A library of cell-surface and secreted proteins fromSchistosoma mansoniidentifies early serological markers of infection

AbstractSchistosomiasis is a major global health problem caused by blood-dwelling parasitic worms and is currently treated by the mass administration of the drug praziquantel. Appropriate drug treatment strategies are informed by diagnostics that establish the prevalence and intensity of infection, which, in regions of low transmission should be highly sensitive. To identify sensitive new serological markers ofSchistosoma mansoniinfections, we have compiled a recombinant protein library of 115 parasite cell surface and secreted proteins expressed in mammalian cells. The vast majority of them were shown to be immunoreactive and to contain heat-labile conformational epitopes when tested against pooled human sera from endemic regions. After probing the library against a time series of sera samples from experimental infections in mice, we identified several markers of infection, the majority of which belong to the saposin-domain-containing and cathepsin families of proteins. These new markers will be a valuable tool to detect ongoing and previousS. mansoniinfections, including in regions of low transmission. We envisage that this new recombinant protein resource will be used in a wide range of cellular and molecular assays to further our understanding ofSchistosomabiology.

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Screening of a Library of Recombinant Schistosoma mansoni Proteins With Sera From Murine and Human Controlled Infections Identifies Early Serological Markers

The Journal of Infectious Diseases ◽

10.1093/infdis/jiaa329 ◽

2020 ◽

Author(s):

Cécile Crosnier ◽

Cornelis H Hokke ◽

Anna V Protasio ◽

Cordelia Brandt ◽

Gabriel Rinaldi ◽

...

Keyword(s):

Schistosoma Mansoni ◽

Mammalian Cells ◽

Treatment Strategies ◽

Serological Markers ◽

Intensity Of Infection ◽

Protein Library ◽

Highly Sensitive ◽

Parasitic Worms ◽

Global Health Problem ◽

Sera Samples

Abstract Background Schistosomiasis is a major global health problem caused by blood-dwelling parasitic worms, which is currently tackled primarily by mass administration of the drug praziquantel. Appropriate drug treatment strategies are informed by diagnostics that establish the prevalence and intensity of infection, which, in regions of low transmission, should be highly sensitive. Methods To identify sensitive new serological markers of Schistosoma mansoni infections, we have compiled a recombinant protein library of parasite cell-surface and secreted proteins expressed in mammalian cells. Results Together with a time series of sera samples from volunteers experimentally infected with a defined number of male parasites, we probed this protein library to identify several markers that can detect primary infections with as low as 10 parasites and as early as 5 weeks postinfection. Conclusions These new markers could be further explored as valuable tools to detect ongoing and previous S mansoni infections, including in endemic regions where transmission is low.

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Reticulon 4a promotes exocytosis in mammalian cells

Molecular Biology of the Cell ◽

10.1091/mbc.e19-03-0159 ◽

2019 ◽

Vol 30 (18) ◽

pp. 2349-2357 ◽

Cited By ~ 2

Author(s):

Richik Nilay Mukherjee ◽

Daniel L. Levy

Keyword(s):

Cell Surface ◽

Protein Trafficking ◽

Mammalian Cells ◽

Cell Types ◽

Vesicular Trafficking ◽

Secreted Proteins ◽

Mammalian Cell Lines ◽

Rough Er ◽

Different Cell Types ◽

Functional Output

Endoplasmic reticulum (ER) tubules and sheets conventionally correspond to smooth and rough ER, respectively. The ratio of ER tubules-to-sheets varies in different cell types and changes in response to cellular conditions, potentially impacting the functional output of the ER. To directly test whether ER morphology impacts vesicular trafficking, we increased the tubule-to-sheet ratio in three different ways, by overexpressing Rtn4a, Rtn4b, or REEP5. Only Rtn4a overexpression increased exocytosis, but not overall levels, of several cell surface and secreted proteins. Furthermore, Rtn4a depletion reduced cell surface trafficking without affecting ER morphology. Similar results were observed in three different mammalian cell lines, suggesting that Rtn4a generally enhances exocytosis independently of changes in ER morphology. Finally, we show that Rtn4a levels modulate cell adhesion, possibly by regulating trafficking of integrins to the cell surface. Taking the results together, we find that altering ER morphology does not necessarily affect protein trafficking, but that Rtn4a specifically enhances exocytosis.

