As SARS-CoV-2 continues to spread and evolve, detecting emerging variants early is critical for public health interventions. Inferring lineage prevalence by clinical testing is infeasible at scale, especially in areas with limited resources, participation, or testing and/or sequencing capacity, which can also introduce biases1–3. SARS-CoV-2 RNA concentration in wastewater successfully tracks regional infection dynamics and provides less biased abundance estimates than clinical testing4,5. Tracking virus genomic sequences in wastewater would improve community prevalence estimates and detect emerging variants. However, two factors limit wastewater-based genomic surveillance: low-quality sequence data and inability to estimate relative lineage abundance in mixed samples. Here we resolve these critical issues to perform a high-resolution, 295-day wastewater and clinical sequencing effort, in the controlled environment of a large university campus and the broader context of the surrounding county. We developed and deployed improved virus concentration protocols and deconvolution software that fully resolve multiple virus strains from wastewater. We detected emerging variants of concern up to 14 days earlier in wastewater samples, and identified multiple instances of virus spread not captured by clinical genomic surveillance. Our study provides a scalable solution for wastewater genomic surveillance that allows early detection of SARS-CoV-2 variants and identification of cryptic transmission.

Background. Since postcastration progression of tumors to an androgen‐independent state appears to be linked to the cessation of androgen‐induced differentiation of tumorigenic stem cells, the authors hypothesized that the replacement of androgens at the end of a period of apoptotic regression might result in the regeneration of differentiated tumor cells with further apoptotic potential. Methods and Results. To determine the effect of intermittent exposure of androgens on the androgen‐dependent Shionogi carcinoma, the tumor was transplanted into a succession of male mice, each of which was castrated when the estimated tumor weight became about 3 g. After the tumor had regressed to 30% of the original weight, it was transplanted into the next noncastrated male. This cycle of transplantation and castration‐induced apoptosis was repeated successfully four times before growth became androgen‐independent during the fifth cycle. In four of Stage C and three of Stage D patients with prostate cancer, androgen withdrawal was initiated with cyproterone acetate (100 mg/d) and diethylstilbestrol (0.1 mg/d) and then maintained with cyproterone acetate in combination with the luteinizing hormone‐releasing hormone agonist, goserelin acetate (3.6 mg/month). After 6 or more months of suppression of serum prostate‐specific antigen (PSA) into the normal range, treatment was interrupted for 2 to 11 months. After recovery of testicular function, androgen‐withdrawal therapy was resumed when serum PSA increased to a level of about 20 μg/l. This cycle was repeated sequentially to a total of two to four times over treatment periods of 21 to 47 months with no loss of androgen dependence. Conclusions. These results demonstrate that intermittent androgen suppression can be used to induce multiple apoptotic regressions of a tumor; they also suggest that the cyclic effects of such treatment on prostate cancer can be followed by the sequential measurement of serum PSA levels.

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Ultrasonography (US) of the carotid arteries is a common imaging study performed for diagnosis of carotid artery disease. In the United States, carotid US may be the only diagnostic imaging modality performed before carotid endarterectomy. Therefore, the information obtained with carotid US must be reliable and reproducible. Technical parameters that can affect the accuracy of carotid US results include the Doppler angle, sample volume box, color Doppler sampling window, color velocity scale, and color gain. Important factors in diagnosis of atherosclerotic disease of the extracranial carotid arteries are the intima-media thickness, plaque morphology, criteria for grading stenosis, limiting factors such as the presence of dissection or cardiac abnormalities, distinction between near occlusion and total occlusion, and the presence of a subclavian steal. Challenges to the consistency of carotid US results may include lack of a standard protocol, poor Doppler technique, inexperience in interpretation of hemodynamic changes reflected in the Doppler waveform, artifacts, and physical challenges. Hindrances in the classification of problematic carotid artery stenoses may be overcome by following a standard protocol and optimizing scanning techniques and Doppler settings.