LC-MS-MS-MS3 for the determination and quantification of ∆9-tetrahydrocannabinol and metabolites in blood samples.

Due to the high prevalence of cannabinoids in forensic toxicology analysis, it is crucial to have an efficient method that allows the use of a small sample amount and that requires a minimal sample preparation for the determination and quantification of low concentrations. A simple, highly selective and high throughput liquid chromatography-tandem mass spectrometry methodology (LC-MS-MS-MS3) was developed for the determination and quantification of ∆9-tetrahydrocannabinol (THC), 11-hydroxy-∆9- tetrahydrocannabinol (THC-OH) and 11-nor-9-carboxy-∆9-tetrahydrocannabinol (THC-COOH) in blood samples. Chromatographic analysis of THC, THC-OH and THC-COOH and their deuterated internal standards was preceded by protein precipitation (PPT) of 0.1 mL of blood samples with acetonitrile. Chromatographic separation was achieved by use of an Acquity UPLC® HHS T3 (100 mm × 2.1 mm i.d., 1.8 µm) reversed-phase column, using a gradient elution of 2 mM aqueous ammonium formate, 0.1% formic acid and methanol at a flow rate of 0.4 mL/min, with a run time of 10 min. For the MS-MS-MS3 analysis, a SCIEX QTRAP® 6500+ triple quadrupole linear ion trap mass spectrometer was used via electrospray ionization (ESI), operated in multiple reaction monitoring (MRM) and linear ion trap mode (MS3). The method was validated in accordance with internationally accepted criteria and guidelines, and proved to be selective and linear between 0.5 and 100 ng/mL (r2 > 0.995). The lower limits of quantification (LLOQ) corresponded to the lowest concentrations used for the calibration curves. The coefficients of variation obtained for accuracy and precision were <15%. The mean recoveries were between 88.0% and 117.2% for the studied concentration levels (1 ng/mL, 5 ng/mL and 50 ng/mL). No significant interfering compounds, matrix effects or carryover were observed. The validated method provides a sensitive, efficient and robust procedure for the quantification of cannabinoids in blood, using LC-MS-MS-MS3 and a sample volume of 0.1 mL. This work is also a proof of concept for using LC-MS3 technique to determine drugs in biological samples.

Conditions of Confinement in U.S. Carceral Facilities During COVID-19: Individuals Speak-Incarcerated During the COVID-19 Epidemic.

Objectives: We aimed to describe conditions of confinement among people incarcerated in the United States during the coronavirus disease 2019 (COVID-19) pandemic using a community-science data collection approach. Methods: We developed a web-based survey with community partners to collect information on confinement conditions (COVID-19 safety, basic needs, support). Formerly incarcerated adults released after March 1, 2020, or nonincarcerated adults in communication with an incarcerated person (proxy) were recruited through social media from July 25, 2020 to March 27, 2021. Descriptive statistics were estimated in aggregate and separately by proxy or formerly incarcerated status. Responses between proxy and formerly incarcerated respondents were compared using Chi-square or Fisher's exact tests based on α=0.05. Results: Of 378 responses, 94% were by proxy, and 76% reflected state prison conditions. Participants reported inability to physically distance (≥6 ft at all times; 92%), inadequate access to soap (89%), water (46%), toilet paper (49%), and showers (68%) for incarcerated people. Among those receiving prepandemic mental health care, 75% reported reduced care for incarcerated people. Responses were consistent between formerly incarcerated and proxy respondents, although responses by formerly incarcerated people were limited. Conclusions: Our findings suggest that a web-based community-science data collection approach through nonincarcerated community members is feasible; however, recruitment of recently released individuals may require additional resources. Our data obtained primarily through individuals in communication with an incarcerated person suggest COVID-19 safety and basic needs were not sufficiently addressed within some carceral settings in 2020-2021. The perspectives of incarcerated individuals should be leveraged in assessing crisis-response strategies.

Macroalgae-Fortified Sausages: Nutritional and Quality Aspects Influenced by Non-Thermal High-Pressure Processing.

