Spectroscopy Technique Speeds Detection of Bacterial Growth in Contained Blood Samples

An international team of researchers at Zhejiang Normal University (Jinhua, China) and Umeå University (Sweden) has developed a technique based on a spectroscopy method to monitor bacterial growth in contained blood samples quickly, accurately, and noninvasively.

Microorganism growth is driven by many factors, so it is critical to be able to assess the quality of blood samples quickly and accurately. Without this ability, samples might need to be discarded or, alternatively, result in or worsen illnesses. Although bacterial blood contamination is rare, it does occur and has led to deaths. A rapid screening method could mean that a larger percentage of blood could be directly tested for bacteria.

Jie Shao, associate professor at the Institute of Information Optics at Zhejiang Normal University, explains that microorganism growth is always associated with the production of carbon dioxide (CO2), so assessing CO2 levels within, for example, closed compartments like bottles or bags would allow them to detect this growth quickly.

Several detection techniques are currently capable of rapid and accurate measurements of gas compositions. Those based on optical spectroscopy are most appealing because they're noninvasive, boast high sensitivity, provide instant responses, and are potentially useful for assessment of bacterial growth. So, the researchers developed an instrument based on a technique called tunable diode laser absorption spectroscopy (TDLAS), which combines all of these properties with ease of use and low cost, Shao says. What's more, it can achieve low detection limits on the order of parts per billion, and measure temperature, pressure, velocity, and mass flux.

The research team used their instrument—which includes a tunable diode laser as the light source, beam shaping optics, a sample to be investigated, receiving optics, and one or more detectors—to assess bacterial growth of various types of samples under a variety of conditions. By applying the technique to transparent containers of organic substances such as food items or medical samples, bacterial growth can be quickly evaluated.

In contrast with conventional and more invasive techniques that require contact with the tested items, the TDLAS method is truly noninvasive for monitoring—in real time—the status of food and medical supplies, or as a tool to determine under which environmental conditions bacterial growth is expected to be severe.

Next, the researchers plan to enhance the technique to allow for assessments of microbial growth in a large variety of samples, expanding beyond food items and medical supplies, Shao says.

Full details of the work appear in the journal Applied Optics.

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Breakthrough Rapid Method for Patients with HIV-Associated Tuberculosis

A rapid, and simple to use, low-cost urine sample to test for TB at the bedside of HIV patients both able and unable to produce sputum, reduced the TB death-rate of patients with advanced HIV. TB treatment can be initiated more quickly than if diagnosed using current diagnostic tools.

The most common cause of death

These are the findings of a multi-centre study led by principle investigator Prof Keertan Dheda of the University of Cape Town (UCT) and colleagues from South Africa, Zimbabwe, Zambia, and Tanzania. The study, a randomised controlled trial using the LAM urine test, evaluated the usefulness of using the simple urine-based TB diagnostic test (similar to a urine pregnancy-like test) in HIV-infected patients with suspected TB who had advanced HIV. It showed that the treatment reduced the TB death rate in hospitals by almost 20%.

TB is the most common cause of death in South Africa, has substantial negative consequences on the economy, and has a substantial mortality and morbidity. It is also the most common cause of death in HIV-infected persons in South Africa. Hospitals in South Africa, and in Africa in general, are inundated with severely ill patients with HIV who also have suspected TB.

The TB is often difficult to diagnose because these patients cannot produce sputum samples, and often the TB is "hiding" in organs such as the liver, lymph glands, or brain where it is difficult to access. Diagnosis is therefore challenging and investigation prolonged.

The new LAM urine test enables a diagnosis in approximately 20 minutes by simply putting a few drops of urine on a low cost strip test. It can be conducted by a minimally trained healthcare worker at the bedside and therefore an answer is available immediately.

“It was unclear till now whether testing makes any difference to treatment-related outcomes, for example death rates because there are other TB diagnostic tests available, and many patients get treated on a ‘doctor best guess',” says Prof Dheda. “But this study showed that the LAM test when used to guide treatment, compared to existing tests and approaches that we use right now, reduced the TB death rate in hospitals by almost 20%. Significantly, these were results obtained using a rapid, and simple to use, low cost bed-side test.”

