Anelloviruses are the most common human infectious viruses and are considered essentially harmles... more Anelloviruses are the most common human infectious viruses and are considered essentially harmless. Compared to other human viruses, they are characterized by enormous diversity, and recombination is suggested to play an important role in their diversification and evolution.
Human blood metagenomics has revealed the presence of different types of viruses in apparently he... more Human blood metagenomics has revealed the presence of different types of viruses in apparently healthy subjects. By far, anelloviruses constitute the viral family that is more frequently found in human blood, although amplification biases and contaminations pose a major challenge in this field. To investigate this further, we subjected pooled plasma samples from 120 healthy donors in Spain to high-speed centrifugation, RNA and DNA extraction, random amplification, and massive parallel sequencing. Our results confirm the abundance of anelloviruses in such samples, which represented >98% of the total viral sequence reads obtained. We assembled 114 different viral genomes belonging to this family, revealing remarkable diversity. Phylogenetic analysis of ORF1 suggested 28 potentially novel anellovirus species, 24 of which were validated by Sanger sequencing to discard artifacts. These findings underscore the importance of implementing more efficient purification procedures that enric...
BACKGROUND Plasma of patients taking anti-CD38 monoclonal antibodies (MoAbs) leads to panagglutin... more BACKGROUND Plasma of patients taking anti-CD38 monoclonal antibodies (MoAbs) leads to panagglutination in the indirect antiglobulin test (IAT), that can mask clinically significant alloantibodies. of test RBCs is the more widespread method for avoiding this interference. Current DTT 0.2 mol/L method is time consuming and damages several red blood groups antigens. This study aims to evaluate low concentration DTT treatment of RBCs adapted for gel testing. MATERIALS AND METHODS Four DTT concentrations (0.01, 0.02, 0.03, and 0.04 mol/L), and three gel test brands were evaluated on six DARA patient's samples. Briefly, the method consists of pipetting 50 μL of 0.8% RBCs on AHG micro columns, followed by 25 μL of DTT, thoroughly mixing and 15 min incubation at 37°C. Then, 25 μL of serum/plasma is added to proceed to IAT. In order to asses the effect of DTT 0.04 mol/L on different blood group antigens, serial dilutions of sera containing anti-K, -k, -Kpb, -Lub, -Yta and anti-JMH antibo...
The transmission of microbial infection through tissue allografts is one of the main risks that m... more The transmission of microbial infection through tissue allografts is one of the main risks that must be controlled in tissue banks. Therefore, microbiological monitoring controls and validated protocols for the decontamination of tissues during processing have been implemented. This study is based on the evaluation of data from microbiological cultures of arteries (mainly long peripheral arteries) processed in the tissue bank of Valencia (Spain). Donors' profile, pre- and post-disinfection tissue samples were assessed. The presence of residual antibiotics in disinfected tissues was determined and the antimicrobial potential of these tissues was tested. Our overall contamination rate was 23.69%, with a disinfection rate (after antibiotic incubation) of 87.5%. Most (76.09%) of the microbial contaminants were identified as Gram positive. Arterial allografts collected from body sites affected by prior organ removal showed higher risk of contamination. Only vancomycin was detected as tissue release. The antimicrobial effect on Candida albicans was lower than that for bacterial species. Risk assessment for microbial contamination suggested the donor's skin and the environment during tissue collection as the main sources for allograft contamination. Antibiotic-disinfected arterial allografts showed antimicrobial potential.
Metagenomics is greatly improving our ability to discover new viruses, as well as their possible ... more Metagenomics is greatly improving our ability to discover new viruses, as well as their possible associations with disease. However, metagenomics has also changed our understanding of viruses in general. The vast expansion of currently known viral diversity has revealed a large fraction of non-pathogenic viruses, and offers a new perspective in which viruses function as important components of many ecosystems. In this vein, studies of the human blood virome are often motivated by the search for new viral diseases, especially those associated with blood transfusions. However, these studies have revealed the common presence of apparently non-pathogenic viruses in blood, in particular human anelloviruses and, to a lower extent, human pegiviruses (HPgV). To shed light on the diversity of the human blood virome, we subjected pooled plasma samples from 587 healthy donors in Spain to a viral enrichment protocol followed by massive parallel sequencing. This showed that anelloviruses were cl...
