Dear friends, I am looking for informational resources on the applicability of bio fuels in the Indian economic scenario. I would be very grateful for any inputs on the same, from academics as well as industry persons dealing with this. Thanks in advance!
information resources for biofuels in India
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Floral dip transformation - student question
Hi everybody,
I'm following an agricultural biotechnology course and I'm wondering if anyone can help me:
We planted floral dip transformed Arabidopsis seeds on a medium containing kanamycin. The transformation (if succeeded) inserted the binairy pXK2FS7 vector, a plasmid containing kanamycin resistance (selection) and a pC aMV 35S - Egfp - gus promotor-gen construct (2 reportergenes). The 35s promotor is flanked by attB1 and attB2 gatewaysites.
If the seeds grow on kanamycin containing medium I come to the conclusion the transformation was succesful. I would expect gfp activity but this was not the case. After two weeks, none of the seedlings were fluorescent under the influence of UV-light. This monday we will check the gus-activity so I don't know about that reportergene yet.
Now I'm wondering why my expectations don't meet reality. Does anybody know why there is no reportergene activity? Are the seedlings to young? Is it the 35S promotor that is not active? If so, why? And how can this be because it is supposed to be strong and constitutive. Or is the promotor active but is there some problem with the mRNA?
Thank a lot for any suggestions,
the biotechnology student
I'm following an agricultural biotechnology course and I'm wondering if anyone can help me:
We planted floral dip transformed Arabidopsis seeds on a medium containing kanamycin. The transformation (if succeeded) inserted the binairy pXK2FS7 vector, a plasmid containing kanamycin resistance (selection) and a pC aMV 35S - Egfp - gus promotor-gen construct (2 reportergenes). The 35s promotor is flanked by attB1 and attB2 gatewaysites.
If the seeds grow on kanamycin containing medium I come to the conclusion the transformation was succesful. I would expect gfp activity but this was not the case. After two weeks, none of the seedlings were fluorescent under the influence of UV-light. This monday we will check the gus-activity so I don't know about that reportergene yet.
Now I'm wondering why my expectations don't meet reality. Does anybody know why there is no reportergene activity? Are the seedlings to young? Is it the 35S promotor that is not active? If so, why? And how can this be because it is supposed to be strong and constitutive. Or is the promotor active but is there some problem with the mRNA?
Thank a lot for any suggestions,
the biotechnology student
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College Comparison and Study Abroad
I am currently doing +2 in PCMB group. I'm planning to do my B. Tech in Biotechnology and i have a couple of choices for college
1. Give me a comparision of Biotechnology PESIT College, Bangalore and PSG Tech, Coimbatore? which is best?
2. If i do B Tech Biotechnology in India, can i do my masters in Molecular biology, abroad?
1. Give me a comparision of Biotechnology PESIT College, Bangalore and PSG Tech, Coimbatore? which is best?
2. If i do B Tech Biotechnology in India, can i do my masters in Molecular biology, abroad?
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BSL3 Lab exhaust installation
We are installing a BSL3 lab in a Institute in India.
Working of the system is: Fresh air is cooled in an Air Handling Unit filtered in 3 stages, 10micron, 5micron & 0.3micron (HEPA) and supplied to the room. Room air is exhausted into the atmosphere after HEPA filtration. The exhaust quantity is controlled by a VFD based on the Negative pressure to be maintained in the room. This system works fine.
There are 2Nos Biosafety Cabinets(BSC) (TypeA2) in the room that exhaust 30% air after HEPA filtration. The ducts are conneted by Thimble connection and connected to the same duct that exhausts the room air.
Is this correct or should we connect the BSC exhaust to dedicated exhaust blower? Even our exhaust system is a dedicated exhaust system for this room.
In our arrangement we have a few advantages viz. additional stage of HEPA filtration of BSC in the exhaust AHU; proper exhaust of air from the BSC through thimble connection, as power of room exhaust suction is more than that of individual blower for the BSC; the preset room negative pressure controls the quantity of air exhausted from the room, there is better control when air is exhausted through one blower; finally the room exhaust blower has a UPS backup to maintain the room at negative pressure for 15minutes or so even when there is power outing, in our case the BSCs also get this advantage (during sudden power outage air from the BSC workarea can come into the Lab and contaminate the room & occupants)
Kindly clarify if the arrangement we have done is good enough or should we go for separate exhaust system for the BSCs.
