Recovery of chronic motor neuropathy due to acute intermittent porphyria after givosiran treatment in a young boy: a case report.

BACKGROUND: We describe the first case of a pediatric patient with acute intermittent porphyria and severe chronic porphyric neuropathy treated with givosiran, a small-interfering RNA that drastically decreases delta-aminolevulinic acid production and reduces porphyric attacks' recurrence. CASE REPORT: A 12-year-old male patient with refractory acute intermittent porphyria and severe porphyric neuropathy was followed prospectively for 12 months after givosiran initiation (subcutaneous, 2.5 mg/kg monthly). Serial neurological, structural, and resting-state functional magnetic resonance imaging (MRI) evaluations were performed, including clinical scales and neurophysiological tests. Delta-aminolevulinic acid urinary levels dropped drastically during treatment. In parallel, all the administered neurological rating scales and neurophysiological assessments showed improvement in all domains. Moreover, an improvement in central motor conduction parameters and resting-state functional connectivity in the sensory-motor network was noticed. At the end of the follow-up, the patient could walk unaided after using a wheelchair for 5 years. CONCLUSIONS: A clear beneficial effect of givosiran was demonstrated in our patient with both clinical and peripheral nerve neurophysiologic outcome measures. Moreover, we first reported a potential role of givosiran in recovering central motor network impairment in acute intermittent porphyria (AIP), which was previously unknown. This study provides Class IV evidence that givosiran improves chronic porphyric neuropathy.

Astrocytic trans-Differentiation Completes a Multicellular Paracrine Feedback Loop Required for Medulloblastoma Tumor Growth.

The tumor microenvironment (TME) is critical for tumor progression. However, the establishment and function of the TME remain obscure because of its complex cellular composition. Using a mouse genetic system called mosaic analysis with double markers (MADMs), we delineated TME evolution at single-cell resolution in sonic hedgehog (SHH)-activated medulloblastomas that originate from unipotent granule neuron progenitors in the brain. First, we found that astrocytes within the TME (TuAstrocytes) were trans-differentiated from tumor granule neuron precursors (GNPs), which normally never differentiate into astrocytes. Second, we identified that TME-derived IGF1 promotes tumor progression. Third, we uncovered that insulin-like growth factor 1 (IGF1) is produced by tumor-associated microglia in response to interleukin-4 (IL-4) stimulation. Finally, we found that IL-4 is secreted by TuAstrocytes. Collectively, our studies reveal an evolutionary process that produces a multi-lateral network within the TME of medulloblastoma: a fraction of tumor cells trans-differentiate into TuAstrocytes, which, in turn, produce IL-4 that stimulates microglia to produce IGF1 to promote tumor progression.

MANF regulates metabolic and immune homeostasis in ageing and protects against liver damage.

Aging is accompanied by altered intercellular communication, deregulated metabolic function, and inflammation. Interventions that restore a youthful state delay or reverse these processes, prompting the search for systemic regulators of metabolic and immune homeostasis. Here we identify MANF, a secreted stress-response protein with immune modulatory properties, as an evolutionarily conserved regulator of systemic and in particular liver metabolic homeostasis. We show that MANF levels decline with age in flies, mice and humans, and MANF overexpression extends lifespan in flies. MANF deficient flies exhibit enhanced inflammation and shorter lifespans, and MANF heterozygous mice exhibit inflammatory phenotypes in various tissues, as well as progressive liver damage, fibrosis, and steatosis. We show that immune cell-derived MANF protects against liver inflammation and fibrosis, while hepatocyte-derived MANF prevents hepatosteatosis. Liver rejuvenation by heterochronic parabiosis in mice further depends on MANF, while MANF supplementation ameliorates several hallmarks of liver aging, prevents hepatosteatosis induced by diet, and improves age-related metabolic dysfunction. Our findings identify MANF as a systemic regulator of homeostasis in young animals, suggesting a therapeutic application for MANF in age-related metabolic diseases.

