data provide evidence that a stressor-specific activation of the BSC and BPI axes may occur in Sparus aurata.

Vertebrates have greatly elaborated the basic chordate body plan and evolved highly distinctive genomes that have been sculpted by two whole-genome duplications. Here we sequence the genome of the Mediterranean amphioxus (Branchiostoma lanceolatum) and characterize DNA methylation, chromatin accessibility, histone modifications and transcriptomes across multiple developmental stages and adult tissues to investigate the evolution of the regulation of the chordate genome. Comparisons with vertebrates identify an intermediate stage in the evolution of differentially methylated enhancers, and a high conservation of gene expression and its cis-regulatory logic between amphioxus and vertebrates that occurs maximally at an earlier mid-embryonic phylotypic period. We analyse regulatory evolution after whole-genome duplications, and find that-in vertebrates-over 80% of broadly expressed gene families with multiple paralogues derived from whole-genome duplications have members that restricted their ancestral expression, and underwent specialization rather than subfunctionalization. Counter-intuitively, paralogues that restricted their expression increased the complexity of their regulatory landscapes. These data pave the way for a better understanding of the regulatory principles that underlie key vertebrate innovations.

homeostasis and possibly contributing to the sensing of sodicity stress.

Iron oxide nanoparticles (IONPs) have emerged as a promising alternative to conventional contrast agents (CAs) for magnetic resonance imaging (MRI). They have been extensively investigated as CAs due to their high biocompatibility and excellent magnetic properties. Furthermore, the ease of functionalization of their surfaces with different types of ligands (antibodies, peptides, sugars, etc.) opens up the possibility of carrying out molecular MRI. Thus, IONPs functionalized with epithelial growth factor receptor antibodies, short peptides, like RGD, or aptamers, among others, have been proposed for the diagnosis of various types of cancer, including breast, stomach, colon, kidney, liver or brain cancer. In addition to cancer diagnosis, different types of IONPs have been developed for other applications, such as the detection of brain inflammation or the early diagnosis of thrombosis. This review addresses key aspects in the development of IONPs for MRI applications, namely, synthesis of the inorganic core, functionalization processes to make IONPs biocompatible and also to target them to specific tissues or cells, and finally in vivo studies in animal models, with special emphasis on tumor models.

Study Design. A retrospective study of 16 patients who underwent the modified Robinson anterior cervical discectomy and fusion at three operative levels. Objectives. To provide long-term follow-up data on the surgical success and patient outcome of three-level anterior cervical discectomies and fusions. Summary of Background Data. The success of arthrodesis for anterior cervical fusion depends on several factors, including the number of surgical levels. To the authors' knowledge, there are no long-term follow-up reports to describe the arthrodesis rate and outcome for patients having specifically three-level discectomy and fusion procedures. Methods. Sixteen patients, with an average age of 59 years, were followed for an average of 37 months. All had an anterior discectomy, burring of the endplates, and placement of an autogenous tricortical iliac crest graft at three levels. All patients had follow-up office visits with examinations and radiographs. Radiographic union, postoperative pain relief, and neurologic recovery were evaluated. Results. Only 9 (56%) of the 16 patients went on to achieve solid arthrodesis at all three levels. Of the seven patients with pseudarthrosis, two had severe pain and required revision; two had moderate pain and three no pain. Of the nine with the solid fusion, three had mild pain and six no pain, a statistically significant difference in comparing the two outcomes (P < 0.01). All patients with preoperative motor deficit recovered, but two patients in whom a pseudarthrosis had developed had limited improvement in function until the nonunion was surgically repaired. Conclusions. A three-level modified Robinson cervical discectomy and fusion results in an unacceptably high rate of pseudarthrosis. Although not all pseudar-throses are painful, these data suggest that those with a successful fusion have a better outcome. It is recommended that these patients undergo additional or alternative measures to achieve arthrodesis consistently.

STUDY DESIGN: A cross-sectional study design was used. OBJECTIVE: To determine the sensitivity and specificity of the Spurling test for cervical radiculopathy. SUMMARY OF BACKGROUND DATA: The Spurling test is an accepted physical examination test, but there is little data on its sensitivity or specificity. METHODS: From 1988 to 1993, 255 consecutive patients were referred for electrodiagnosis of upper extremity nerve disorders. A Spurling test administered before other testing was performed. The Spurling test was scored as positive if it caused pain or tingling that started in the shoulder and radiated distally to the elbow. After the electrodiagnostic examination, a score was given to each diagnosis in the differential diagnosis according to the likelihood of its presence. To determine the odds ratio, sensitivity, and specificity, chi2 analysis was used. Also, the percentage of subjects with positive results from the Spurling test was calculated for several nerve disease diagnoses. RESULTS: The Spurling test had a sensitivity of 6/20 (30%) and a specificity of 160/172 (93%). The results were positive in 16.6% of the normal group, in 3.4% of the group with nerve disorders other than a radiculopathy, in 25% of the group with an abnormality not consistent with any specific diagnosis group, in 37.5% of the group with possible radiculopathy, and in 40% of the group with certain radiculopathy. CONCLUSIONS: The Spurling test is not very sensitive, but it is specific for cervical radiculopathy diagnosed by electromyography. Therefore, it is not useful as a screening test, but it is clinically useful in helping to confirm a cervical radiculopathy.

