Synthesis and In Vitro Antimycobacterial and Antibacterial Activity of 8-OMe Ciprofloxacin-Hydrozone/Azole Hybrids.

A series of novel 8-OMe ciprofloxacin (CPFX)-hydrazone/azole hybrids were designed, synthesized, and evaluated for their in vitro biological activities. Our results reveal that all of the hydrozone-containing hybrids (except for 7) show potency against Mycobacterium tuberculosis (MTB) H37Rv (minimum inhibitory concentration (MIC): <0.5 µM), which is better than the parent drug CPFX, and comparable to moxifloxacin and isoniazid, some of the tested Gram-positive strains (MIC: 0.06-4 µg/mL), and most Gram-negative strains (MIC: ≤0.03-4 µg/mL).

A molecular and serological survey in Taiwan to determine the true risk of babesiosis in dogs not receiving regular tick prevention.

Canine babesiosis is an important tick-borne disease worldwide. The prevalence varies between regions and countries; however, the incidence of tick infection is associated with the status of preventive tick control measures by the owner. To date, no studies have investigated the incidence of canine babesiosis and the condition of tick prevention in Taiwan. Therefore, the true risk of babesiosis could be underestimated in dogs that are not receiving tick prophylaxis. Samples were collected at 51 hospitals around Taiwan from 265 dogs not receiving regular tick prophylaxis. Diagnostic real-time PCR was performed, and 28 dogs (10.6%) were positive for Babesia spp., including B. gibsoni (26/28) and B. vogeli (2/28). Thirty-nine dogs (14.7%) were seropositive to B. gibsoni. Take the real-time PCR positive as the Babesia infected case, the positive and negative predictive value of serological assay were 64.1% and 98.7%, respectively. The seropositivity of B. gibsoni was significantly associated with real-time PCR positivity for Babesia spp. and vice versa (p < 0.001). The odds of seropositive representing real-time PCR positivity was 132.7 times greater than the seronegative (OR: 132.731, 95% CI 35.683-493.728). Risk factors in the population identified included: dogs with a short-haired coat; intact dogs; dogs from multi-dog households; dogs with more than 10 ticks and fleas on the skin; dogs that go outdoors more than 9 times per week; and dogs with an abnormal blood test result that included anemia, thrombocytopenia, and leukopenia. However, the dogs were not tested for other co-infections, therefore, these hematological risk factors should be carefully interpreted and confirmed by further diagnostic tests. In conclusion, when dogs present with abnormal blood test results and share the risk factors listed above, babesiosis should be seriously considered and followed up with molecular and serological testing. The serological assay used in this study can provide valuable information in diagnosing babesiosis in dogs in Taiwan.

USP7 facilitates SMAD3 autoregulation to repress cancer progression in p53-deficient lung cancer.

USP7, one of the most abundant ubiquitin-specific proteases (USP), plays multifaceted roles in many cellular events, including oncogenic pathways. Accumulated studies have suggested that USP7, through modulating the MDM2/MDMX-p53 pathway, is a promising target for cancer treatment; however, little is known about the function of USP7 in p53-deficient tumors. Here we report that USP7 regulates the autoregulation of SMAD3, a key regulator of transforming growth factor ß (TGFß) signaling, that represses the cell progression of p53-deficient lung cancer. CRISPR/Cas9-mediated inactivation of USP7 in p53-deficient lung cancer H1299 line resulted in advanced cell proliferation in vitro and in xenograft tumor in vivo. Genome-wide analyses (ChIP-seq and RNA-seq) of USP7 KO H1299 cells reveal a dramatic reduction of SMAD3 autoregulation, including decreased gene expression and blunted function of associated super-enhancer (SE). Furthermore, biochemical assays show that SMAD3 is conjugated by mono-ubiquitin, which negatively regulates the DNA-binding function of SMAD3, in USP7 KO cells. In addition, cell-free and cell-based analyses further demonstrate that the deubiquitinase activity of USP7 mediates the removal of mono-ubiquitin from SMAD3 and facilitates the DNA-binding of SMAD3-SMAD4 dimer at SMAD3 locus, and thus enhance the autoregulation of SMAD3. Collectively, our study identified a novel mechanism by which USP7, through catalyzing the SMAD3 de-monoubiquitination, facilitates the positive autoregulation of SMAD3, and represses the cancer progression of p53-deficient lung cancer.