Rhodococcus equi Pneumonia in Kidney Transplant Recipient Affected by Acute Intermittent Porphyria: A Case Report.

Rhodococcus equi is a gram-positive coccobacillus responsible for severe infections in patients with weakened immune systems. R equi generally causes pnumonia that may evolve into fatal systemic infection if left untreated. Here, we present a case of a 67-year-old woman affected by acute intermittent porphyria (AIP) who developed R equi pneumonia 7 months after kidney transplantation. Although clinical features at presentation were nonspecific, lung computed tomography showed right perihilar consolidation with a mass-like appearance causing bronchial obstruction. Appropriate antibiotic including intravenous meropenem and oral azithromycin that was then switched to oral levofloxacin and oral azithromycin along with reduction of immunosuppressive therapy resolved pneumonia without provoking an acute attack of porphyria. AIP limited the choice of antibiotics for the treatment of R equi infection because some potentially porphyrinogenic antibacterial agents were avoided. Based on this experience, azithromycin and meropenem can be safely administered for the treatment of R Equi infection in patients with AIP.

Cnidarian Primary Cell Culture as a Tool to Investigate the Effect of Thermal Stress at Cellular Level.

In the context of global change, symbiotic cnidarians are largely affected by seawater temperature elevation leading to symbiosis breakdown. This process, also called bleaching, is triggered by the dysfunction of the symbiont photosystems causing an oxidative stress and cell death to both symbiont and host cells. In our study, we wanted to elucidate the intrinsic capacity of isolated animal cells to deal with thermal stress in the absence of symbiont. In that aim, we have characterized an animal primary cell culture form regenerating tentacles of the temperate sea anemone Anemonia viridis. We first compared the potential of whole tissue tentacle or separated epidermal or gastrodermal monolayers as tissue sources to settle animal cell cultures. Interestingly, only isolated cells extracted from whole tentacles allowed establishing a viable and proliferative primary cell culture throughout 31 days. The analysis of the expression of tissue-specific and pluripotency markers defined cultivated cells as differentiated cells with gastrodermal origin. The characterization of the animal primary cell culture allowed us to submit the obtained gastrodermal cells to hyperthermal stress (+ 5 and + 8 °C) during 1 and 7 days. Though cell viability was not affected at both hyperthermal stress conditions, cell growth drastically decreased. In addition, only a + 8 °C hyperthermia induced a transient increase of antioxidant defences at 1 day but no ubiquitin or carbonylation protein damages. These results demonstrated an intrinsic resistance of cnidarian gastrodermal cells to hyperthermal stress and then confirmed the role of symbionts in the hyperthermia sensitivity leading to bleaching.

Abordaje cerrado en patología de vía aérea superior con el uso de microelectrodos: nuestra experiencia / Approach closed upper airway pathology using microelectrodes: our experience

El láser como alternativa a la cirugía abierta de la vía aérea superior ha venido a modificar la forma de abordaje de las patologías en esta área, pero no deja de ser un procedimiento costoso que no está al alcance de todos los servicios. Por este motivo se han reinventado nuevas formas de abordaje que cumplan los mismos requisitos tanto de la cirugía abierta como con láser pero con un menor coste. Presentamos una serie de 30 casos realizados en un período de 6 años por motivos tanto tumorales como no, en los que se realizaron abordajes cerrados a través de microcirugía con disección mediante microelectrodos. Obteniendo pocas complicaciones y una disminución de la estancia hospitalaria significativa. Con lo cual nos parece una técnica eficiente para abordajes de este tipo.

The laser as an alternative to open surgery of the upper airway has come to change the form of approaching the disease in this area, but it is still an expensive procedure that is not available to all services. For this reason a new ways of approach to meet the same requirements both open as laser but at a lower cost surgery. We present a series of 30 cases performed over a period of 6 years for reasons as much tumor, which closed approaches through microsurgical dissection were performed using microelectrodes. Obtaining few complications and significant decreased hospital stay. Our considerations is it seems an efficient technique for such approaches.

