Evolution of asexual and sexual reproduction in the aspergilli.

Aspergillus nidulans has long-been used as a model organism to gain insights into the genetic basis of asexual and sexual developmental processes both in other members of the genus Aspergillus, and filamentous fungi in general. Paradigms have been established concerning the regulatory mechanisms of conidial development. However, recent studies have shown considerable genome divergence in the fungal kingdom, questioning the general applicability of findings from Aspergillus, and certain longstanding evolutionary theories have been questioned. The phylogenetic distribution of key regulatory elements of asexual reproduction in A. nidulans was investigated in a broad taxonomic range of fungi. This revealed that some proteins were well conserved in the Pezizomycotina (e.g. AbaA, FlbA, FluG, NsdD, MedA, and some velvet proteins), suggesting similar developmental roles. However, other elements (e.g. BrlA) had a more restricted distribution solely in the Eurotiomycetes, and it appears that the genetic control of sporulation seems to be more complex in the aspergilli than in some other taxonomic groups of the Pezizomycotina. The evolution of the velvet protein family is discussed based on the history of expansion and contraction events in the early divergent fungi. Heterologous expression of the A. nidulans abaA gene in Monascus ruber failed to induce development of complete conidiophores as seen in the aspergilli, but did result in increased conidial production. The absence of many components of the asexual developmental pathway from members of the Saccharomycotina supports the hypothesis that differences in the complexity of their spore formation is due in part to the increased diversity of the sporulation machinery evident in the Pezizomycotina. Investigations were also made into the evolution of sex and sexuality in the aspergilli. MAT loci were identified from the heterothallic Aspergillus (Emericella) heterothallicus and Aspergillus (Neosartorya) fennelliae and the homothallic Aspergillus pseudoglaucus (=Eurotium repens). A consistent architecture of the MAT locus was seen in these and other heterothallic aspergilli whereas much variation was seen in the arrangement of MAT loci in homothallic aspergilli. This suggested that it is most likely that the common ancestor of the aspergilli exhibited a heterothallic breeding system. Finally, the supposed prevalence of asexuality in the aspergilli was examined. Investigations were made using A. clavatus as a representative 'asexual' species. It was possible to induce a sexual cycle in A. clavatus given the correct MAT1-1 and MAT1-2 partners and environmental conditions, with recombination confirmed utilising molecular markers. This indicated that sexual reproduction might be possible in many supposedly asexual aspergilli and beyond, providing general insights into the nature of asexuality in fungi.

Frequency and outcome of total anterior circulation strokes without intracranial large-vessel occlusion.

BACKGROUND AND PURPOSE: The percentage of patients with clinical total anterior circulation infarct (TACI) syndrome treated with reperfusion therapies in the absence of intracranial large-vessel occlusion (ILVO) was determined and their characteristics and outcome are described. METHODS: Data from a population-based, prospective, externally audited registry of all stroke patients treated with intravenous thrombolysis (IVT) and endovascular therapies in Catalonia from January 2011 to December 2013 were used. Patients with a baseline TACI and initial stroke severity measured by the National Institute of Health Stroke Scale (NIHSS) ≥ 8, evaluated less than 4.5 h post-onset, for whom a vascular study prior to treatment was available (n = 1070) were selected. Clinical characteristics, outcome and radiological data for patients treated with IVT alone (n = 605) were compared between those with detected ILVO (n = 474) and non-ILVO patients (n = 131). RESULTS: A total of 1070 patients met study criteria; non-ILVO was found in 131 (12.2%). Analysing the 605 patients treated only with IVT, no significant differences were found between non-ILVO and ILVO patients in age, sex, risk factors, time-to-treatment and type of radiological studies performed. Although non-ILVO patients had lower initial stroke severity (P < 0.001) and a better prognosis (P = 0.001), 51.3% had a poor outcome and 16% were deceased at 90 days. In 66.4% of patients without ILVO, a recent anterior territorial infarct was detected. CONCLUSIONS: Intracranial artery patency was observed in 12.2% of TACI patients evaluated within 4.5 h. Although absence of ILVO was associated with slightly better prognosis, more than half had a poor outcome at 3 months.

Vitamin D promotes human extravillous trophoblast invasion in vitro.

