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Pharmacophores such as hydroxyethylamine (HEA) and phthalimide (PHT) have been identified as potential synthons for the development of compounds against various parasitic infections. In order to further advance our progress, we conducted an experiment utilising a collection of PHT and HEA derivatives through phenotypic screening against a diverse set of protist parasites. This approach led to the identification of a number of compounds that exhibited significant effects on the survival of Entamoeba histolytica, Trypanosoma brucei, and multiple life-cycle stages of Leishmania spp. The Leishmania hits were pursued due to the pressing necessity to expand our repertoire of reliable, cost-effective, and efficient medications for the treatment of leishmaniases. Antileishmanials must possess the essential capability to efficiently penetrate the host cells and their compartments in the disease context, to effectively eliminate the intracellular parasite. Hence, we performed a study to assess the effectiveness of eradicating L. infantum intracellular amastigotes in a model of macrophage infection. Among eleven L. infantum growth inhibitors with low-micromolar potency, PHT-39, which carries a trifluoromethyl substitution, demonstrated the highest efficacy in the intramacrophage assay, with an EC50 of 1.2 +/- 3.2 µM. Cytotoxicity testing of PHT-39 in HepG2 cells indicated a promising selectivity of over 90-fold. A chemogenomic profiling approach was conducted using an orthology-based method to elucidate the mode of action of PHT-39. This genome-wide RNA interference library of T. brucei identified sensitivity determinants for PHT-39, which included a P-type ATPase that is crucial for the uptake of miltefosine and amphotericin, strongly indicating a shared route for cellular entry. Notwithstanding the favourable properties and demonstrated efficacy in the Plasmodium berghei infection model, PHT-39 was unable to eradicate L. major infection in a murine infection model of cutaneous leishmaniasis. Currently, PHT-39 is undergoing derivatization to optimize its pharmacological characteristics.
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Antiprotozoários , Leishmania infantum , Leishmania , Leishmaniose Cutânea , Humanos , Animais , Camundongos , Antiprotozoários/farmacologia , Antiprotozoários/uso terapêutico , Anfotericina B/uso terapêutico , Leishmaniose Cutânea/parasitologia , Ftalimidas/farmacologia , Ftalimidas/uso terapêuticoRESUMO
With gradual fall in autopsy all over the world in recent years, the present study aimed to assess the accuracy of clinical diagnosis and efficacy of needle autopsy from the emergency department. Fifty deceased patients, who died in the emergency department during a period of 1 year, were subjected to needle autopsy of the major viscera, using spring-loaded automated biopsy gun, and the findings were correlated with clinical diagnosis. The deceased patients were in the age range of 12 to 80 years (mean [SD], 50.48 [18.41] years). The tissues yielded from various organs were as follows: lungs, 90%; liver, 82%; kidney, 48%; heart, 28%; spleen, 22%; and pancreas, 18%. Before death, 86 clinical diagnoses were recorded, of which 21 (24%) (eg, metabolic encephalopathy, cardiac arrhythmia, diabetic ketoacidosis) were impossible to verify on needle autopsy. A total of 48 new diagnoses, missed by physicians, were revealed by needle autopsy. The most frequently missed diagnoses were liver fatty change (19 patients) and pneumonitis (11 patients). Other frequently missed diagnoses were chronic hepatitis (3 patients) and cancer (2 patients: 1 lung squamous cell carcinoma and 1 lung adenocarcinoma). Major diagnostic errors (Goldman classes I and II) were noted in 16 (32%) of 50 cases. Needle autopsy can be a better alternative in the absence of conventional autopsy.
