The ultrastructure of infective larvae (L3) of Wuchereria bancrofti after treatment with diethylcarbamazine

Although the large use of diethylcarbamazine (DEC), as the major anti-filaricide drug, its mechanism of action remains a matter of controversy. Several authors defend the hypothesis that DEC has no direct effect on nematodes. This study demonstrated that infective larvae (L3) of Wuchereria bancrofti treated in vitro with DEC presented several behaviour and morphological changes. The first alteration produced by treatment for 2 h with 3, 5, 10 microg/ml of DEC was the reduction of motility. Larvae treated with 5, 10 microg/ml DEC showed severely affected organelles, formation of several vacuoles, mainly in neurocytes and in the muscle cells, and dissolution of cytoplasm. Some larvae showed extreme cellular disorganization with abundance of large and dense mitochondria and numerous large vacuoles containing residual organelles. Lamellar bodies, probably related to an assembly of hipodermal membranes, were also observed in some damaged larvae. Thus, undoubtedly in vitro treatment with concentrations of DEC similar to therapeutic conditions, which are 1-5 microg/ml (Hawking, 1979), had a direct effect on infective larvae of W. bancrofti by causing, primarily neuromuscular alterations with subsequent damage to organelles.     

Some morphological aspects of Wuchereria bancrofti uterus after treatment with diethylcarbamazine

Confocal and EM analyses revealed that some female Wuchereria bancrofti, obtained from volunteers that received recommended diethylcarbamazine dose regimens, showed few or no embryos. Furthermore, inside the gravid uterus of female W. bancrofti treated with DEC we observed a finely granular, electron-dense material organised as strings of pearls, approximately 70 nm in maximal length surrounding intra-uterine microfilariae and apparently secreted by the embryo. Over the eggshells a similar material was also observed, possibly secreted by the uterine wall. The surface of intra-uterine microfilariae presented a material with identical electron-density to the scattered structures observed inside the egg. Similarly, the sheath of blood microfilariae of W. bancrofti also showed electron-dense projections, with shape and size similar to that observed inside the uterus.     

高功率毫米波辐照对秀丽线虫行为学和细胞形态学的影响

 采用角锥喇叭天线,使用不同输出平均功率的毫米波激光辐照秀丽线虫,采用实时显微摄像系统记录线虫行为学变化,并使用体视显微镜观察线虫细胞形态学变化。受照射后自由活动的秀丽线虫能够快速逃离高功率毫米波的辐照区域。当受输出平均功率为10 W和12 W辐照后,线虫先后出现运动速度增加、运动速度减慢直至身体僵直不动的变化过程。当输出平均功率为5 W时,线虫出现热耐受现象。细胞形态学观察结果显示:辐照后的线虫腹中卵细胞核结构出现不同程度的异常。高功率毫米波辐照可显著影响线虫的行为学特征及细胞形态学,同时说明秀丽线虫是可适用于高功率毫米波生物效应研究的模式生物之一。     

Molecular and genomic characterization of pathogenic traits of group A Streptococcus pyogenes

Group A streptococcus (GAS) or Streptococcus pyogenes causes various diseases ranging from self-limiting sore throat to deadly invasive diseases. The genome size of GAS is 1.85–1.9 Mb, and genomic rearrangement has been demonstrated. GAS possesses various surface-associated substances such as hyaluronic capsule, M proteins, and fibronectin/laminin/immunoglobulin-binding proteins. These are related to the virulence and play multifaceted and mutually reflected roles in the pathogenesis of GAS infections. Invasion of GAS into epithelial cells and deeper tissues provokes immune and non-immune defense or inflammatory responses including the recruitment of neutrophils, macrophages, and dendritic cells in hosts. GAS frequently evades host defense mechanisms by using its virulence factors. Extracellular products of GAS may perturb cellular and subcellular functions and degrade tissues enzymatically, which leads to the aggravation of local and/or systemic disorders in the host. In this review, we summarize some important cellular and extracellular substances that may affect pathogenic processes during GAS infections, and the host responses to these.     