INTRODUCTION: Incomplete human extravillous trophoblast (EVT) invasion of the decidua and maternal spiral arteries is characteristic of pre-eclampsia, a condition linked to low maternal vitamin D status. It is hypothesized that dysregulated vitamin D action in uteroplacental tissues disrupts EVT invasion leading to malplacentation. METHODS: This study assessed the effects of the active vitamin D metabolite, 1,25-dihydroxyvitamin D3 (1,25-D3), and its precursor, 25-hydroxyvitamin D3 (25-D3), on primary human EVT isolated from first trimester pregnancies. Expression of EVT markers (cytokeratin-7, HLA-G), the vitamin D-activating enzyme (CYP27B1) and 1,25-D3 receptor (VDR) was assessed by immunocytochemistry. EVT responses following in vitro treatment with 1,25-D3 (0-10 nM) or 25-D3 (0-100 nM) for 48-60 h were assessed using quantitative RT-PCR (qRT-PCR) analysis of key target genes. Effects on EVT invasion through Matrigel(®) were quantified alongside zymographic analysis of secreted matrix metalloproteinases (MMPs). Effects on cell viability were assessed by measurement of MTT. RESULTS: EVT co-expressed mRNA and protein for CYP27B1 and VDR, and demonstrated induction of mRNA encoding vitamin D-responsive genes, 24-hydroxylase (CYP24A1) and cathelicidin following 1,25-D3 treatment. EVT could respond to 1,25-D3 and 25-D3, both of which significantly increased EVT invasion, with maximal effect at 1 nM 1,25-D3 (1.9-fold; p < 0.01) and 100 nM 25-D3 (2.2-fold; p < 0.05) respectively compared with untreated controls. This was accompanied by increased pro-MMP2 and pro-MMP9 secretion. The invasion was independent of cell viability, which remained unchanged. DISCUSSION: These data support a role for vitamin D in EVT invasion during human placentation and suggest that vitamin D-deficiency may contribute to impaired EVT invasion and pre-eclampsia.

How predictors and patterns of stroke recurrence after a TIA differ during the first year of follow-up.

The highest risk of subsequent stroke after a TIA occurs within the first week after the index event. However, the risk of stroke recurrence (SR) remains high during the first year of follow-up. We studied the temporal pattern and predictors of SR (at 7 days and from 7 days to 1-year follow-up). Between April 2008 and December 2009, we included 1,255 consecutive TIA patients from 30 Spanish stroke centers (PROMAPA study). We determined the short-term (at 7 days) and long-term (from 8 days to 1 year) risk of SR. Patients who underwent short-term recurrence and long-term recurrence were compared with regard to clinical findings, vascular territories, and etiology. Enough information (clinical variables and extracranial vascular imaging) was assessed in 1,137 (90.6 %) patients. The 7-day stroke risk was 2.6 %. 32 (3.0 %) patients had an SR after 7-day follow-up. Multiple TIA (HR 3.50, 1.67-7.35, p = 0.001) and large artery atherosclerosis (HR 2.51, 1.17-5.37, p = 0.018) were independent predictors of early SR, whereas previous stroke (HR 1.40, 1.03-1.92, p = 0.034) and coronary heart disease (2.65, 1.28-5.50, p = 0.009) were independent predictors of late SR. Notoriously, 80 % of SR happened in the same territory of the index TIA at 7-day follow-up, whereas only 38 % during the long-term follow-up (p < 0.001). Different predictors of SR were identified throughout the follow-up period. Moreover, the ischemic mechanism differed in early and late stroke recurrences.

Predictive value of brain and vascular imaging including intracranial vessels in transient ischaemic attack patients: external validation of the ABCD3-I score.

BACKGROUND AND PURPOSE: Recently, brain and vascular imaging have been added to clinical variables to identify patients with transient ischaemic attack (TIA) with a high risk of stroke recurrence. The aim of our study was to externally validate the ABCD3-I score and the same score taking into account intracranial circulation. METHODS: We analyzed data from 1137 patients with TIA from the PROMAPA study who underwent diffusion-weighted magnetic resonance imaging (DWI) within 7 days of symptom onset. Clinical variables and diagnostic work-up were recorded prospectively. The end-points were subsequent stroke at 7 and 90 days follow-up. RESULTS: A total of 463 (40.7%) subjects fulfilled all inclusion criteria. During follow-up, eight patients (1.7%) had a stroke within 7 days, and 14 (3.1%) had a stroke within 3 months. In the Cox proportional hazard multivariate analyses, the combination of large-artery atherosclerosis and positive DWI remained as independent predictors of stroke recurrence at 7- and 90-day follow-up [HR 8.23, 95% confidence interval (CI) 2.89-23.46, P < 0.001]. The ABCD3-I score was a powerful predictor of subsequent stroke. The area under the receiver operating characteristic curve was 0.83 (95% CI 0.72-0.93) at 7 days and 0.69 (95% CI 0.53-0.85) at 90 days. When we include intracranial vessel disease in the score, the area under the curve increases but the difference observed was non-significant. CONCLUSION: The inclusion of vascular and neuroimaging information to clinical scales (ABCD3-I score) provides important prognostic information and also helps management decisions, although it cannot give a complete distinction between high-risk and low-risk groups.

