RESUMO
Objective: To observe the treatment response of a two-dose regimen of inotuzumab ozogamicin (inotuzumab), a monoclonal antibody targeting CD22, for patients with heavily treated relapsed/refractory B-cell acute lymphoblastic leukemia (R/R B-ALL), including those failed or relapsed after chimeric antigen receptor (CAR) -T-cell therapy. Methods: Pediatric and adult patients who received two doses of inotuzumab and who were evaluated after inotuzumab treatment were included. Antibody infusions were performed between March 2020 and September 2022. All patients expressed CD22 antigen as detected by flow cytometry (>80% leukemic cells displaying CD22) before treatment. For adults, the maximum dosage per administration was 1 mg (with a total of two administrations). For children, the maximum dosage per administration was 0.85 mg/m(2) (no more than 1 mg/dose; total of two administrations). The total dosage administered to each patient was less than the standard dosage of 1.8 mg/m(2). Results: Twenty-one patients with R/R B-ALL were included, including five children (<18 years old) and sixteen adults. Seventeen patients presented with 5.0% -99.0% leukemic blasts in the bone marrow/peripheral blood or with extramedullary disease, and four patients were minimal residual disease (MRD) -positive. Fourteen patients underwent both CD19 and CD22 CAR-T-cell therapy, four underwent CD19 CAR-T-cell therapy, and three underwent blinatumomab therapy. Eleven patients underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT). After inotuzumab treatment, 14 of 21 patients (66.7% ) achieved a complete response (CR, one was MRD-positive CR), and all four MRD-positive patients turned MRD-negative. Four of six patients who failed recent CD22 CAR-T-cell therapy achieved a CR after subsequent inotuzumab treatment. Seven patients (33.3% ) demonstrated no response. Grade 1-3 hepatotoxicity occurred in five patients (23.8% ), one child with no response experienced hepatic veno-occlusive disease (HVOD) during salvage transplantation and recovered completely. Conclusion: For patients with heavily treated R/R B-ALL, including those who had undergone allo-HSCT and CD19/CD22 CAR-T-cell therapy, the two-dose regimen of inotuzumab resulted in a CR rate of 66.7%, and the frequency of hepatotoxicity and HVOD was low.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas , Leucemia-Linfoma Linfoblástico de Células Precursoras , Receptores de Antígenos Quiméricos , Adulto , Humanos , Criança , Adolescente , Inotuzumab Ozogamicina , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Anticorpos Monoclonais , Proteínas Adaptadoras de Transdução de Sinal , Antígenos CD19RESUMO
Two components, (Z)-8-dodecenyl acetate (Z8-12:Ac) and (Z)-8-dodecenol (Z8-12:OH), were isolated from sex pheromone glands of the carambola fruit borer, Eucosma notanthes, and were identified by GC, and GC-MS, chemical derivatization, and comparison of retention times. The ratio of the alcohol to acetate in the sex pheromone extracts was 2.7. However, synthetic mixtures (1 mg) in ratios ranging from 0.5 to 1.5 were more effective than other blends in trapping male moths in field tests.
Assuntos
Lepidópteros/química , Atrativos Sexuais/isolamento & purificação , Atrativos Sexuais/farmacologia , Animais , Comportamento Animal , Cromatografia Gasosa , Glândulas Exócrinas/química , Cromatografia Gasosa-Espectrometria de Massas , Larva/química , MovimentoRESUMO
Transovarial transmission of a yeast-like symbiote (YLS) in the brown planthopper, Nilaparvata lugens Stal, was observed with light and electron microscopy. Light micrographs showed that there was no YLS in testes and spermathecae of the mated females, indicating that sperm is not involved in the transovarial transmission of the symbiote. Both light and electron micrographs showed the processes of YLS transmission from fat body to the oocyte. In females, the symbiotes in mycetocytes moved out of the syncytium, which is formed from a layer of fat body cells, by exocytosis, and released into hemocoel. Then, the free YLS in hemolymph approached to the ovarioles near pedicel and were enclosed by follicle cells. They entered the follicle cells around the primary oocyte by endocytosis at epithelial plug of the ovariole. The YLS aggregated at the posterior end of the mature egg after entering, and finally formed a symbiote ball.