Working of the system is: Fresh air is cooled in an Air Handling Unit filtered in 3 stages, 10micron, 5micron & 0.3micron (HEPA) and supplied to the room. Room air is exhausted into the atmosphere after HEPA filtration. The exhaust quantity is controlled by a VFD based on the Negative pressure to be maintained in the room. This system works fine.
There are 2Nos Biosafety Cabinets(BSC) (TypeA2) in the room that exhaust 30% air after HEPA filtration. The ducts are conneted by Thimble connection and connected to the same duct that exhausts the room air.
Is this correct or should we connect the BSC exhaust to dedicated exhaust blower? Even our exhaust system is a dedicated exhaust system for this room.
In our arrangement we have a few advantages viz. additional stage of HEPA filtration of BSC in the exhaust AHU; proper exhaust of air from the BSC through thimble connection, as power of room exhaust suction is more than that of individual blower for the BSC; the preset room negative pressure controls the quantity of air exhausted from the room, there is better control when air is exhausted through one blower; finally the room exhaust blower has a UPS backup to maintain the room at negative pressure for 15minutes or so even when there is power outing, in our case the BSCs also get this advantage (during sudden power outage air from the BSC workarea can come into the Lab and contaminate the room & occupants)
Kindly clarify if the arrangement we have done is good enough or should we go for separate exhaust system for the BSCs.
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phd in nus through gate score
hello!!! my name is k rajsekhar... i am doing M.tech in IIT roorkee in hydraulics engg(civil).....my gate percentile is 99.35, mtech current cgpa is 8.00 and hope to increase upto 8.4, btech cgpa is 7.83...... i want to know what are the chances of me getting a seat in NUS for phd. am applying for january 2016 intake.
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Interview Query!!!
I am an Mtech Student of IIT Madras. I am going for Mtech project to South Korea from Jan- Mar end, I have applied for NUS (Aug intake), the shortlisted students will have interview in March during which I will be in Korea.
I wanted to ask if the Interview can be postponed OR will they conduct interview through skype/ online?
In the application, they have mentioned : "For foreign applicants who are not residing in P. R. China or India Where possible, the University may arrange an interview for you at the Singapore Embassy of the country in which you are residing. "
So If I want to give interview in South Korea during this time, will it be allowed with help of Singapore embassy there?
Hope the question is clear.
Thank you.
![.pdf]( "Adobe Acrobat PDF")
RApplFormAug14Jan15.pdf (Size: 429.93 KB / Downloads: 0)
I wanted to ask if the Interview can be postponed OR will they conduct interview through skype/ online?
In the application, they have mentioned : "For foreign applicants who are not residing in P. R. China or India Where possible, the University may arrange an interview for you at the Singapore Embassy of the country in which you are residing. "
So If I want to give interview in South Korea during this time, will it be allowed with help of Singapore embassy there?
Hope the question is clear.
Thank you.
RApplFormAug14Jan15.pdf (Size: 429.93 KB / Downloads: 0)
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job - organic farming
i am going to start a biotech industry - its an agro based
production of organic fruits and vegetables in that biotech lab will be there to quality control, production and packing
disease control and production of microbes in large scale. etc.
also research will be carried out.
i need a person who can handle entire farm and biotech lab - entire operation.
first payment will be less only after the business pickup
as the candidate ability payment will be resumed
if any if one interested please contact
by
dr.m.vijay antony
drmvijayantony@gmail.com
production of organic fruits and vegetables in that biotech lab will be there to quality control, production and packing
disease control and production of microbes in large scale. etc.
also research will be carried out.
i need a person who can handle entire farm and biotech lab - entire operation.
first payment will be less only after the business pickup
as the candidate ability payment will be resumed
if any if one interested please contact
by
dr.m.vijay antony
drmvijayantony@gmail.com
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Biotechnology for Beginners
So I've recently gotten interested in Biotechnology, and I was wondering if anyone had some tips on learning more, books, where to get more information, how to find out what I might be interested in more specifically, and those sort of things? Or should I just check out all the books from the library, cause that's all I've done so far!
~
Thanks for reading!
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Thanks for reading!
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Gene Probes?
I'm currently learning about gene probes, but everything still seems confusing. ![Huh]( "Huh")
Can someone help explain it to me?
Can someone help explain it to me?
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application in chem engg for masters in NUS
Hello I am now in 4th yr of my undergrad program in chemical engg... I will be giving GATE on 31st Jan,2015 and results will be declared in the month of March...I want to apply for masters in NUS and I heard that they take admissions through GATE score...now I want to ask that whether they have any extended deadlines for Indian students for applying through GATE for Fall'2015?
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Re: CNG?