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Towards a comprehensive Plasmodium falciparum merozoite cell surface and secreted recombinant protein library

Malaria Journal ◽

10.1186/1475-2875-13-93 ◽

2014 ◽

Vol 13 (1) ◽

pp. 93 ◽

Cited By ~ 19

Author(s):

Zenon A Zenonos ◽

Julian C Rayner ◽

Gavin J Wright

Keyword(s):

Plasmodium Falciparum ◽

Recombinant Protein ◽

Cell Surface ◽

Protein Library ◽

Falciparum Merozoite

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Expression and Functional Characterization of Bluetongue Virus VP2 Protein: Role in Cell Entry

Journal of Virology ◽

10.1128/jvi.73.12.9832-9842.1999 ◽

1999 ◽

Vol 73 (12) ◽

pp. 9832-9842 ◽

Cited By ~ 86

Author(s):

Sharifah S. Hassan ◽

PoLly Roy

Keyword(s):

Recombinant Protein ◽

Cell Surface ◽

Bluetongue Virus ◽

Mammalian Cells ◽

Functional Characterization ◽

Recombinant Baculovirus ◽

Protein Molecule ◽

Blood Feeding ◽

Rnase A ◽

Cell Surface Molecule

ABSTRACT Segment 2 of bluetongue virus (BTV) serotype 10, which encodes the outer capsid protein VP2, was tagged with the S-peptide fragment of RNase A and expressed by a recombinant baculovirus. The recombinant protein was subsequently purified to homogeneity by virtue of the S tag, and the oligomeric nature of the purified protein was determined. The data obtained indicated that the majority of the protein forms a dimer and, to a lesser extent, some trimer. The recombinant protein was used to determine various biological functions of VP2. The purified VP2 was shown to have virus hemagglutinin activity and was antigenically indistinguishable from the VP2 of the virion. Whether VP2 is responsible for BTV entry into permissive cells was subsequently assessed by cell surface attachment and internalization studies with an immunofluorescence assay system. The results demonstrated that VP2 alone is responsible for virus entry into mammalian cells. By competition assay, it appeared that both VP2 and the BTV virion attached to the same cell surface molecule(s). The purified VP2 also had a strong affinity for binding to glycophorin A, a sialoglycoprotein component of erythrocytes, indicating that VP2 may be responsible for BTV transmission by the Culicoides vector to vertebrate hosts during blood feeding. Further, by various enzymatic treatments of BTV-permissive L929 cells, preliminary data have been obtained which indicated that the BTV receptor molecule(s) is likely to be a glycoprotein and that either the protein moiety of the glycoprotein or a second protein molecule could also serve as a coreceptor for BTV infection.

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Evaluation of the nucleopolyhedrovirus of Anticarsia gemmatalis as a vector for gene therapy in mammals

Current Gene Therapy ◽

10.2174/1566523220999201217155945 ◽

2020 ◽

Vol 20 ◽

Author(s):

Cintia N. Parsza ◽

Diego L. Mengual Gómez ◽

Jorge Alejandro Simonin ◽

Mariano Nicolás Belaich ◽

Pablo Daniel Ghiringhelli

Keyword(s):

Gene Therapy ◽

Mammalian Cells ◽

Insect Cells ◽

Autographa Californica ◽

Anticarsia Gemmatalis ◽

Agricultural Pests ◽

Mammalian Cell Lines ◽

Delivery Vector ◽

Wide Range ◽

Insect Pathogens

Background: Baculoviruses are insect pathogens with important biotechnological applications that transcend their use as biological controllers of agricultural pests. One species, Autographa californica multiple nucleopolhyedrovirus (AcMNPV) has been extensively exploited as a molecular platform to produce recombinant proteins and as a delivery vector for genes in mammals, because it can transduce a wide range of mammalian cells and tissues without replicating or producing progeny. Objective/Method: To investigate if the budded virions of Anticarsia gemmatalis multiple nucleopolhyedrovirus (AgMNPV) species has the same ability, the viral genome was modified by homologous recombination into susceptible insect cells to integrate reporter genes and then it was evaluated on mammalian cell lines in comparative form with respect to equivalent viruses derived from AcMNPV. Besides, the replicative capacity of AgMNPV´s virions in mammals was determined. Results: The experiments carried out showed that the recombinant variant of AgMNPV transduces and support the expression of delivered genes but not replicates in mammalian cells. Conclusion: Consequently, this insect pathogen is proposed as an alternative of non-infectious viruses in humans to explore new approaches in gene therapy and other applications based on the use of mammalian cells.

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Advances and Applications of Water Phytoremediation: A Potential Biotechnological Approach for the Treatment of Heavy Metals from Contaminated Water

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph18105215 ◽

2021 ◽

Vol 18 (10) ◽

pp. 5215

Author(s):

Cristián Raziel Delgado-González ◽

Alfredo Madariaga-Navarrete ◽

José Miguel Fernández-Cortés ◽

Margarita Islas-Pelcastre ◽

Goldie Oza ◽

...