Fluorescence resonance energy transfer: A promising tool for investigation of the interaction between 1-anthracene sulphonate and serum albumins

This present investigation has revealed that steady state as well as time-resolved fluorescence techniques can serve as highly sensitive monitors for exploring the interaction of fluorescent probe 1-anthracene sulphonate (1-AS) with model transport proteins, bovine serum albumin (BSA) and human serum albumin (HSA).We have focused on fluorescence resonance energy transfer (FRET) between excited tryptophan in transport proteins to 1-AS, for the study of relaxation dynamics of biological molecules.     

Immunolocalization of an osteopontin-like protein in dense granules of Toxoplasma gondii tachyzoites and its association with the parasitophorous vacuole

Toxoplasma gondii is an apicomplexan parasite infecting a broad host range, including humans. The parasite invades host cell by active penetration with the participation of its secretory organelles proteins during this process. Until now, only a limited number of secretory proteins have been discovered, and the effectors molecules involved in parasite invasion and survival are not well understood. Osteopontin (OPN) is a multifunctional glycophosphoprotein, secreted by different cell types, which is involved in various physiological and pathological events including cell signaling and survival. For the first time we demonstrated in this work by immunofluorescence and immunoelectron microscopy approaches the localization of an OPN-like protein in dense granules of extracellular T. gondii tachyzoites. Western blotting and RT-PCR confirmed this protein expression by the parasites. Our results also showed, after macrophage invasion, an intense positive labeling for OPN-like protein at the sub-apical portion of tachyzoites, the site of dense granules secretion, and the localization of this protein at the parasitophorous vacuole membrane. These data suggest that dense granules secrete an OPN-like protein, and we speculate that this protein participates during the parasite interaction process with host cells and parasitophorous vacuole formation.     

Fish host-cestode parasite stable isotope enrichment patterns in marine, estuarine and freshwater fishes from Northern Canada

Cestode parasites from freshwater (threespine stickleback, Gasterosteus aculeatus), estuarine (brook charr, Salvelinus fontinalis) and marine (Greenland cod, Gadus ogac) fish from northern Quebec, Canada, were used to investigate the hypotheses that cestode parasites are (13)C and (15)N enriched relative to host food sources, but (15)N depleted with respect to host muscle tissue as a result of differential enrichment during the assimilation of common nutrient sources. Cestode parasites and fish were generally similarly enriched in (13)C with respect to common food sources and, in the case of Greenland cod, cestode parasites were (13)C enriched relative to host tissue. Cestode parasites were also generally (15)N enriched with respect to mean host dietary signatures, but depleted with respect to host muscle tissue. In the case of Greenland cod cestode parasites, no significant (15)N enrichment relative to host dietary signature was observed. Cestode parasites appear generally to experience smaller (15)N enrichment than hosts as a result of trophic transfer of common dietary sources. Differential (15)N enrichment patterns in parasites and fish may be attributed to differences in parasite and host metabolism, particularly the anaerobic and aerobic natures of their respective metabolisms. Results imply that isotope enrichment paradigms developed for the study of aquatic foodwebs cannot be routinely applied to quantitatively assess the role of parasites in aquatic foodwebs and that reference to host muscle tissue measures will not allow accurate characterization of parasite foodweb position. Appropriate reference to assimilated food sources is required to accurately estimate parasite isotopic enrichment patterns and to determine parasite trophic position relative to the host.     

Intramolecular motions in proteins and model polymers on Moessbauer Spectroscopy and Rayleigh Scattering of Moessbauer Radiation data

In the survey, the scales and correlation times of motion on different levels are considered: iron atoms in active centre of myoglobin and hemoglobin, iron containing core in iron storage proteins and model polymer systems, and the average fragmental motion of a protein globule of myoglobin and human serum albumin. The models of intramolecular protein mobility have been drawn.     

Laser fluorimetry of proteins containing one and two tryptophan residues

The true molecular photophysical parameters’ values of tryptophan residues in single-tryptophan-containing and two-tryptophan-containing proteins (by the example of human serum albumin and bovine serum albumin) have been determined for the first time with the use of the authors’ suggested algorithm. The algorthm is based on the simultaneous use of nonlinear and kinetic-laser fluorimetry. The obtained result opens up new approaches in the monitoring of living systems and other objects, containing single fluorophores and localized pairs of fluorophores.     

