Fourier transform infrared spectroscopy as a non-destructive method for analysing herbarium specimens.

Dried plant specimens stored in herbaria are an untapped treasure chest of information on environmental conditions, plant evolution and change over many hundreds of years. Owing to their delicate nature and irreplaceability, there is limited access for analysis to these sensitive samples, particularly where chemical data are obtained using destructive techniques. Fourier transform infrared (FTIR) spectroscopy is a chemical analysis technique which can be applied non-destructively to understand chemical bonding information and, therefore, functional groups within the sample. This provides the potential for understanding geographical, spatial and species-specific variation in plant biochemistry. Here, we demonstrate the use of mid-FTIR microspectroscopy for the chemical analysis of Drosera rotundifolia herbarium specimens, which were collected 100 years apart from different locations. Principal component and hierarchical clustering analysis enabled differentiation between three main regions on the plant (lamina, tentacle stalk and tentacle head), and between the different specimens. Lipids and protein spectral regions were particularly sensitive differentiators of plant tissues. Differences between the different sets of specimens were smaller. This study demonstrates that relevant information can be extracted from herbarium specimens using FTIR, with little impact on the specimens. FTIR, therefore, has the potential to be a powerful tool to unlock historic information within herbaria.

Molecular subtyping reveals immune alterations associated with progression of bronchial premalignant lesions.

Bronchial premalignant lesions (PMLs) are precursors of lung squamous cell carcinoma, but have variable outcome, and we lack tools to identify and treat PMLs at risk for progression to cancer. Here we report the identification of four molecular subtypes of PMLs with distinct differences in epithelial and immune processes based on RNA-Seq profiling of endobronchial biopsies from high-risk smokers. The Proliferative subtype is enriched with bronchial dysplasia and exhibits up-regulation of metabolic and cell cycle pathways. A Proliferative subtype-associated gene signature identifies subjects with Proliferative PMLs from normal-appearing uninvolved large airway brushings with high specificity. In progressive/persistent Proliferative lesions expression of interferon signaling and antigen processing/presentation pathways decrease and immunofluorescence indicates a depletion of innate and adaptive immune cells compared with regressive lesions. Molecular biomarkers measured in PMLs or the uninvolved airway can enhance histopathological grading and suggest immunoprevention strategies for intercepting the progression of PMLs to lung cancer.

Vibrational spectroscopic monitoring and biochemical analysis of pericellular matrix formation and maturation in a 3-dimensional chondrocyte culture model.

Isolated and monolayer expanded chondrocytes are not the ideal cell form to produce a cartilage matrix. In articular cartilage, each chondrocyte is surrounded by a 2-4 µm thick collagen VI-rich pericellular matrix (PCM) forming a chondron. Freshly extracted chondrons form a more cartilage-like extracellular matrix (ECM) than chondrocytes and their surrounding PCM is thought to maintain the chondrocyte phenotype. To regenerate articular cartilage, preserving and/or regenerating a functional PCM is essential. In this study, a highly biomimicking hyaluronic acid (HA) hydrogel was used as a 3-dimensional system to culture freshly isolated bovine chondrons (with an intact PCM) and chondrocytes (without a PCM) for up to 21 days. We assessed the HA hydrogel's capacity to maintain and potentially re-generate PCM formation by both biochemical and immunological analyses of the key components of the PCM. For the first time, synchrotron based Fourier transform infrared (SR-FTIR) microspectroscopy was utilised to reveal the dynamic process of PCM re-generation. At day 1, highly specific collagen VI staining was visible within chondron containing HA hydrogels. In contrast, collagen VI was absent at day 1 but punctate, focal staining increased during the culture period of chondrocyte containing HA hydrogels. Chondron containing HA hydrogels produced more collagen II and GAGs than the chondrocyte containing HA hydrogels. Principal component analysis (PCA) of spectra in fingerprint regions of the chondrocyte-containing constructs at day 7, 14 and 21 culturing showed clear spectral differences. The clusters of day 14 and day 21 samples were closer to the chondron samples, while the day 7 samples were closer to chondrocytes. PCA scores in the lipid region revealed no major differences between chondrocyte and chondron samples, but showed that the cultured chondrocyte samples at day 7, day 14 and day 21 clustered together. These data would indicate that SR-FTIR microspectroscopy can help to better understand the PCM formation and maturation in tissue engineered models, which involves subtle changes in collagen and aggrecan.