Prediction of early stroke recurrence in transient ischemic attack patients from the PROMAPA study: a comparison of prognostic risk scores.

BACKGROUND: Several clinical scales have been developed for predicting stroke recurrence. These clinical scores could be extremely useful to guide triage decisions. Our goal was to compare the very early predictive accuracy of the most relevant clinical scores [age, blood pressure, clinical features and duration of symptoms (ABCD) score, ABCD and diabetes (ABCD2) score, ABCD and brain infarction on imaging score, ABCD2 and brain infarction on imaging score, ABCD and prior TIA within 1 week of the index event (ABCD3) score, California Risk Score, Essen Stroke Risk Score and Stroke Prognosis Instrument II] in consecutive transient ischemic attack (TIA) patients. METHODS: Between April 2008 and December 2009, we included 1,255 consecutive TIA patients from 30 Spanish stroke centers (PROMAPA study). A neurologist treated all patients within the first 48 h after symptom onset. The duration and typology of clinical symptoms, vascular risk factors and etiological work-ups were prospectively recorded in a case report form in order to calculate established prognostic scores. We determined the early short-term risk of stroke (at 7 and 90 days). To evaluate the performance of each model, we calculated the area under the receiver operating characteristic curve. Cox proportional hazards multivariate analyses determining independent predictors of stroke recurrence using the different components of all clinical scores were calculated. RESULTS: We calculated clinical scales for 1,137 patients (90.6%). Seven-day and 90-day stroke risks were 2.6 and 3.8%, respectively. Large-artery atherosclerosis (LAA) was observed in 190 patients (16.7%). We could confirm the predictive value of the ABCD3 score for stroke recurrence at the 7-day follow-up [0.66, 95% confidence interval (CI) 0.54-0.77] and 90-day follow-up (0.61, 95% CI 0.52-0.70), which improved when we added vascular imaging information and derived ABCD3V scores by assigning 2 points for at least 50% symptomatic stenosis on carotid or intracranial imaging (0.69, 95% CI 0.57-0.81, and 0.63, 95% CI 0.51-0.69, respectively). When we evaluated each component of all clinical scores using Cox regression analyses, we observed that prior TIA and LAA were independent predictors of stroke recurrence at the 7-day follow-up [hazard ratio (HR) 3.97, 95% CI 1.91-8.26, p < 0.001, and HR 3.11, 95% CI 1.47-6.58, p = 0.003, respectively] and 90-day follow-up (HR 2.35, 95% CI 1.28-4.31, p = 0.006, and HR 2.20, 95% CI 1.15-4.21, p = 0.018, respectively). CONCLUSION: All published scores that do not take into account vascular imaging or prior TIA when identifying stroke risk after TIA failed to predict risk when applied by neurologists. Clinical scores were not able to replace extensive emergent diagnostic evaluations such as vascular imaging, and they should take into account unstable patients with recent prior transient episodes.

Clinical outcome of spontaneous non-aneurysmal subarachnoid hemorrhage in 108 patients.

BACKGROUND: The cause of spontaneous subarachnoid hemorrhage (SAH) is unknown in 15% of cases; idiopathic SAH has a better prognosis than aneurysmal SAH. When bleeding is confined to the perimesencephalic cisterns, SAH has an especially benign course. METHODS: We retrospectively studied 108 patients admitted for spontaneous non-aneurysmal SAH between 1991 and 2004. We divided patients into two groups according to the bleeding pattern at cranial CT: perimesencephalic pattern (n=60) and aneurysmal pattern (n=48). We included only patients in whom no source of bleeding was detected at angiography; patients with aneurysmal pattern underwent at least two angiographic examinations. Mean follow-up was 5.5years; follow-up consisted of telephone interview in 84.7% of patients. RESULTS: All but one patient with perimesencephalic pattern were classified as grade I or II on the Hunt and Hess scale; the exception was the only patient in this group with a complication (hydrocephalus), who was classified as grade IV. Three-quarters of the patients with aneurysmal pattern were classified as grade I or II on the Hunt and Hess scale; 5 patients presented with hydrocephalus that required drainage and 2 with vasospasms without repercussions. No rebleeding or long-term complications were observed in either group. CONCLUSIONS: Non-aneurysmal SAH with a perimesencephalic pattern of bleeding has a benign course and excellent short-term and long-term prognosis. Patients with non-aneurysmal SAH with an aneurysmal pattern of bleeding have more complications, and the initial clinical situation has a significant impact on their prognosis.