Investigating passive immunodiffusion reactions in the channels of a microfluidic device

The passive immunodiffusion reaction in a microfluidic system is investigated via the interaction between fluorescein human albumin conjugate and monoclonal antibodies. The topology of a microfluidic device suitable for this reaction was designed, and the device was fabricated using LIGA technology. A system for transitioning from the macro- to the microlevel is developed and produced. The dissociation constants and specificity of interaction in complexes formed as a result of the reaction between fluorescein human albumin conjugates and monoclonal antibodies are determined. The results from our study reveal that systems based on combinations of passive immunodiffusion reactions and microfluidic technologies are applicable for detecting antigens and antibodies in the analyzed fluids.     

An elegant nano-injection machinery for sabotaging the host: Role of Type III secretion system in virulence of different human and animal pathogenic bacteria

Various Gram-negative bacteria possess a specialized membrane-bound protein secretion system known as the Type III secretion system (T3SS), which transports the bacterial effector proteins into the host cytosol thereby helping in bacterial pathogenesis. The T3SS has a special needle-like translocon that can sense the contact with the host cell membrane and translocate effectors. The export apparatus of T3SS recognizes these effector proteins bound to chaperones and translocates them into the host cell. Once in the host cell cytoplasm, these effector proteins result in modulation of the host system and promote bacterial localization and infection. Using molecular biology, bioinformatics, genetic techniques, electron microscopic studies, and mathematical modeling, the structure and function of the T3SS and the corresponding effector proteins in various bacteria have been studied. The strategies used by different human pathogenic bacteria to modulate the host system and thereby enhance their virulence mechanism using T3SS have also been well studied. Here we review the history, evolution, and general structure of the T3SS, highlighting the details of its comparison with the flagellar export machinery. Also, this article provides mechanistic details about the common role of T3SS in subversion and manipulation of host cellular processes. Additionally, this review describes specific T3SS apparatus and the role of their specific effectors in bacterial pathogenesis by considering several human and animal pathogenic bacteria.     

Fluorescence and Ultraviolet Spectroscopic Evaluation of Phenolic Compounds,Antioxidant and Binding Activities in Some Kiwi Fruit Cultivars

Spectrophotometric assays (Folin–Ciocalteu, ferric-reducing/antioxidant power, cupric-reducing antioxidant capacity, and β-carotene linoleate model system) have been used to evaluate the total phenolics and flavonoids in kiwi fruit cultivars in ethanol and water extracts. Fluorescent and infrared measurements were correlated with the obtained spectroscopic data. It was found that the contents of the bioactive compounds and the level of antioxidant activity in different extracts differ significantly (p < 0.05). Bioactive compounds and antioxidant activities were significantly higher in two kiwi fruit cultivars (“Bidan” and “SKK12”) than in other studied samples. To our knowledge this is the first report showing the differences and similarities in new kiwi fruit cultivars, using these spectroscopic data. The ethanol extracts of these cultivars exhibited high binding properties with human serum albumin compared with rutin. In conclusion, the applied analytical methods showed the main compounds in the kiwi fruit cultivars and can be used for determination of these compounds in any plants. The relative knowledge would contribute to the pharmaceutical development and clinical application of extracts of kiwi fruit extracts.     

Metabolite mapping of human filarial parasite, Brugia malayi with nuclear magnetic resonance