The use of autologous bone flap for cranioplasty after decompressive craniectomy is a widely used... more The use of autologous bone flap for cranioplasty after decompressive craniectomy is a widely used strategy that allows alleviating health expenses. When the patient has recovered from the primary insult, the cranioplasty restores protection and cosmesis, recovering fluid dynamics and improving neurological status. During this time, the bone flap must be stored, but there is a lack of standardization of tissue banking practices for this aim. In this work, we have reviewed the literature on tissue processing and storage practices. Most of the published articles are focused from a strictly clinical and surgical point of view, paying less attention to issues related to tissue manipulation. When bone resorption is avoided and the risk of infection is controlled, the autograft represents the most efficient choice, with the lowest risk of complication. Otherwise, depending on the degree of involvement, the patient may have to undergo new surgery, assuming further risks and higher healthcare costs. Therefore, tissue banks must implement protocols to provide products with the highest possible clinical effectiveness, without compromising safety. With a centralised management of tissue banking practices there may be a more uniform approach, thus facilitating the standardization of procedures and guidelines.
BACKGROUND COVID-19 convalescent plasma (CCP) is an experimental treatment against SARS-CoV-2. Al... more BACKGROUND COVID-19 convalescent plasma (CCP) is an experimental treatment against SARS-CoV-2. Although there has so far been no evidence of transmission through transfusion, pathogen reduction technologies (PRT) have been applied to CCP to mitigate risk of infectious disease. This study aims to assess the impact of methylene blue (MB) plus visible light PRT on the virus-neutralising activity of the specific antibodies against SARS-CoV-2. MATERIAL AND METHODS Thirty-five plasma doses collected by plasmapheresis from COVID-19 convalescent donors were subjected to MB plus visible light PRT. Anti-SARS-CoV-2 RBD S1 epitope IgGs antibodies were quantified by ELISA. Titres of SARS-CoV-2 neutralising antibodies (NtAbs) were measured before and after the PRT process. A Spearman's correlation was run to determine the relationship between antibody neutralisation ability and SARS-CoV-2 IgG ELISA ratio. Pre- and post-inactivation neutralising antibody titres were evaluated using a Wilcoxon ...
Platelet concentrates (PCs), stored at room temperature, have the highest risk of bacterial conta... more Platelet concentrates (PCs), stored at room temperature, have the highest risk of bacterial contamination among labile blood components. We report the case of an apheresis platelet product contaminated with Klebsiella oxytoca showing an amorphous mass on the platelet's bag. A 66-year-old patient with myelodysplastic syndrome under prophylactic platelet transfusion support experienced malaise, chills, and a sudden drop in blood pressure 15 minutes after starting the transfusion of a single donor platelet's unit. The platelet unit had not been pathogen reduced or screened for bacterial contamination. The patient needed admission to the critical care ward due to septic shock. Microbiological cultures were performed on the patient's blood and the transfused bag. Bacterial cultures were positive on both samples, and K. oxytoca was isolated. It was reported to our blood center for further investigation. The involved component corresponded to a double-dose donation and was transfused on day 3 after collection. The remaining untransfused sister unit was located in another hospital, and it was returned to the blood center where a large white amorphous mass was noted (Figures 1 and 2). K. oxytoca was also isolated in this second portion. Antibiograms from the three isolates (patient and PCs) were the same. Therefore, the mass was likely to be a platelet aggregate caused by the growth of bacteria. The source of
The possibility to use CCR5-∆32 umbilical cord blood to cure HIV infection in patients in need of... more The possibility to use CCR5-∆32 umbilical cord blood to cure HIV infection in patients in need of a hematopoietic transplant has been suggested. The less stringent HLA compatibility needed in this type of transplant facilitates the search of a suitable donor having the CCR5-∆32 mutation. To achieve an inventory of CCR5-∆32 cord blood units, the 20,236 best cell quality units of the Spanish Registry were genotyped. Furthermore, their CD34and total nucleated cells counts, blood type, gender, HLA and donor's geographical and ancestral origin were analyzed. The results showed 130 (0.64%) units homozygous for the deletion, 2,646 (13.08%) heterozygous and 17,460 (86.28%) did not present the mutation. Interestingly, a significant lower amount of CD34cells was found in the CCR5-∆32 homozygous units. In addition, a significant association was found among donor's ancestral origin and the mutation, with a higher percentage of CCR5-∆32 units with a European ancestry. In summary, identif...