Resistance of uveal melanoma to the interstrand cross-linking agent mitomycin C is associated with reduced expression of CYP450R.

BACKGROUND: Uveal melanoma (UM) is the most common primary intraocular tumour of adults, frequently metastasising to the liver. Hepatic metastases are difficult to treat and are mainly unresponsive to chemotherapy. To investigate why UM are so chemo-resistant we explored the effect of interstrand cross-linking agents mitomycin C (MMC) and cisplatin in comparison with hydroxyurea (HU). METHODS: Sensitivity to MMC, cisplatin and HU was tested in established UM cell lines using clonogenic assays. The response of UM to MMC was confirmed in MTT assays using short-term cultures of primary UM. The expression of cytochrome P450 reductase (CYP450R) was analysed by western blotting, and DNA cross-linking was assessed using COMET analysis supported by γ-H2AX foci formation. RESULTS: Both established cell lines and primary cultures of UM were resistant to the cross-linking agent MMC (in each case P<0.001 in Student's t-test compared with controls). In two established UM cell lines, DNA cross-link damage was not induced by MMC (in both cases P<0.05 in Students's t-test compared with damage induced in controls). In all, 6 out of 6 UMs tested displayed reduced expression of the metabolising enzyme CYP450R and transient expression of CYP450R increased MMC sensitivity of UM. CONCLUSION: We suggest that reduced expression of CYP450R is responsible for MMC resistance of UM, through a lack of bioactivation, which can be reversed by complementing UM cell lines with CYP450R.

Colitis por citomegalovirus / No disponible

No disponible

Neuropsychological profile of bilateral paramedian infarctions: three cases.

INTRODUCTION: The thalamus is one of the strategic diencephalic structures of the human brain. The artery of Percheron, an asymmetrical common trunk arising from a P1 segment of the posterior cerebral artery, is a peculiar presentation of the three variants involved in the irrigation of the paramedian thalamic territory. Occlusion of this artery results in bilateral median thalamic infarction. The paramedian syndrome includes an acute loss or reduction of consciousness, often associated with oculomotor and neuropsychological disturbances. PATIENTS AND METHODS: We present three cases of bilateral paramedian thalamic infarction with onset of acute coma, followed by fluctuations in the level of consciousness, memory, and behavioural alterations. A neuroradiological study with MRI identified individual thalamic nuclei, and a complete neuropsychological study was performed one month after onset of ictus. RESULTS: One of the patients showed severe memory and executive function impairments without improvement of vertical gaze palsy. The other two patients presented with mild executive dysfunction with complete resolution of neurological symptoms. Neuroimaging results showed a bilateral lesion of the dorsomedial nuclei in the three patients. CONCLUSIONS: Severe amnesia has been associated with an affection of the structures of the paramedian thalamic territory. Presently, the role of the dorsomedial nucleus remains controversial, with the suggestion that memory deficits observed in this type of lesion could be secondary to executive function deficits. In our case, the patient with the most severe dysexecutive deficit presented the most severe memory impairments.

Enzymatic function of multiple origins regulates the progression of colorectal cancer and the development of metastases.

Enzymes play a crucial role in the progression of colorectal cancer and the development of metastases. They facilitate malignant cell invasion through the degradation of the extracellular matrix, the rupture of the basement membrane and the derangement of cell-cell adhesion. Furthermore, they promote tumour cell migration and support the evolution of metastatic lesions in the liver and other organs, through multiple molecular mechanisms, including growth factor release and angiogenesis. Urokinase plasminogen activator system, matrix metalloproteinases, heparanase and autocrine motility factor constitute important enzymatic complexes which assist colorectal cancer growth, with potential clinical applications in the diagnosis and treatment of the disease.