Fourier transform infrared spectra of cells on glass coverslips. A further step in spectral pathology.

Over the last few years, great effort has been placed on developing Fourier Transform Infrared (FTIR) microspectroscopy as a tool to help in the histopathological diagnosis of cancer. The ever increasing workload in pathology departments is calling for a technique that could identify the presence of cancer cells in cytology and tissue samples in an objective, fast and automated way. However, pathologists use glass slides which absorb infrared (IR) radiation thus removing important mid-IR spectral data in the fingerprint region (proteins, DNA, RNA; 1800 cm-1 to 900 cm-1). To this purpose, we hypothesised whether using thinner glass slides, i.e., glass coverslips, would allow us to obtain spectral data not only from the lipid region (3100 cm-1 to 2700 cm-1) but also from the fingerprint region. To this purpose, we studied peripheral blood mononuclear cells (PBMC), a leukaemia cell line (K562) and a lung cancer cell line (CALU-1). Cells were placed on DAKO coverslips and their FTIR spectra obtained at MIRAS beamline, Alba synchrotron light source (Barcelona, Catalonia). The data presented here not only shows for the first time that it is possible to obtain spectral data from most of the amide I region (1800 cm-1 to 1570 cm-1) of cells placed on glass coverslips but more important, principal component analysis was able to separate between the three types of cells for both the lipid and the amide I regions. The methodology here described is a further step in the application of FTIR microspectroscopy in histopathology departments.

Profile of BAFF and its receptors' expression in lupus nephritis is associated with pathological classes.

Background/Objective B-cell activating factor (BAFF) plays an important role in the pathogenesis of systemic lupus erythematosus. However, the role of BAFF in lupus nephritis (LN) is not understood. Our aim was to evaluate the expression of BAFF and its three receptors in renal biopsy samples from patients with LN and investigate a relationship with pathological class. Methods We conducted a prospective descriptive study (2011-2014) on 52 kidney biopsy samples from patients with LN. Immunohistochemistry for BAFF, its receptors (transmembrane activator and calcium modulator and cyclophilin ligand interaction (TACI), protein maturation of B cells (BCMA), and BAFF-receptor (BAFF-R)), and CD20 expression was performed. Samples were scored according to the percentage of cells with positive expression. Results In class II LN, BAFF-R and TACI were not expressed, whereas BCMA and BAFF were lowly expressed in the interstitial inflammatory infiltrates. Proliferative class III/IV had elevated BAFF expression in the glomeruli, and TACI was expressed in interstitial inflammatory infiltrates and the glomeruli. Interestingly, the class IV cases with vasculopathy ( n = 4) had endothelial BAFF expression, which was not visible in thrombotic microangiopathy ( n = 4). Class V was characterized by low BAFF expression in interstitial inflammatory infiltrates and by BAFF, TACI, and BCMA expression in the glomeruli. BAFF expression was associated with inflammatory scores and CD20 positive infiltrates, mainly in class IV. Conclusions Expression patterns of BAFF and its receptors differ according to LN class. Our study provides evidence that BAFF could be used as a routine marker in LN biopsies and to determine which patients will benefit from anti-BAFF therapy.

Detecting the Presence and Progression of Premalignant Lung Lesions via Airway Gene Expression.