Assuntos
Insetos/microbiologia , Simbiose , Leveduras/ultraestrutura , Animais , Microscopia EletrônicaRESUMO
The wound-healing processes in the mosquito Armigeres subalbatus (Coquillett) were observed with electron microscopy. The initial reaction involved wound contractions and aggregation of injured surface tissues, cell debris and movement of granulocytes toward the wound. Granulocytes first aggregated around the surface of the wound and many filamentous filopodia protruded to connect with cytoplasmic strands. These strands were then interconnected to form a network coagulum resulting in wound closure to prevent body fluid loss. Granulocytes lysed on the wound-site and released granular materials around the wound, inducing localized clot formation. These results suggested that wound-healing in this mosquito species involved both humoral and cellular reactions. The latter reaction involved the movement of plasmatocytes to the basement membrane of the epidermis beneath the wound-site and epithelial cells regeneration. Our observations revealed that wound-healing in A. subalbatus involves the wound contraction, formation of a temporary cellular clump, scar formation, basement membrane formation, and reepithelialization. The larvae neither discarded the wound scar nor secreted a new cuticle until the next molting. Based on the ultrastructural observations, it is suggested that the wound-healing reaction in A. subalbatus was probably a typical response employed by other members of the family Culicidae.
Assuntos
Culicidae/ultraestrutura , Animais , Larva/ultraestrutura , Microscopia Eletrônica/métodosRESUMO
ABSTRACT A heteroduplex mobility assay (HMA) was used to analyze the variability among five isolates of Zucchini yellow mosaic virus (ZYMV; TW-TC1, TW-CY2, TW-TN3, TW-TNML1, and TW-NT1) collected from cucurbit fields in different areas of Taiwan. A cDNA fragment of 760 bp covering the variable region of the N terminal half of the coat protein (CP) gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and subsequently subjected to HMA analysis for sequence variation. When TW-NT1 combined with any of the other Taiwan isolates, the heteroduplexes obtained migrated much more slowly than did the heteroduplexes obtained in combinations among the other four Taiwan isolates, indicating that TW-TC1, TW-CY2, TW-TN3, and TW-TNML1 share a high degree of sequence homology, while the TW-NT1 isolate is more distinct. The complete nucleotide sequences of the CP genes and the 3' noncoding regions of the five isolates were determined from RT-PCR-derived cDNA clones. A phylogenetic tree derived from the actual sequences of the 760-bp fragments of the five Taiwan and another six ZYMV isolates from different geographic areas revealed four genotypes. TW-TNML1, TW-TC1, TWC-Y2, and TW-TN3 were in genotype I, while TW-NT1 and U.S. isolates were in genotype II. The Singapore and Reunion Island isolates were separated into genotypes III and IV, respectively. Comparison of the CP genes of the five Taiwan isolates indicated that they share 92.8 to 98.7% nucleotide identities and 96.4 to 99.3% amino acid identities. The amino acid positions 73, 102, 109, and 149 of the CP gene, where lysine, serine, arginine, and aspartic acid reside, respectively, were uniquely conserved for genotype I Taiwan isolates. Thus, results of HMA agreed well with those of phylogenetic analysis based on the sequence data of the five Taiwan ZYMV isolates. These five ZYMV isolates of known sequence can be used as reference strains for HMA to analyze the variability of ZYMV in Taiwan.
RESUMO
The encapsulation of microfilariae in the haemocoels of mosquitoes combines both humoral and cellular reactions: the microfilariae are first encased in an acellular layer of melanin, followed by a cellular encapsulation by plasmatocytes. In this study, we demonstrated that cellular encapsulation of Brugia pahangi microfilariae in the haemocoel of the mosquito Anopheles quadrimaculatus was terminated by the formation of a basement membrane-like structure on the outermost surface of the cellular capsule. This structure occurred in the early stage of cellular encapsulation and was evident on the exterior surface of the plasmatocyte, when the active haemocytes were attaching to the already melanized microfilariae. The termination structure appears to be laid down by releasing the vesicle inclusions of haemocytes and has similarities in ultrastructure and cationic colloidal gold staining properties with that of mosquito basement membranes.