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Autopsia/métodos , Biópsia por Agulha , Serviço Hospitalar de Emergência , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Erros de Diagnóstico , Fígado Gorduroso/diagnóstico , Feminino , Patologia Legal , Hepatite Crônica/diagnóstico , Hospitais de Ensino , Humanos , Índia , Rim/patologia , Fígado/patologia , Pulmão/patologia , Neoplasias Pulmonares/diagnóstico , Masculino , Pessoa de Meia-Idade , Miocárdio/patologia , Pâncreas/patologia , Pneumonia/diagnóstico , Pneumonia/microbiologia , Estudos Prospectivos , Baço/patologia , Centros de Atenção Terciária , Tuberculose Pulmonar/diagnóstico , Adulto JovemRESUMO
Acquired amegakaryocytic thrombocytopenia (AAMT) is a rare disorder of the bone marrow characterized by a lack of megakaryocytes and preservation of other cell lines. It can occur due to an intrinsic stem cell defect or secondary to viral infections, autoimmune disorders, lymphoproliferative disorders or environmental toxins. With time, it can progress to aplastic anaemia (AA) and can have a poor prognosis. No standard guidelines exist for the treatment of AAMT progressing to AA. Herein, we report a rare case of AAMT leading to AA and review the handful of cases previously published in the literature. LEARNING POINTS: Acquired amegakaryocytic thrombocytopenia can present as isolated severe thrombocytopenia which can initially be misdiagnosed as immune thrombocytopenia.Lack of response to steroids and intravenous immunoglobulin should raise suspicion for acquired amegakaryocytic thrombocytopenia.Over time, acquired amegakaryocytic thrombocytopenia can progress to aplastic anaemia, which confers a worse prognosis.
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Previously, our group had demonstrated long term stabilization of protein biomarkers using BioCaRGOS, a silica sol-gel technology. Herein, we describe workflow modifications to allow for extraction of cell free DNA (cfDNA) from primary samples containing working concentrations of BioCaRGOS, as well as the compatibility of BioCaRGOS with droplet digital PCR (ddPCR) analysis for pancreatic cancer biomarkers i.e., KRAS circulating tumor DNA (ctDNA). Preliminary attempts to extract ctDNA from BioCaRGOS containing samples demonstrated interference in the extraction of primary samples and the interference with ddPCR analysis when BioCaRGOS was directly introduced to stabilize sample extracts. In our modified technique, we have minimized the interference caused by methanol with ddPCR by complete removal of methanol from the activated BioCaRGOS formulation prior to addition to the biospecimen or ctDNA extract. Interference of the silica matrix present in BioCaRGOS with ctDNA extraction was eliminated through the introduction of invert filtration of the sample prior to extraction. These modifications to the workflow of BioCaRGOS containing samples allow for use of BioCaRGOS for stabilization of trace quantities of nucleic acid biomarkers such as plasma ctDNA, while retaining the capability to extract the biomarker and quantify based on ddPCR.
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Constant emergence of drug-resistant Plasmodium falciparum warrants urgent need for effective and inexpensive drugs. Herein, phthalimide (Pht) analogs possessing the bioactive scaffolds, benzimidazole and 1,2,3-triazole, were evaluated for in vitro and in vivo anti-plasmodial activity without any apparent hemolysis, or cytotoxicity. Analogs 4(a-e) inhibited the growth of 3D7 and RKL-9 strains at submicromolar concentrations. Defects were observed during parasite egress from or invasion of the red blood cells. Mitochondrial membrane depolarization was measured as one of the causes of cell death. Phts 4(a-e) in combination with artemisinin exhibited two-to three-fold increased efficacy. Biophysical and biochemical analysis suggest that Pht analogs mediate plasmodial growth inhibition by interacting with tubulin protein of the parasite. Lastly, Phts 4(a-e) significantly decreased parasitemia and extended host survival in murine model Plasmodium berghei ANKA infection. Combined, the data indicate that Pht analogs should be further explored, which could offer novel value to the antimalarial drug development pipeline.
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Antimaláricos , Malária , Animais , Antimaláricos/química , Malária/tratamento farmacológico , Malária/parasitologia , Camundongos , Ftalimidas/química , Ftalimidas/farmacologia , Plasmodium berghei , Plasmodium falciparum , Tubulina (Proteína)RESUMO
Malaria remains one of the world's most life-threatening diseases and, thus, it is a major public health concern all around the world. The disease can become devastating if not treated with proper medication in a timely manner. Currently, the number of viable treatment therapies is in continuous decline due to compromised effectiveness, probably owing to the complex life cycle of Plasmodium falciparum. The factors responsible for the unclear status of malaria eradication programmes include ever-developing parasite resistance to the most effective treatments used on the frontline (i.e., artemisinin derivatives) and the paucity of new effective therapeutics. Due to these circumstances, the development of novel effective drug candidates with unique modes of action is essential for overcoming the listed obstacles. As such, the discovery of novel chemical compounds based on validated pharmacophores remains an unmet need in the field of medicinal chemistry. In this area, functionalized phthalimide (Pht) analogs have been explored as potential candidates against various diseases, including malaria. Pht presents a promising bioactive scaffold that can be easily functionalized and thus utilized as a starting point for the development of new antimalarial candidates suitable for preclinical and clinical studies. In this short review, we highlight a wide range of Pht analogs that have been investigated for their activity against various strains of Plasmodium falciparum.
