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PLOS Global Public Health 2023Measurement of HIV-1 viral load (VL) is essential for monitoring antiretroviral treatment (ART) efficacy. The preferred specimen type for VL is plasma, but in remote...
Measurement of HIV-1 viral load (VL) is essential for monitoring antiretroviral treatment (ART) efficacy. The preferred specimen type for VL is plasma, but in remote settings where collection and preservation of plasma many not be possible, dried blood spots (DBS) are often used instead. A new specimen collection matrix, the cobas plasma separation card (PSC, Roche Diagnostics Solutions), enables specimen preparation from a finger prick or venous blood, using a multi-layer absorption and filtration design that results in a specimen similar to dried plasma. We sought to confirm the correlation between VL results obtained using PSC prepared from venous blood to those from plasma or DBS, as well as PSC prepared with capillary blood from a finger prick. PSC, DBS and plasma were prepared with blood from HIV-1 infected persons attending a primary care clinic in Kampala, Uganda. VL in PSC and plasma was measured using cobas HIV-1 (Roche Diagnostics), while VL in DBS was measured with RealTime HIV-1 (Abbott Diagnostics). The correlation between VL from plasma and PSC made from capillary or venous blood was high (regression coefficient of determination r2 between 0.87 and 0.91), and there was good agreement based on mean bias (-0.14 to 0.24 log10 copies/mL) and classification of VL above or below 1000 copies/mL (91.4% agreement). In contrast, VL from DBS was lower than plasma or PSC (mean bias 0.51 to 0.63 log10 copies/mL) and not as well correlated (r2 0.78 to 0.81, 75.1-80.5% agreement). These results confirm the utility of PSC as an alternative specimen type for HIV-1 viral load measurement in areas where preparation and optimal storage or shipment of plasma is an obstacle to provision of treatment and care of HIV-1 infected people.
PubMed: 37379313
DOI: 10.1371/journal.pgph.0002099 -
Pharmaceuticals (Basel, Switzerland) Jun 2023Opioids are considered the cornerstone of pain management: they show good efficacy as a first-line therapy for moderate to severe cancer pain. Since...
BACKGROUND
Opioids are considered the cornerstone of pain management: they show good efficacy as a first-line therapy for moderate to severe cancer pain. Since pharmacokinetic/pharmacodynamic information about the tissue-specific effect and toxicity of opioids is still scarce, their quantification in post-mortem autoptic specimens could give interesting insights.
METHODS
We describe an ultra-high-performance liquid chromatography coupled with tandem mass spectrometry method for the simultaneous quantification of methadone, morphine, oxycodone, hydrocodone, oxymorphone, hydromorphone and fentanyl in several tissues: liver, brain, kidney, abdominal adipose tissue, lung and blood plasma. The presented method has been applied on 28 autoptic samples from different organs obtained from four deceased PLWH who used opioids for palliative care during terminal disease.
RESULTS
Sample preparation was based on tissue weighing, disruption, sonication with drug extraction medium and a protein precipitation protocol. The extracts were then dried, reconstituted and injected onto the LX50 QSight 220 (Perkin Elmer, Milan, Italy) system. Separation was obtained by a 7 min gradient run at 40 °C with a Kinetex Biphenyl 2.6 µm, 2.1 × 100 mm. Concerning the analyzed samples, higher opioids concentrations were observed in tissues than in plasma. Particularly, O-MOR and O-COD showed higher concentrations in kidney and liver than other tissues (>15-20 times greater) and blood plasma (>100 times greater).
CONCLUSIONS
Results in terms of linearity, accuracy, precision, recovery and matrix effect fitted the recommendations of FDA and EMA guidelines, and the sensitivity was high enough to allow successful application on human autoptic specimens from an ethically approved clinical study, confirming its eligibility for post-mortem pharmacological/toxicological studies.
PubMed: 37375850
DOI: 10.3390/ph16060903 -
Bioengineering (Basel, Switzerland) Jun 2023The aim of this study was to compare concentrations of endogenous N-acylethanolamine (NAE) lipid mediators-palmitoylethanolamide (PEA), oleoylethanolamide (OEA), and...
