Enhancing chlamydospore production in Duddingtonia flagrans on solid substrate: The impact of mannitol and varied cultivation conditions.

Duddingtonia flagrans is a nematophagous fungus which has shown promising results as a non-chemical parasitic control tool. The fungus disrupts the parasite's life cycle by trapping larvae in the environment through the networks generated from chlamydospores, thus preventing the reinfection of animals. One barrier to the development of a commercial product using this tool is the need to increase chlamydospore production in the laboratory for its administration to livestock. The purpose of this study was to evaluate the addition of mannitol to an enriched culture medium and the effect of adverse cultivation conditions on chlamydospore production. D. flagrans was cultivated on Petri dishes with corn agar for 4 weeks at 27 °C and 70% relative humidity (RH). Four groups were then formed, all with Sabouraud agar as a base, to which different growth inducers were added: GSA (glucose Sabouraud agar), GSA-MI (glucose Sabouraud agar + meso inositol), GSA-E (enriched glucose Sabouraud agar), and AE-M (enriched agar + mannitol). After 4 weeks, chlamydospores were recovered by washing the surface of each plate with distilled water and then quantified. The medium that yielded the highest amount of chlamydospores was subjected to different cultivation conditions: NC (normal conditions): 70% RH and 27 °C, AC (adverse conditions) 1: 20% RH and 40 °C, CA2: 60% RH and 27 °C, and CA3: 55% RH and 24 °C. It was determined that mannitol increases chlamydospore production (65x106 chlamydospores/plate), and when reducing humidity by 10% under cultivation conditions it resulted in an approximately 10% increase in chlamydospore production compared to the control group. These results suggest that the addition of polyols, as well as its cultivation under certain environmental conditions, can improve chlamydospore production on a laboratory scale.

Oral microbiome and systemic antineoplastics in cancer treatment: A systematic review.

BACKGROUND: Oral mucositis is one of the most common side effects in cancer patients receiving systemic antineoplastics. However, the underlying biological mechanisms leading to this condition are still unclear. For this reason, it has been hypothesised that systemic antineoplastics may cause an imbalance on the oral microbiota that subsequently triggers oral mucosa damage. MATERIAL AND METHODS: A systematic review was performed following the PRISMA protocol and the PICO question established was: patients diagnosed with cancer, who are candidates for receiving systemic antineoplastics (P=Patients), that undergo oral microbiome determinations (I=Intervention), before and after systemic antineoplastics administration (C=Comparison), to analyse changes in the oral microbiome composition (O=Outcome). The bibliographic search was carried out in PubMed and other scientific repositories. RESULTS: Out of 166 obtained articles, only 5 met eligibility criteria. Acute myeloid leukaemia (AML) was the most frequent type of cancer (40 %) among the participants. Only one of the studies included a control group of healthy subjects. Heterogeneity in the protocols and approaches of the included studies hindered a detailed comparison of the outcomes. However, it was stated that a decrease in bacteria α diversity is often associated with oral mucositis. On the other hand, fungal diversity was not associated with oral mucositis although α diversity was lower at baseline on patients developing oral candidiasis. CONCLUSIONS: There is insufficient scientific evidence of oral microbiological changes in patients undergoing systemic antineoplastics. Further investigations ought to be carried out to identify microorganisms that might play a key role in the pathogenesis of oral mucosa damage in patients undergoing systemic antineoplastics.

Increased intraocular pressure secondary to retinal detachment.

Four clinical cases of patients with rhegmatogenous retinal detachment (RRD) associated with elevated intraocular pressure (IOP) are described. All the patients were men who came to the emergency service with floaters, all presented mild to moderate inflammation in the anterior chamber and increased intraocular pressure greater than 30 mmHg. All were diagnosed of RRD. This demonstrates that a RRD does not always present low IOP. In these cases of clinical retinal pathology and high IOP, we must carefully examine the anterior chamber and the fundus of the eye so that an associated retinal detachment does not remain unnoticed, and can be treated immediately. This association of elevated IOP and RRD is called as Schwartz-Matsuo Syndrome.1.

Electronic excitations of C60 aggregates.