Transfusion and apheresis science : official journal of the World Apheresis Association : official journal of the European Society for Haemapheresis, Jan 22, 2017
Diverse variables are involved in apheresis platelet collection, processing and storage. This sur... more Diverse variables are involved in apheresis platelet collection, processing and storage. This survey shows how these are realized in Spain. An analysis of collected data was performed in a questionnaire completed by ten Transfusion Centers (TC) which perform between 50 and 520 apheresis procedures per month. This information comprises the procedures used to collect, inspect and store apheresis platelet concentrates (PC), and quality control data. Macroscopic inspection of PC is performed in all TC, especially during the first few hours post-collection and before distribution. The type of processor, duration of post-collection resting periods and temperature from the time of collection until distribution are similar in all TC. In 80% of TC, PC with small and scarce aggregates are distributed to transfusion services. The presence of clumps is influenced by type of processor, female donor, cold ambient temperature and collection of hyperconcentrated platelets, and is often recurrent in...
Anelloviruses are the most common human infectious viruses and are considered essentially harmles... more Anelloviruses are the most common human infectious viruses and are considered essentially harmless. Compared to other human viruses, they are characterized by enormous diversity, and recombination is suggested to play an important role in their diversification and evolution.
Human blood metagenomics has revealed the presence of different types of viruses in apparently he... more Human blood metagenomics has revealed the presence of different types of viruses in apparently healthy subjects. By far, anelloviruses constitute the viral family that is more frequently found in human blood, although amplification biases and contaminations pose a major challenge in this field. To investigate this further, we subjected pooled plasma samples from 120 healthy donors in Spain to high-speed centrifugation, RNA and DNA extraction, random amplification, and massive parallel sequencing. Our results confirm the abundance of anelloviruses in such samples, which represented >98% of the total viral sequence reads obtained. We assembled 114 different viral genomes belonging to this family, revealing remarkable diversity. Phylogenetic analysis of ORF1 suggested 28 potentially novel anellovirus species, 24 of which were validated by Sanger sequencing to discard artifacts. These findings underscore the importance of implementing more efficient purification procedures that enric...
BACKGROUND Plasma of patients taking anti-CD38 monoclonal antibodies (MoAbs) leads to panagglutin... more BACKGROUND Plasma of patients taking anti-CD38 monoclonal antibodies (MoAbs) leads to panagglutination in the indirect antiglobulin test (IAT), that can mask clinically significant alloantibodies. of test RBCs is the more widespread method for avoiding this interference. Current DTT 0.2 mol/L method is time consuming and damages several red blood groups antigens. This study aims to evaluate low concentration DTT treatment of RBCs adapted for gel testing. MATERIALS AND METHODS Four DTT concentrations (0.01, 0.02, 0.03, and 0.04 mol/L), and three gel test brands were evaluated on six DARA patient's samples. Briefly, the method consists of pipetting 50 μL of 0.8% RBCs on AHG micro columns, followed by 25 μL of DTT, thoroughly mixing and 15 min incubation at 37°C. Then, 25 μL of serum/plasma is added to proceed to IAT. In order to asses the effect of DTT 0.04 mol/L on different blood group antigens, serial dilutions of sera containing anti-K, -k, -Kpb, -Lub, -Yta and anti-JMH antibo...