The aim of this study was to compare concentrations of endogenous N-acylethanolamine (NAE) lipid mediators-palmitoylethanolamide (PEA), oleoylethanolamide (OEA), and anandamide (AEA)-in fresh, decontaminated, cryopreserved, and freeze-dried amniotic membrane (AM) allografts, thereby determining whether AM's analgesic and anti-inflammatory efficiency related to NAEs persists during storage. The concentrations of NAEs were measured using ultra-high-performance liquid chromatography-tandem mass spectrometry. Indirect fluorescent immunohistochemistry was used to detect the PEA PPAR-α receptor. The concentrations of PEA, OEA, and AEA were significantly higher after decontamination. A significant decrease was found in cryopreserved AM compared to decontaminated tissue for PEA but not for OEA and AEA. However, significantly higher values for all NAEs were detected in cryopreserved samples compared to fresh tissue before decontamination. The freeze-dried AM had similar values to decontaminated AM with no statistically significant difference. The nuclear staining of the PPAR-α receptor was clearly visible in all specimens. The stability of NAEs in AM after cryopreservation was demonstrated under tissue bank storage conditions. However, a significant decrease, but still higher concentration of PEA compared to fresh not decontaminated tissue, was found in cryopreserved, but not freeze-dried, AM. Results indicate that NAEs persist during storage in levels sufficient for the analgesic and anti-inflammatory effects. This means that cryopreserved AM allografts released for transplant purposes before the expected expiration (usually 3-5 years) will still show a strong analgesic effect. The same situation was confirmed for AM lyophilized after one year of storage. This work thus contributed to the clarification of the analgesic effect of NAEs in AM allografts.
PubMed: 37370671
DOI: 10.3390/bioengineering10060740 -
Annals of Clinical and Translational... Aug 2023Duchenne muscular dystrophy (DMD) is an X-linked disorder resulting in progressive muscle weakness and atrophy, cardiomyopathy, and in late stages, cardiorespiratory...
OBJECTIVE
Duchenne muscular dystrophy (DMD) is an X-linked disorder resulting in progressive muscle weakness and atrophy, cardiomyopathy, and in late stages, cardiorespiratory impairment, and death. As treatments for DMD have expanded, a DMD newborn screening (NBS) pilot study was conducted in New York State to evaluate the feasibility and benefit of NBS for DMD and to provide an early pre-symptomatic diagnosis.
METHODS
At participating hospitals, newborns were recruited to the pilot study, and consent was obtained to screen the newborn for DMD. The first-tier screen measured creatine kinase-MM (CK-MM) in dried blood spot specimens submitted for routine NBS. Newborns with elevated CK-MM were referred for genetic counseling and genetic testing. The latter included deletion/duplication analysis and next-generation sequencing (NGS) of the DMD gene followed by NGS for a panel of neuromuscular conditions if no pathogenic variants were detected in the DMD gene.
RESULTS
In the two-year pilot study, 36,781 newborns were screened with CK-MM. Forty-two newborns (25 male and 17 female) were screen positive and referred for genetic testing. Deletions or duplications in the DMD gene were detected in four male infants consistent with DMD or Becker muscular dystrophy. One female DMD carrier was identified.
INTERPRETATION
This study demonstrated that the state NBS program infrastructure and screening technologies we used are feasible to perform NBS for DMD. With an increasing number of treatment options, the clinical utility of early identification for affected newborns and their families lends support for NBS for this severe disease.
Topics: Infant; Humans; Male; Infant, Newborn; Female; Muscular Dystrophy, Duchenne; Neonatal Screening; Pilot Projects; Genetic Testing; High-Throughput Nucleotide Sequencing
PubMed: 37350320
DOI: 10.1002/acn3.51829 -
Journal of Orthopaedic Surgery (Hong... 2023Previous studies lacked adequate quantitative data on sustentaculum tali (ST), especially in Chinese population. The aims of this study are to explore the quantitative...
BACKGROUND
Previous studies lacked adequate quantitative data on sustentaculum tali (ST), especially in Chinese population. The aims of this study are to explore the quantitative morphology of ST in dried bone specimens, and to discuss its implications related to ST screw fixation, talar articular facet variation, as well as subtalar coalitions.
METHODS
A total of 965 dried intact calcanei from Chinese adult donors were evaluated. All linear parameters were measured by two observers with a digital sliding vernier caliper.
RESULTS
Most parts of ST body can accommodate a commonly-used 4-mm-diameter screw, but the minimum height of anterior ST is only 4.02 mm. The shapes of the STs are slightly affected by left-right, subtalar facet, but the subtalar coalition may potentially increase the sizes of STs. The incidence of tarsal coalition is 14.09%. Among the osseous connection, there are 58.8% of type A articular surface and 76.5% of middle and posterior talar facet (MTF and PTF) involvement. ROC curve shows that subtalar coalition will be detected when ST length is greater than 16.815 mm.
CONCLUSIONS
Theoretically, all the STs can accommodate 4 mm diameter screw, but a 3.5 mm diameter screw is recommended to be placed in the middle or posterior of the small ST for safety. The shapes of the STs are greatly influenced by the subtalar coalition, while they are less affected by left-right, subtalar facet. The osseous connection is common in type A articular surface and always involved in the MTF and PTF. The cut-off value of the length of STs was confirmed as 16.815 mm for predicting subtalar coalition.