The other benefits of the study, including being able to rapidly and easily diagnose TB in HIV-infected hospitalised patients unable to produce sputum, who could more rapidly be put onto anti-TB treatment, were an “added value” of the study, according to Prof Dheda. The greatest benefit will be derived in hospitals where diagnostic resources are most limited and where patients present with severe illness, advanced immunosuppression, and an inability to produce sputum.

Low cost bedside test

The data from this and previous studies by Prof Dheda and colleagues have informed clinical practice and the roll out of new diagnostic modalities in TB and HIV endemic settings, resulting in the urine LAM test being endorsed by the World Health Organisation (WHO).

“Policy makers should consider implementation of this low cost bedside TB test in hospitalised patients in resource-limited settings with high TB/HIV burdens whilst further data accumulate”, says Prof Dheda. This study clears the way for the roll-out and implementation of the urine LAM test in hospitals in South Africa and Africa in general.

The paper was published in the Lancet. Co-investigators from the UCT team, based within the Division of Pulmonology, included Dr Jonny Peter, Dr Grant Theron, Dr Greg Calligaro, and Dr Phindile Gina.

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Lasers Help Speed Up the Detection of Bacterial Growth in Packaged Food and Other Environments

It’s important to know how microorganisms — particularly pathogenic microbes — grow under various conditions. Certain bacteria can cause food poisoning when eaten and bacterial growth in medical blood supplies, while rare, might necessitate discarding the blood.

Now a group of researchers from Zhejiang Normal University in China and Umeå University in Sweden report a fast, accurate, and noninvasive technique for monitoring bacterial growth. They report the results in Applied Optics, a journal of The Optical Society (OSA).

Microorganism growth is driven by many factors, which make it far from easy to accurately estimate the amount of bacteria within food containers or blood samples at any given time.

To avoid the risk that any particular packaged food item will go bad and cause illness, it’s given an unnecessarily short shelf life. In short, a better understanding of the growth process of microorganisms could reduce food waste and prevent people from being sickened by food poisoning — or both.

Within the medical realm, it’s critical to be able to assess the quality of blood samples quickly and accurately. Without this ability, samples might need to be discarded or, alternatively, result in or worsen illnesses. Although bacterial blood contamination is rare, it does occur and has led to deaths. A rapid screening method could mean that a larger percentage of blood could be directly tested for bacteria.

“Microorganism growth is always associated with the production of carbon dioxide (CO2),” said Jie Shao, associate professor at the Institute of Information Optics, Zhejiang Normal University, Jinhua, China. “By assessing the level of CO2 within a given closed compartment — bottle or bag — it’s possible to assess the microbial growth.”

Several detection techniques are currently capable of rapid and accurate measurements of gas compositions. Those based on optical spectrometry are most appealing because they’re noninvasive, boast high sensitivity, provide instant responses, and are potentially useful for assessment of bacterial growth.

“A technique referred to as ‘tunable diode laser absorption spectroscopy’ (TDLAS) is particularly suitable because it combines all of these properties with an ease of use and low cost,” Shao said.

So the group decided to develop an easy-to-use instrument based on TDLAS to assess bacterial growth of various types of samples under a variety of conditions.

TDLAS is by far the most common laser-based absorption technique for quantitative assessments of species within a gas phase. It can be used to measure the concentration of specific gaseous species — carbon monoxide, CO2, water, or methane, to name a handful — within gaseous mixtures by using absorption spectrometry based on tunable diode lasers.

“One major advantage TDLAS offers is its ability to achieve very low detection limits, on the order of parts per billion,” Shao said. “Apart from concentration, it’s also possible to determine other properties of the gas under observation — temperature, pressure, velocity and mass flux.”

The group’s basic setup simply involves a tunable diode laser as the light source, beam shaping optics, a sample to be investigated, receiving optics, and one or more detectors.

“The emission wavelength of the laser is tuned over a characteristic absorption line transition — of the species within the gas being assessed,” Shao explained. “This causes a reduction of the measured signal intensity, which we can use to determine the gas concentration.”

When the wavelength is rapidly tuned across the transition in a specific manner, it can be combined with a modulation technique called “wavelength modulation” (WM), which gives the TDLAS technique an enhanced sensitivity. It’s referred to as “WM-TDLAS.”

By applying the technique to transparent containers of organic substances such as food items or medical samples, bacterial growth can be quickly evaluated. “Although we anticipated that the WM-TDLAS technique would be suitable for assessing bacterial growth, we didn’t expect this level of accuracy,” Shao noted.