MADM-ML, a mouse genetic mosaic system with increased clonal efficiency.

Mosaic Analysis with Double Markers (MADM) is a mouse genetic system that allows simultaneous gene knockout and fluorescent labeling of sparse, clonally-related cells within an otherwise normal mouse, thereby circumventing embryonic lethality problems and providing single-cell resolution for phenotypic analysis in vivo. The clonal efficiency of MADM is intrinsically low because it relies on Cre/loxP-mediated mitotic recombination between two homologous chromosomes rather than within the same chromosome, as in the case of conditional knockout (CKO). Although sparse labeling enhances in vivo resolution, the original MADM labels too few or even no cells when a low-expressing Cre transgene is used or a small population of cells is studied. Recently, we described the usage of a new system, MADM-ML, which contains three mutually exclusive, self-recognizing loxP variant sites as opposed to a single loxP site present in the original MADM system (referred to as MADM-SL in this paper). Here we carefully compared the recombination efficiency between MADM-SL and MADM-ML using the same Cre transgene, and found that the new system labels significantly more cells than the original system does. When we established mouse medulloblastoma models with both the original and the new MADM systems, we found that, while the MADM-SL model suffered from varied tumor progression and incomplete penetrance, the MADM-ML model had consistent tumor progression and full penetrance of tumor formation. Therefore MADM-ML, with its higher recombination efficiency, will broaden the applicability of MADM for studying many biological questions including normal development and disease modeling at cellular resolution in vivo.

Mouse TU tagging: a chemical/genetic intersectional method for purifying cell type-specific nascent RNA.

Transcriptional profiling is a powerful approach for understanding development and disease. Current cell type-specific RNA purification methods have limitations, including cell dissociation trauma or inability to identify all RNA species. Here, we describe "mouse thiouracil (TU) tagging," a genetic and chemical intersectional method for covalent labeling and purification of cell type-specific RNA in vivo. Cre-induced expression of uracil phosphoribosyltransferase (UPRT) provides spatial specificity; injection of 4-thiouracil (4TU) provides temporal specificity. Only UPRT(+) cells exposed to 4TU produce thio-RNA, which is then purified for RNA sequencing (RNA-seq). This method can purify transcripts from spatially complex and rare (<5%) cells, such as Tie2:Cre(+) brain endothelia/microglia (76% validated by expression pattern), or temporally dynamic transcripts, such as those acutely induced by lipopolysaccharide (LPS) injection. Moreover, generating chimeric mice via UPRT(+) bone marrow transplants identifies immune versus niche spleen RNA. TU tagging provides a novel method for identifying actively transcribed genes in specific cells at specific times within intact mice.

Fibromixoma odontogénico en el maxilar superior: reporte de un caso clínico y revisión de la literatura / Fibromixoma odontogenic in the maxilla: report of a case and literture review

El Fibromixoma Odontogénico es una variante del Mixoma Odontogénico. Se describe como una lesión intraósea agresiva derivada del tejido conjuntivo embrionario asociada con la odontogénesis1 constituida principalmente por grandes cantidades de tejido fibroso celular maduro. Su origen es controvertido, aparece en el esqueleto facial, afectando con mayor frecuencia a la mandíbula2. A continuación se presenta el caso clínico de un paciente de sexo femenino de 36 años de edad que presentó un aumento de volumen a nivel del ápice de diente 1.6 ,en la que se realizó un curetaje logrando la completa resección de la lesión, el resultado del informe patológico da el diagnóstico de Fibromixoma de origen Odontogénico

The Odontogenic Fibromyxoma is a variant of the Odontogenic Myxoma. It is described as an agressive intraoseous lesion that derives from the embrionary connective tissue associated with the odontogenesis, constituted by great amounts of celular mature fibrous tissue. It has a controverted origin, appears in the facial skeleton affecting more frecuently the mandible. We present a case of a 36 year old female who consulted with an increase of volume in relation to the 1.6 theet where we practiced a curetaje obtaining a complete resection of the lesion, the results of the patologic inform gives the diagnostic of odontogenic fibromyxoma

Fast frequency sweeping in resonance-tracking SPM for high-resolution AFAM and PFM imaging.