Not just the students, but companies like Nokia, Schneider Electric and STMicroelectronics have been involved with it........!!!!!!!!!!!!
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How well can you focus? Identification of neural network in visual attention
A new study on the lateral prefrontal cortex (LPFC) brain area of macaques has identified a network of neurons that interact to rapidly process visual information while ignoring distractions. In humans, this ability is key to effective performance of tasks from driving a car to performing brain surgery. The study, published in the current issue of the journal Neuron, comes from researchers based in Canada and Germany and is the first convincing demonstration that an ensemble of neurons in the LPFC reliably generates attentional signals in a way that is resilient to distraction. The results of the study have implications for people with some neurological diseases and for paralysed people.
In the current study, the researchers recorded the brain activity of the macaques as they moved their eyes to watch objects on a computer screen while ignoring visual distractions. The recorded signals were processed by a decoder on a personal computer mimicking the type of computations that the brain carries out while focussing. Lead author Julio Martinez-Trujillo, of McGill University in Montreal explains the results obtained: "The decoder was able to predict very consistently and within a few milliseconds where the macaques were covertly focusing attention even before they looked in that direction…We were also able to predict whether the monkey would be distracted by some intrusive stimulus even before the onset of that distraction."
Furthermore, the researchers were able to manipulate the neuronal interactions that had been recorded and thereby alter the ability of the computer to ‘focus’. They could thus induce both ‘focused’ and ‘distracted’ states in the computer. First author Sébastien Tremblay of McGill University explains the significance of this result in terms of understanding of and intervention in diseases: "This suggests that we are tapping into the mechanisms responsible for the quality of the attentional focus, and might shed light into the reasons why this process fails in certain neurological diseases such as ADHD, autism and schizophrenia…Being able to extract and read the neuronal code from higher-level areas of the brain could also lead to important breakthroughs in the emerging field of neural prosthetics, where people who are paralysed use their thoughts to control objects in their environment."
Reference: Tremblay S, Pieper F, Sachs A, Martinez-Trujillo J. Attentional Filtering of Visual Information by Neuronal Ensembles in the Primate Lateral Prefrontal Cortex. Neuron 85(1): 202–215, 7 January 2015. DOI: http://dx.doi.org/10.1016/j.neuron.2014.11.021
Press release: McGill University; available at http://www.eurekalert.org/pub_releases/2015-01/mu-hah010615.php
In the current study, the researchers recorded the brain activity of the macaques as they moved their eyes to watch objects on a computer screen while ignoring visual distractions. The recorded signals were processed by a decoder on a personal computer mimicking the type of computations that the brain carries out while focussing. Lead author Julio Martinez-Trujillo, of McGill University in Montreal explains the results obtained: "The decoder was able to predict very consistently and within a few milliseconds where the macaques were covertly focusing attention even before they looked in that direction…We were also able to predict whether the monkey would be distracted by some intrusive stimulus even before the onset of that distraction."
Furthermore, the researchers were able to manipulate the neuronal interactions that had been recorded and thereby alter the ability of the computer to ‘focus’. They could thus induce both ‘focused’ and ‘distracted’ states in the computer. First author Sébastien Tremblay of McGill University explains the significance of this result in terms of understanding of and intervention in diseases: "This suggests that we are tapping into the mechanisms responsible for the quality of the attentional focus, and might shed light into the reasons why this process fails in certain neurological diseases such as ADHD, autism and schizophrenia…Being able to extract and read the neuronal code from higher-level areas of the brain could also lead to important breakthroughs in the emerging field of neural prosthetics, where people who are paralysed use their thoughts to control objects in their environment."
Reference: Tremblay S, Pieper F, Sachs A, Martinez-Trujillo J. Attentional Filtering of Visual Information by Neuronal Ensembles in the Primate Lateral Prefrontal Cortex. Neuron 85(1): 202–215, 7 January 2015. DOI: http://dx.doi.org/10.1016/j.neuron.2014.11.021
Press release: McGill University; available at http://www.eurekalert.org/pub_releases/2015-01/mu-hah010615.php
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Cocoa flavanols are good for your brain
Dietary supplementation with cocoa flavanols improves aspects of cognitive function in older adults and also confer cardiometabolic advantages. These are the major findings of a new paper in the American Journal of Clinical Nutrition on older people without cognitive impairment. It is the second instalment of a study from Italy's University of L'Aquila and Mars, Incorporated, the first of which, in the journal Hypertension, demonstrated similar benefits in older adults with mild cognitive impairment.