Assuntos
Anopheles/parasitologia , Brugia pahangi , Animais , Anopheles/imunologia , Anopheles/ultraestrutura , Membrana Basal/citologia , Membrana Basal/ultraestrutura , Brugia pahangi/imunologia , Interações Hospedeiro-Parasita , Imunidade Celular , Microfilárias/imunologia , Microscopia Eletrônica , Ratos , Fatores de TempoRESUMO
Dengue 2 virus was found by transmission electron microscopy to be present in the reproductive tissues of male Aedes aegypti (L) 14 d after intrathoracic inoculation. Dengue 2 particles were detected in the matrix, epithelial cells, and the peripheral fat body of the testes; secretory droplets of columnar cells of the accessory glands; and the epithelial and muscle cells of the seminal vesicles. However, none was found in the germ cells (i.e., spermatogonia, spermatocytes, spermatid, or spermatozoa). These observations indicate that fluid transfer may be the mechanism of venereal transmission of dengue 2 virus by Ae. aegypti.
Assuntos
Aedes/virologia , Vírus da Dengue , Aedes/ultraestrutura , Animais , Linhagem Celular , Genitália Masculina/ultraestrutura , Genitália Masculina/virologia , MasculinoRESUMO
Differential viral gene expression during both productive and persistent infections of Hz-1 virus in insect cells was elucidated. Despite more than 100 viral transcripts being expressed during productive viral infection, massive viral gene shutoff was observed during viral persistency, leaving the 2.9-kb persistence-associated transcript 1 (PAT1) as the only detectable viral RNA. Persistence-associated gene 1 (pag1), which encodes PAT1, was cloned and found to contain no significant open reading frames. PAT1 is not associated with the cellular translation machinery and is located exclusively in the nucleus. Further experiments showed that PAT1 is functional in the establishment of persistent Hz-1 viral infection in the cells. All the evidence collectively indicates that PAT1 is a novel nuclear transcript of viral origin. Our results showed that although PAT1 and XIST RNA, a mammalian X-inactive specific transcript, are transcribed by different genes, they have interesting similarities.
Assuntos
Baculoviridae/genética , Genes Virais , RNA Viral , Proteínas Virais/genética , Latência Viral , Animais , Baculoviridae/fisiologia , Sequência de Bases , Sítios de Ligação , Linhagem Celular , Núcleo Celular/metabolismo , DNA Viral , Genoma Viral , Dados de Sequência Molecular , Mariposas/citologia , Família Multigênica , Fases de Leitura Aberta , Iniciação Traducional da Cadeia Peptídica , Regiões Promotoras Genéticas , Sequências Repetitivas de Ácido Nucleico , Ribossomos , Spodoptera/citologia , Transcrição GênicaRESUMO
The melanization of Dirofilaria immitis microfilariae in Armigeres subalbatus haemolymph in vitro is a two-step process. Firstly, the microfilariae are encased in a transparent capsule, then the capsule material is melanized later. Benzamadine HC1 and p-amidinophenylmethanesulfonyl fluoride, both serine protease inhibitors, inhibited the deposition of the transparent capsule material and melanization. Diethyldithiocarbamate, a phenoloxidase inhibitor, did not prevent the deposition of the transparent capsule material but it did inhibit melanization. m-Hydroxybenzylhydrazine, a dopa decarboxylase inhibitor, had no inhibitory action on the deposition of the transparent capsule material. However, the time for the transparent capsule to become melanized was delayed due to the presence of m-hydroxybenzylhydrazine in the test system. Immmunocytochemical localization showed that propheoloxidase and/or phenoloxidase was present in the transparent capsule material. These results suggest (1) that prophenoloxidase cascade is essential in the melanization of microfilariae in mosquitoes and its putative activation behaviour is similar to that of other insects and (2) that the deposition of transparent capsule material is the result of the activation of prophenoloxidase by serine proteases and the melanization of the transparent capsule material is due to the presence of active phenoloxidase in capsule.