Excitation properties of the isolated C(60) and (C(60))(N) model clusters (N = 2, 3, 4, 6 and 13) are studied using an a priori parameterized and self-consistent Hamiltonian, the Complete Neglect of Differential Overlap considering the l azimuthal quantum number method. This method properly describes electron excitations of the isolated C(60) after the configuration interaction of singles (CIS) procedure, when those are compared with experimental data in n-hexane solution and in a molecular beam. Geometry models of (C(60))(N) clusters to model the effect of aggregation were obtained from the fullerene fcc crystal. Some peaks in the low energy edge of the absorption spectrum appear corresponding to clustering effects, as well as small increases of bandwidths in the strong bands at the UV region. An analysis of the theoretical absorption spectrum for dimer models has been carried out, taking into account the influence of the distance between fullerene centers. The density of states of CIS for fullerene clusters in the range from 2.0 to 6.5 eV shows the possibility of electron transitions as functions of the size of the clusters.

Expression of the chemokine N51/KC in the thymus and epidermis of transgenic mice results in marked infiltration of a single class of inflammatory cells.

Transgenic mice expressing the chemokine N51/KC in thymus, skin, and tongue showed a marked infiltration of a single class of inflammatory cells (neutrophils) in the sites of transgene expression. In the thymus, neutrophils were most numerous in the cortex and juxta-medullary regions, often forming aggregates or clusters. A similar, but less intense, neutrophilic infiltrate occurred in close proximity to the epidermal basal layer of the tongue and skin. No morphologic evidence of injury was observed in the thymus, skin, or tongue of these transgenic mice, indicating that N51/KC expression induces recruitment but not inflammatory activation of neutrophils. The lack of activation in the thymus resulted in a large senescent neutrophilic population that was phagocytosed by thymic macrophages and epithelial-reticular cells. These results indicate that N51/KC is a neutrophil chemoattractant in vivo and establish these transgenic mice as effective models to study the phenomena of recruitment and clearance of neutrophils, events that are critical for the initiation and resolution of the inflammatory response.

Control of acetylcholinesterase gene expression during myogenesis.

Acetylcholinesterase (AChE) and the nicotinic acetylcholine receptor show enhanced expression upon differentiation of C2-C12 myoblasts into myotubes. Northern blots and RNAase protection show substantial increases in the mRNAs encoding both proteins. In contrast to transcriptional control of gene expression of acetylcholine receptor subunits during differentiation, Ache mRNA levels increase through stabilization of the transcript. The transcriptional rate remains unchanged during Ache mRNA accumulation, whereas transcription rate increase for the receptor subunits. Inhibition of protein synthesis produces a dramatic superinduction of Ache mRNA in undifferentiated cells without affecting transcription rates, suggesting the presence of labile proteins that uniquely destabilize the mRNA. Hence, Ache mRNA in myoblasts shows rapid turnover, and upon differentiation, its level is increased by slowing the rate of mRNA degradation.

Neutrophil infiltration, glial reaction, and neurological disease in transgenic mice expressing the chemokine N51/KC in oligodendrocytes.

Chemokines (pro-inflammatory chemoattractant cytokines) are expressed in pathological conditions of the central nervous system (CNS). Previous studies suggested that the CNS is relatively resistant to leukocyte diapedesis after chemokine injection, leaving their functional role unresolved. The CNS function of N51/KC, a neutrophil-selective chemokine, was addressed by expressing N51/KC under control of the myelin basic protein (MBP) promoter in transgenic (tg) mice (MBP-N51/KC mice). CNS-specific N51/KC expression produced remarkable neutrophil infiltration into perivascular, meningeal, and parenchymal sites, demonstrating that this chemokine exerts the multiple functions in vivo required to recruit leukocytes into the CNS. MBP-N5 1/KC mice represent an incisive model for the molecular dissection of neutrophil entry into the CNS. Unexpectedly, MBP-N51/KC mice developed a neurological syndrome of pronounced postural instability and rigidity at high frequency beginning at 40 days of age, well after peak chemokine expression. 68/182 mice in one tg fine were found dead before one year of age, with prominent neurological symptoms premortem in 26 (38%). Florid microglial activation and blood-brain barrier disruption without dysmyelination were the major neuropathological alterations. Late-onset neurological symptoms in MBP-N51/KC mice may indicate unanticipated consequences of CNS chemokine expression.

The proteoglycan decorin is synthesized and secreted by differentiated myotubes.