The present work evaluated the nutritional impact of macroalgae flours used as new ingredients in fermented sausages and the feasibility of using high-pressure processing (HPP) as a non-thermal pasteurization methodology to keep the quality attributes of the new food products. A commercial macroalgae mix was used in the formulation of new macroalgae-fortified meat frankfurter sausages (F-MFS), macroalgae-fortified vegetable frankfurter sausages (F-VFS) and in macroalgae-fortified traditional Portuguese sausage "chouriço" (F-TPS), overall incrementing the contents of Mg, K, Ca, Mn and Fe and decreasing the Na/K ratio. The application of HPP allowed extending the shelf-life of frankfurters by about 3-fold and improved the safety of "chouriço" along 180 days of storage, keeping its microbial load below the detection limit. The prevention of microbial growth in F-MFS and F-VFS was accompanied by pH stability of the products. In addition, no significant detriment on surface color and fatty acids was observed between pressurized and non-pressurized sausages, allowing consolidating the suitability of HPP in seaweed-fortified fermented sausages.

Deconstruction of banana peel for carbohydrate fractionation.

The deconstruction of banana peel for carbohydrate recovery was performed by sequential treatment (acid, alkaline, and enzymatic). The pretreatment with citric acid promoted the extraction of pectin, resulting in a yield of 8%. In addition, xylose and XOS, 348.5 and 17.3 mg/g xylan, respectively, were also quantified in acidic liquor as a result of partial depolymerization of hemicellulose. The spent solid was pretreated with alkaline solution (NaOH or KOH) for delignification and release of residual carbohydrates from the hemicellulose. The yields of xylose and arabinose (225.2 and 174.0 mg/g hemicellulose) were approximately 40% higher in the pretreatment with KOH, while pretreatment with NaOH promoted higher delignification (67%), XOS yield (32.6 mg/g xylan), and preservation of cellulosic fraction. Finally, the spent alkaline solid, rich in cellulose (76%), was treated enzymatically to release glucose, reaching the final concentration of 28.2 g/L. The mass balance showed that through sequential treatment, 9.9 g of xylose, 0.5 g of XOS, and 8.2 g of glucose were obtained from 100 g of raw banana peels, representing 65.8% and 46.5% conversion of hemicellulose and cellulose, respectively. The study of the fractionation of carbohydrates in banana peel proved to be a useful tool for valorization, mainly of the hemicellulose fraction for the production of XOS and xylose with high value applications in the food industry.

Hydrothermal treatment on depolymerization of hemicellulose of mango seed shell for the production of xylooligosaccharides.

Hydrothermal processing is an interesting biorefinery technology for converting lignocellulosic biomass into biofuels and biocompounds. This process is based on the selective solubilization and depolymerization of hemicellulose fraction (xylan) and may be considered beneficial, due to the possibility of obtaining xylooligosaccharides (XOS) with a degree of polymerization (DP) suitable for prebiotic applications. This study evaluated the effect of pressure (2.5 and 10 MPa) in a kinetic study (30 min) of hydrothermal treatment (180 °C) to optimize the extraction of XOS from mango seed shell. Total reducing sugars (TRS) values were close to the maximum in 15 min showing a slower rate for both pressures after this time, but at 10 MPa the value was 20 % lower than at 2.5 MPa. Based on these results, a new extraction was performed at 2.5 MPa and 15 min, and the extracted XOS were quantified, yielding 393.44 mg XOS/g xylan. XOS with a degree of polymerization between X2-X6 corresponded to 82.24 mg/g and XOS with X > 6 (or soluble xylan) corresponded to 311.20 mg/g. A low amount of xylose (8.81 mg/g xylan) was released, resulting in a hemicellulose conversion of 40.2 %. In general, approximately 8.1 kg of total XOS was produced from 100 kg of dried mango seed shell (X2-X6-1.7 kg and X > 6-6.4 kg).

Development and Validation of an Analytical Method for Volatiles with Endogenous Production in Putrefaction and Submersion Situations.