In contrast with conventional and more invasive techniques that require contact with the tested items, the WM-TDLAS method is truly noninvasive, making it ideal for monitoring the status of food and medical supplies, or as a tool to determine under which environmental conditions bacterial growth is expected to be severe. “It can provide real-time analysis,” Shao said.

Next, the researchers plan to enhance the technique to “allow for assessments of microbial growth in a large variety of samples — expanding beyond food items and medical supplies,” Shao added.

Paper: Jie Shao, Jindong Xiang, Ove Axner, and Chaofu Ying, "Wavelength-modulated tunable diode-laser absorption spectrometry for real-time monitoring of microbial growth," Appl. Opt. 55, 2339-2345 (2016).

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Hong Kong Polytechnic University Develops Novel Nano Biosensor for Rapid Detection of Flu Virus

The Department of Applied Physics (AP) and Interdisciplinary Division of Biomedical Engineering (BME) of The Hong Kong Polytechnic University (PolyU) have jointly developed a novel nano biosensor for rapid detection of flu and other viruses. PolyU's new invention utilizes an optical method called upconversion luminescence resonance energy transfer (LRET) process for ultrasensitive virus detection. It involves simple operational procedures, significantly reducing its testing duration from around 1-3 days to 2-3 hours, making it more than 10 times quicker than traditional clinical methods. Its cost is around HK$20 per sample, which is 80% lower than traditional testing methods. The technology can be widely used for the detection of different types of viruses, shedding new light on the development of low-cost, rapid and ultrasensitive detection of different viruses.

Traditional biological methods for flu virus detection include genetic analysis -- reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) used in immunology. However, RT-PCR is expensive and time-consuming while the sensitivity for ELISA is relatively low. Such limitations make them difficult for clinical use as a front-line and on-site diagnostic tool for virus detection, paving the way for PolyU's development of the new upconversion nanoparticle biosensor which utilizes luminescent technique in virus detection.

PolyU's researchers have developed a biosensor based on luminescent technique which operates like two matching pieces of magnet with attraction force. It involves the development of upconversion nanoparticles (UCNPs) conjugated with a probe oligo whose DNA base pairs are complementary with that of the gold nanoparticles (AuNPs) flu virus oligo. Given the complementary nature of the DNA base pairs of the UCNPs probe oligo and AuNPs flu virus oligo, they work like two matching pieces of magnet, which would be drawn together due to attraction force. This process is also called oligo hybridization. Upon being illuminated by a portable near-infrared laser pen, the UCNPs emit eye-visible green light while the AuNPs would absorb the green light. One can easily quantify the concentration of the targeted flu virus by measuring the decrease in the green light intensity.

Initially, PolyU researchers have utilized upconversion LRET for ultrasensitive virus detection in liquid phase system. The research team has further improved the sensitivity of the luminescent detection method by utilizing a solid phased nanoporous membrane system (NAAO) for virus detection. As NAAO membrane consists of many hollow channels, they allow more space for oligo hybridization to take place, significantly increasing its sensitivity by more than 10 folds compared to the liquid phase system, proven by clinical detection using inactivated virus samples.

Not only is the design and operation of PolyU's invention simple, it does not require expensive instruments and sophisticated operational skills, with its sensitivity comparable to traditional clinical methods. In comparison to conventional downconversion luminescent technique, it causes low damage to genetic materials and does not induce background fluorescence. In addition, since each virus has a unique genetic sequence, researchers would be able to design a complementary probe once the genetic sequence of the targeted virus is known. In other words, the upconversion LRET technology can be widely used for the detection of different types of viruses simply by modifying the UCNPs capture probe.

The related results have been recently published in ACS Nano and Small, two leading journals in nano material research. With the support from the Innovation and Technology Support Programme, the research team will continue to enhance the nano biosensor for rapid virus detection, which includes increasing its sensitivity and specificity, and developing a matrix for detection of multiple flu viruses on a single testing platform.

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Miacom Diagnostics and MetaSystems Partner to Provide Rapid Pathogen identification

Miacom diagnostics GmbH, a company focused on improving therapy and outcomes for patients suffering from acute infectious diseases, today announced a new partnership with MetaSystems, an industry leading company in computerized imaging. The companies will join forces to offer microbiologists all the advantages of FISH technology together with the accuracy and speed of a fully automated reading and data analysis platform.