Proteoglycans (PGs) are important components of the skeletal muscle extracellular matrix (ECM). Skeletal muscles are composed of muscle fibers and mononucleated cells. The latter are known to synthesize and secrete several PGs. Rat skeletal muscle ECM contains a chondrotin/dermatan sulfate PG which was immunoprecipitated by antibodies against rat decorin. The synthesis and secretion of PGs by a mouse cell line was analyzed during in vitro differentiation. PGs were characterized by biochemical and immunological techniques including immunocytolocalization experiments. At least three different PGs are synthesized and secreted by differentiated myotubes: a 220 to 460 kDa heparan sulfate, a 250 to 310 kDa chondroitin/dermatan sulfate, and a 75 to 130 kDa chondroitin/dermatan sulfate. This latter PG was specifically immunoprecipitated with antibodies against rat fibroblast decorin. Indirect immunocytolocalization analysis revealed that decorin was localized inside the cells, with a strong reaction around the nuclei. During differentiation the relative proportions of some PGs changed. Thus, a decrease in the relative proportion of the heparan sulfate PG was observed, whereas a significant increase in the relative proportion of decorin was detected. No change in the large chondroitin/dermatan PG was seen during the differentiation process. The possible cell sources of decorin found in rat skeletal muscle ECM are discussed.

Amphiphilic behavior of a brain tetrameric acetylcholinesterase form lacking the plasma membrane anchoring domain.

We have studied the behavior of a mammalian brain tetrameric acetylcholinesterase (AChE) form released by proteinase K from a crude membrane fraction of bovine caudate nucleus. The solubilization of active AChE indicated the presence of a protease-sensitive site in the anchored protein. Unexpectedly, the solubilized AChE maintained its capacity to form aggregates in detergent-free gradients. We demonstrate here that this property was due neither to the presence of the hydrophobic membrane-anchoring domain still linked to the enzyme, nor to the presence of AChE activity trapped in small plasma membrane vesicles. Moreover, we found that the proteinase K-treated extract, devoid of AChE activity, induced the aggregation of purified hydrophilic AChE which usually does not form aggregates. Our results suggest the presence of an AChE aggregating factor in bovine brain extracts prepared in the presence of proteinase K. It is possible that this aggregation may reflect a process of AChE clustering on neurons.

Monomeric amphiphilic forms of acetylcholinesterase appear early during brain development and may correspond to biosynthetic precursors of the amphiphilic G4 forms.

We have studied the development of mouse brain acetylcholinesterase (AChE). Only tetrameric (G4) and monomeric (G1) forms were detected both in vivo and in vitro. The amphiphilic G4 form increased continuously during development, whereas an amphiphilic G1 form appears transiently around embryonic day 17. A causal relationship between the monomers and tetramers was established using pulse-chase experiments with paraoxon, a reversible AChE inhibitor. We report here, for the first time, the presence of an amphiphilic monomer possibly involved in the assembly of the amphiphilic G4 AChE form during mouse brain development.

A membrane-associated dimer of acetylcholinesterase from Xenopus skeletal muscle is solubilized by phosphatidylinositol-specific phospholipase C.

The susceptibility to phosphatidylinositol-specific phospholipase C of the membrane associated acetylcholinesterase (AChE) forms of Xenopus laevis skeletal muscle was examined. This treatment released almost all the detergent-soluble AChE species from muscle homogenates. Sucrose gradient analysis showed that the released acetylcholinesterase form corresponds to a hydrophilic G2 dimer, indicating that this dimer has a glycolipid anchoring domain which contains phosphatidylinositol.

Structure and regulation of expression of the acetylcholinesterase gene.

Acetylcholinesterase, an enzyme essential for the termination of the action of acetylcholine, is encoded by a single gene. Alternative mRNA processing gives rise to the expression of enzyme forms with three distinct carboxyl-termini. These structural differences govern the cellular disposition of the expressed enzyme but do not influence catalytic activity. Alternative polyadenylation signals give rise to distinct 3' non-coding regions which are likely to affect mRNA stability. Alternative splicing also occurs at the 5' end of the gene where two promoter regions can be identified. Hence, regulation of expression of the gene occurs at 3 levels, transcriptional through alternative promoters, translational by affecting mRNA stability and processing of distinct mRNAs and post-translationally by giving rise to distinct peptide chains which are processed differently. Recombinant DNA studies have also been extended to modifying protein structure through site-specific mutagenesis and studying the function of the mutant enzymes.