A group of 16 volatile substances (ethyl acetate, 2-propanol, 1-propanol, methanol, acetone, ethanol, acetaldehyde, diethyl ether, methyl ethyl ketone, 1-butanol, 2-butanol, t-butanol, isobutanol, 2-methyl-1-butanol, 3-methyl-1-butanol and 1-pentanol) were qualitatively and quantitatively analyzed through a method developed for volatiles with endogenous production in putrefaction and submersion situations. The method was validated for blood, urine and vitreous humor, using a gas chromatograph (Varian 450-GC) with a flame ionization detector coupled to a headspace injector (HS-GC-FID). The vials were prepared by diluting 100 µL of the sample of interest in 1 mL of internal standard (acetonitrile 100 mg/L), using two capillary columns (VF-624ms and VF-5ms) with different polarities to ensure that all test compounds would be properly identified and undoubtedly distinguished from the rest. All volatiles were studied in a range of 50 to 2,000 mg/L in terms of selectivity/specificity, detection and quantification limits, linearity and calibration model, precision, accuracy, bias, robustness and stability according to the Scientific Working Group for Forensic Toxicology. Detection and quantification limits were between 1 to 8 mg/L and 4 to 24 mg/L, respectively, with coefficient of variation values under 10% in bias studies and in intermediate precision studies for most substances. The developed method was applied to real cases to test the method.

Identification and Quantification of Antipsychotics in Blood Samples by LC-MS-MS: Case Reports and Data from Three Years of Routine Analysis.

Antipsychotic drugs (AP) are widely prescribed for the treatment of schizophrenia and psychosis. The pharmacological treatment of schizophrenia is often performed with the simultaneous use of two or more antipsychotic agents to achieve the desired control of psychotic symptoms Available AP include both conventional (typical) and new (atypical) antipsychotic medications. Atypical AP, such as quetiapine, now account for the vast majority of AP prescriptions. In forensic toxicology, AP are of considerable interest because of their potential abuse and their involvement in intoxications and suicides. The authors retrospectively examined AP positive cases detected in samples collected during autopsies performed in the Forensic Clinical and Pathology Service of National Institute of Legal Medicine and Forensic Sciences Centre Branch or in other autopsies carried out in the central region of Portugal, between January 2016 and December 2018. A quantitative liquid chromatography-tandem mass spectrometry assay was developed for the simultaneous determination of 16 AP (amisulpride, aripiprazole, chlorpromazine, clozapine, cyamemazine, fluphenazine, haloperidol, levomepromazine, melperone, olanzapine, paliperidone, promethazine, quetiapine, risperidone, sulpiride and ziprasidone) in blood samples of postmortem cases. The Laboratory of Forensic Chemistry and Toxicology received 3,588 requests for toxicological analysis: 1,413 cases were positive for drugs from which 351 (24.8%) cases were positive for AP, 60.1% from male individuals and 39.9% from female. Quetiapine was the most prevalent AP (36.5%) followed by olanzapine (20.8%). During this period, there were 25 postmortem cases with AP blood concentrations above therapeutic range, in which 36% of those are in agreement with the information received (psychological history or acute intoxication suspicion) and the manner of death was suicide. Our results point that antipsychotics are an increasingly prevalent class of drugs. AP must be measured not only in toxic concentrations but also in therapeutic levels in postmortem cases; therefore, it is important to come up with a sensitive method to cover the low therapeutic range in which AP are usually present.

Exame físico na enfermagem: avaliação do conhecimento teórico-prático / Examen físico en enfermería: evaluación del conocimiento teórico-práctico / Physical examination in nursing: evaluation of theoretical-practical knowledge