Purpose: Lung cancer is the leading cause of cancer-related death in the United States. The molecular events preceding the onset of disease are poorly understood, and no effective tools exist to identify smokers with premalignant lesions (PMLs) that will progress to invasive cancer. Prior work identified molecular alterations in the smoke-exposed airway field of injury associated with lung cancer. Here, we focus on an earlier stage in the disease process leveraging the airway field of injury to study PMLs and its utility in lung cancer chemoprevention.Experimental Design: Bronchial epithelial cells from normal appearing bronchial mucosa were profiled by mRNA-Seq from subjects with (n = 50) and without (n = 25) PMLs. Using surrogate variable and gene set enrichment analysis, we identified genes, pathways, and lung cancer-related gene sets differentially expressed between subjects with and without PMLs. A computational pipeline was developed to build and test a chemoprevention-relevant biomarker.Results: We identified 280 genes in the airway field associated with the presence of PMLs. Among the upregulated genes, oxidative phosphorylation was strongly enriched, and IHC and bioenergetics studies confirmed pathway findings in PMLs. The relationship between PMLs and squamous cell carcinomas (SCC) was also confirmed using published lung cancer datasets. The biomarker performed well predicting the presence of PMLs (AUC = 0.92, n = 17), and changes in the biomarker score associated with progression/stability versus regression of PMLs (AUC = 0.75, n = 51).Conclusions: Transcriptomic alterations in the airway field of smokers with PMLs reflect metabolic and early lung SCC alterations and may be leveraged to stratify smokers at high risk for PML progression and monitor outcome in chemoprevention trials. Clin Cancer Res; 23(17); 5091-100. ©2017 AACR.

Oncogenic Characterization and Pharmacologic Sensitivity of Activating Fibroblast Growth Factor Receptor (FGFR) Genetic Alterations to the Selective FGFR Inhibitor Erdafitinib.

Fibroblast growth factor receptor (FGFR) genetic alterations are frequently observed in cancer, suggesting that FGFR inhibition may be a promising therapy in patients harboring these lesions. Identification of predictive and pharmacodynamic biomarkers to select and monitor patients most likely to respond to FGFR inhibition will be the key to clinical development of this class of agents. Sensitivity to FGFR inhibition and correlation with FGFR pathway activation status were determined in molecularly annotated panels of cancer cell lines and xenograft models. Pathway inhibition in response to FGFR inhibitor treatment was assessed in cell lines (both in vitro and in vivo) and in samples from patients treated with the FGFR inhibitor JNJ-42756493 (erdafitinib). Frequency of FGFR aberrations was assessed in a panel of NSCLC, breast, prostate, ovarian, colorectal, and melanoma human tumor tissue samples. FGFR translocations and gene amplifications present in clinical specimens were shown to display potent transforming activity associated with constitutive pathway activation. Tumor cells expressing these FGFR activating mutants displayed sensitivity to the selective FGFR inhibitor erdafitinib and resulted in suppression of FGFR phosphorylation and downstream signal transduction. Clinically, patients receiving erdafitinib showed decreased Erk phosphorylation in tumor biopsies and elevation of serum phosphate. In a phase I study, a heavily pretreated bladder cancer patient with an FGFR3-TACC3 translocation experienced a partial response when treated with erdafitinib. This preclinical study confirmed pharmacodynamics and identified new predictive biomarkers to FGFR inhibition with erdafitinib and supports further clinical evaluation of this compound in patients with FGFR genetic alterations. Mol Cancer Ther; 16(8); 1717-26. ©2017 AACR.

Discovery and Pharmacological Characterization of JNJ-42756493 (Erdafitinib), a Functionally Selective Small-Molecule FGFR Family Inhibitor.