“This joint venture will bring something truly unique to the market” stated Dr. Mirko Stange, CEO Miacom diagnostics. “During the last year MetaSystems and our team have succeeded in utilizing the full power of the MetaSystems platform to automatically handle and analyse Miacom processed patient samples. We are now able to automatically identify bacteria directly from respiratory secretions in just minutes. The benefits are tremendous - laboratories can report the infection causing pathogen within the timeframe of a Gram stain and enable ICUs to immediately improve antibiotic treatment.” For customers from the pharmaceutical industry and larger hospital chains the “Rapid Pathogen Identification” or RPI System comes with a significant option. A small RPI System was specifically developed to enable decentralized sample processing while the resulting data can be stored and analyzed centrally anywhere in the world. “Particularly for antibiotic trials this system facilitates patient enrollment while reducing overall cost and reducing risk for trial participants. A trained physician in Boston, for example, can make a diagnosis of a patient sample being tested in New Delhi – this occurs in real-time, so that within 30min the correct pathogen ID can be confirmed and the decision can be made to enroll the patient in a trial.” explains Dr. Andreas Plesch, CEO of MetaSystems. To fit into modern laboratory processes the RPI System insures that all results are digitally transmitted and stored, and that they are fully compatible with LIM systems.

With the incorporation of MetaSystems’ technology, Miacom gets one step closer to fully automate its assays. Microbiologists now have a cost effective ID alternative at hand, which can be easily integrated into the workflow of all laboratories. Current assays include an FDA cleared multiplex assay for Sepsis and assays detecting and quantifying pneumonia-related pathogens directly from sputum or other pulmonary secretions. The small scale system as well as an RPI designed for high-throughput applications will be officially launched at the ECCMID conference in Amsterdam in the beginning of April. Customers are invited to observe a live demo of the RPI-System and learn about the exciting pipeline of new assays to be run on this platform at the Miacom booth (No 63).

Miacom diagnostics specializes in in vitro diagnostic tests for pathogens causing acute systemic diseases. Miacom will change critical patient care by providing rapid molecular diagnostics tools designed for routine use to promote more efficient antibiotic therapies and help healthcare organizations reduce costs. Miacom’s current “tool box” includes the world’s only molecular diagnostics (MDx) test that identifies all relevant pneumonia-related bacteria directly from sputum in only 30 minutes as well as rapid multiplex solutions for sepsis patients.

MetaSystems is a leader in automated microscopy and slide screening and a provider of fluorescent DNA probes for genetic diagnostics in cytogenetics, hematology, and pathology. Imaging solutions range from manual or semi-automated acquisition stations to fully automated walk-away systems comprising a robotic slide feeder for unattended screening and analysis of up to 800 slides.

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Rapid Lab-on-a-Drone PCR Enabled With Convective Flow Thermal Cycling

In low-resource settings with poor infrastructure, setting up and staffing remote lab sites during an infectious disease outbreak is a huge challenge.

Researchers have long sought ways to deliver molecular diagnostic testing capabilities to far-flung areas. Now, scientists at Texas A&M University have developed an inexpensive way to perform rapid molecular testing using quadcopter drones and a rapid PCR-based system.

The technique, published in Analytical Chemistry last month, relies on a thermal cycling method that is unlike typical spatial or temporal thermal cycling, in which either the sample moves between heat sources or the sample is stationary and the heat source fluctuates between melting and annealing temperatures.

Convective flow thermal cycling is actually isothermal, in so far as there is only a single temperature applied to the sample tube. The heat source is set against one face of a small reaction column, such that the opposite face is cooled by convection.

With just the right height and width, a convective flow pattern will become established within the column. Nucleic acids melt at the bottom of the column, but as they become hotter and less dense they rise to the top, passing through lower temperature zones and elongating along the way. They then cool, becoming more dense, annealing, and sinking back to the bottom to melt again.

Thus, the method is spatial PCR in that the sample is traveling between heat sources. It is also isothermal — with respect to the instrument, not the PCR reaction — in the sense that only a single temperature is applied, and this enables very low electrical power consumption.

Lead investigator Victor Ugaz, a chemical engineer at TAMU, said he had been working for over 10 years on the convective flow method, beginning as a post-doc at the University of Michigan.