A new resonance-tracking (RT) method using fast frequency sweeping excitation was developed for quantitative scanning probe microscopy (SPM) imaging. This method allows quantitative imaging of elastic properties and ferroelectrical domains with nanoscale resolution at high data acquisition rates. It consists of a commercial AFM system combined with a high-frequency lock-in amplifier, a programmed function generator and a fast data acquisition card. The resonance-tracking method was applied to the atomic force acoustic microscopy (AFAM) and to the piezoresponse force microscopy (PFM) modes. Plots of amplitude versus time and phase versus time for resonant spectra working with different sweeping frequencies were obtained to evaluate the response speed of the lock-in amplifier. It was proved that this resonance-tracking method allows suitable spectral acquisition at a rate of about 5 ms/pixel, which is useful for SPM imaging in a practical scanning time. In order to demonstrate the system performance, images of RT-AFAM for TiN films and RT-PFM for GeTe are shown.

Axonal loss and cognitive deficits in term infants with normal umbilical artery Doppler born small-for-gestational age.

OBJECTIVE: To assess cognitive outcomes and structural changes in the central nervous system, the latter using a novel approach to examine changes in neuronal integrity of the optic nerve, in children at 5-6½ years of age who were born small-for-gestational age (SGA) at term having shown normal umbilical artery (UA) Doppler. METHODS: We compared neuronal damage, cognitive deficits and visuospatial perception in two cohorts of infants, one born SGA (n = 40) and one born appropriate-for-gestational age (AGA) (n = 39) in weight. Neuronal damage was evaluated using optical coherence tomography (OCT) of the optic nerve. Cognitive deficits were assessed with the Wechsler Preschool and Primary Scale of Intelligence (WPPSI) test. Visuospatial perception was evaluated with Rey-Osterreich Complex Figure (ROCF) tasks. RESULTS: Children from the SGA group had a significantly thinner average retinal nerve fiber layer (RNFL) compared with those from the AGA group (98.2 vs 104.5 µm, P = 0.012). Children from the SGA group exhibited impaired performance in copy tasks on the ROCF (3.27 vs 3.56, P = 0.036) and a higher rate of suboptimal WPPSI test performance intelligence quotient scores (15% vs 0%; P = 0.025) compared with those from the AGA group. CONCLUSION: Term infants with normal UA Doppler born SGA are at increased risk for cognitive deficits and axonal loss in the RNFL at the age of 5-6½ years.

Maternal active smoking and newborn body composition.

OBJECTIVE: Maternal smoking during pregnancy is associated with a reduction in birth size but very few studies have collated changes in neonatal anthropometry. Our aims were both to assess body composition differences by anthropometry between new-borns from smoking mothers and those from non-smoking mothers, and to show whether these differences affect proportional body mass distribution. METHODS: Caucasian mothers and their full term singleton new-borns (N=1216) were selected during 2009. A structured questionnaire was completed regarding obstetric and demographic data, as well as tobacco consumption. Women were categorized, according to their smoking habits, into a non-smoking group (never smoked or stopped smoking prior to pregnancy) and a smoking group (smoked throughout pregnancy). RESULTS: 22.1% of mothers smoked during pregnancy (median: 6 cigarettes/day, range: l-40). Smoking mothers were significantly younger than non-smoking mothers but there were no differences regarding other aspects which could affect infant weight. Infants from non-smoking mothers were heavier, longer, and body circumferences were all larger than those from smoking mothers (p<0.001), but the Ponderal Index showed no statistical differences. Skinfold thicknesses were significantly lower in new-borns from smoking mothers but these differences were less evident than those from body size. Subcutaneous fat distribution did not show statistical differences between the two groups. After gestational age, to smoke during gestation is the second main determinant of birth weight. CONCLUSIONS: Smoking during pregnancy involves a generalized reduction of most axiological parameters as a result of proportionate fetal growth impairment. In those infants born from mothers who smoked during gestation, neonatal lean body mass appears to be more affected than body fat, and distribution of subcutaneous fat is not different.