Objetivo: Verificar o conhecimento teórico-prático dos enfermeiros sobre a técnica do exame físico céfalo-caudal em um hospital universitário da cidade de São Paulo. Método: Estudo descritivo-exploratório de abordagem quantitativa realizado com 51 enfermeiros de unidades de internação e terapia intensiva adulto, submetidos a um questionário estruturado. A análise dos resultados foi feita por meio da estatística descritiva e análise inferencial por associação entre as variáveis de interesse. Resultados: Os dados revelaram índices de acertos acima de 70%. A graduação foi considerada o período de maior aquisição de conhecimento sobre o exame físico (p=0,039). O tempo de formação e atuação foram associados a maior escolaridade (p<0,05), a qual foi determinante para prática do exame físico (73,8%). Conclusão: os achados desta investigação reforçam a importância da educação permanente junto aos enfermeiros para a prática do exame físico com objetivo de manter uma assistência de qualidade orientada para a segurança do paciente.(AU)

Objective: To verify the theoretical and practical knowledge of nurses on the technique of cephalo-caudal physical examination at a university hospital in the city of São Paulo. Method: Descriptive-exploratory study with a quantitative approach carried out with 51 nurses from inpatient and adult intensive care units, submitted to a structured questionnaire. The analysis of the results was performed using descriptive statistics and inferential analysis using evidence of association between the variables of interest. Results: The data revealed success rates above 70%. Graduation was considered the period of the greatest acquisition of knowledge about the physical examination (p = 0.039). The time since graduation was associated with higher education (p <0.05), which was decisive for the practice of physical examination (73.8%). Conclusion: the findings of this investigation reinforce the importance of continuing education with nurses for the practice of physical examination in order to maintain quality care oriented to patient safety. (AU)

Objetivo: verificar los conocimientos teóricos y prácticos de las enfermeras sobre la técnica del examen físico cefalo-caudal en un hospital universitario de la ciudad de São Paulo. Método: estudio descriptivo-exploratorio con enfoque cuantitativo realizado con 51 enfermeras de unidades de cuidados intensivos para pacientes hospitalizados y adultos, sometidas a un cuestionario estructurado. El análisis de los resultados se realizó mediante estadística descriptiva y análisis inferencial utilizando evidencia de asociación entre las variables de interés. Resultados: Los datos revelaron tasas de éxito superiores al 70%. La graduación se consideró el período de mayor adquisición de conocimiento sobre el examen físico (p = 0.039). El tiempo transcurrido desde la graduación se asoció con la educación superior (p <0.05), lo cual fue decisivo para la práctica del examen físico (73.8%). Conclusión: los resultados de esta investigación refuerzan la importancia de la educación continua con enfermeras para la práctica del examen físico a fin de mantener una atención de calidad orientada a la seguridad del paciente. (AU)

Magnésio para perda auditiva: sinopse baseada em evidências / Magnesium for hearing loss: evidence-based synopsis

Contexto: O magnésio age no transporte iônico na cóclea e sua deficiência é relacionada ao aumento do estresse oxidativo na orelha interna, com consequente surgimento de perda auditiva e sintomas otoneurológicos, como tontura e zumbido. Alguns pesquisadores têm preconizado sua suplementação para tratamento de perda auditiva sensorioneural. O estudo objetivou avaliar as evidências na literatura relativas ao uso de magnésio para tratamento da perda auditiva em humanos. Desenho de estudo: Trata-se de sinopse de evidências. Métodos: Procedeu-se à busca em quatro bases eletrônicas de dados: Cochrane Central Register of Controlled Trials (2019), PUBMED (1966-2019), EMBASE (1974-2019) e Portal BVS (1982-2019), além do "megabuscador" de evidências Trip Database (2019). Não houve restrição geográfica e de idioma, sendo utilizados descritores e termos do Decs (Descritores em Ciências da Saúde). O método de síntese envolveu a combinação de estudos semelhantes em uma revisão narrativa. Resultados: Foram identificadas 75 citações e 4 estudos foram incluídos. A amostragem nos estudos, em geral, foi pequena e as intervenções não permitem afirmar a efetividade do magnésio na prevenção e no tratamento da perda auditiva sensorioneural. Discussão: A evidência é baixa e não permite responder à questão com os estudos atuais, sendo recomendada a realização de novos ensaios clínicos de qualidade. Conclusões: Não há suporte atualmente na literatura para uso do magnésio na prevenção e no tratamento da perda auditiva sensorioneural, sendo que sua utilização clínica é eminentemente empírica.