Metabolite mapping of human filarial parasite, Brugia malayi was carried out in vitro as well as in situ in host Mastomys coucha by ³¹P nuclear magnetic resonance (NMR) spectroscopy. Detection of parasites by visualizing contrast spots due to pathologic changes was observed by ¹H magnetic resonance imaging (MRI). Major metabolites of adult B. malayi observed by ³¹P-NMR spectroscopy were of sugar phosphates (SP), phosphomonoesters (PME), glycerophosphoryl-ethanolamine (GPE), -choline (GPC), phosphoenolpyruvate (PEP), inorganic phosphate (Pi), nucleoside diphosphosugar and nucleotides-mono, -di and -tri phosphates. PEP and GPC were present in high concentration; PEP being the major energy reservoir and GPC the major phospholipid in this species of filaria. The ³¹P NMR spectra of testis of mastomys, showed seven major peaks of SP, PME, phosphocreatine (PCr), phosphodiesters (PDE), Pi, and nucleotides di- and tri-phosphates. The ³¹P-NMR spectra of testis of B. malayi infected animal also consisted of seven major peaks with significant decrease in the SP and PME peak showing changes in the carbohydrate and lipid metabolism of filaria infected testis. Thus, in vivo ³¹P MRS provided a non-invasive assessment of tissue bioenergetics and phospholipid metabolism.     

Luminescencence determination of ecotoxicants in protein-based media

The structural changes in the protein macromolecules caused by polycyclic aromatic hydrocarbon (PAH) ecotoxicants were studied using the data on intrinsic fluorescence of proteins and fluorescence of PAH molecules introduced into proteins. A luminescence method for PAH determination in proteins was developed and used to study the interaction of two PAHs (pyrene and anthracene) with proteins of two types (bovine serum albumin and human serum albumin). The results were interpreted using the Stern–Volmer fluorescence quenching model. The association constants and the number of binding sites in the protein–ligand complexes were calculated. The binding of PAHs with proteins was described based on the static version of quenching with formation of nonfluorescent complexes of protein fluorophores with PAHs.     

Trichobilharzia regenti (Schistosomatidae): 3D imaging techniques in characterization of larval migration through the CNS of vertebrates

Migration of parasitic worms through the host tissues, which may occasionally result in fatal damage to the internal organs, represents one of the major risks associated with helminthoses. In order to track the parasites, traditionally used 2D imaging techniques such as histology or squash preparation do not always provide sufficient data to describe worm location/behavior in the host. On the other hand, 3D imaging methods are widely used in cell biology, medical radiology, osteology or cancer research, but their use in parasitological research is currently occasional. Thus, we aimed at the evaluation of suitability of selected 3D methods to monitor migration of the neuropathogenic avian schistosome Trichobilharzia regenti in extracted spinal cord of experimental vertebrate hosts. All investigated methods, two of them based on tracking of fluorescently stained larvae with or without previous chemical clearing of tissue and one based on X-ray micro-CT, exhibit certain limits for in vivo observation. Nevertheless, our study shows that the tested methods as ultramicroscopy (used for the first time in parasitology) and micro-CT represent promising tool for precise analyzing of parasite larvae in the CNS. Synthesis of these 3D imaging techniques can provide more comprehensive look at the course of infection, host immune response and pathology caused by migrating parasites within entire tissue samples, which would not be possible with traditional approaches.     

Zipf's law in gene expression

Using data from gene expression databases on various organisms and tissues, including yeast, nematodes, human normal and cancer tissues, and embryonic stem cells, we found that the abundances of expressed genes exhibit a power-law distribution with an exponent close to -1; i.e., they obey Zipf's law. Furthermore, by simulations of a simple model with an intracellular reaction network, we found that Zipf's law of chemical abundance is a universal feature of cells where such a network optimizes the efficiency and faithfulness of self-reproduction. These findings provide novel insights into the nature of the organization of reaction dynamics in living cells.     

Carotenes and carotenoids in natural biological samples: a Raman spectroscopic analysis

Raman spectroscopic studies of a range of naturally occurring carotenoids in over 50 specimens of plant tissue and a range of standard extracts have been undertaken, and the characteristic bands of CC and C C stretching and C CH bending have been recorded. Comparison of the spectroscopic data with the chemical assignment of the carotenoids from chemical extraction of the plant tissues reveals that there is a problem in the interpretation of the spectroscopic data which can be attributed to significant wavenumber shifts, particularly in the CC stretching band wavenumber, for carotenoids in the organic tissues arising from molecular interactions between the carotenoid and its host matrix. The simple identification of carotenoids in biological tissues on the basis of comparison with the standard spectra of extracted material must be made with caution; the progressive shift in wavenumber of the CC stretching band in the conjugated polyene chain of carotenoids with the number of CC groups, and hence the identification of the carotenoid, cannot be unambiguously interpreted for the range of materials studied here. Copyright © 2009 John Wiley & Sons, Ltd.     

