Monitoring of COVID-19 in wastewater across the Eastern Upper Peninsula of Michigan.

Wastewater-based epidemiology is being used as a tool to monitor the spread of COVID-19 and provide an early warning for the presence or increase of clinical cases in a community. The majority of wastewater-based epidemiology for COVID-19 tracking has been utilized in sewersheds that service populations in the tens-to-hundreds of thousands. Few studies have been conducted to assess the usefulness of wastewater in predicting COVID-19 clinical cases specifically in rural areas. This study collected samples from 16 locations across the Eastern Upper Peninsula of Michigan from June to December 2021. Sampling locations included 12 rural municipalities, a Tribal housing community and casino, a public university, three municipalities that also contained a prison, and a small island with heavy tourist traffic. Samples were analyzed for SARS-CoV-2 N1, N2, and variant gene copies using reverse transcriptase droplet digital polymerase chain reaction (RT-ddPCR). Wastewater N1 and N2 gene copies and clinical case counts were correlated to determine if wastewater results were predictive of clinical cases. Significant correlation between N1 and N2 gene copies and clinical cases was found for all sites (⍴= 0.89 to 0.48). N1 and N2 wastewater results were predictive of clinical case trends within 0-7 days. The Delta variant was detected in the Pickford and St. Ignace samples more than 12-days prior to the first reported Delta clinical cases in their respective counties. Locations with low correlation could be attributed to their high rates of tourism. This is further supported by the high correlation seen in the public university, which is a closed population. Long-term wastewater monitoring over a large, rural geographic area is useful for informing the public of potential outbreaks in the community regardless of asymptomatic cases and access to clinical testing.

Adult-onset mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes (MELAS) syndrome with progressive sensorineural hearing loss: A case report.

Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome is one of the most common maternally inherited mitochondrial disorders, with no specific treatment available. We report a case of a 34-year-old female in whom symptoms of MELAS were initially misdiagnosed as herpes simplex encephalitis (HSE). Her clinical course was marked by an acute episode of consciousness disturbance with newly developed lesions on brain MRI five years after disease onset and followed by progressive sensorineural hearing loss. Brain imaging sequences throughout the seven years of her illness are presented. The final diagnosis of MELAS syndrome was confirmed by m.3243A>G mitochondrial mutation. In conclusion, understanding the overlapping imaging features between MELAS syndrome and other mimickers, such as HSE or ischemic stroke, is crucial to help establish early diagnosis and initiate appropriate treatment.

Detection of HPAI H5N1 viruses in ducks sampled from live bird markets in Vietnam.

In Vietnam, highly pathogenic avian influenza (HPAI) H5N1 infections in poultry often occur without concomitant clinical signs and outbreaks are not consistently reported. Live bird markets represent a convenient site for surveillance that does not rely on farmers' notifications. Two H5N1 surveys were conducted at live bird markets/slaughter points in 39 districts (five provinces) in the Red River, Mekong delta, and central Vietnam during January and May 2011. Oropharyngeal and rectal swab samples from 12 480 ducks were tested for H5N1 by reverse transcription-polymerase chain reaction in pools of five. Traders and stallholders were interviewed using standardized questionnaires; 3·3% of pools tested positive. The highest prevalence (6·6%) corresponded to the Mekong delta, and no H5N1 was detected in the two Red River provinces. The surveys identified key risk behaviours of traders and stallholders. It is recommended that market surveys are implemented over time as a tool to evaluate progress in HPAI control in Vietnam.

The ethics of a clinical trial when the protocol clashes with international guidelines.

Due to their nature and complexity, clinical trials often take some time to launch after the protocol has been designed and ethics approval obtained. During this time, there may be changes in international treatment guidelines and recommendations that result in a conflict between study protocol and recommended international best practice. Here, we describe the situation that arose in a pharmacokinetic study on the use of two different doses of rifabutin in patients with human immunodeficiency virus-associated tuberculosis who initiated antiretroviral therapy (ART) with a lopinavir-ritonavir-based regimen in South Africa and Viet Nam. The study protocol specified that ART should be started 10 weeks after the start of anti-tuberculosis treatment. The study in South Africa was approved in June 2008, went ahead as scheduled and was completed in August 2010. The study in Viet Nam was approved in October 2008 and was started in June 2010. A few weeks later, the World Health Organization released their 2010 guidelines for adult ART; one of its strong recommendations (with moderate quality of evidence) was that ART should be started 2-8 weeks after the start of anti-tuberculosis treatment. Emerging scientific evidence also supported this recommendation. The investigators felt that the Viet Nam study protocol was in conflict with recommended international best practice, and the trial was stopped in October 2010. An amended study protocol in which ART was started at 2 weeks was developed and implemented. The ethics issues around this decision and the need to change the study protocol are discussed in this article.