Assuntos
Inibidores das Descarboxilases de Aminoácidos Aromáticos , Culicidae/enzimologia , Dirofilaria immitis/metabolismo , Hemolinfa/enzimologia , Melaninas/metabolismo , Monofenol Mono-Oxigenase/antagonistas & inibidores , Inibidores de Serina Proteinase/farmacologia , Animais , Benzamidinas/farmacologia , Culicidae/parasitologia , Ditiocarb/farmacologia , Dopa Descarboxilase/metabolismo , Inibidores Enzimáticos/metabolismo , Inibidores Enzimáticos/farmacologia , Interações Hospedeiro-Parasita , Hidrazinas/farmacologia , Microfilárias/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Fluoreto de Fenilmetilsulfonil/análogos & derivados , Fluoreto de Fenilmetilsulfonil/farmacologiaRESUMO
Sequence analysis of the p10 genes of three Bombyx mori nuclear polyhedrosis virus (BmNPV) isolates collected in Taiwan (Ta) and Japan (T3 and D1) showed that all possessed a deletion of an adenine residue, 210 bp downstream from the first base of the initiation codon when compared to the p10 gene of Autographa californica (multinucleocapsid) NPV (AcMNPV). This deletion caused a downstream termination codon to come inframe with the coding sequence of p10, so that the p10 gene of BmNPV encoded a protein of 70 amino acid residues with an M(r) of 7700. This is considerably shorter than the 10,000 M(r) protein encoded by the closely related AcMNPV.
Assuntos
Bombyx/microbiologia , Genes Virais/genética , Nucleopoliedrovírus/genética , Proteínas Virais/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Mutação da Fase de Leitura , Dados de Sequência Molecular , Deleção de Sequência , Homologia de Sequência do Ácido Nucleico , Especificidade da EspécieRESUMO
A background level of sister-chromatid exchange (SCE) is found in mammalian cells. However, in the somatic cells of Drosophila melanogaster, there was no spontaneous SCE if a minimum concentration of bromodeoxyuridine (BrdU) was used. In this communication we report that in a mosquito (Aedes albopictus) cell line (C6/36), some cells also have no background SCE (0-SCE). Subclones of high (44%) or low (12%) frequency of 0-SCE cells were obtained, but none of them contained 100% 0-SCE cells. Increasing frequency of SCE/cell concomitant with decreasing frequency of 0-SCE cells was observed by raising the BrdU concentration in the culture medium during the first cell cycle, culturing cells at lower density and depleting reduced glutathione (GSH) with buthionine sulfoximine. Since these mosquito cells contain much higher level of GSH than Chinese hamster ovary (CHO) cells, and feeding cells with BrdU increased GSH in mosquito cells but decreased GSH in CHO cells, we speculate that GSH may play a role in the low or non-existent background SCE and in the high resistance to many DNA damaging agents of mosquito cells.
Assuntos
Aedes/genética , Bromodesoxiuridina/toxicidade , Testes de Mutagenicidade/métodos , Mutagênicos/toxicidade , Troca de Cromátide Irmã , Animais , Butionina Sulfoximina , Células CHO , Células Cultivadas , Células Clonais , Cricetinae , Relação Dose-Resposta a Droga , Drosophila melanogaster , Glutationa/metabolismo , Metionina Sulfoximina/análogos & derivados , Metionina Sulfoximina/farmacologia , Mitose , Troca de Cromátide Irmã/efeitos dos fármacosRESUMO
Both Bacillus thuringiensis subsp. isrealensis (Bti) and Bacillus sphaericus are common bacteria found in a variety of soil and aquatic habitats. Some strains of them can produce a protein crystal, called parasporal body (or crystal) beside the spore in the cell during sporulation, and it is toxic if eaten by mosquito larvae. As the parasporal bodies are ingested into a mosquito digestive tract, they will be digested and activated by certain enzymes in alkaline digestive juice. The activated toxins can destroy midgut epithelium and other tissues, and then the mosquitoes will be intoxicated and killed. There are some factors affecting efficiency of the toxins produced by these two mosquitocidal bacteria. First, the toxicity of the toxins will decrease as larval age and density increase. Second, difference in mosquito species will demonstrate a different tolerance to both toxins. However, larvae of Anopheles are more tolerant to Bti than those of Aedes and Culex; whereas larvae of Aedes are more tolerant to B. sphaericus than Culex and Anopheles. Environmental factors are as important as the mosquito hosts in influencing the efficiency of the toxins. For examples, raising temperature under suitable ranges may increase the toxicity of the toxins. The UV light of sunlight could inhibit activity of the toxins and therefore decrease their efficiency. Quality and depth of the water are also important environmental factors affecting toxicity of the toxins to the mosquito larvae. The precipitation of the crystal and organic materials absorbing the toxins decreases their toxicity to the mosquito larvae as well.