Fibroblast growth factor (FGF) signaling plays critical roles in key biological processes ranging from embryogenesis to wound healing and has strong links to several hallmarks of cancer. Genetic alterations in FGF receptor (FGFR) family members are associated with increased tumor growth, metastasis, angiogenesis, and decreased survival. JNJ-42756493, erdafitinib, is an orally active small molecule with potent tyrosine kinase inhibitory activity against all four FGFR family members and selectivity versus other highly related kinases. JNJ-42756493 shows rapid uptake into the lysosomal compartment of cells in culture, which is associated with prolonged inhibition of FGFR signaling, possibly due to sustained release of the inhibitor. In xenografts from human tumor cell lines or patient-derived tumor tissue with activating FGFR alterations, JNJ-42756493 administration results in potent and dose-dependent antitumor activity accompanied by pharmacodynamic modulation of phospho-FGFR and phospho-ERK in tumors. The results of the current study provide a strong rationale for the clinical investigation of JNJ-42756493 in patients with tumors harboring FGFR pathway alterations. Mol Cancer Ther; 16(6); 1010-20. ©2017 AACR.

Effects of nilotinib on leukaemia cells using vibrational microspectroscopy and cell cloning.

Over the last few years, both synchrotron-based FTIR (S-FTIR) and Raman microspectroscopies have helped to better understand the effects of drugs on cancer cells. However, cancer is a mixture of cells with different sensitivity/resistance to drugs. Furthermore, the effects of drugs on cells produce both chemical and morphological changes, the latter could affect the spectra of cells incubated with drugs. Here, we successfully cloned sensitive and resistant leukaemia cells to nilotinib, a drug used in the management of leukaemia. This allowed both the study of a more uniform population and the study of sensitive and resistant cells prior to the addition of the drug with both S-FTIR and Raman microspectroscopies. The incubation with nilotinib produced changes in the S-FTIR and Raman spectra of both sensitive and resistant clones to nilotinib. Principal component analysis was able to distinguish between cells incubated in the absence or presence of the drug, even in the case of resistant clones. The latter would confirm that the spectral differences between the so-called resistant clonal cells prior to and after adding a drug might reside on those more or less sensitive cells that have been able to remain alive when they were collected to be studied with S-FTIR or Raman microspectroscopies. The data presented here indicate that the methodology of cell cloning can be applied to different types of malignant cells. This should facilitate the identification of spectral biomarkers of sensitivity/resistance to drugs. The next step would be a better assessment of sensitivity/resistance of leukaemia cells from patients which could guide clinicians to better tailor treatments to each individual patient.

Worsening of calcinosis cutis with teriparatide treatment in two osteoporotic patients.

We present two cases of patients with systemic autoimmune diseases (one with dermatomyositis and one with CREST syndrome) who presented with a worsening of calcinosis cutis after treatment of osteoporosis with teriparatide. To our knowledge, this association is not described in the literature and might be considered in the spectrum of adverse reactions to teriparatide.

Phase I Dose-Escalation Study of JNJ-42756493, an Oral Pan-Fibroblast Growth Factor Receptor Inhibitor, in Patients With Advanced Solid Tumors.

PURPOSE: JNJ-42756493 is an orally administered pan-fibroblast growth factor receptor (FGFR) tyrosine kinase inhibitor. This first-in-human study evaluates the safety, pharmacokinetics, and pharmacodynamics and defines the recommended phase II dose (RP2D) of JNJ-42756493. PATIENTS AND METHODS: Eligible patients with advanced solid tumors received escalating doses of JNJ-42756493 from 0.5 to 12 mg administered continuously daily or JNJ-42756493 10 or 12 mg administered intermittently (7 days on/7 days off). RESULTS: Sixty-five patients were enrolled. The most common treatment-emergent adverse events included hyperphosphatemia (65%), asthenia (55%), dry mouth (45%), nail toxicity (35%), constipation (34%), decreased appetite (32%), and dysgeusia (31%). Twenty-seven patients (42%) experienced grade ≥ 3 treatment-emergent adverse events, and one dose-limiting toxicity of grade 3 ALT elevation was observed at 12 mg daily. Maximum-tolerated dose was not defined. Nine milligrams daily was considered as the initial RP2D; however, tolerability was improved with intermittent schedules, and 10 mg administered on a 7-days-on/7-days-off schedule was considered the final RP2D. Pharmacokinetics were linear, dose proportional, and predictable, with a half-life of 50 to 60 hours. Dose-dependent elevations in serum phosphate, a manifestation of pharmacodynamic effect, occurred in all patients starting at 4 mg daily. Among 23 response-evaluable patients with tumor FGFR pathway alterations, four confirmed responses and one unconfirmed partial response were observed in patients with glioblastoma and urothelial and endometrial cancer (all with FGFR2 or FGFR3 translocations); 16 patients had stable disease. CONCLUSION: JNJ-42756493 administered at 10 mg on a 7-days-on/7-days-off schedule achieved exposures at which clinical responses were observed, demonstrated pharmacodynamic biomarker activity, and had a manageable safety profile.