While some deterioration in cognitive function is normal with aging, the increasing life expectancy of the global population means that deferring and reducing such deterioration is more important than ever. In this double-blind, controlled, parallel-arm study, 90 elderly individuals aged 61-85 years who showed no evidence of cognitive impairment were randomly assigned to daily consumption of drinks containing different levels of cocoa flavanol, either high flavanol (HF; 993 mg), intermediate flavanol (IF; 520mg) or low flavanol (LF; 48 mg). The study was performed over an eight-week period. These drinks were nutritionally matched and specially prepared. The HF and IF cocoa drinks were produced using Mars' patented Cocoapro® process, while the LF drink was made from highly processed, alkalized cocoa powder. Normal diets and regular lifestyle were maintained throughout the study period apart from the supplementation of the diet with the drinks. Cognitive function was assessed at the beginning and the end of the eight-week study period using three tests, namely the Mini-Mental State Examination (MMSE), the Trail Making Test (TMT) A and B, and the Verbal Fluency Test (VFT).
The results of the study showed no differences were observed between groups for the MMSE. However, for the TMT, the improvement in the mean time to complete tests A and B after consumption of the drinks was significantly greater in the HF and IF groups compared to the LF group. For the VFT, performance improved in all three groups, however the magnitude of the improvement was significantly greater in the HF compared to either the IF or the LF groups. Beyond the improvements in cognitive function, the results of the study also indicated significant improvement in cardiometabolic outcomes, including insulin resistance, blood pressure and lipid peroxidation.
The mechanism behind the improvement in cognitive function in response to cocoa flavanols is not fully understood but the study's authors suggest that the improvements in insulin resistance and blood pressure may be significant in this regard. Lead author Dr. Giovambattista Desideri explains: "Earlier studies suggest a central role for insulin resistance in brain aging…These results could therefore provide some insight into a possible mechanism of action for the cognitive improvements we have observed." Improvements in vascular function as a result of cocoa flavanol consumption are indicated in various studies over the past decade. Co-author Dr. Catherine Kwik-Uribe, human health and nutrition director at Mars, Incorporated, says, "Since the brain is a heavily vascularized tissue, we might also be looking at vascular improvements as underlying the observed improvements in cognitive function."
While these results on cocoa flavanols are encouraging, there is a note of caution to be sounded to those who might be reaching for the chocolate. The trial was carried out with a special cocoa flavanol test product, which is designed to deliver a standardized amount of flavanols within a nutritionally suitable drink. Unfortunately, commercially available chocolate has a variable flavanol content and, given its macronutrient profile, is not recommended as a health food.
References:
Mastroiacovo D et al. Cocoa flavanol consumption improves cognitive function, blood pressure control, and metabolic profile in elderly subjects: the Cocoa, Cognition, and Aging (CoCoA) Study—a randomized controlled trial. Am J Clin Nutr doi: 10.3945/ajcn.114.092189
Desideri G et al. Benefits in Cognitive Function, Blood Pressure, and Insulin Resistance Through Cocoa Flavanol Consumption in Elderly Subjects With Mild Cognitive Impairment: The Cocoa, Cognition, and Aging (CoCoA) Study. Hypertension 60: 794-801; doi:10.1161/HYPERTENSIONAHA.112.193060
Press release: Mars Symbioscience; available at http://www.eurekalert.org/pub_releases/2015-01/ms-med010615.php
While some deterioration in cognitive function is normal with aging, the increasing life expectancy of the global population means that deferring and reducing such deterioration is more important than ever. In this double-blind, controlled, parallel-arm study, 90 elderly individuals aged 61-85 years who showed no evidence of cognitive impairment were randomly assigned to daily consumption of drinks containing different levels of cocoa flavanol, either high flavanol (HF; 993 mg), intermediate flavanol (IF; 520mg) or low flavanol (LF; 48 mg). The study was performed over an eight-week period. These drinks were nutritionally matched and specially prepared. The HF and IF cocoa drinks were produced using Mars' patented Cocoapro® process, while the LF drink was made from highly processed, alkalized cocoa powder. Normal diets and regular lifestyle were maintained throughout the study period apart from the supplementation of the diet with the drinks. Cognitive function was assessed at the beginning and the end of the eight-week study period using three tests, namely the Mini-Mental State Examination (MMSE), the Trail Making Test (TMT) A and B, and the Verbal Fluency Test (VFT).