The transmission of microbial infection through tissue allografts is one of the main risks that m... more The transmission of microbial infection through tissue allografts is one of the main risks that must be controlled in tissue banks. Therefore, microbiological monitoring controls and validated protocols for the decontamination of tissues during processing have been implemented. This study is based on the evaluation of data from microbiological cultures of arteries (mainly long peripheral arteries) processed in the tissue bank of Valencia (Spain). Donors' profile, pre- and post-disinfection tissue samples were assessed. The presence of residual antibiotics in disinfected tissues was determined and the antimicrobial potential of these tissues was tested. Our overall contamination rate was 23.69%, with a disinfection rate (after antibiotic incubation) of 87.5%. Most (76.09%) of the microbial contaminants were identified as Gram positive. Arterial allografts collected from body sites affected by prior organ removal showed higher risk of contamination. Only vancomycin was detected as tissue release. The antimicrobial effect on Candida albicans was lower than that for bacterial species. Risk assessment for microbial contamination suggested the donor's skin and the environment during tissue collection as the main sources for allograft contamination. Antibiotic-disinfected arterial allografts showed antimicrobial potential.
Metagenomics is greatly improving our ability to discover new viruses, as well as their possible ... more Metagenomics is greatly improving our ability to discover new viruses, as well as their possible associations with disease. However, metagenomics has also changed our understanding of viruses in general. The vast expansion of currently known viral diversity has revealed a large fraction of non-pathogenic viruses, and offers a new perspective in which viruses function as important components of many ecosystems. In this vein, studies of the human blood virome are often motivated by the search for new viral diseases, especially those associated with blood transfusions. However, these studies have revealed the common presence of apparently non-pathogenic viruses in blood, in particular human anelloviruses and, to a lower extent, human pegiviruses (HPgV). To shed light on the diversity of the human blood virome, we subjected pooled plasma samples from 587 healthy donors in Spain to a viral enrichment protocol followed by massive parallel sequencing. This showed that anelloviruses were cl...
The use of autologous bone flap for cranioplasty after decompressive craniectomy is a widely used... more The use of autologous bone flap for cranioplasty after decompressive craniectomy is a widely used strategy that allows alleviating health expenses. When the patient has recovered from the primary insult, the cranioplasty restores protection and cosmesis, recovering fluid dynamics and improving neurological status. During this time, the bone flap must be stored, but there is a lack of standardization of tissue banking practices for this aim. In this work, we have reviewed the literature on tissue processing and storage practices. Most of the published articles are focused from a strictly clinical and surgical point of view, paying less attention to issues related to tissue manipulation. When bone resorption is avoided and the risk of infection is controlled, the autograft represents the most efficient choice, with the lowest risk of complication. Otherwise, depending on the degree of involvement, the patient may have to undergo new surgery, assuming further risks and higher healthcare costs. Therefore, tissue banks must implement protocols to provide products with the highest possible clinical effectiveness, without compromising safety. With a centralised management of tissue banking practices there may be a more uniform approach, thus facilitating the standardization of procedures and guidelines.