BACKGROUND: The ideal technique for the use of AlloDerm (LifeCell Corp) in complicated ventral hernia repair has not been defined. The expense of these products mandates careful evaluation to justify their widespread use. We compared two techniques of fascial bridging versus fascial reinforcement repair with regard to their longterm recurrence rates using AlloDerm. STUDY DESIGN: We retrospectively studied patients with abdominal defects repaired with AlloDerm at our institution. RESULTS: Thirty-seven patients with abdominal wall repairs using AlloDerm were identified between January 2004 and December 2005. Eleven patients underwent bridged fascial repair; 26 patients had reinforced fascial repair. There was no statistical significance between the 2 groups in terms of average age (57 versus 52 years), body mass index (35 versus 29 kg/m(2)), American Society of Anesthesiologists score (2.9 versus 2.5), or number of earlier abdominal operations (3.4 versus 3.5). The average sizes of AlloDerm used were 175 cm(2) for bridged and 89 cm(2) for reinforced repair (p=0.005). In patients with reinforced closure, primary repair was achieved with lateral component separation in 22 of 26 patients. Mean followup was 21.4 months (range 15 to 36 months). In the bridged group, 1 patient died on postoperative day 20. Of the remaining 10 patients, 8 patients (80%) developed recurrences. Seven patients required reoperation, but one patient refused repair. In the reinforced group, four patients were lost to followup and two patients died. Four of the remaining 20 patients (20%) developed recurrences that required repair; this was significantly different from the recurrence rate in the bridged group (p=0.009). CONCLUSIONS: This study demonstrated that the method in which AlloDerm is used in abdominal wall reconstruction has a significant impact on recurrence rates. Based on our findings, AlloDerm should be used only as a reinforcement after primary fascial reappoximation.

The influence of nonlinear (fourth-order) terms in the Lagrangians of some classical relativistic field theories is investigated, independently of the mass and coupling-constant parameters. In the spinor case, stable localized configurations with a lowest energy state are shown to exist always for positive values of the coupling constant.

Long interspersed element-1 (L1) retrotransposons compose ∼20% of the mammalian genome, and ongoing L1 retrotransposition events can impact genetic diversity by various mechanisms. Previous studies have demonstrated that endogenous L1 retrotransposition can occur in the germ line and during early embryonic development. In addition, recent data indicate that engineered human L1s can undergo somatic retrotransposition in human neural progenitor cells and that an increase in human-specific L1 DNA content can be detected in the brains of normal controls, as well as in Rett syndrome patients. Here, we demonstrate an increase in the retrotransposition efficiency of engineered human L1s in cells that lack or contain severely reduced levels of ataxia telangiectasia mutated, a serine/threonine kinase involved in DNA damage signaling and neurodegenerative disease. We demonstrate that the increase in L1 retrotransposition in ataxia telangiectasia mutated-deficient cells most likely occurs by conventional target-site primed reverse transcription and generate either longer, or perhaps more, L1 retrotransposition events per cell. Finally, we provide evidence suggesting an increase in human-specific L1 DNA copy number in postmortem brain tissue derived from ataxia telangiectasia patients compared with healthy controls. Together, these data suggest that cellular proteins involved in the DNA damage response may modulate L1 retrotransposition.