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BACKGROUND: Premature infants are born with lower iron stores and are at risk for iron deficiency during early infancy. To prevent iron deficiency, premature infants are routinely supplemented with 2 mg/kg/day oral elemental iron. Reticulocyte hemoglobin content (RET-He), a measure of iron deficiency, has not been well evaluated prior to discharge in premature infants. OBJECTIVES: Our objectives were to evaluate RET-He and its correlation with serum ferritin (SF), an index of iron stores, at 35-36 weeks postmenstrual age (PMA) in ≤32 weeks gestational age (GA) infants. METHODS: We performed a prospective nested study involving 24-32 weeks GA infants who were receiving 2 mg/kg/day oral elemental iron with full enteral feedings at 35-36 weeks PMA. Infants with the following conditions were excluded: craniofacial malformation, chromosomal disorders, TORCH (toxoplasmosis, other infections, rubella, cytomegalovirus, and herpes simplex), culture-proven sepsis, C-reactive protein >5 mg/l within 10 days of iron status assessment, or erythropoietin therapy. SF and RET-He were measured at 35-36 weeks PMA using chemiluminescence immunoassay and Sysmex XN hematology analyzer, respectively. RET-He <27 pg was deemed indicative of iron deficiency. RESULTS: Ninety-eight infants were studied, of which 21 infants had RET-He <27 pg. There was a positive correlation between RET-He and SF (coefficient 0.22, p = .03) that remained significant after controlling for GA (coefficient 0.21, p = .03) and frequency of prior erythrocyte transfusions (coefficient 0.21, p = .03). On stratified analysis, there was a positive correlation between SF and RET-He in females (N = 52, coefficient 0.23, p = .02), but not in males (N = 46, coefficient 0.05). CONCLUSIONS: Most premature infants receiving 2 mg/kg/day oral elemental iron are iron replete for erythropoiesis at 35-36 weeks PMA. RET-He increases with an increase in iron stores, suggesting that additional iron supplementation prior to discharge to very premature infants with borderline low RET-He may help prevent iron deficiency during early infancy.
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Anemia Ferropriva , Reticulócitos , Feminino , Hemoglobinas/análise , Humanos , Lactente , Recém-Nascido , Recém-Nascido Prematuro , Ferro , Masculino , Estudos Prospectivos , Reticulócitos/químicaRESUMO
Room temperature biospecimen storage for prolonged periods is essential to eliminate energy consumption by ultra-low freezing or refrigeration-based storage techniques. State of the art practices that sufficiently minimize the direct or hidden costs associated with cold-chain logistics include ambient temperature storage of biospecimens (i.e., DNA, RNA, proteins, lipids) in the dry state. However, the biospecimens are still well-exposed to the stress associated with drying and reconstitution cycles, which augments the pre-analytical degradation of biospecimens prior to their downstream processing. An aqueous storage solution that can eliminate these stresses which are correlated to several cycles of drying/rehydration or freezing of biospecimens, is yet to be achieved by any current technology. In our study, we have addressed this room temperature biospecimen-protection challenge using aqueous capture and release gels for optimized storage (Bio-CaRGOS) of biospecimens. Herein, we have demonstrated a single-step â¼95% recovery of a metalloprotein hemoglobin at room temperature using a cost-effective standard microwave-based aqueous formulation of Bio-CaRGOS. Although hemoglobin samples are currently stored at sub-zero or under refrigeration (4 °C) conditions to avoid loss of integrity and an unpredictable diagnosis during their downstream assays, our results have displayed an unprecedented room temperature integrity preservation of hemoglobin. Bio-CaRGOS formulations efficiently preserve hemoglobin in its native state, with single-step protein recovery of â¼95% at ambient conditions (1 month) and â¼96% (7 months) under refrigeration conditions. In contrast, two-thirds of the control samples degrade under ambient (1 month) and refrigeration (7 months) settings.