BACKGROUND COVID-19 convalescent plasma (CCP) is an experimental treatment against SARS-CoV-2. Al... more BACKGROUND COVID-19 convalescent plasma (CCP) is an experimental treatment against SARS-CoV-2. Although there has so far been no evidence of transmission through transfusion, pathogen reduction technologies (PRT) have been applied to CCP to mitigate risk of infectious disease. This study aims to assess the impact of methylene blue (MB) plus visible light PRT on the virus-neutralising activity of the specific antibodies against SARS-CoV-2. MATERIAL AND METHODS Thirty-five plasma doses collected by plasmapheresis from COVID-19 convalescent donors were subjected to MB plus visible light PRT. Anti-SARS-CoV-2 RBD S1 epitope IgGs antibodies were quantified by ELISA. Titres of SARS-CoV-2 neutralising antibodies (NtAbs) were measured before and after the PRT process. A Spearman's correlation was run to determine the relationship between antibody neutralisation ability and SARS-CoV-2 IgG ELISA ratio. Pre- and post-inactivation neutralising antibody titres were evaluated using a Wilcoxon ...
Platelet concentrates (PCs), stored at room temperature, have the highest risk of bacterial conta... more Platelet concentrates (PCs), stored at room temperature, have the highest risk of bacterial contamination among labile blood components. We report the case of an apheresis platelet product contaminated with Klebsiella oxytoca showing an amorphous mass on the platelet's bag. A 66-year-old patient with myelodysplastic syndrome under prophylactic platelet transfusion support experienced malaise, chills, and a sudden drop in blood pressure 15 minutes after starting the transfusion of a single donor platelet's unit. The platelet unit had not been pathogen reduced or screened for bacterial contamination. The patient needed admission to the critical care ward due to septic shock. Microbiological cultures were performed on the patient's blood and the transfused bag. Bacterial cultures were positive on both samples, and K. oxytoca was isolated. It was reported to our blood center for further investigation. The involved component corresponded to a double-dose donation and was transfused on day 3 after collection. The remaining untransfused sister unit was located in another hospital, and it was returned to the blood center where a large white amorphous mass was noted (Figures 1 and 2). K. oxytoca was also isolated in this second portion. Antibiograms from the three isolates (patient and PCs) were the same. Therefore, the mass was likely to be a platelet aggregate caused by the growth of bacteria. The source of
The possibility to use CCR5-∆32 umbilical cord blood to cure HIV infection in patients in need of... more The possibility to use CCR5-∆32 umbilical cord blood to cure HIV infection in patients in need of a hematopoietic transplant has been suggested. The less stringent HLA compatibility needed in this type of transplant facilitates the search of a suitable donor having the CCR5-∆32 mutation. To achieve an inventory of CCR5-∆32 cord blood units, the 20,236 best cell quality units of the Spanish Registry were genotyped. Furthermore, their CD34and total nucleated cells counts, blood type, gender, HLA and donor's geographical and ancestral origin were analyzed. The results showed 130 (0.64%) units homozygous for the deletion, 2,646 (13.08%) heterozygous and 17,460 (86.28%) did not present the mutation. Interestingly, a significant lower amount of CD34cells was found in the CCR5-∆32 homozygous units. In addition, a significant association was found among donor's ancestral origin and the mutation, with a higher percentage of CCR5-∆32 units with a European ancestry. In summary, identif...
Transfusion and apheresis science : official journal of the World Apheresis Association : official journal of the European Society for Haemapheresis, Jan 22, 2017
Diverse variables are involved in apheresis platelet collection, processing and storage. This sur... more Diverse variables are involved in apheresis platelet collection, processing and storage. This survey shows how these are realized in Spain. An analysis of collected data was performed in a questionnaire completed by ten Transfusion Centers (TC) which perform between 50 and 520 apheresis procedures per month. This information comprises the procedures used to collect, inspect and store apheresis platelet concentrates (PC), and quality control data. Macroscopic inspection of PC is performed in all TC, especially during the first few hours post-collection and before distribution. The type of processor, duration of post-collection resting periods and temperature from the time of collection until distribution are similar in all TC. In 80% of TC, PC with small and scarce aggregates are distributed to transfusion services. The presence of clumps is influenced by type of processor, female donor, cold ambient temperature and collection of hyperconcentrated platelets, and is often recurrent in...
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Papers by LUIS LARREA