Topics: Adult; Humans; Bone Screws; Calcaneus; Clinical Relevance; East Asian People; Lower Extremity
PubMed: 37341523
DOI: 10.1177/10225536231178354 -
Environment International Jul 2023Human exposure to mercury can have serious health effects, especially in vulnerable groups such as children and fetuses. The use of dried blood spot (DBS) samples to...
Human exposure to mercury can have serious health effects, especially in vulnerable groups such as children and fetuses. The use of dried blood spot (DBS) samples to collect capillary blood greatly facilitates sample collection and fieldwork, being a less invasive alternative to blood collection by venipuncture, needing a small volume of sample, and does not require specialized medical staff. Moreover, DBS sampling reduces logistical and financial barriers related to transport and storage of blood samples. We propose here a novel method to analyze total mercury in DBS samples in a Direct Mercury Analyzer (DMA) that allow the control of the volume of the DBS samples. This method has shown good results in terms of precision (<6% error), accuracy (<10% coefficient of variation) and recovery (75-106%). The applicability of the method in human biomonitoring (HBM) was demonstrated in a pilot study involving 41 adults aged 18-65. Mercury concentrations of DBS samples from capillary blood collected by finger prick (real DBS samples) were determined in the DMA and compared with those determined in whole blood (venous blood) by ICP-MS, the method usually used in HBM. The sampling procedure was also validated by comparison of real DBS samples and DBS generated artificially in the laboratory by depositing venous samples in cellulose cards (laboratory DBS). There were no statistically significant differences in the results obtained using both methodologies (DMA: Geometric Mean (confidence interval 95%) = 3.87 (3.12-4.79) µg/L; ICP-MS: Geometric Mean (confidence interval 95%) = 3.46 (2.80-4.27) µg/L). The proposed method is an excellent alternative to be applied in clinical settings as screening methodology for assessing mercury exposure in vulnerable groups, such us pregnant woman, babies and children.
Topics: Adult; Pregnancy; Female; Child; Humans; Biological Monitoring; Pilot Projects; Blood Specimen Collection; Mercury
PubMed: 37285712
DOI: 10.1016/j.envint.2023.107958 -
Iranian Journal of Basic Medical... 2023Neuroinflammation and microglial activation are pathological features in central nervous system disorders. Excess levels of reactive oxygen species (ROS) and...
OBJECTIVES
Neuroinflammation and microglial activation are pathological features in central nervous system disorders. Excess levels of reactive oxygen species (ROS) and pro-inflammatory cytokines have been implicated in exacerbation of neuronal damage during chronic activation of microglial cells. , a brown macroalga, has been demonstrated to have various pharmacological properties such as anti-neuroinflammatory activity. However, the underlying mechanism mediating the anti-neuroinflammatory potential of remains poorly understood. We explored the use of Malaysian in attenuating lipopolysaccharide (LPS)-stimulated neuroinflammation in BV2 microglial cells.
MATERIALS AND METHODS
Fresh specimens of were freeze-dried and subjected to ethanol extraction. The ethanol extract (PAEE) was evaluated for its protective effects against 1 µg/ml LPS-stimulated neuroinflammation in BV2 microglial cells.
RESULTS
LPS reduced the viability of BV2 microglia cells and increased the levels of nitric oxide (NO), prostaglandin E (PGE), intracellular reactive oxygen species (ROS), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6). However, the neuroinflammatory response was reversed by 0.5-2.0 mg/ml PAEE in a dose-dependent manner. Analysis of liquid chromatography-mass spectrometry (LC-MS) of PAEE subfractions revealed five compounds; methyl α-eleostearate, ethyl α-eleostearate, niacinamide, stearamide, and linoleic acid.
CONCLUSION
The protective effects of PAEE against LPS-stimulated neuroinflammation in BV2 microglial cells were found to be mediated by the suppression of excess levels of intracellular ROS and pro-inflammatory mediators and cytokines, denoting the protective role of in combating continuous neuroinflammation. Our findings support the use of as a possible therapeutic for neuroinflammatory and neurodegenerative diseases.
PubMed: 37275754
DOI: 10.22038/IJBMS.2023.67835.14842 -
PhytoKeys 2023was confirmed as polyphyletic by recent phylogenetic analyses, with Chinese species of distantly related to those from Japan. Among the Chinese species of , is a...
was confirmed as polyphyletic by recent phylogenetic analyses, with Chinese species of distantly related to those from Japan. Among the Chinese species of , is a morphologically unique as well as taxonomically problematic species endemic to South China, of which the generic designation is still uncertain. Molecular analyses based on both plastid and nuclear genomic data demonstrated that this species is closest to the recently published genus . Morphologically, the two are somewhat similar to each other in flowering branches developing at the nodes of every order of branches, raceme-like units of inflorescence with 3-5 short spikelets, each spikelet with few florets including a rudimentary one at the apex, and each floret with 3 stamens and 2 stigmas. However, is very different from species in many reproductive and vegetative characters, such as the morphology of paracladia (lateral spikelet "pedicels"), the absence or existence of pulvinus at the base of paracladia, the relative length of the upper glume and the lowest lemma, the shape of lodicules and primary culm buds, the branch complement, the morphology of nodes, culm leaves and dried foliage leaf blades, and the number of foliage leaves per ultimate branchlet. The morphological and molecular evidence warrants recognition of a new genus to accommodate this unique species, which is here named . After consulting related literature and examination of herbarium specimens or specimen photos, a taxonomic revision of and its synonyms was made, and it was confirmed that four names, viz. , , and , should be merged with , while and are distinct species.