"It's like a mini lava lamp," Ugaz said. "You heat reagents from below, and if you do it correctly you can generate a flow pattern by imposing a temperature gradient."

But unlike its conceptual cousin, which perhaps evokes a more meditative and tranquil mindset, convective flow PCR is based on some hardcore physics and is quite rapid, with 35-second equivalent cycle times, demonstrating 20-minute in-flight replication of Staphylococcus aureus and λ-phage DNA targets. It is also quite small, with reaction volumes of about 20 microliters.

A previous iteration of the system published in the Journal of Visualized Experiments showed the relationship between the aspect ratio of the column and the circulatory flow. The previous method, however, used two hot plates, while the current method uses only one. The system has also now been upgraded so that the shape of the PCR column allows the same system to run reactions with different melting and annealing temperatures and is optimized by adopting a chaotic flow pattern.

The convective method was initially described in a 2002 Science paper. "Since then, it's been like a curiosity; we hadn't found the right niche," Ugaz said. However, the current incarnation has proven to be remarkably rugged.

Case in point: it can run on a flying quadcopter drone.

The work was inspired in part by a graduate student, who showed Ugaz a video illustrating the portability of a 3D printer by having it print something while flying on a drone.

"That gave us an idea. ... We should be able to attach [our system] as a payload and do the reaction during flight," Ugaz said.

But ultimately the drone project was taken on by a team of undergrads participating in an engineering challenge at TAMU. Ugaz mentored the team, with the challenge of creating a prototype commercial product from ongoing research.

The team then made the first prototype of the device, winning second place in the "AggiE-Challenge" for the project. All 14 members are authors on the Analytical Chemistry study.

The PCR is performed with heat from two resistors, requiring a 5-volt USB power source. It can also be run with a battery, or using solar or hand crank power. And, although the quadcopter drone can cost around $500, the PCR system costs only about $50 to make. It is also very "simple to produce," and weighs only about 300 grams, or roughly the same as the GoPro cameras that are standard payloads for quadcopter drones.

Additionally, the method uses fluorescent detection with a smartphone camera and an app called PCR to Go, developed by grad student and co-author Aashish Priye.

The app takes a sequence of photos and analyzes intensity of a region of interest to derive fluorescence changes over time. For now, this data has to be corrected, thresholded, and plotted offline to get a real-time curve, but the group is working to build these functions into a future app.

The group also collaborated with Season Wong of TAMU and AI Biosciences. Wong, who has recently developed a low-resource method of PCR using thermoses, collaborated to help apply the device for specific diagnostics tests.

Wong has "been working on a lot of low-cost, diagnostic gadgets," Ugaz said, and also saw that the motors of the quadcopter could be repurposed for centrifuging spin columns. He and a colleague at AI Biosciences 3D-printed attachments that enabled centrifugation at speeds of 10,000 RPM.

"Sample prep is another ongoing need for portable analysis," Ugaz explained, noting other low-power solutions, such as using a salad-spinner or bicycle wheel, are good for separating plasma from whole blood but can't achieve the speeds needed for conventional sample prep spin columns.

"The drone has four motors, so we got the idea we could use the drone itself as a lab instrument to do part of the workflow," Ugaz said. And from a mobile PCR platform with centrifuging capabilities, several scenarios could arise, he said, such as deploying the drone to be used as a lab instrument at a remote site. "Or you could imagine sending more than one, with one used as a centrifuge and another used for transportation," Ugaz said.

Ultimately, the ability to send out easily-modifiable networks of diagnostic modules, rather than laboratorians themselves, could alter public health approaches to outbreaks and infectious disease emergencies.

"You have to kind of change the way you think about deploying things," Ugaz said. "Conventional ways are in a set sequence of events, but maybe there are other ways that could be enabled if you have a low-cost way to deploy many things, in many different ways."

The convection flow thermal cycling method is currently under patent protection, and the lab is seeking additional funding. The group is also interested in collaboration, said Ugaz.

"We're excited about this work because we're hoping it can really help us connect with an industrial partner, or anyone, and that this could get on the radar screen and maybe lead to a practical application," Ugaz said, adding, "I think that's what anybody in science would love to see — their discovery actually translate into a product that can really help people."