First report of Neotrombicula inopinata infestation in domestic cats from Portugal.

Trombiculids parasitize a wide variety of terrestrial vertebrates, including domestic animals, throughout the world. They are parasites only during their larval stages, causing several dermatological lesions on their hosts, such as acute dermatitis, erythema, excoriation, erosion, papules, crusts and alopecia on the ear margins, face, interdigital spaces and abdomen. Neotrombicula is one of the several genera in Trombiculidae family, which cause trombiculosis. The most common species implicated in clinical cases is Neotrombicula autumnalis. However, several reports have shown that Neotrombicula inopinata (Oudemans, 1909) can also play a role in trombiculosis. Here, we describe the first case of N. inopinata infestation in domestic cats from mainland Portugal. Since nucleic acids of Anaplasma phagocytophilum and Borrelia burgdorferi have been found in Neotrombicula autumnalis and Rickettsia spp. in Neotrombicula inopinata, a correct taxonomical identification is essential to understand the role of these mite species as possible vectors of pathogens.

Traumatic Brain Injury-Related Symptoms Reported by Parents: Clinical, Imaging, and Host Predictors in Children with Impairments in Consciousness Less than 24 Hours.

This study examined the relationship between acute neuroimaging, host and injury factors, and parent-reported traumatic brain injury (TBI)-related symptoms in children with noncritical head injury at two weeks and three months after injury. Data were collected prospectively on 45 subjects aged three to 16 years old enrolled in the Transforming Research and Clinical Knowledge in Traumatic Brain Injury (TRACK-TBI) study. Subjects had rapid recovery of mental status (Glasgow Coma Score [GCS] = 15 within 24 h), and had no clinical need for neurosurgical intervention. Intra- or extra-axial magnetic resonance imaging (MRI) lesions were categorized using Common Data Elements (CDE) definitions. Host and acute injury factors including neurobehavioral history, race, extracranial injuries, loss of consciousness (LOC), and GCS were analyzed while controlling for pre-injury symptoms, age, sex, and socioeconomic status. Parent-reported cognitive and somatic symptoms were measured by the Health and Behavior Inventory (HBI). Forty-nine percent of children had MRI lesions, most of which were relatively small. LOC predicted increased cognitive and somatic symptoms at two weeks. At three months, pre-injury neurobehavioral history predicted increased cognitive and somatic symptoms. Neuroimaging findings did not predict parent-reported symptom severity, except at three months where extra-axial lesions were associated with less severe cognitive symptoms. While structural MRI lesions do not predict increased parent-reported symptoms in this population, age-specific child performance measures may be more sensitive outcome measures and require further study. Children with pre-injury neurobehavioral problems have more severe symptoms at three months and thus may benefit from longer follow-up and monitoring after traumatic brain injury.

Acute intraparenchymal cerebral haemorrhage in an Iberian golden eagle - a case report.

BACKGROUND: In birds there are reports of intracranial lesions but not of the clinical, computed tomographic and histopathologic features of acute intraparenchymal cerebral haemorrhage in Iberian golden eagle. CASE PRESENTATION: The following report describes a case of a 30-year-old Iberian golden eagle (Aquila chrysaetos homeyeri) with no history of trauma, presented with acute opisthotonus, left head tilt and circling, anisocoria, positional nystagmus, and ataxia. The main differential diagnosis were hypovitaminosis B or E and intracranial disease due to trauma, infection, toxins or masses. A computed tomography (CT) of the head was performed with an 8-slices scanner and evidenced a hyperdense (63-65 HU) non-enhancing homogeneous well delineated round area in the midbrain, with 6 mm in its highest diameter. The attenuation values and the non-enhancing nature of the lesion strongly suggested the diagnosis of acute intraparenchymal haemorrhage, which was histologically confirmed after necropsy. CONCLUSIONS: In birds with a central neurological dysfunction, the diagnosis of acute brain haemorrhage should be considered when the CT evidences a non-enhancing, homogeneous, well circumscribed hyperattenuated round area.