PubMed: 37250355
DOI: 10.3897/phytokeys.221.98920 -
Influenza and Other Respiratory Viruses May 2023We estimated combined protection conferred by prior SARS-CoV-2 infection and COVID-19 vaccination against COVID-19-associated acute respiratory illness (ARI).
BACKGROUND
We estimated combined protection conferred by prior SARS-CoV-2 infection and COVID-19 vaccination against COVID-19-associated acute respiratory illness (ARI).
METHODS
During SARS-CoV-2 Delta (B.1.617.2) and Omicron (B.1.1.529) variant circulation between October 2021 and April 2022, prospectively enrolled adult patients with outpatient ARI had respiratory and filter paper blood specimens collected for SARS-CoV-2 molecular testing and serology. Dried blood spots were tested for immunoglobulin-G antibodies against SARS-CoV-2 nucleocapsid (NP) and spike protein receptor binding domain antigen using a validated multiplex bead assay. Evidence of prior SARS-CoV-2 infection also included documented or self-reported laboratory-confirmed COVID-19. We used documented COVID-19 vaccination status to estimate vaccine effectiveness (VE) by multivariable logistic regression by prior infection status.
RESULTS
Four hundred fifty-five (29%) of 1577 participants tested positive for SARS-CoV-2 infection at enrollment; 209 (46%) case-patients and 637 (57%) test-negative patients were NP seropositive, had documented previous laboratory-confirmed COVID-19, or self-reported prior infection. Among previously uninfected patients, three-dose VE was 97% (95% confidence interval [CI], 60%-99%) against Delta, but not statistically significant against Omicron. Among previously infected patients, three-dose VE was 57% (CI, 20%-76%) against Omicron; VE against Delta could not be estimated.
CONCLUSIONS
Three mRNA COVID-19 vaccine doses provided additional protection against SARS-CoV-2 Omicron variant-associated illness among previously infected participants.
Topics: Adult; Humans; COVID-19 Vaccines; COVID-19; SARS-CoV-2; Outpatients; Vaccine Efficacy; Influenza Vaccines
PubMed: 37246146
DOI: 10.1111/irv.13143 -
Fujita Medical Journal May 2023To establish a point-of-care test for coronavirus disease 2019 (COVID-19), we developed a dry loop-mediated isothermal amplification (LAMP) method to detect severe acute...
OBJECTIVES
To establish a point-of-care test for coronavirus disease 2019 (COVID-19), we developed a dry loop-mediated isothermal amplification (LAMP) method to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA.
METHODS
We carried out reverse transcription (RT)-LAMP using the Loopamp SARS-CoV-2 Detection kit (Eiken Chemical, Tokyo, Japan). The entire mixture, except for the primers, is dried and immobilized inside the tube lid.
RESULTS
To determine the specificity of the kit, 22 viruses associated with respiratory infections, including SARS-CoV-2, were tested. The sensitivity of this assay, determined by either a real-time turbidity assay or colorimetric change of the reaction mixture, as evaluated by the naked eye or under illumination with ultraviolet light, was 10 copies/reaction. No LAMP product was detected in reactions performed with RNA from any pathogens other than SARS-CoV-2. After completing an initial validation analysis, we analyzed 24 nasopharyngeal swab specimens collected from patients suspected to have COVID-19. Of the 24 samples, 19 (79.2%) were determined by real-time RT-PCR analysis as being positive for SARS-CoV-2 RNA. Using the Loopamp SARS-CoV-2 Detection kit, we detected SARS-CoV-2 RNA in 15 (62.5%) of the 24 samples. Thus, the sensitivity, specificity, positive predictive value, and negative predictive values of the Loopamp 2019-CoV-2 detection reagent kit were 78.9%, 100%, 100%, and 55.6%, respectively.
CONCLUSIONS
The dry LAMP method for detecting SARS-CoV-2 RNA is fast and easy to use, and its reagents can be stored at 4°C, solving the cold chain problem; thus, it represents a promising tool for COVID-19 diagnosis in developing countries.
PubMed: 37234399
DOI: 10.20407/fmj.2022-003