New and emerging technologies for the diagnosis of urinary tract infections

Urinary tract infections affect 150 million people worldwide, yet the diagnosis of this common infection is not straightforward. Misdiagnoses and incorrect prescriptions are frequent in the treatment of urinary tract infections; this also contributes to the increase in antibiotic resistance among pathogens. Present diagnostic practices take 2–3 days for pathogen identification and antibiotic susceptibility testing. New technologies are urgently needed for improved patient care as well as to promote antibiotic stewardship. An ideal new diagnostic technology will test clinical urine samples directly and identify the pathogen and determine its antibiotic susceptibilities within a few hours such that the patient can be prescribed the appropriate antibiotic treatment the same day. Screening tools, such as flow cytometers and new dipstick assays, can help with rapidly identifying negative samples and improving workflow and reducing costs. Several groups have made progress in optimizing mass spectrometry methods for direct urine processing, and there are also new multiplex PCR panels that are specific for UTI pathogens and antibiotic resistance. We also discuss several emerging technologies – microfluidics, biosensors, real-time microscopy systems, and sequence-based diagnostics – that show huge potential in delivering rapid results.

Urinary cell mRNA Profiling of Kidney Allograft Recipients: A Systematic Investigation of a Filtration Based Protocol for the Simplification of Urine Processing

Background. Kidney transplantation is a life-restorative therapy, but immune rejection undermines allograft survival. Urinary cell mRNA profiles offer a noninvasive means of diagnosing kidney allograft rejection, but urine processing protocols have logistical constraints. We aimed to determine whether the centrifugation-based method for urinary cell mRNA profiling could be replaced with a simpler filtration-based method without undermining quality. Methods. We isolated RNA from urine collected from kidney allograft recipients using the Cornell centrifugation-based protocol (CCBP) or the Zymo filter-based protocol (ZFBP) and compared RNA purity and yield using a spectrophotometer or a fluorometer and measured absolute copy number of transcripts using customized real-time quantitative PCR assays. We investigated the performance characteristics of RNA isolated using ZFBP and stored either at -80oC or at ambient temperature for 2 to 4 days and also when shipped to our Gene Expression Monitoring (GEM) Core at ambient temperature. We examined the feasibility of initial processing of urine samples by kidney allograft recipients trained by the GEM Core staff and the diagnostic utility for acute rejection, of urine processed using the ZFBP. Results. RNA purity (P=0.0007, Wilcoxon matched paired signed-ranks test ) and yield (P<0.0001) were higher with ZFBP vs. CCBP, and absolute copy number of 18S rRNA was similar (P=0.79) following normalization of RNA yield by reverse transcribing a constant amount of RNA isolated using either protocol. RNA purity, yield, and absolute copy numbers of 18S rRNA, TGF-β1 mRNA and microRNA-26a were not different (P>0.05) in the filtrates containing RNA stored either at -800C or at ambient temperature for 2 to 4 days or shipped overnight at ambient temperature. RNA purity, yield, and absolute copy numbers of 18S rRNA and TGF-β1 mRNA were also not different (P>0.05) between home processed and laboratory processed urine filtrates. Urinary cell levels of mRNA for granzyme B (P=0.01) and perforin (P=0.0002) in the filtrates were diagnostic of acute rejection in human kidney allografts. Conclusions. Urinary cell mRNA profiling was simplified using the ZFBP without undermining RNA quality or diagnostic utility. Home processing by the kidney allograft recipients, the stability of RNA containing filtrates at ambient temperature, and the elimination of the need for centrifuges and freezers represent some of the advantages of ZFBP over the CCBP for urinary cell mRNA profiling.

Urine Treatment on the International Space Station: Current Practice and Novel Approaches

A reliable, robust, and resilient water recovery system is of paramount importance on board the International Space Station (ISS). Such a system must be able to treat all sources of water, thereby reducing resupply costs and allowing for longer-term space missions. As such, technologies able to dewater urine in microgravity have been investigated by different space agencies. However, despite over 50 years of research and advancements on water extraction from human urine, the Urine Processing Assembly (UPA) and the Water Processor Assembly (WPA) now operating on the ISS still achieve suboptimal water recovery rates and require periodic consumables resupply. Additionally, urine brine from the treatment is collected for disposal and not yet reused. These factors, combined with the need for a life support system capable of tolerating even dormant periods of up to one year, make the research in this field ever more critical. As such, in the last decade, extensive research was conducted on the adaptation of existing or emerging technologies for the ISS context. In virtue of having a strong chemical resistance, small footprint, tuneable selectivity and versatility, novel membrane-based processes have been in focus for treating human urine. Their hybridisation with thermal and biological processes as well as the combination with new nanomaterials have been particularly investigated. This article critically reviews the UPA and WPA processes currently in operation on the ISS, summarising the research directions and needs, highlighted by major space agencies, necessary for allowing life support for missions outside the Low Earth Orbit (LEO). Additionally, it reviews the technologies recently proposed to improve the performance of the system as well as new concepts to allow for the valorisation of the nutrients in urine or the brine after urine dewatering.

Karakteristik Program dan Kelompok serta Kendala Pelaksanaan Program Simantri di Kabupaten Badung

Characteristics of Program and Group and Implementation Obstacle Simantri Program in Badung regency Integrated Farming System or Simantri is a breakthrough effort to accelerate the adoption of agricultural technology, Simantri Program development goal is to support integrated diversification of farming to local potentials, to increase income to alleviate poverty, to integrate crop and livestock farming, and to pioneer the development of integrated agriculture in a sustainable manner. The purpose of this research is to investigate the Characteristic of Program and Characteristic of Group, and Constraints of Simantri Program Implementation in the Badung Regency.. The results showed that the Characteristics of Simantri Program is quite good with average achievement score of 3.8, Characteristics of Simantri group is good with average achievement score of 4.1, constraints faced in the characteristics of the program i.e. optimal cattle maintenance management, cattle maintenance, inadequate cage quality and many are not functioningproperly, cow urine channel and bio-urine processing tools are easily damaged, bio-gas was also easily damaged, and the location of Simantri makes the gas bio channel unable to reach the houses of the Simantri's group, group members rarely processed manure because of their other activities, members were reluctant to maintain and care for planted demonstration plots.

Assessing the Reusability of 3D-Printed Photopolymer Microfluidic Chips for Urine Processing

Three-dimensional (3D) printing is emerging as a method for microfluidic device fabrication boasting facile and low-cost fabrication, as compared to conventional fabrication approaches, such as photolithography, for poly(dimethylsiloxane) (PDMS) counterparts. Additionally, there is an increasing trend in the development and implementation of miniaturized and automatized devices for health monitoring. While nonspecific protein adsorption by PDMS has been studied as a limitation for reusability, the protein adsorption characteristics of 3D-printed materials have not been well-studied or characterized. With these rationales in mind, we study the reusability of 3D-printed microfluidics chips. Herein, a 3D-printed cleaning chip, consisting of inlets for the sample, cleaning solution, and air, and a universal outlet, is presented to assess the reusability of a 3D-printed microfluidic device. Bovine serum albumin (BSA) was used a representative urinary protein and phosphate-buffered solution (PBS) was chosen as the cleaning agent. Using the 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) fluorescence detection method, the protein cross-contamination between samples and the protein uptake of the cleaning chip were assessed, demonstrating a feasible 3D-printed chip design and cleaning procedure to enable reusable microfluidic devices. The performance of the 3D-printed cleaning chip for real urine sample handling was then validated using a commercial dipstick assay.