Keyword(s):

Water Quality ◽

Plant Species ◽

Treatment Strategies ◽

Pollution Sources ◽

Water Contaminants ◽

Biotechnological Approach ◽

Wide Range ◽

Great Relevance ◽

Multidisciplinary Perspective ◽

Viable Method

Potable and good-quality drinking water availability is a serious global concern, since several pollution sources significantly contribute to low water quality. Amongst these pollution sources, several are releasing an array of hazardous agents into various environmental and water matrices. Unfortunately, there are not very many ecologically friendly systems available to treat the contaminated environment exclusively. Consequently, heavy metal water contamination leads to many diseases in humans, such as cardiopulmonary diseases and cytotoxicity, among others. To solve this problem, there are a plethora of emerging technologies that play an important role in defining treatment strategies. Phytoremediation, the usage of plants to remove contaminants, is a technology that has been widely used to remediate pollution in soils, with particular reference to toxic elements. Thus, hydroponic systems coupled with bioremediation for the removal of water contaminants have shown great relevance. In this review, we addressed several studies that support the development of phytoremediation systems in water. We cover the importance of applied science and environmental engineering to generate sustainable strategies to improve water quality. In this context, the phytoremediation capabilities of different plant species and possible obstacles that phytoremediation systems may encounter are discussed with suitable examples by comparing different mechanistic processes. According to the presented data, there are a wide range of plant species with water phytoremediation potential that need to be studied from a multidisciplinary perspective to make water phytoremediation a viable method.

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Autophagy Deficiency by Atg4B Loss Leads to Metabolomic Alterations in Mice

Metabolites ◽

10.3390/metabo11080481 ◽

2021 ◽

Vol 11 (8) ◽

pp. 481

Author(s):

Gemma G. Martínez-García ◽

Raúl F. Pérez ◽

Álvaro F. Fernández ◽

Sylvere Durand ◽

Guido Kroemer ◽

...

Keyword(s):

Skeletal Muscle ◽

Amino Acids ◽

Mammalian Cells ◽

Tissue Homeostasis ◽

In Vivo Studies ◽

Protective Mechanism ◽

Cardiac Damage ◽

Wide Range ◽

First Time

Autophagy is an essential protective mechanism that allows mammalian cells to cope with a variety of stressors and contributes to maintaining cellular and tissue homeostasis. Due to these crucial roles and also to the fact that autophagy malfunction has been described in a wide range of pathologies, an increasing number of in vivo studies involving animal models targeting autophagy genes have been developed. In mammals, total autophagy inactivation is lethal, and constitutive knockout models lacking effectors of this route are not viable, which has hindered so far the analysis of the consequences of a systemic autophagy decline. Here, we take advantage of atg4b−/− mice, an autophagy-deficient model with only partial disruption of the process, to assess the effects of systemic reduction of autophagy on the metabolome. We describe for the first time the metabolic footprint of systemic autophagy decline, showing that impaired autophagy results in highly tissue-dependent alterations that are more accentuated in the skeletal muscle and plasma. These changes, which include changes in the levels of amino-acids, lipids, or nucleosides, sometimes resemble those that are frequently described in conditions like aging, obesity, or cardiac damage. We also discuss different hypotheses on how impaired autophagy may affect the metabolism of several tissues in mammals.

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Prevalence of Hand Malformations in Patients With Moebius Syndrome and Their Management

Hand ◽

10.1177/1558944721994265 ◽

2021 ◽

pp. 155894472199426

Author(s):

Jose E. Telich-Tarriba ◽

David F. Navarro-Barquin ◽

Karol Verdezoto-Gaibor ◽

Alexander Cardenas-Mejia

Keyword(s):

Upper Extremity ◽

Treatment Strategies ◽

Abducens Nerve ◽

Poland Syndrome ◽

Statistical Association ◽

Moebius Syndrome ◽

Cross Sectional ◽

Reconstructive Procedures ◽

Nerve Paralysis ◽

Wide Range

Background: Moebius syndrome is a disorder characterized by facial and abducens nerve paralysis. Patients can present a wide range of upper extremity malformations. Literature focused on orthopedic manifestations of Moebius syndrome shows variability in the prevalence and clinical presentation of upper extremity anomalies. The aim of this work is to evaluate the prevalence of upper extremity malformations in patients with Moebius syndrome, clarify its various clinical presentations, and present treatment strategies for their management. Methods: This is a retrospective, cross-sectional study including patients with Moebius syndrome and upper extremity malformations between 2012 and 2019. Data include demographic characteristics, Moebius syndrome subtype, type of malformation, affected extremity, and surgical procedures underwent. Quantitative data were recorded as mean (standard deviation [SD]), and qualitative data were expressed in terms of totals and percentages. Statistical association between Moebius syndrome subtype and development of upper extremity anomalies was evaluated using binary logistic regression. Results: Twenty-five out of 153 patients (16.3%) presented upper extremity malformations (48% male). Mean age of presentation was 9.08 ± 9.43 years. Sixty-eight percent of the malformations were unilateral. The most common presentations included Poland syndrome and simple syndactyly with 8 cases each (32%), followed by 5 cases of brachysyndactyly (20%), 3 cases of amniotic band syndrome (12%), and 1 case of cleft hand (4%). No statistical association was found between Moebius syndrome subtype and odds ratio for development of upper extremity anomalies. Thirteen patients (52%) underwent reconstructive procedures. Conclusion: Poland syndrome and syndactyly are the most common anomalies in patients with Moebius syndrome. Patients may present with a wide range of hand malformations, each patient should be carefully evaluated in order to determine whether surgical treatment is needed and to optimize rehabilitation protocols.

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