Electric lead field for a piecewise homogeneous volume conductor model of the head.

A new method is presented for computing the electric lead field of a realistic head shape model which has piecewise homogeneous conductivity. The basic formulae are derived using the well-known reciprocity theorem. Previously described methods are also based upon this theorem, but these first calculate the electric potential inside the head by a scalar boundary element method (BEM), and then approximate the ohmic current density by some sort of gradient. In contrast, this paper proposes the direct evaluation of the ohmic current density by discretizing the vector Green's second identity which leads to a vector version of BEM. This approach also allows the derivation of the same equations for the three concentric spheres model as obtained by Rush and Driscoll [8]. The results of simulations on a spherical head model indicate that the use of a vector BEM leads to an improvement of accuracy in the computation of the ohmic current density with respect to those reported previously, in term of different measures of error.

[Feasibility and safety of intracoronary ultrasound. Experience of a single center]. / Viabilidad y seguridad del empleo de la ecografía intracoronaria. Experiencia de un centro.

BACKGROUND AND OBJECTIVES: Intracoronary ultrasound provides a number of advantages in the quantification and characterization of coronary stenoses with regard to contrast angiography. However, previous studies have reported a 3.5 to 11% complication rate, and a 10-30% failure rate in performing this technique. The purpose of the study is to analyze the feasibility of performing intracoronary ultrasound and the incidence of complications associated with the use of contemporary, state of the art equipment. MATERIAL AND METHODS: The feasibility of performing intracoronary ultrasound, analyzed as the percentage of successes and failures in performing the examination was reviewed, as well as the complication rate associated with the technique in all the procedures carried out between July 1, 1994 and February 29, 1996 in which intravascular ultrasound was attempted. Complications were categorized as related, non-related and uncertainly related to the ultrasound study. RESULTS: 239 vessels were studied with intravascular ultrasound in 209 procedures (74% interventional) performed on 139 patients. Ultrasound examination was feasible in all the diagnostic studies and in 96% of the interventional procedures. The major and minor procedural complication rate was 2.4 and 10.5% respectively. No major complication was related to the ultrasound examination. Three patients experienced minor complications (1.4%) related to the ultrasound study. All three complications occurred in baseline studies during interventional procedures. CONCLUSIONS: Intracoronary ultrasound is feasible and safe in the vast majority of the procedures. Improvements in smaller catheter size and design and larger operator expertise have significantly reduced the complication rate, particularly the most frequent coronary spasm so far. Complications are associated with baseline studies during interventional procedures and with less operator expertise.

[Comparison of optic nerve head measurements obtained with the slit lamp and with CSLO]. / Comparación de las medidas de la papila obtenidas con biomicroscopia y con oftalmoscopia de láser confocal.

PURPOSE: To evaluate the accuracy of disk size and cup/disk ratio (C/D) estimation by measuring the vertical diameter and cup/disk ratio at the slit lamp. METHODS: Two hundred and thirty eight eyes of 185 people, 82 glaucomas and 156 normal subjects were included. All underwent slit lamp examination, tonometry, optic nerve photographs, scanning laser tomography (HRT) and standard perimetry (Humphrey). Correlation between slit lamp measurements (vertical diameter and C/D ratio) and HRT parameters was evaluated. RESULTS: There is a significant correlation between the disk diameter measured at the slit lamp and the disk area measured with HRT (Pearson Test, r: 0.52, p<0.001). Additionally, there is significant correlation between the semi-quantitative estimation of C/D ratio at the slit lamp and the C/D area ratio from the HRT (r: 0.83, p<0.001). CONCLUSION: Measuring the vertical diameter and the C/D ratio at the slit lamp is a useful method for estimating the real disk area and C/D ratio. It is highly recommended that disk size be estimated when attempting to interpret clinical findings in the optic nerve head.