The results of the study showed no differences were observed between groups for the MMSE. However, for the TMT, the improvement in the mean time to complete tests A and B after consumption of the drinks was significantly greater in the HF and IF groups compared to the LF group. For the VFT, performance improved in all three groups, however the magnitude of the improvement was significantly greater in the HF compared to either the IF or the LF groups. Beyond the improvements in cognitive function, the results of the study also indicated significant improvement in cardiometabolic outcomes, including insulin resistance, blood pressure and lipid peroxidation.
The mechanism behind the improvement in cognitive function in response to cocoa flavanols is not fully understood but the study's authors suggest that the improvements in insulin resistance and blood pressure may be significant in this regard. Lead author Dr. Giovambattista Desideri explains: "Earlier studies suggest a central role for insulin resistance in brain aging…These results could therefore provide some insight into a possible mechanism of action for the cognitive improvements we have observed." Improvements in vascular function as a result of cocoa flavanol consumption are indicated in various studies over the past decade. Co-author Dr. Catherine Kwik-Uribe, human health and nutrition director at Mars, Incorporated, says, "Since the brain is a heavily vascularized tissue, we might also be looking at vascular improvements as underlying the observed improvements in cognitive function."
While these results on cocoa flavanols are encouraging, there is a note of caution to be sounded to those who might be reaching for the chocolate. The trial was carried out with a special cocoa flavanol test product, which is designed to deliver a standardized amount of flavanols within a nutritionally suitable drink. Unfortunately, commercially available chocolate has a variable flavanol content and, given its macronutrient profile, is not recommended as a health food.
References:
Mastroiacovo D et al. Cocoa flavanol consumption improves cognitive function, blood pressure control, and metabolic profile in elderly subjects: the Cocoa, Cognition, and Aging (CoCoA) Study—a randomized controlled trial. Am J Clin Nutr doi: 10.3945/ajcn.114.092189
Desideri G et al. Benefits in Cognitive Function, Blood Pressure, and Insulin Resistance Through Cocoa Flavanol Consumption in Elderly Subjects With Mild Cognitive Impairment: The Cocoa, Cognition, and Aging (CoCoA) Study. Hypertension 60: 794-801; doi:10.1161/HYPERTENSIONAHA.112.193060
Press release: Mars Symbioscience; available at http://www.eurekalert.org/pub_releases/2015-01/ms-med010615.php
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U.S. brain drain of young scientific talent needs to be stemmed
A ‘brain drain’ of young talent is threatening the future of scientific research in the U.S. That’s according to a new article by Johns Hopkins University President Ronald J. Daniels, published this week in the online Early Edition of the journal Proceedings of the National Academy of Sciences. The article shows that grants for young scientists in the U.S. have been declining for more than a generation.
One striking figure highlighted in the article shows that the percentage of principal investigators with a leading National Institutes of Health grant aged 36 years old or younger fell from 18 % in 1983 to only 3 % in 2010. The average age for a scientist with a medical degree to obtain their first of these grants has risen from less than 38 years old in 1980 to over 45 in 2013. Prof Daniels points out the implications of these facts for younger scientists in the article: "Without their own funding, young researchers are prevented from starting their own laboratories, pursuing their own research, and advancing their own careers in academic science. It is not surprising that many of our youngest minds are choosing to leave their positions."
Prof Daniels goes on to point out that if the exodus of young researchers from academia continues, it will have implications including gradual evaporation of new discoveries, loss of future leaders and mentors, reduced diversity in the workforce and the loss of scientists just as they reach what should be a pivotal point in their career. The article identifies the three main reasons behind the reduction in research funding for younger researchers as longer training periods, a grant system that may favour incumbents and increases in cost of research to universities, which may result in them sticking with established researchers rather than taking a chance on younger candidates with no established funding stream.
In the article, Prof Daniels suggests policy reforms to address the position of young scientists. These include more robust funding of the NIH, with more money set aside for new talent, and refining of the peer review process to better accommodate inexperienced scientists and more daring proposals. He also suggests that a standing body should be established to review the issue on an ongoing basis and assess effectiveness of any interventions, as well as to drive stakeholders such as the NIH, Congress and the universities to take effective action.
Prof Daniels says that the U.S. can learn from other countries: "The inability to staunch -- if not reverse -- the above trends stands as an urgent and compelling policy challenge. The current stewards of the U.S. research enterprise bear a responsibility to sustain and safeguard that enterprise so that it can provide a platform for the scientists and the science of generations to come…Other countries are marshalling the will and resources to invest in the next generation of young scientists. A comparable solution in the United States will require a comparable commitment on the part of all actors in the biomedical science ecosystem. ... Our next generation of scientists, and indeed our next generation of science, demands nothing less."