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Novel coronavirus disease 2019 (COVID-19) has significantly altered the socio-economic status of countries. Although vaccines are now available against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a causative agent for COVID-19, it continues to transmit and newer variants of concern have been consistently emerging world-wide. Computational strategies involving drug repurposing offer a viable opportunity to choose a medication from a rundown of affirmed drugs against distinct diseases including COVID-19. While pandemics impede the healthcare systems, drug repurposing or repositioning represents a hopeful approach in which existing drugs can be remodeled and employed to treat newer diseases. In this review, we summarize the diverse computational approaches attempted for developing drugs through drug repurposing or repositioning against COVID-19 and discuss their advantages and limitations. To this end, we have outlined studies that utilized computational techniques such as molecular docking, molecular dynamic simulation, disease-disease association, drug-drug interaction, integrated biological network, artificial intelligence, machine learning and network medicine to accelerate creation of smart and safe drugs against COVID-19.
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Storage of biospecimens in their near native environment at room temperature can have a transformative global impact, however, this remains an arduous challenge to date due to the rapid degradation of biospecimens over time. Currently, most isolated biospecimens are refrigerated for short-term storage and frozen (-20 °C, -80 °C, liquid nitrogen) for long-term storage. Recent advances in room temperature storage of purified biomolecules utilize anhydrobiosis. However, a near aqueous storage solution that can preserve the biospecimen nearly "as is" has not yet been achieved by any current technology. Here, we demonstrate an aqueous silica sol-gel matrix for optimized storage of biospecimens. Our technique is facile, reproducible, and has previously demonstrated stabilization of DNA and proteins, within a few minutes using a standard benchtop microwave. Herein, we demonstrate complete integrity of miRNA 21, a highly sensitive molecule at 4, 25, and 40 °C over a period of â¼3 months. In contrast, the control samples completely degrade in less than 1 week. We attribute excellent stability to entrapment of miRNA within silica-gel matrices.
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Nucleophilic ring opening reactions of epoxides with aromatic amines are in the forefront of the synthetic organic chemistry research to build new bioactive scaffolds. Here, convenient, green, and highly efficient regioselective ring opening reactions of sterically hindered (2R,3S)-3-(N-Boc-amino)-1-oxirane-4-phenylbutane with various poorly reactive aromatic amines are accomplished under microwave irradiation in nitromethane. All the reactions effectively implemented for various aromatic amines involve the reuse of nitromethane that supports its dual role as a solvent and catalyst. The corresponding new ß-alcohol analogs of hydroxyethylamine (HEA) are isolated in 41-98% yields. The reactions proceed under mild conditions for a broad range of less reactive and sterically hindered aromatic amines. Proton NMR experiments suggest that the nucleophilicity of amines is influenced by nitromethane, which is substantiated by the extensive computational studies. Overall, this methodology elucidates the first-time use of nitromethane as a solvent for the ring opening reactions under microwave conditions involving an equimolar ratio of epoxide and aromatic amine without any catalyst, facile ring opening of complex epoxide by less reactive aromatic amines, low reaction time, less energy consumption, recycling of the solvent, and simple workup procedures.
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Malaria, a global threat to the human population, remains a challenge partly due to the fast-growing drug-resistant strains of Plasmodium species. New therapeutics acting against the pathogenic asexual and sexual stages, including liver-stage malarial infection, have now attained more attention in achieving malaria eradication efforts. In this paper, two previously identified potent antiplasmodial hydroxyethylamine (HEA) compounds were investigated for their activity against the malaria parasite's multiple life stages. The compounds exhibited notable activity against the artemisinin-resistant strain of P. falciparum blood-stage culture with 50% inhibitory concentrations (IC50) in the low micromolar range. The compounds' cytotoxicity on HEK293, HepG2 and Huh-7 cells exhibited selective killing activity with IC50 values > 170 µM. The in vivo efficacy was studied in mice infected with P. berghei NK65, which showed a significant reduction in the blood parasite load. Notably, the compounds were active against liver-stage infection, mainly compound 1 with an IC50 value of 1.89 µM. Mice infected with P. berghei sporozoites treated with compound 1 at 50 mg kg-1 dose had markedly reduced liver stage infection. Moreover, both compounds prevented ookinete maturation and affected the developmental progression of gametocytes. Further, systematic in silico studies suggested both the compounds have a high affinity towards plasmepsin II with favorable pharmacological properties. Overall, the findings demonstrated that HEA and piperidine possessing compounds have immense potential in treating malarial infection by acting as multistage inhibitors.