Source: genomeweb

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Cepheid Announces First FDA-Cleared Test for Detection of Carbapenem Resistance Genes in Isolates of Multidrug Resistant 'Superbugs'

Cepheid today announced that it has received clearance from the U.S. Food and Drug Administration (FDA) to market Xpert® Carba-R, a qualitative in vitro diagnostic test for fast, accurate, and reproducible identification of 5 distinct families of carbapenem resistance genes that together represent the most common carbapenemases identified globally, including KPC, NDM, VIM, OXA-48 and IMP.  Xpert Carba-R is the first FDA-cleared test for detection and differentiation of carbapenemase genes in pure bacterial isolates, previously shown to be non-susceptible to carbapenem antibiotics, which may be cultured from a wide range of clinical specimens, including blood cultures, urine, respiratory samples, abscesses and swab surveillance specimens.

"The emergence of carbapenemase producing organisms, known as CPOs, represents a significant global healthcare risk since these bacteria are resistant to many of the beta-lactam antibiotics used for empiric therapy for gram negative infections.  These resistance genes are highly transmissible from one bacterial species to another, leading to well-documented outbreaks associated with high morbidity and mortality," said David Persing, M.D., Ph.D., Cepheid's Chief Medical and Technology Officer.  "Carbapenem resistance mechanisms can be extremely difficult to characterize with conventional laboratory procedures.  Now, with a bacterial isolate and less than a minute of hands-on time, Xpert Carba-R delivers a result in under an hour, allowing timely detection and differentiation of the most prevalent carbapenemase genes associated with CPO outbreaks to support infection control efforts."

"The availability of a rapid and accurate molecular test for the most prevalent mechanisms of carbapenem resistance represents an important addition to the limited tools the laboratory has available to fight the emergence of multidrug resistant organisms, like CPOs," said Paul Schreckenberger, Ph.D., Director of Microbiology at Loyola University Medical Center.  "Knowing which carbapenemase gene is present in a resistant isolate helps the hospital identify and manage outbreaks and can also be valuable for monitoring the spread of multi-resistant bacteria among hospital populations considered most at risk."

Xpert Carba-R will begin shipping in the United States later this month, and is the 20th US-IVD test available to run on Cepheid's GeneXpert® System, the world's leading molecular diagnostic platform with more than 10,000 systems installed globally.  Xpert Carba-R is the latest diagnostic solution from Cepheid to help infection control professionals fight antimicrobial resistance, building on a portfolio that already includes MRSA, C. difficile and VRE.  The product's availability in the United States is particularly timely, following publication of a study last week from the Centers for Disease Control and Prevention citing carbapenem-resistant Enterobacteriaceae as an urgent threat and emphasizing the prevention of antibiotic-resistant healthcare-acquired infections as an important strategy for reducing the impact of antibiotic-resistant bacteria on human health, including the prevention of sepsis and death.

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MD Biosciences Launches Rapid Assay to Detect Zika Virus

MD Biosciences, Inc. Clinical and Diagnostic Services Laboratory announces the release of its rapid assay to detect the virus in human blood and urine samples. The nucleic acid test can be performed in a few hours in blood, plasma, serum or urine samples. The test is specific for the Zika virus and was shown not to cross react with other infectious viruses such as Dengue, West Nile or Chikunguya. The laboratory-developed test is available immediately.

The World Health Organization states that the spread of the mosquito-borne virus will grow explosively throughout the Americas. According to the CDC, the virus has also been shown to be transmitted through sexual contact. Upwards of four million infections are expected in the region. Close to 80% of infections are asymptomatic while others suffer from fever, rash, joint aches and other hallmarks. A steep increase in birth defects and Guillain-Barre syndrome has correlated to the spread of the virus. The explosive spread of the virus, along with major crowd and traveller-related events such as this year's Olympic Games in Brazil, have created the need for steps in fighting the virus, including the ability to rapidly and specifically identify the infection.

Eddie Moradian, Chief Executive Officer at MD Biosciences states, "We are extremely proud of our CLIA diagnostics laboratory's ability to provide a reliable, accurate and rapid method of testing for Zika virus. This demonstrates our commitment in placing our expertise, knowledge and infrastructure for the betterment of health, in the US and throughout the world, whether through the development of more advanced diagnostics or by advancing newer, better and more efficient therapies."

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