Du fait de la nature et de la complexité des études cliniques, leur mise en œuvre est souvent longue après l'élaboration du protocole et son approbation éthique. Pendant cette période, il peut y avoir un changement des lignes directrices internationales de traitement et des recommandations qui provoquent un conflit entre le protocole et les meilleures pratiques recommandées internationalement. Nous décrivons ici la situation apparue dans une étude pharmacocinétique portant sur l'utilisation de deux doses différentes de rifabutin chez des patients atteints de tuberculose (TB) associée au virus de l'immunodéficience humaine et commençant un traitement antirétroviral (ART) à base de lopinavir-ritonavir en Afrique du Sud et au Viet Nam. Le protocole de l'étude spécifiait de commencer l'ART 10 semaines après le début de la thérapie antituberculeuse. En Afrique du Sud, l'étude a été approuvée en juin 2008, s'est déroulée comme prévu et a été achevée en juin 2010. Au Viet Nam, l'étude a été approuvée en octobre 2008 et a démarré en juin 2010. Quelques semaines après, l'Organisation Mondiale de la Santé a publié ses lignes directrices de 2010 pour l'utilisation de l'ART chez les adultes, dont l'une des vives recommandations (basée sur des données de qualité modérée) était de commencer l'ART entre 2 et 8 semaines après le début du traitement de la TB. L'arrivée de nouvelles preuves scientifiques est aussi venue à l'appui de cette recommandation. Les investigateurs ont eu le sentiment que le protocole d'étude au Viet Nam était en conflit avec les meilleures pratiques internationales et l'étude a été arrêtée en octobre 2010. Un protocole d'étude amendé a été développé et mis en œuvre. Les problèmes éthiques entourant cette décision et la nécessité de changer le protocole sont discutés dans ce papier.

Los ensayos clínicos, dadas sus características y su complejidad, suelen exigir mucho tiempo desde la elaboración del protocolo y la aprobación por parte del comité de ética hasta su realización. Durante este lapso, pueden surgir modificaciones en las recomendaciones y las directrices terapéuticas internacionales, lo cual genera un conflicto entre el protocolo del estudio y las prácticas óptimas recomendadas. A continuación se describe la situación que se presentó en Suráfrica y Viet Nam durante un estudio de farmacocinética sobre el uso de dos dosificaciones diferentes de rifabutina, en pacientes aquejados de tuberculosis (TB) asociada con la infección por el virus de la inmunodeficiencia humana (HIV), quienes habían comenzado el tratamiento antirretrovírico (ART) con un régimen basado en la asociación lopinavir y ritonavir. El protocolo del estudio precisaba que el ART se debía comenzar 10 semanas después de haber iniciado el tratamiento antituberculoso. En Suráfrica, el estudio recibió la aprobación en junio del 2008, comenzó en el tiempo previsto y se completó en agosto del 2010. En Viet Nam, se obtuvo la aprobación del estudio en octubre del 2008 y se comenzó en junio del 2010. A las pocas semanas, la Organización Mundial de Salud publicó sus directrices del ART en los adultos del 2010, una de cuyas recomendaciones más firmes consistía en que el ART se debía iniciar entre 2 y 8 semanas después de haber comenzado el tratamiento antituberculoso (con una calidad probatoria moderada). Algunos resultados científicos de aparición reciente respaldaban igualmente esta recomendación. Los investigadores consideraron que el protocolo del estudio en Viet Nam entraba en conflicto con las prácticas óptimas internacionales recomendadas e interrumpieron su realización en octubre del 2010. Se introdujeron modificaciones al protocolo, según las cuales el ART se comenzaría a las 2 semanas y se puso en práctica el estudio. En el presente artículo se analizan los aspectos éticos en torno a esta decisión y a la necesidad de modificar el protocolo del estudio.

Radioprotection by hymenialdisine-derived checkpoint kinase 2 inhibitors.

