Failure of PCR to Detect Treponema pallidum ssp. pertenue DNA in Blood in Latent Yaws

Yaws, caused by Treponema pallidum ssp. pertenue, is a neglected tropical disease closely related to venereal syphilis and is targeted for eradication by 2020. Latent yaws represents a diagnostic challenge, and current tools cannot adequately distinguish between individuals with true latent infection and individuals who are serofast following successful treatment. PCR on blood has previously been shown to detect T. pallidum DNA in patients with syphilis, suggesting that this approach may be of value in yaws. We performed real-time PCR for Treponema pallidum ssp. pertenue on blood samples from 140 children with positive T. pallidum Particle Agglutination (TPPA) and Rapid Plasma Reagin (RPR) tests and 7 controls (negative serology), all collected as part of a prospective study of yaws in the Solomon Islands. All samples were also tested by a nested PCR for T. pallidum. 12 patients had clinical evidence of active yaws whilst 128 were considered to have latent yaws. 43 children had high titre rapid plasma reagins (RPRs) of ≥1:32. PCR testing with both assays gave negative results in all cases. It is possible that the failure to detect T. pallidum ssp. pertenue in blood reflects lower loads of organism in latent yaws compared to those in latent infection with T. pallidum ssp. pertenue, and/or a lower propensity for haematogenous dissemination in yaws than in syphilis. As the goal of the yaws control programme is eradication, a tool that can differentiate true latent infection from individuals who are serofast would be of value; however, PCR of blood is not that tool.     

Impact of Community Mass Treatment with Azithromycin for Trachoma Elimination on the Prevalence of Yaws

Background

Community mass treatment with 30mg/kg azithromycin is central to the new WHO strategy for eradicating yaws. Both yaws and trachoma— which is earmarked for elimination by 2020 using a strategy that includes mass treatment with 20mg/kg azithromycin—are endemic in the Pacific, raising the possibility of an integrated approach to disease control. Community mass treatment with azithromycin for trachoma elimination was conducted in the Solomon Islands in 2014.

Methods

We conducted a study to assess the impact of mass treatment with 20mg/kg azithromycin on yaws. We examined children aged 5-14 years and took blood and lesion samples for yaws diagnosis.

Results

We recruited 897 children, 6 months after mass treatment. There were no cases of active yaws. Serological evidence of current infection was found in 3.6% (95% CI= 2.5-5.0%). This differed significantly between individuals who had and had not received azithromycin (2.8% vs 6.5%, p=0.015); the prevalence of positive serology in 5-14 year-olds had been 21.7% (95% CI=14.6%-30.9%) 6 months prior to mass treatment. Not receiving azithromycin was associated with an odds of 3.9 for infection (p=0.001). National figures showed a 57% reduction in reported cases of yaws following mass treatment.

Discussion

Following a single round of treatment we did not identify any cases of active yaws in a previously endemic population. We found a significant reduction in latent infection. Our data support expansion of the WHO eradication strategy and suggest an integrated approach to the control of yaws and trachoma in the Pacific may be viable.     

A BLOC-1–AP-3 super-complex sorts a cis-SNARE complex into endosome-derived tubular transport carriers

Membrane transport carriers fuse with target membranes through engagement of cognate vSNAREs and tSNAREs on each membrane. How vSNAREs are sorted into transport carriers is incompletely understood. Here we show that VAMP7, the vSNARE for fusing endosome-derived tubular transport carriers with maturing melanosomes in melanocytes, is sorted into transport carriers in complex with the tSNARE component STX13. Sorting requires either recognition of VAMP7 by the AP-3δ subunit of AP-3 or of STX13 by the pallidin subunit of BLOC-1, but not both. Consequently, melanocytes expressing both AP-3δ and pallidin variants that cannot bind their respective SNARE proteins are hypopigmented and fail to sort BLOC-1–dependent cargo, STX13, or VAMP7 into transport carriers. However, SNARE binding does not influence BLOC-1 function in generating tubular transport carriers. These data reveal a novel mechanism of vSNARE sorting by recognition of redundant sorting determinants on a SNARE complex by an AP-3–BLOC-1 super-complex.     