Vibrational spectroscopy in stem cell characterisation: is there a niche?

Vibrational spectroscopy using both infrared and Raman spectroscopies has been used in recent years with the aim to aid clinicians in disease diagnosis. More recently, these techniques have been applied to study stem cell differentiation and to determine stem cell presence in tissues. These studies have demonstrated the potential of these techniques in better characterising stem cell differentiation phenotypes with potential applications in tissue engineering strategies. However, before the translation of vibrational spectroscopy into clinical practice becomes a reality, several issues still need to be addressed. We describe here an overview of the work carried out so far and the problems that might be encountered when using vibrational spectroscopy.

Paciente pediátrico crítico. Cuidados Intensivos Pediátricos, Hospital Infantil La Paz / Critical pediatric patient. Pediatric intensive cares, Hospital Infantil La Paz

Los Cuidados Intensivos Pediátricos (CIPs) iniciaron su actividad en el Hospital Infantil La Paz en el año 1074. El servicio actual dispone de 16 camas, cuatro de cuidados medios, ocho de cuidados intensivos y cuatro de aislamiento con exclusa y filtros HEPA (High Efficiency Particulate Air). En los últimos 6 años (2006-2010) se han atendido 3.674 pacientes, media de 612 pacientes año, con una ocupación media anual del 83,4% y mortalidad media anual del 4,9%. Al ser considerado un servicio polivalente, se ha establecido una estrecha colaboración prácticamente con la totalidad de los servicios y especialidades pediátricas médico-quirúrgicas del hospital, con mayor o menor frecuencia según las patologías asistidas, destacando entre las patologías asistidas, destacado entre las patologías quirúrgicas los pacientes con asistencia ventricular (Berlin Hear), ECMO (Extracorporeal Membrane Oxygenation) y trasplante cardiaco; así como tratamiento psotoepratorio de los trasplantes hepáticos, renales, intestinales y multiviscerales. Entre las líneas de trabajo, hay que destacar el tratamiento intracoronario con células progenitoras autólogas de médula ósea en pacientes con miocardiopatía dilatada e insuficiencia cardíaca; creación de un equipo mixto, médico y de enfermería para el abordaje guiado por ecografía, de accesos venosos centrales insertados periféricamente; registro de una patente europea de válvula de cierre ultrarrápido y sin fugas para respiradores; utilización a nivel pediátrico de la tecnología ventilatoria NAVA (Neurally Adjusted Ventilatory Asist); monitorización hemodinámica con el monitor PiCCO2 (Pulsion Medical System); la promoción de la ecuación médica basada en la simulación de ata fidelidad y la implantación de la asistencia ventricular externa en Pediatría (AU)