Reference: Daniels RJ. A generation at risk: Young investigators and the future of the biomedical workforce. PNAS doi: 10.1073/pnas.1418761112
Press release: Johns Hopkins University; available at http://www.eurekalert.org/pub_releases/2015-01/jhu-sar010715.php
One striking figure highlighted in the article shows that the percentage of principal investigators with a leading National Institutes of Health grant aged 36 years old or younger fell from 18 % in 1983 to only 3 % in 2010. The average age for a scientist with a medical degree to obtain their first of these grants has risen from less than 38 years old in 1980 to over 45 in 2013. Prof Daniels points out the implications of these facts for younger scientists in the article: "Without their own funding, young researchers are prevented from starting their own laboratories, pursuing their own research, and advancing their own careers in academic science. It is not surprising that many of our youngest minds are choosing to leave their positions."
Prof Daniels goes on to point out that if the exodus of young researchers from academia continues, it will have implications including gradual evaporation of new discoveries, loss of future leaders and mentors, reduced diversity in the workforce and the loss of scientists just as they reach what should be a pivotal point in their career. The article identifies the three main reasons behind the reduction in research funding for younger researchers as longer training periods, a grant system that may favour incumbents and increases in cost of research to universities, which may result in them sticking with established researchers rather than taking a chance on younger candidates with no established funding stream.
In the article, Prof Daniels suggests policy reforms to address the position of young scientists. These include more robust funding of the NIH, with more money set aside for new talent, and refining of the peer review process to better accommodate inexperienced scientists and more daring proposals. He also suggests that a standing body should be established to review the issue on an ongoing basis and assess effectiveness of any interventions, as well as to drive stakeholders such as the NIH, Congress and the universities to take effective action.
Prof Daniels says that the U.S. can learn from other countries: "The inability to staunch -- if not reverse -- the above trends stands as an urgent and compelling policy challenge. The current stewards of the U.S. research enterprise bear a responsibility to sustain and safeguard that enterprise so that it can provide a platform for the scientists and the science of generations to come…Other countries are marshalling the will and resources to invest in the next generation of young scientists. A comparable solution in the United States will require a comparable commitment on the part of all actors in the biomedical science ecosystem. ... Our next generation of scientists, and indeed our next generation of science, demands nothing less."
Reference: Daniels RJ. A generation at risk: Young investigators and the future of the biomedical workforce. PNAS doi: 10.1073/pnas.1418761112
Press release: Johns Hopkins University; available at http://www.eurekalert.org/pub_releases/2015-01/jhu-sar010715.php
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Cell Line Development & Engineering | 9–12 February 2015| Dublin, Ireland|
Cell Line Development & Engineering 2015
Organisers: Informa Life Sciences
Dates: 9th–12th February 2015
Location: Radisson Blu Royal Hotel, Dublin, Ireland
Website: http://www.informa-ls.com/event/Cellline
The website gives all the necessary information on abstract submission, fees, registration, conference agenda, hotel, exhibitions and other important facts. The attached programme also gives details on all the scientific and social programme as well as the conference abstracts.
Purpose of the conference:
This is designed to be the ‘go-to’ European conference to benchmark against leading industry case studies on the latest in novel technologies and strategic approaches in cell banking, quality control, automation, high throughput clone selection, cell line development, cell line engineering and the application of the 'omics for improved bioprocessing. It will feature presentations of latest industry data, interactive focus groups, panel discussions and dedicated networking time, and is intended to ensure streamlined processes and strategies to get products to the market quickly.
Session topics:
A detailed programme is attached, containing all the information on individual speakers and presentations. To summarise the outline of the event:
Features:
Who should attend?
Heads of Department, Directors, Managers, Team Leaders, Associates, Consultants and Coordinators from:
![.pdf]( "Adobe Acrobat PDF")
CQ3542_Cell_Line_Development_and_Engineering_2015_(2).pdf (Size: 1.35 MB / Downloads: 0)
Organisers: Informa Life Sciences
Dates: 9th–12th February 2015
Location: Radisson Blu Royal Hotel, Dublin, Ireland
Website: http://www.informa-ls.com/event/Cellline
The website gives all the necessary information on abstract submission, fees, registration, conference agenda, hotel, exhibitions and other important facts. The attached programme also gives details on all the scientific and social programme as well as the conference abstracts.
Purpose of the conference:
This is designed to be the ‘go-to’ European conference to benchmark against leading industry case studies on the latest in novel technologies and strategic approaches in cell banking, quality control, automation, high throughput clone selection, cell line development, cell line engineering and the application of the 'omics for improved bioprocessing. It will feature presentations of latest industry data, interactive focus groups, panel discussions and dedicated networking time, and is intended to ensure streamlined processes and strategies to get products to the market quickly.