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Differentiating between malignant phyllodes tumors and metaplastic spindle cell carcinomas could be problematic, especially on core biopsies. Immunohistochemical staining for cytokeratin cocktail and p63 has been utilized to differentiate between these tumor types. Forty-three phyllodes tumors (27 benign, 6 borderline, and 10 malignant) and 22 metaplastic carcinomas, consisting at least 80% of spindle cells, were identified. At least 4 tissue blocks from each phyllodes tumor were subjected to immunohistochemical staining for cytokeratin cocktail and p63. The immunohistochemical profiles for the spindle cells in metaplastic carcinoma were reviewed. Phyllodes tumor was diagnosed in the younger age group (mean age 41 y) with a larger tumor size (mean size 6.6 cm), compared with metaplastic spindle cell carcinoma (mean age 62.7 y, mean size 3.4 cm). Focal expression (5% of the tumor cells) of cytokeratin cocktail and p63 was identified in the stroma of 2 of 10 malignant phyllodes tumors in a scattered/patchy pattern. The stroma of benign and borderline phyllodes tumors was negative for these markers. In metaplastic spindle cell carcinomas, cytokeratin cocktail was negative in 2 of 15 cases and very focally positive in another 3 cases, whereas p63 was negative in one case and focally positive in another case. There can be anomalous, focal expression of cytokeratin and p63 in the stroma of malignant phyllodes tumors, whereas metaplastic spindle cell carcinoma can occasionally have cytokeratin and/or p63-negative staining or have very focal positivity. Caution should be exercised when relying on these markers for confirming a diagnosis, especially on core biopsies.
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Carcinoma/metabolismo , Queratinas/metabolismo , Proteínas de Membrana/metabolismo , Tumor Filoide/metabolismo , Adolescente , Adulto , Idoso de 80 Anos ou mais , Carcinoma/fisiopatologia , Diagnóstico Diferencial , Humanos , Imuno-Histoquímica , Limite de Detecção , Metaplasia , Pessoa de Meia-Idade , Tumor Filoide/fisiopatologiaRESUMO
Non-tuberculous mycobacteria are increasingly recognized as a cause of infection in both immunocompromised and immunocompetent hosts. Mycobacterium heraklionense is a recently described member of the Mycobacterium terrae complex. Herein we report a case of M. heraklionense chronic flexor tenosynovitis in the hand, managed with surgery and antibiotics.
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Mãos/patologia , Infecções por Mycobacterium não Tuberculosas/diagnóstico , Infecções por Mycobacterium não Tuberculosas/patologia , Micobactérias não Tuberculosas/isolamento & purificação , Tenossinovite/etiologia , Tenossinovite/patologia , Idoso , Antibacterianos/uso terapêutico , Mãos/cirurgia , Humanos , Masculino , Infecções por Mycobacterium não Tuberculosas/microbiologia , Infecções por Mycobacterium não Tuberculosas/terapia , Micobactérias não Tuberculosas/classificação , Procedimentos Cirúrgicos Operatórios , Tenossinovite/microbiologia , Tenossinovite/terapia , Resultado do TratamentoRESUMO
In X-ray fluorescence, the earlier derived matrix effects from fundamental relations of intensities of analyte/matrix elements with basic atomic and experimental setup parameters and tested on synthetic known samples were found empirically related to analyte/matrix elemental amounts. The present study involves the application of these relations on potassium and calcium macronutrients of maize saplings treated with different fertilizers. The novelty of work involves a determination of an element in the presence of its secondary excitation rather than avoiding the secondary fluorescence. Therefore, the possible utility of this process is in studying the absorption for some intermediate samples in a lot of a category of samples with close Z interfering constituents (just like Ca and K). Once the absorption and enhancement terms are fitted to elemental amounts and fitted coefficients are determined, with the absorption terms from the fit and an enhancer element amount known from its selective excitation, the next iterative elemental amount can be directly evaluated from the relations.