Central adiposity in children born small and large for gestational age.

OBJECTIVE: To evaluate body composition differences between children that were born small (SGA) or large for gestational age (LGA) compared with their counterparts born adequate for gestational age (AGA). METHODS: Body composition was assessed in 124 healthy Caucasian children (50% girls) aged 6-10, classified according to their birth weight for gestational age as AGA, SGA and LGA. Fat mass (FM), percentage of FM, lean mass (LM), bone mineral content (BMC) and bone mineral density were measured by dual-energy X-ray absorptiometry (DXA) in the whole body and at different body regions. RESULTS: LM (adjusted for age and sex) and total BMC (adjusted for age, sex and weight) were both significantly higher in LGA children and lower in SGA when compared with those born AGA. After adjustments for height, LM and BMC differences between groups were not significant. In SGA children, truncal (P<0.05) and abdominal fatness (P<0.01) were higher when compared with both AGA and LGA children, after adjustments for age, sex and height. There were no differences in the percentage of total and central FM between children born LGA and AGA. CONCLUSIONS: During childhood, children born SGA had higher central adiposity regardless of their body size. Children born LGA seem to have a higher body size but with harmonic body composition and adequate body fat distribution. Small size for gestational age at birth could programme excess abdominal fat deposition in children, which is a major factor for the clustering of cardiovascular disease risk factors defining the metabolic syndrome.

Development of a method for the purification and culture of rodent astrocytes.

The inability to purify and culture astrocytes has long hindered studies of their function. Whereas astrocyte progenitor cells can be cultured from neonatal brain, culture of mature astrocytes from postnatal brain has not been possible. Here, we report a new method to prospectively purify astrocytes by immunopanning. These astrocytes undergo apoptosis in culture, but vascular cells and HBEGF promote their survival in serum-free culture. We found that some developing astrocytes normally undergo apoptosis in vivo and that the vast majority of astrocytes contact blood vessels, suggesting that astrocytes are matched to blood vessels by competing for vascular-derived trophic factors such as HBEGF. Compared to traditional astrocyte cultures, the gene profiles of the cultured purified postnatal astrocytes much more closely resemble those of in vivo astrocytes. Although these astrocytes strongly promote synapse formation and function, they do not secrete glutamate in response to stimulation.

[Acute abdominal pain in elderly patients evaluated in the emergency department at a tertiary level]. / Dolor abdominal agudo en pacientes geriátricos evaluados en el servicio de urgencias en un hospital de tercer nivel.

BACKGROUND: Life expectancy in México has increased in the last decades with a remarkable increase in geriatric population. Acute abdominal pain (AAP) in elderly people compared with young people has different clinical presentation because of the concomitant chronic diseases, the use of medications, history of abdominal surgeries and decrease in perception of pain and immunity. OBJECTIVE: To know the cause and associated mortality of acute abdominal pain in geriatric patients who attend the emergency room. METHODS: Geriatric patients' files with acute abdominal pain admitted from January 2004 to December 2008 were retrospectively reviewed. Age, gender, presence of chronic diseases, use of medications, history of surgical procedures, definitive diagnosis causative of the symptoms and the associated mortality were recorded. RESULTS: 17 524 patients were admitted, of whom 324 (1.8%) were geriatric patients with AAP: 110 were men (36.9) and 214 were women (66%), with a mean age of 78 years (range 60 to 102 years). The most common causes of AAP were acute cholecystitis in 49 patients (15.1%), irritable bowel syndrome in 42 (12.9%), ulcerative syndrome in 40 (12.3%), intestinal obstruction in 35 (10.8%) and diverticulitis in 23 (10.8%). Nine patients died (2.7%). CONCLUSIONS: In our hospital the most common cause of AAP in geriatric patients is related to biliary disease followed by functional gastrointestinal disorder and ulcerative syndrome. Mortality is low.