Session topics:
A detailed programme is attached, containing all the information on individual speakers and presentations. To summarise the outline of the event:
- Day 1: Product Quality & Cell Line Development Strategies for Difficult to Express Proteins, Non-Antibody Proteins and Biosimilars
- Day 2A: Automation, Screening, Clone Selection, Product Quality and High Throughput Analytics
- Day 2B: Transient Expression Systems
- Day 3D: Cell Line Engineering, Targeted Integration and Cell Culture Optimisation
- Day 3E: Cell Banking: Characterisation, Contamination Control and Regulatory Compliance
- Day 4: The 'Omics and Cell Engineering
Features:
- There is a 1 Day Pass available to facilitate flexible attendance
- A parallel stream format is available to maximise time out of the office
- There will be 50 case studies and practical advice
- There will be essential insight on the application of targeted integration and genome editing technologies including CRISPR Cas9 to improve cell line engineering
- There will be information on how to integrate NGS and early high throughput analytics to enhance product quality and shorten development timeframes
- There will be advice on gaining practical strategies for translating next generation ‘omics data into knowledge
- There will be information on how to reduce time from product candidate to the clinic.
Who should attend?
Heads of Department, Directors, Managers, Team Leaders, Associates, Consultants and Coordinators from:
- Cell Line Development
- Cell Line Engineering
- Cell Culture Technologies/Development
- Process Development
- R&D
CQ3542_Cell_Line_Development_and_Engineering_2015_(2).pdf (Size: 1.35 MB / Downloads: 0)
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An avocado a day keeps the heart doctor away?
Incorporating one fresh avocado per day into a moderate fat diet results in a greater improvement of high density lipoprotein cholesterol (HDL; ‘good’) to low density lipoprotein (LDL; ‘bad’) cholesterol ratio and total LDL reduction than a moderate or low fat diet without avocado in obese or overweight adults. This is the main finding of a new study in the Journal of the American Heart Association.
Elevated blood cholesterol has been associated with increased cardiovascular disease risk factors. It is recommended that a ‘heart healthy’ diet be adopted to keep cholesterol levels in the normal range and in particular to improve the LDL to HDL cholesterol ratio and reduce overall levels of LDL cholesterol. To this end, public health advice such as the 2010 Dietary Guidelines for Americans recommends limitation of saturated fat in the diet and replacement with unsaturated fats and increased fruits and vegetables intake.
In the current randomized, cross-over, controlled feeding trial, 45 healthy overweight or obese men and women were recruited. To establish a baseline prior to the study diet, subjects followed a typical American high saturated fat diet for two weeks. Participants then followed three different calorie controlled diets for five weeks each, in which 6-7% of energy from the saturated fat in the American diet was differentially replaced. In the low fat diet, it was replaced with complex carbohydrates mainly from grains and in the moderate fat diet it was replaced with monounsaturated fat mainly from high oleic acid oils (sunflower oil and canola oil) and some low-fat dairy products. Finally the avocado diet was similar to the moderate fat diet but the 6-7% of energy was replaced by monounsaturated fat, mainly from one fresh avocado daily (~ 136 g, without skin and seeds). The diets were matched for calories, macronutrients and fatty acids but not for fibre, phytosterols, or other bioactives.
The results of the study showed that the reduction in both LDL and non‐ HDL cholesterol on the avocado diet was significantly greater than for the MF or LF diets. Also, only the avocado diet significantly decreased LDL particle number, small dense LDL cholesterol and the ratio of LDL/HDL. Possible reasons why the avocado element in the moderate fat diet was beneficial include the unique combination of vitamins, minerals, fibre, phytosterols, and other dietary bioactives from the avocados that was missing in the other two diets. It is noteworthy that the avocado diet provided 35% more fibre, for example, than the other diets. These results were based on daily consumption of one whole avocado. Additional research should determine whether similar results are achieved with the recommended daily serving size of 1/5 of an avocado.
Lead author on the study Dr Penny Kris-Etherton of Pennsylvania State University says: "The results of this study suggest that the monounsaturated fat, fibre, phytosterols and other dietary bioactives in avocados may provide greater benefits to cardiovascular disease risk factors compared to a calorie matched low fat diet...Furthermore, using novel advanced lipid testing methods, this study demonstrated that consumption of one avocado a day may affect atherogenic lipoprotein particle numbers (APL)."