Asociación de carcinoma renal con neuromiotonía y afectación de neurona motora inferior / Association of renal carcinoma with neuromyotonia and involvement of inferior motor neuron

La neuromiotonía adquirida, definida como actividad muscular continua originada en el nervio periférico debida a alteraciones de la membrana del axón motor, se ha considerado una manifestación paraneoplásica en relación a tumores relacionados con el sistema inmunitario, principalmente linfomas. De forma similar, la afectación de neurona motora ha sido descrita como una complicación paraneoplásica infrecuente también en relación con linfomas. Aportamos un caso excepcional, dada la presencia de dos manifestaciones paraneoplásicas infrecuentes como la neuromiotonía y la enfermedad de neurona motora secundarias a carcinoma renal de células claras

Acquired neuromyotonia is defined as continuous muscle activity originated in peripheral nerve due to alterations of the motor axon membrane. It has been considered a paraneoplastic syndrome in patients with neoplasms of the immune system, mainly lymphomas. Similarly, involvement of the motor neuron has been described as an uncommon paraneoplastic complication, also in relationship to lymphomas. We report a rare case, given the presence of two uncommon paraneoplastic manifestations such as neuromyotonia and reversible paraneoplastic lower motor neuronopathy secondary to clear cell renal carcinoma

[Association of renal carcinoma with neuromyotonia and involvement of inferior motor neuron]. / Asociación de carcinoma renal con neuromiotonía y afectación de neurona motora inferior.

Acquired neuromyotonia is defined as continuous muscle activity originated in peripheral nerve due to alterations of the motor axon membrane. It has been considered a paraneoplastic syndrome in patients with neoplasms of the immune system, mainly lymphomas. Similarly, involvement of the motor neuron has been described as an uncommon paraneoplastic complication, also in relationship to lymphomas. We report a rare case, given the presence of two uncommon paraneoplastic manifestations such as neuromyotonia and reversible paraneoplastic lower motor neuronopathy secondary to clear cell renal carcinoma.

Extravasación de contraste tras stent carotídeo con clínica de infarto cerebral / Extravasation of contrast medium after carotid stent with brain infarction symptoms

La angioplastia con o sin colocación de stent como técnica de revascularización carotídea se ha convertido en un tratamiento alternativo para la estenosis carotídea, siendo considerado en ocasiones el tratamiento de elección. El principal temor de dicha técnica es la embolización distal durante el procedimiento, habiéndose descrito otras complicaciones menos frecuentes como la disección, vasoespasmo e incluso la rotura arterial. Entre las complicaciones infrecuentes destacan el síndrome de hiperperfusión y más raramente la encefalopatía por extravasación de contraste. Presentamos a una paciente que fue sometida a angioplastia con colocación de stent por estenosis carotídea con signos de presencia cálcica que sufrió un cuadro semejante a un ictus de la arteria cerebral media derecha como consecuencia de extravasación del contraste tras rotura del balón de angioplastia, hecho no descrito previamente en la literatura

Angioplasty, with or without stent placement, as a carotid revascularisation technique, has become an alternative treatment for carotid stenosis, it being considered under certain circumstances the treatment of choice. The main concern about this technique is the distal embolisation during the procedure, while other, less frequent, complícations have also been described such as dissection, vasospasm or even arterial rupture. Some of the infrequent complications include the hyperperfusion syndrome, and, more exceptionally, cases of extravasation of the contrast medium. We present the case of a patient who was subjected to angioplasty and stent insertion due tocarotid stenosis with signs of calcification who suffered a stroke-like disorder in her right medial brain artery as a consequence of the extravasation of contrast medium after the rupture of the angioplasty balloon, a situation wich has never been described in the literature befote

[Extravasation of contrast medium after carotid stent with brain infarction symptoms]. / Extravasación de contraste tras stent carotídeo con clínica de infarto cerebral.

Angioplasty, with or without stent placement, as a carotid revascularisation technique, has become an alternative treatment for carotid stenosis, it being considered under certain circumstances the treatment of choice. The main concern about this technique is the distal embolisation during the procedure, while other, less frequent, complícations have also been described such as dissection, vasospasm or even arterial rupture. Some of the infrequent complications include the hyperperfusion syndrome, and, more exceptionally, cases of extravasation of the contrast medium. We present the case of a patient who was subjected to angioplasty and stent insertion due to carotid stenosis with signs of calcification who suffered a stroke-like disorder in her right medial brain artery as a consequence of the extravasation of contrast medium after the rupture of the angioplasty balloon, a situation which has never been described in the literature before.