Ultrasound assisted cyanotoxin extraction for nematode inhibition in soil

Root-knot nematodes are one of the plant damaging nematodes in agriculture causing a projected annual yield loss of ∼12 % (∼$160 billion) worldwide. Conventional solutions to control these plant-parasitic nematodes involve chemical nematicides. To reduce the use of harmful chemicals, microalgal extracts can be used as greener alternatives for nematode management. Microalgae produce valuable metabolites, including cyanotoxins which can aid in nematode suppression.In this study, two microalgae species, Trichormus variabilis and Nostoc punctiforme, were treated with ultrasound for intensified recovery of secondary metabolites. Ultrasound results in cell wall disruption of the microalgal species, thus resulting in enhanced release of secondary metabolites. Microalgal biomass was treated with an ultrasound probe at 50 % amplitude, 20 kHz frequency, using water as the extraction medium, for 5–30 min. The extraction efficiency was determined in terms of the total chlorophyll (Chl) content of the extract. Microscopic images of the treated cells were also investigated to gain insight into the effect of the ultrasonication time on the cell morphology. Our results suggest that ultrasonication resulted in the intensified release of secondary metabolites, as established through the total chlorophyll content of the ultrasonicated microalgal samples as well as the microscopic images of the ruptured cells. The best extraction for Trichormus variabilis was achieved with 15 min extraction time where the Total Chl content increased by 29 times (compared to the non-ultrasonicated sample), and for the Nostoc punctiforme, 30 min extraction time gave the highest metabolite recovery of 6.4 times higher than the non-ultrasonicated sample.Ultrasonicated algal extracts were then tested for their nematicidal potential against root-knot nematode, Meloidogyne hapla, in infested field soil samples. Experimental study was conducted using different concentrations of each microalga, Trichormus sp. and Nostoc sp., individually, as well as in combination. The nematode count for the treated soil was compared with that of the control (untreated soil). Ultrasonicated microalgal extracts showed 66% to 100% inhibition on root-knot nematodes in the soil samples tested.     

Molecular basis of the structure and function of H1 hemagglutinin of influenza virus

Influenza virus hemagglutinin (HA) contains antigenic sites recognized by the host immune system, cleavage sites cleaved by host proteases, receptor binding sites attaching to sialyl receptors on the target cell, and fusion peptides mediating membrane fusion. Change in an amino acid(s) in these sites may affect the potential of virus infection and spread within and between hosts. Influenza viruses with H1 HA infect birds, pigs and humans and have caused two of the four pandemics in the past 100 years: 1918 pandemic that killed 21-50 million people and 2009 pandemic that caused more than 18,000 deaths. Understanding the relationship between antigenic structure and immune specificity, the receptor binding specificity in virus transmission, how the cleavage site controls pathogenicity, and how the fusion peptide causes membrane fusion for the entry of influenza virus into the host cell should provide information to find more effective ways to prevent and control influenza.     

Preparation and water relaxation properties of proteins labeled with paramagnetic metal chelates

The proteins bovine serum albumin (BSA) and bovine immunoglobulin (IgG) have been labeled with paramagnetic gadolinium (III) and manganese (II) complexes using the bifunctional chelate approach. Diethylenetriaminepentaacetic acid (DTPA) and ethylenediaminetetraacetic acid (EDTA) were attached to several free amino groups on the proteins using cyclic anhydride forms of these ligands. The incorporation of the metal ions Gd+3 and Mn+2 into the chelating groups yielded highly paramagnetic proteins. The water relaxation ability (or relaxivity) of the protein-bound chelates at 20 MHz was found to be superior to that of the free metal complexes. Differences in relaxivity between the DTPA and EDTA conjugates could largely be accounted for by differences in the metal ion exposure to water. This labeling technique can be used in the preparation of intravascular NMR contrast agents (like paramagnetically-labeled human serum albumin) or target-specific agents (labeled monoclonal antibodies or fibrinogen).