bRapid real-time PCR (RT-PCR) can be performed in a community hospital setting to identify Coccidioides species using the new Becton Dickinson molecular instrument BD Max. Following sample preparation, DNA extraction and PCR were performed on the BD Max using the BD Max extraction kit ExK-DNA-1 test strip and a master mix prepared by BioGX (Birmingham, AL). Sample preparation took 2 h, and testing on the BD Max took an additional 2 h. Method sensitivity and specificity were evaluated along with the limits of detection to confirm that this convenient method would provide medically useful information. Using serial dilutions, the lower limit of detection was determined to be 1 CFU/l. Testing with this method was validated using samples from various body sites, including bronchial alveolar lavage (BAL) fluid; sputum and lung tissue samples; and pleural and spinal fluids. Safety protocols were established, and specimen preparation processes were developed for the various types of specimens. The range for the cycle threshold (C T ) indicating adequate fluorescent signal to signify a positive result was established along with the acceptable range for the internal standard. Positive controls run with each batch were prepared by spiking a pooled BAL fluid specimen with a known dilution of Coccidioides immitis organism. Our experience with testing >330 patient samples shows that clinically relevant information can be available within 4 h using an RT-PCR method on the BD Max to identify Coccidioides spp., with sensitivity equivalent to culture. V alley fever caused by the dimorphic fungi Coccidioides immitis in the Central Valley of California and Coccidioides posadasiiin other arid areas of the southwestern United States continues to be an important illness in those areas (1-3). According to the Centers for Disease Control and Prevention, 4,431 cases of Valley fever were reported in California in 2012 (4). For most healthy residents of the Central Valley who contract this infection, Valley fever causes a rather mild influenza-like illness. However, the illness can be severe in some individuals, especially those with a compromised immune system. Without sensitive testing methods, it is usually not possible to distinguish the early symptoms of coccidioidomycosis from other causes of community-acquired pneumonia, which may lead to delayed or improper treatment (5).Current laboratory testing methods rely on lengthy, labor-intensive protocols using experienced and highly skilled laboratory personnel. Methods include traditional serology testing for immunodiffusion tube precipitin (TP) or complement fixation (CF) antibody, enzyme immunoassays, culture, and histopathology (6). Serology testing for Coccidioides is helpful but has limitations for diagnosing current, active infections, since antibodies may be slow to increase to detectable levels, especially in immunocompromised patients (7, 8). Culture-based methods require several days or weeks to grow sufficient fungi for identification by molecular methods, posing additional sa...

Targeted protein degradation and stabilization are promising therapeutic modalities due to their potency and versatility. However, only few E3 ligases and deubiquitinases have been harnessed for this purpose. Moreover, there may be other protein classes that could be exploited for protein stabilization or degradation. Here, we used a proteome-scale platform to identify hundreds of human proteins that can promote the degradation or stabilization of a target protein in a proximity-dependent manner. This allowed us to comprehensively compare the activities of human E3s and deubiquitinases, characterize non-canonical protein degraders and stabilizers, and establish that effectors have vastly different activities against diverse targets. Notably, the top degraders were more potent against multiple therapeutically relevant targets than the currently used E3s CBRN and VHL. Our study provides a functional catalogue of effectors for targeted protein degradation and stabilization and highlights the potential of induced proximity screens for discovery of novel proximity-dependent protein modulators.

Background Sperm storage plays a key role in the reproductive success of many sexually-reproducing organisms, and the capacity of long-term sperm storage varies across species. While there are theoretical explanations for why such variation exists, to date there are no controlled empirical tests of the reproductive consequences of additional long-term sperm storage. While Dipterans ancestrally have three long-term sperm organs, known as the spermathecae, Drosophila contain only two. Results We identified a candidate gene, which we call spermathreecae (sp3), in which a disruption cause the development of three functional spermathecae rather than the usual two in Drosophila. We used this disruption to test the reproductive consequences of having an additional long-term sperm storage organ. Compared to females with two spermathecae, females with three spermathecae store a greater total number of sperm and can produce offspring a greater length of time. However, they did not produce a greater total number of offspring. Conclusions Thus, additional long-term sperm storage in insects may increase female fitness through extending the range of conditions where she produces offspring, or through increasing the quality of offspring via enhanced local sperm competition at fertilization.

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