DNA damage induced by ionizing radiation activates the ataxia telangiectasia mutated pathway, resulting in apoptosis or DNA repair. The serine/threonine checkpoint kinase (Chk2) is an important transducer of this DNA damage signaling pathway and mediates the ultimate fate of the cell. Chk2 is an advantageous target for the development of adjuvant drugs for cancer therapy, because inhibition of Chk2 allows normal cells to enter cell cycle arrest and DNA repair, whereas many tumors bypass cell cycle checkpoints. Chk2 inhibitors may thus have a radioprotective effect on normal cells. We report herein a class of natural product derived Chk2 inhibitors, exemplified by indoloazepine 1, that elicit a strong ATM-dependent Chk2-mediated radioprotection effect in normal cells and p53 wt cells, but not p53 mutant cells (>50% of all cancers). This study represents the first example of a radioprotective effect in human cells other than T-cells and implicates a functional ATM pathway as a requirement for IR-induced radioprotection by this class of Chk2 inhibitors. Several of the hymenialdisine-derived analogues inhibit Chk2 at nanomolar concentrations, inhibit autophosphorylation of Chk2 at Ser516 in cells, and increase the survival of normal cells following ionizing radiation.

Characterization of a deoxyguanosine adduct of tetrachlorobenzoquinone: dichlorobenzoquinone-1,N2-etheno-2'-deoxyguanosine.

Pentachlorophenol (PCP), a widespread environmental pollutant that is possibly carcinogenic to humans, is metabolically oxidized to tetrachloroquinone. DNA adducts attributable to tetrachloroquinone have been observed previously in vitro and detected in vivo. In addition, an unidentified adduct in these studies coeluted with the product of the reaction of deoxyguanosine (dG) and tetrachlorobenzoquinone (Cl4BQ). We have synthesized, isolated, purified, and characterized the predominant adduct formed from the reaction of dG and Cl4BQ. The preparation of a 13C-labeled version of this adduct facilitated its structural characterization. On the basis of 1H NMR, 13C NMR, MS, IR, UV, and cyclic voltammetry, we propose that the adduct is a dichlorobenzoquinone nucleoside in which two chlorine atoms in Cl4BQ have been displaced by reaction at the 1- and N2-positions of dG. The 1H and 13C NMR chemical shifts are consistent with the dichlorobenzoquinone assignment. In contrast, under standard analytical conditions, LC-MS data are consistent with a reduced hydroquinone structure, similar to what may be expected based on results from other chloroquinones. Data from the present study indicate that this reduction could be occurring in the electrospray ionization source and that the initial product of the reaction of dG and Cl4BQ is a dichlorobenzoquinone. The results of this study contribute to the hypothesis that direct reactions between chlorophenols and DNA may play a role in the toxic effects of chlorophenols and indicate a potential difference in reactivity and biological influence between PCP and other less substituted chlorophenols or phenols.

Value of MR imaging in evaluating focal fatty infiltration of the liver: preliminary study.

Focal fatty infiltration of the liver is a well-known entity that occasionally mimics metastatic disease on ultrasonographic (US) and computed tomographic (CT) scans and requires biopsy for diagnosis. To determine if high-field-strength magnetic resonance (MR) imaging might be useful in the differential diagnosis of the lesions, the authors compared US, CT, and MR findings in three patients with biopsy-proved fatty hepatic lesions. Areas of focal fatty infiltration were hyperechoic on US scans and had low attenuation on CT scans. No mass effect of the lesions on vascular structures or liver contours was observed, particularly on contrast material-enhanced CT scans. For all three patients, MR findings suggested the correct diagnosis by demonstrating focal high signal intensity on spin-echo T1- and T2-weighted images. On the basis of these preliminary findings, it appears that focal fatty infiltration of the liver may be differentiated from metastatic disease by means of high-field-strength MR imaging.

[Lymphographic evaluation of cervico-uterine carcinoma (author's transl)]. / Bilan lymphographique des cancers cervico-utérins.

Bilateral pedial lymphography is a technique which has already been long employed and which had aroused great hopes in the detection of metastatic node involvement. The method has nevertheless failed to form an invariable part of the pretherapeutic assessment of cervico-uterine carcinomas. This attitude may be easily explained by the difficulties encountered in the interpretation of lymphographies as indicated in the first publications. New data nevertheless have made it possible to refine the method. Whilst false negatives remain inevitable in almost half the cases with histological evidence of involvement, false positives may be reduced to a minimum by very strict criteria of interpretation. Lymphography, a harmless technique, can then provide valuable information which enables the surgeon to confirm the success of his lymph node dissection and the radiotherapist to define areas requiring treatment. It is of major prognostic value. It may be used to guide transparietal lymph node biopsies and secondary detect lumbo-aortic nodes.