Infectious Postpartum Sacroiliitis: The Importance and Difficulty of Early Diagnosis.

The sacroiliitis accounts for about 1.5% - 10% of all cases of septic arthritis and it is strongly associated with gynaecological infections, pelvic trauma or drug abuse (3.4% - 12.8% of cases occur during the postpartum period). Early diagnosis is difficult because the symptoms are nonspecific in pregnancy and in the postpartum period, making the delay of treatment a serious risk of irreversible damage to the joint and development of post-infectious complications. The authors describe the case of a 37-year-old puerperal woman presented to hospital, weeks after urgent caesarean section, with endometritis, post-anesthetic epidural hematoma and secondary infectious postpartum sacroiliitis. The diagnosis of sacroiliac joints pathology during pregnancy and puerperium is challenging. The pathogenesis of infectious sacroiliitis results from local contamination by contiguous infection or hematogenous spread of bacterial infections. The prognosis is usually favorable and depends on early diagnosis and treatment.

A sacroileíte representa 1,5% - 10% de todos os casos de artrite séptica e está fortemente associada a infecções ginecológicas, trauma pélvico e a abuso de drogas (3,4% - 12,8% dos casos ocorrem durante o puerpério). O diagnóstico precoce é difícil, pois os sintomas são inespecíficos na gravidez e no puerpério, retardando o tratamento, podendo assim causar danos irreversíveis à articulação e desenvolvimento de complicações pós-infecciosas. Os autores descrevem o caso de uma puérpera de 37 anos que duas semanas após realização de cesariana urgente iniciou quadro de endometrite, hematoma epidural secundário a complicação anestésica e sacroileíte infecciosa secundária. O diagnóstico de patologia da articulação sacroilíaca durante a gravidez e puerpério constitui um desafio. A patogénese da artrite séptica resulta da contaminação local por infecção contígua ou da disseminação hematogénea. O prognóstico da sacroileíte infecciosa do pós-parto é geralmente favorável e depende do diagnóstico e tratamento precoces.

Temporal windows of reproductive opportunity reinforce species barriers in a marine broadcast spawning assemblage.

Prezygotic isolating mechanisms act to limit hybridization and maintain the genetic identity of closely-related species. While synchronous intraspecific spawning is a common phenomenon amongst marine organisms and plays an important role in reproductive success, asynchronous spawning between potentially hybridizing lineages may also be important in maintaining species boundaries. We tested this hypothesis by comparing reproductive synchrony over daily to hourly timescales in a sympatric assemblage of intertidal fucoid algae containing selfing hermaphroditic (Fucus spiralis and Fucus guiryi) and dioecious (Fucus vesiculosus and Fucus serratus) species. Our results confirm that gametes are released on semi-lunar cycles in all species. However, sister species with different mating systems showed asynchronous spawning at finer circadian timescales, thus providing evidence for a partial reproductive barrier between hermaphroditic and dioecious species. Finally, our data also emphasize the ecological, developmental, and/or physiological constraints that operate to restrict reproduction to narrow temporal windows of opportunity in the intertidal zone and more generally the role of ecological factors in marine speciation.

Reproductive investment, synchrony and recruitment success in marine broadcast spawners: Effects of mating system and habitat (exposed shore versus estuary).

The timing and synchrony of gamete release in broadcast spawners have important implications for fertilization success, recruitment and to explain differences in reproductive success under distinct reproductive modes in sympatry. Our objective was to compare the reproductive timing and investment for sister species with contrasting mating systems; Fucus guiryi (selfing hermaphroditic) and Fucus vesiculosus (dioecious) in habitats with different wave exposures (exposed shore and estuary). Over two months, daily gamete release, recruitment and population structure were recorded. Our results show spawning synchrony between species and habitats, but release events in hermaphrodites occupied broader temporal windows in estuarine than exposed shore habitats. On the exposed shore both species increased the synchrony of release and amount of eggs. In the estuary, hermaphrodites relied on broader temporal spawning windows and a larger canopy, and the dioecious species had higher recruitment success, important factors determining persistence.