Elevación de la presión intraocular secundaria a desprendimiento de retina / Increased intraocular pressure secondary to retinal detachment

Se describen 4 casos clínicos de pacientes con desprendimiento de retina regmatógeno (DRR) asociado a elevación de presión intraocular (PIO) en el momento del diagnóstico del DRR. Todos los pacientes eran hombres que acudieron a urgencias por miodesopsias, presentaban inflamación leve a moderada en cámara anterior y elevación de la presión intraocular mayor de 30mmHg. En todos ellos se detectó la presencia de DRR. Esto demuestra que los desprendimientos de retina no siempre presentan la PIO baja, y es compatible con valores de PIO alta. En esos casos de enfermedad retiniana y PIO elevada, se debe examinar minuciosamente la cámara anterior y el fondo de ojo para que no pase desapercibido un desprendimiento de retina que requiere tratamiento inmediato. Esta asociación de una PIO elevada y un DRR se denomina síndrome de Schwartz-Matsuo (AU)

Four clinical cases of patients with rhegmatogenous retinal detachment (RRD) associated with elevated intraocular pressure (IOP) are described. All the patients were men who came to the emergency service with floaters, all presented mild to moderate inflammation in the anterior chamber and increased intraocular pressure greater than 30mmHg. All were diagnosed of RRD. This demonstrates that a RRD does not always present low IOP. In these cases of clinical retinal pathology and high IOP, we must carefully examine the anterior chamber and the fundus of the eye so that an associated retinal detachment does not remain unnoticed, and can be treated immediately. This association of elevated IOP and RRD is called as Schwartz-Matsuo Syndrome (AU)

Computer simulation of elastic constants of hydroxyapatite and fluorapatite.

Hydroxyapatite (HAP) and fluorapatite (FAP) are essential components of dental enamel and bone. In this paper, we report a computational study of the elastic properties of HAP and FAP using ab initio and force field techniques. We have obtained the HAP and FAP elastic stiffness constants in hexagonal symmetry by fitting the Hooke law for both the energy-strain and stress-strain relations. Our ab initio HAP stiffness constants differ from the results of previous calculations, but follow similar trends. The HAP and FAP stiffness constants calculated with the ab initio method are very similar, although FAP is slightly stiffer than HAP in the hexagonal plane, and more compliant along the hexagonal axis. The pseudo-single-crystal HAP experimental stiffness constants in current use are critically reviewed. Combining the data from the ab initio simulations with the experimental FAP stiffness constants, several alternative sets of HAP stiffness constants are proposed. The mismatch in properties between HAP and FAP is evidently too small to assume it to be directly responsible for dental enamel mechanical degradation with fluorosis disease.

Expressions for the fractional modification in different monocyclic enzyme cascade systems: analysis of their validity tested by numerical integration.

This paper presents the derivation, under a minimal set of assumptions, of a general expression for the steady-state fractional modification of an interconvertible protein involved in four different schemes of monocyclic enzyme cascade systems. From this general expression we derive, as particular cases, other, simpler expressions by applying additional assumptions and which have, therefore, a smaller range of validity. Some of these particular expressions coincide with those already obtained in previous contributions on individualised analyses. We discuss the relationships between the kinetic parameters and the concentrations needed for the fulfilment of the additional assumptions. The goodness of the analysis was tested by reference to the shape in the steady-state of the simulated time progress curves obtained by numerical integration.

Characterization of a tetrameric G4 form of acetylcholinesterase from bovine brain: a comparison with the dimeric G2 form of the electric organ.

Globular forms (G forms) of acetylcholinesterase (AChE) are formed by monomers, dimers and tetramers of the catalytic subunits (G1, G2 and G4). In this work the hydrophobic G2 and G4 AChE forms were purified to homogeneity from Discopyge electric organ and bovine caudate nucleus and studied from different points of view, including: velocity sedimentation, affinity to lectins and SDS-polyacrylamide gel electrophoresis under reducing and non-reducing conditions. The polypeptide composition of Discopyge electric organ G2 is similar to Torpedo, however the pattern of the brain G4 AChE is much complex. Under non-reducing conditions the catalytic subunit possesses a molecular weight of 65 kDa, however this value increases to 68 kDa after reduction, suggesting that intrachain-disulfide bonds are important in the folding of the catalytic subunits of the AChE. Also it was found that after mild proteolysis; the (125I)-TID-20 kDa fragment decreased its molecular weight to approximately 10 kDa with little loss of AChE activity. Finally, we suggest a model for the organization of the different domains of the hydrophobic anchor fragment of the G4 form.