Depressive and adjustment disorders - some questions about the differential diagnosis: case studies.

BACKGROUND: Diagnosis and treatment of mood disorders in youth are still problematic because in this age the clinical presentation is atypical, and the diagnostic tools and the therapies are the same as that used for the adults. Mood disorders are categorically divided into unipolar disorders (major depressive disorder and dysthymic disorder) and bipolar disorder in Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition, Text Revision), but mood symptoms are also comprised in the diagnostic criteria of the adjustment disorder (AD), which occur in many different psychiatric disorders, and may also be found in some physical conditions. The differential diagnosis is not much addressed in the midst of clinical investigation and so remains the major problem in the clinical practice. AIMS: The associations between some variables and the depressive disorder and AD were analyzed to make considerations about differential diagnosis. PATIENTS AND METHODS: We reported a retrospective study of 60 patients affected by depressive disorder and AD. The analysis has evaluated the association between some variables and the single diagnostic categories. We have considered 10 variables, of which 6 are specific to the disorders, and 4 have been considered related problems. RESULTS: The statistical analysis showed significant results for the associations of 3 variables (prevalent symptoms, treatment, and family history) with the single diagnostic categories. CONCLUSION: The discriminate analysis resulted in statistically significant differences between patients with depressive disorders and those with AD on 3 variables, of which 2 are specific to the disorders, and 1 is included in the related problems. The other variables were weakly associated with the single diagnostic categories without any statistically significant differences. The 3 variables that were associated with the single diagnostic categories support the distinct construct validity of the 2 diagnostic categories, but, to date, it is difficult to establish if these variables can be considered diagnostic predictors. On the other hand, the other variables did not support the distinct construct validity of the 2 diagnostic categories, which suggest an overlapping and dimensional concept. The spectrum approach could unify categorical classification that is essential with a dimensional view. Combination of dimensional and categorical principles for classifying mood disorders may help to reduce the problems of underdiagnosis and undertreatment.

Focal cerebral ischemia induces a multilineage cytogenic response from adult subventricular zone that is predominantly gliogenic.

The purpose of this study was to ascertain the relative contribution of neural stem/progenitor cells (NSPCs) of the subventricular zone (SVZ) to lineages that repopulate the injured striatum following focal ischemia. We utilized a tamoxifen-inducible Cre/loxP system under control of the nestin promoter, which provides permanent YFP labeling of multipotent nestin(+) SVZ-NSPCs prior to ischemic injury and continued YFP expression in all subsequent progeny following stroke. YFP reporter expression was induced in adult male nestin-CreER(T2):R26R-YFP mice by tamoxifen administration (180 mg kg(-1), daily for 5 days). Fourteen days later, mice were subjected to 60-min transient middle cerebral artery occlusion (MCAO) and sacrificed at 2 days, 2 weeks, or 6 weeks post-MCAO for phenotypic fate mapping of YFP(+) cells using lineage-specific markers. Migration of YFP(+) cells from SVZ into the injured striatal parenchyma was apparent at 2 and 6 weeks, but not 2 days, post-MCAO. At 2 weeks post-MCAO, the average percent distribution of YFP(+) cells within the injured striatal parenchyma was as follows: 10% Dcx(+) neuroblasts, 15-20% oligodendrocyte progenitors, 59% GFAP(+) astrocytes, and only rare NeuN(+) postmitotic neurons. A similar phenotypic distribution was observed at 6 weeks, except for an increased average percentage of YFP(+) cells that expressed Dcx(+) (20%) or NeuN (5%). YFP(+) cells did not express endothelial markers, but displayed unique anatomical relationships with striatal vasculature. These results indicate that nestin(+) NSPCs within the SVZ mount a multilineage response to stroke that includes a gliogenic component more predominant than previously appreciated.