The Pediatric Intensive Care Unit of La Paz Universitary Hospital, was founded in 1974. It is now a polyvalent medical-surgical tertiary unit, equipped with 4 intermediate care beds, 8 intensive care beds and 4 isolation beds with airlock and HEPA filters for the treatment of transplanted and immunocompromised patients. In the last 6 years, 3674 patients have been treated (mean 612 patients/year) with a mean occupancy rate of 83,4% and a mortality rate of 4,9&. As the polyvalent unit it is, PICU staff maintains close and daily contact and cooperation with almost all the rest of the medical and surgical departments of the hospital. In this regard Critically ill cardiac patients are admitted in the PICU ( for the treatment with ECMO and ventricular assist devices (EXCOR Berlin Heart), as well as post-operative patients after cardiac, liver, renal, intestinal and multivisceral transplantation. Of all the achievements and current lines of work of the department, it can be highlighted the intracoronary treatment with autologous bone marrow derived progenitor cells in patients with dilated cardiomyopathy, the implantation development of a pediatric ventricular assist program, the echo guided cannulation of peripherally inserted central venous catheter by a especially trained intravenous therapy team, the registration of a European patent for a ultrarapid shutoff respirator valve, the incorporation of NAVA mode for mechanically ventilated patients and the PiCCO2 monitor for the hemodynamic monitorization of patient with cardiovascular dysfunction, and the promotion of medical education based on high-fidelity simulation (AU)

Síndrome de apnea obstructiva del sueño con repercusión sistémica. A propósito de un caso / Obstructive sleep apnea syndrome with systemic repercussion. Case report

Presentamos el caso de un niño de 4 años que ingresa en la unidad de cuidados intensivos pediátricos (UCIP) por mal estado general, dificultad respiratoria severa con débil esfuerzo y ronquido inspiratorio. La auscultación cardiopulmonar muestra hipoventilación y ritmo de galope, tiene hepatomegalia y, desde el punto de vista neurológico, presenta disminución del nivel de conciencia (Glasgow 6/15). En las pruebas complementarias, se observa un daño hipóxico-isquémico generalizado(creatinina sérica de 1,8 mg/dL; GOT de 23.730 UI/L y GPT de 5.771 UI/L; actividad de protrombina del 31% y troponina de1,73 ng/mL). La radiografía de tórax muestra una discreta cardiomegalia y la ecocardiografía hipertensión pulmonar. En la eco-Doppler abdominal se observa una severa hiperecogenicidad cortical renal y hepatomegalia. En el electroencefalograma hay signos de afectación cerebral generalizada, y en la tomografía computarizada (TC) craneal aparecen dos áreas cerebrales sugestivas de infartos isquémicos e hipertrofia adenoidea. A los pocos días del ingreso, se realiza una adenoamigdalectomía. En el momento del alta, los parámetros analíticos son normales, y en la ecocardiografía no se observa hipertensión pulmonar (AU)

We present a case report of a four year old child who is admittedat the PICU due to general bad condition, serious respiratorydifficulty with low effort and inspiratory snoring. The cardiopulmonarauscultation shows hypoventilation and gallop rhythm.He also shows hepatomegaly and awareness reduction at aneurological level (Glasgow 6/15). The complementary testsshow a generalized hypoxic-ischemic injury (serum creatinine of1.8 mg/dL; GOT of 23,730 IUI/L and GPT of 5,771 IU/L; prothrombinactivity 31% and troponin 1.73 ng/mL). Thorax X-rayshows cardiomegaly and the echocardiography pulmonary hypertension.The abdominal echo-doppler shows a serious renalcortical hyperechogenicity and hepatomegaly. At the electroencephalographthere are some signs of generalized cerebraldamage and in the cranial scan there are two cerebral areasthat suggest ischemic infarctions and adenoid hypertrophy. Atonsillectomy is performed some days after admission. At thedischarge, the analytical parameters are normal and there is nopulmonary hypertension (PHT) in the echocardiography (AU)

Dielectric spectroscopy of normal and malignant human lung cells at ultra-high frequencies.