Proteolysis of BB0323 results in two polypeptides that impact physiologic and infectious phenotypes in Borrelia burgdorferi

Borrelia burgdorferi gene product BB0323 is required for cell fission and pathogen persistence in vivo. Here, we show that BB0323, which is conserved among globally prevalent infectious strains, supports normal spirochaete growth and morphology even at early phases of cell division. We demonstrate that native BB0323 undergoes proteolytic processing at the C‐terminus, at a site after the first 202 N‐terminal amino acids. We further identified a periplasmic BB0323 binding protein in B. burgdorferi, annotated as BB0104, having serine protease activity responsible for the primary cleavage of BB0323 to produce discrete N‐ and C‐terminal polypeptides. These two BB0323 polypeptides interact with each other, and either individually or as a complex, are associated with multiple functions in spirochaete biology and infectivity. While N‐terminal BB0323 is adequate to support cell fission, the C‐terminal LysM domain is dispensable for this process, despite its ability to bind B. burgdorferi peptidoglycan. However, the LysM domain or the precisely processed BB0323 product is essential for mammalian infection. As BB0323 is a membrane protein crucial for B. burgdorferi survival in vivo, exploring its function may suggest novel ways to interrupt infection while enhancing our understanding of the intricate spirochaete fission process.     

The Lack of Toxic Effect of High-Power Short-Pulse 101 GHz Millimeter Waves on Healthy Mice

Irradiation of cancer cells by non-ionizing millimeter waves (MMW) causes increased cell mortality. We examined if MMW have toxic effects on healthy mice. To that end, the skin of healthy C57BL/6 mice was irradiated locally at the right flank with 101 GHz MMW in a pulsed (5–10 µs) regime using a free electron laser. Irradiation was performed in a dose-dependent manner, with 20–50 pulses and a power range of 0.5–1.5 kW. Physical, physiological, and pathological parameters as well as behavior were examined before and after irradiation. Our results showed that all parameters were within normal range for all experimental mice groups and for the control group. No significant changes were noted in the physical, physiological, or behavioral status of the mice following irradiation as compared with the control group. In addition, no significant changes were found in locomotor, exploratory behavior, or anxiety of the irradiated mice and no pathological changes were noted following the hematological and biochemical blood analysis. Our results indicate that irradiation of healthy mice with MMW does not cause any general toxic effects. Bioelectromagnetics. © 2020 Bioelectromagnetics Society.     

Formation and morphology of epidermal sclerites from a deep-sea hydrothermal vent solenogaster (Helicoradomenia sp., Solenogastres, Mollusca)

The deep-sea hydrothermal vent solenogaster Helicoradomenia is covered with calcium carbonate sclerites. Light and electron microscopy reveal varying morphologies of these sclerites. Many sclerites have hollow tips and/or are pitted and etched. Bacteria are found on and in sclerites. Initial sclerite formation occurs in an extracellular crystalline chamber formed by the invagination of a cuboidal basal cell of the columnar microvillus mantle epithelium. As the sclerite grows, it fills the crystalline chamber resulting in direct contact with the microvilli of both the basal cell and neighboring secondary sclerite-forming cells. These cells shape a collar around the base of the growing sclerite. As growth continues, the sclerite-forming cells stretch around the sclerite forming a sheath in which the base of the sclerite resides. Mature sclerites grow through the cuticle into the external environment. The erosion pattern of sclerites reveals a less stable inner medullary region and a harder outer cortical region. This points to a secondary character state, where foremost hollow acicular sclerites develop into solid sclerites. This is in agreement with the systematic position of the genus Helicoradomenia within Simrothiellidae, a taxon typically with hollow sclerites.