STUDY DESIGN: A prospective randomized study investigated the radiographic progress of fusion at 6, 12, and 24 months in 42 patients who underwent a single-level anterior lumbar interbody fusion using cylindrical interbody fusion cages. OBJECTIVE: To determine the patterns and rates of osteoinduction associated with the use of recombinant human bone morphogenetic protein type 2 (rhBMP-2) and an absorbable collagen sponge carrier in anterior lumbar interbody fusion with a tapered cylindrical fusion device. SUMMARY OF BACKGROUND DATA: Studies have shown that rhBMP-2 used with allograft dowels increases the rate of interbody fusion by promoting osteoinduction and enhancing incorporation of the allograft. In a small series of human patients undergoing anterior lumbar interbody fusion with a tapered cylindrical fusion cage, rhBMP-2 has been shown to promote osteoinduction and fusion. METHODS: In this prospective nonblinded study, 42 patients were randomly divided into two groups. The investigational group underwent interbody fusion using two tapered cylindrical fusion cages (LT-CAGE) and rhBMP-2 on an absorbable collagen sponge, and a control group underwent the procedure, receiving the devices and autogenous iliac crest bone graft. Plain radiographs and computed tomographic scans were used to evaluate the pattern of osteoinduction in the interbody space and the progression of fusion 6, 12, and 24 months after surgery. RESULTS: All the patients who received rhBMP-2 showed radiographic evidence of osteoinduction in the interbody cages 6 months after surgery. The density in the cages had increased an average of 142 Hounsfield units by 6 months. At 12 months, the increase had reached 228.7 Hounsfield units New bone formation occurred in the disc space outside the cages by 6 months in 18 of the patients in the investigational group (18/22; 82%). By 24 months, all the investigational patients showed new formation outside the cages. In the autograft control group, the density in the cages increased an average of 42 Hounsfield units, and 10 patients (10/20; 50%) showed evidence of bone formation outside the cages. CONCLUSIONS: The use of rhBMP-2 is a promising method for facilitating anterior intervertebral spinal fusion in patients who have undergone anterior lumbar fusion surgery.

BACKGROUND: Flatfish metamorphosis involves major physiological and morphological changes. Due to its importance in aquaculture and as a model for developmental studies, some gene expression studies have focused on the understanding of this process using quantitative real-time PCR (qRT-PCR) technique. Therefore, adequate reference genes for accurate normalization are required. RESULTS: The stability of 12 potential reference genes was examined during larval development in Senegalese sole (Solea senegalensis) and Atlantic halibut (Hippoglossus hippoglossus) to determine the most suitable genes for qRT-PCR analysis. Transcription levels of genes encoding beta-Actin (ACTB), glyceraldehyde-3P-dehydrogenase (GAPDH), annexin A2 (ANXA2), glutathione S-transferase (GST), ornithine decarboxylase (ODC), hypoxanthine phosphoribosyltransferase (HPRT1), ubiquitin (UBQ), elongation factor 1 alpha (eEF1A1), 18S ribosomal RNA, and the ribosomal proteins S4 (RPS4) and L13a (RPL13a) were quantitated. Two paralogous genes for ACTB were analyzed in each of both flatfish species. In addition, two paralogous genes for GAPDH were studied in Senegalese sole. RPL13a represented non-orthologous genes between both flatfish species. GeNorm and NormFinder analyses for expression stability revealed RPS4, UBQ and eEF1A1 as the most stable genes in Senegalese sole, Atlantic halibut and in a combined analysis. In all cases, paralogous genes exhibited differences in expression stability. CONCLUSION: This work suggests RPS4, UBQ, and eEF1A1 genes as useful reference genes for accurate normalization in qRT-PCR studies in Senegalese sole and Atlantic halibut larvae. The congruent results between both species in spite of the drastic differences in larval development suggest that selected housekeeping genes (HKGs) could be useful in other flatfish species. However, the finding of paralogous gene copies differentially expressed during development in some HKGs underscores the necessity to identify orthologous genes.

STUDY DESIGN: A prospective, randomized, and double-blind study comparing high-dose methylprednisolone sodium succinate (MPSS) with placebo, in the treatment of patients with acute cervical spinal cord injury. OBJECTIVES: To evaluate the complications of high-dose MPSS in patients with acute cervical spinal cord injury when administered within 8 hours of injury. SUMMARY OF BACKGROUND DATA: High-dose therapy with MPSS has been demonstrated to improve the recovery of motor function in patients with acute cervical spinal cord injury. However, little is known about the follow-up complications. METHODS: Forty-six patients, 42 men and 4 women (mean age, 60.6 years; range, 18-84), were included in the study: 23 in the MPSS group and 23 in the placebo group. They were treated without surgery for spinal cord injury in the cervical spine, and were enrolled in the trial if a diagnosis had been made and treatment had begun within 8 hours. Complications of high-dose therapy with MPSS were compared with placebo treatment throughout the study period and up to 2 months after injury. RESULTS: The MPSS group had 13 patients (56.5%) with complications, whereas the placebo group had 8 (34.8%). The difference between the two groups was not statistically significant (P = 0.139). There were eight instances of pulmonary complication with MPSS (34.8%) and one instance (4.34%) with placebo (P = 0.009). There were four instances of gastrointestinal complication (17.4%) with MPSS and none with placebo (P = 0.036). Pulmonary (complications were more prevalent in patients aged more than 60 years (P = 0.029). CONCLUSION: Aged patients with cervical spinal injury may be more likely to have pulmonary side effects (P = 0.029) after high-dose therapy with MPSS and thus deserve special care.