Microwave techniques for biomedical applications aimed at cancer treatment or diagnosis, either by imaging or spectroscopy, are promising. Their use relies on knowledge of the dielectric properties of tissues, especially on a detectable difference between malignant and normal tissues. As most studies investigated the dielectric properties of ex vivo tissues, there is a need for better biophysical understanding of human tissues in their living state. As an essential component of tissues, cells represent valuable objects of analysis. The approach developed in this study is an investigation at cell level. Its aim was to compare human lung normal and malignant cells by dielectric spectroscopy in the beginning of the microwave range, where such information is of substantial biomedical importance. These cells were embedded in small and low-conductivity agarose hydrogels and laid on an open-ended coaxial probe connected to a vector network analyser operated from 200 MHz to 2 GHz. The comparison between normal and malignant cells was drawn using the variation of measured dielectric properties and fitting the measurements using the Maxwell-Wagner equation. Both methods revealed slight differences between the two cell lines, which were statistically significant regarding conductivities of composite gels and cells.

[Pneumococcal conjugate vaccine failure in HIV-infected child. Clinical case]. / Fallo de la vacuna antineumocócica conjugada en un niño VIH positivo. A propósito de un caso.

Despite appropriate antimicrobial therapy and vaccination, invasive pneumococcal infections remain associated with significant mortality, especially in selected high-risk groups (asplenic, humoral immunity deficient patients, etc.). We present a 13-year-old caucasian boy with HIV infection (vertical transmission). He received treatment with highly-active antiretroviral therapy (amprenavir, lamivudine and zidovudine) and vaccination with 23-valent vaccine (6 years old) and 7-valent pneumococcal conjugate vaccine (10 years old). His CD4 count and his viral load at these times were 2,063/microl and 13461 cop/ml, when he was 6 years old and 1,315/microl and 32400 cop/ml when he was 10 years old, respectively. The latest CD4 count (1,000/microl) and his viral load (3800 cop/ml) confirmed satisfactory control of the disease. He was referred to our emergency department presenting with fever, head and stomach-ache and vomiting. In the following hours his condition continued to deteriorate and depressed level of consciousness and meningismus were observed. Streptococcus pneumoniae, serotype 18 C, was detected in blood and cerebrospinal fluid cultures. Despite appropriate treatment with antibiotics (cefotaxime and vancomycin) and anti-oedema medications, brain-death was confirmed 24 hours after his admittance.

Fallo de la vacuna antineumocócica conjugada en un niño VIH positivo. A propósito de un caso / Pneumococcal conjugate vaccine failure in HIV-infected child

La infección neumocócica sigue asociada con una importante mortalidad, especialmente en grupos de riesgo (esplenectomizados, déficit de la inmunidad humoral, etc.) a pesar de vacunas y antibióticos adecuados. Presentamos a un niño de 13 años VIH positivo por transmisión vertical tratado con triple terapia (amprenavir, lamivudina y zidovudina). Fue vacunado con vacuna 23-valente a los 6 años y conjugada heptavalente a los 10 años de edad. El recuento de células CD4 y su carga viral a los 6 años eran de 2.063/μl y 13.461 copias/ml, respectivamente. A los 10 años el recuento de CD4 y su carga viral eran de 1.315/μl y 32.400 copias/ml, respectivamente. El último recuento de CD4 (1.000/μl) y la carga viral (3.800 copias/ml) confirmaban un buen control de la enfermedad 15 días antes del ingreso. Acude a urgencias por fiebre, dolor abdominal y vómitos. Hay un progresivo deterioro del nivel de conciencia y signos meníngeos. En el hemocultivo y en el cultivo de LCR crece Streptococcus pneumoniae serotipo 18C, y es tratado con cefotaxima y vancomicina, así como medidas antiedema cerebral, pero evoluciona a muerte cerebral en 24 h (AU)

Despite appropriate antimicrobial therapy and vaccination, invasive pneumococcal infections remain associated with significant mortality, especially in selected high-risk groups (asplenic, humoral immunity deficient patients, etc.). We present a 13-year-old caucasian boy with HIV infection (vertical transmission). He received treatment with highly-active antiretroviral therapy (amprenavir, lamivudine and zidovudine) and vaccination with 23-valent vaccine (6 years old) and 7-valent pneumococcal conjugate vaccine (10 years old). His CD4 count and his viral load at these times were 2,063/μl and 13461 cop/ml, when he was 6 years old and 1,315/μl and 32400 cop/ml when he was 10 years old, respectively. The latest CD4 count (1,000/μl) and his viral load (3800 cop/ml) confirmed satisfactory control of the disease. He was referred to our emergency department presenting with fever, head and stomach-ache and vomiting. In the following hours his condition continued to deteriorate and depressed level of consciousness and meningismus were observed. Streptococcus pneumoniae, serotype 18 C, was detected in blood and cerebrospinal fluid cultures. Despite appropriate treatment with antibiotics (cefotaxime and vancomycin) and anti-oedema medications, brain-death was confirmed 24 hours after his admittance (AU)

Estimating and correcting mie scattering in synchrotron-based microscopic fourier transform infrared spectra by extended multiplicative signal correction.

We present an approach for estimating and correcting Mie scattering occurring in infrared spectra of single cells, at diffraction limited probe size, as in synchrotron based microscopy. The Mie scattering is modeled by extended multiplicative signal correction (EMSC) and subtracted from the vibrational absorption. Because the Mie scattering depends non-linearly on alpha, the product of the radius and the refractive index of the medium/sphere causing it, a new method was developed for estimating the Mie scattering by EMSC for unknown radius and refractive index of the Mie scatterer. The theoretically expected Mie contributions for a range of different alpha values were computed according to the formulae developed by Van de Hulst (1957). The many simulated spectra were then summarized by a six-dimensional subspace model by principal component analysis (PCA). This subspace model was used in EMSC to estimate and correct for Mie scattering, as well as other additive and multiplicative interference effects. The approach was applied to a set of Fourier transform infrared (FT-IR) absorbance spectra measured for individual lung cancer cells in order to remove unwanted interferences and to estimate ranges of important alpha values for each spectrum. The results indicate that several cell components may contribute to the Mie scattering.

Maximal lipidic power in high competitive level triathletes and cyclists.

OBJECTIVE: To describe the fat-oxidation rate in triathlon and different modalities of endurance cycling. METHODS: 34 endurance athletes (15 male triathletes, 4 female triathletes, 11 road cyclists and 4 male mountain bikers) underwent a progressive cycloergometer test until exhaustion. Relative work intensity (VO(2max)), minimal lactate concentration (La(-)(min)), lactic threshold, individual lactic threshold (ILT), maximal fat-oxidation rate (Fat(max), Fat(max) zone) and minimal fat-oxidation rate (Fat(min)) were determined in each of the groups and were compared by means of one-way analysis of variance. RESULTS: No significant differences were found for Fat(max), Fat(min) or for the Fat(max) zone expressed as fat oxidation rate (g/min). Intensities -20%, -10% and -5% Fat(max) were significantly lower for mountain bikers with respect to road cyclists and female triathletes, expressed as % VO(2max). Intensities 20%, 10% and 5% Fat(max) were significantly lower for mountain bikers with respect to male triathletes and female triathletes, and for male triathletes in comparison with female triathletes, expressed as % VO(2max). Lactic threshold and La(-)(min) did not show significant differences with respect to Fat(max). Lactic threshold was found at the same VO(2max) with respect to the higher part of the Fat(max) zone, and La(-)(min) at the same VO(2max) with respect to the lower part of the Fat(max) zone. CONCLUSIONS: The VO(2max) of Fat(max) and the Fat(max) zone may explain the different endurance adaptations of the athletes according to their sporting discipline. Lactic threshold and La(-)(min) were found at different relative work intensities with respect to those of Fat(max) even though they belonged to the Fat(max) zone.