The conclusions from this study are encouraging, although they cannot be generalised to all populations. Nevertheless, the study does support the notion that the monounsaturated fat, fibre, phytosterols and other bioactives in avocados are likely to be beneficial to heart health.
Reference: Wang L, Bordi PL, Fleming JA, Hill AM, Kris‐Etherton PM. Effect of a Moderate Fat Diet With and Without Avocados on Lipoprotein Particle Number, Size and Subclasses in Overweight and Obese Adults: A Randomized, Controlled Trial. JAMA doi: 10.1161/JAHA.114.001355
Press release: FoodMinds LLC; available at http://www.eurekalert.org/pub_releases/2015-01/fl-nre010715.php
Elevated blood cholesterol has been associated with increased cardiovascular disease risk factors. It is recommended that a ‘heart healthy’ diet be adopted to keep cholesterol levels in the normal range and in particular to improve the LDL to HDL cholesterol ratio and reduce overall levels of LDL cholesterol. To this end, public health advice such as the 2010 Dietary Guidelines for Americans recommends limitation of saturated fat in the diet and replacement with unsaturated fats and increased fruits and vegetables intake.
In the current randomized, cross-over, controlled feeding trial, 45 healthy overweight or obese men and women were recruited. To establish a baseline prior to the study diet, subjects followed a typical American high saturated fat diet for two weeks. Participants then followed three different calorie controlled diets for five weeks each, in which 6-7% of energy from the saturated fat in the American diet was differentially replaced. In the low fat diet, it was replaced with complex carbohydrates mainly from grains and in the moderate fat diet it was replaced with monounsaturated fat mainly from high oleic acid oils (sunflower oil and canola oil) and some low-fat dairy products. Finally the avocado diet was similar to the moderate fat diet but the 6-7% of energy was replaced by monounsaturated fat, mainly from one fresh avocado daily (~ 136 g, without skin and seeds). The diets were matched for calories, macronutrients and fatty acids but not for fibre, phytosterols, or other bioactives.
The results of the study showed that the reduction in both LDL and non‐ HDL cholesterol on the avocado diet was significantly greater than for the MF or LF diets. Also, only the avocado diet significantly decreased LDL particle number, small dense LDL cholesterol and the ratio of LDL/HDL. Possible reasons why the avocado element in the moderate fat diet was beneficial include the unique combination of vitamins, minerals, fibre, phytosterols, and other dietary bioactives from the avocados that was missing in the other two diets. It is noteworthy that the avocado diet provided 35% more fibre, for example, than the other diets. These results were based on daily consumption of one whole avocado. Additional research should determine whether similar results are achieved with the recommended daily serving size of 1/5 of an avocado.
Lead author on the study Dr Penny Kris-Etherton of Pennsylvania State University says: "The results of this study suggest that the monounsaturated fat, fibre, phytosterols and other dietary bioactives in avocados may provide greater benefits to cardiovascular disease risk factors compared to a calorie matched low fat diet...Furthermore, using novel advanced lipid testing methods, this study demonstrated that consumption of one avocado a day may affect atherogenic lipoprotein particle numbers (APL)."
The conclusions from this study are encouraging, although they cannot be generalised to all populations. Nevertheless, the study does support the notion that the monounsaturated fat, fibre, phytosterols and other bioactives in avocados are likely to be beneficial to heart health.
Reference: Wang L, Bordi PL, Fleming JA, Hill AM, Kris‐Etherton PM. Effect of a Moderate Fat Diet With and Without Avocados on Lipoprotein Particle Number, Size and Subclasses in Overweight and Obese Adults: A Randomized, Controlled Trial. JAMA doi: 10.1161/JAHA.114.001355
Press release: FoodMinds LLC; available at http://www.eurekalert.org/pub_releases/2015-01/fl-nre010715.php
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query about gate bioprocess engineering questions
goodevening sir, this is mallika pandey. i am persuing B.tech biotechnology from invertis university bareilly. Sir, i am preparing for gate and i had read your articles about gate preparation , they are highly inspiring for gate aspirants.Sir , i have an doubt on one question of bioprocess engineering of gate 2014. Sir, please do me a favour in this.
Question) Consider a continuous culture provided with a sterile feed containing 10 mM glucose. The steady
state cell density and substrate concentration at three different dilution rates are given in the table
below.
Dilution rate (h-1) Cell density (g L-1) Substrate concentration (mM)
0.05 0.248 0.067
0.5 0.208 1.667
5 0 10
The maximum specific growth rate µm (in h−1), will be _____.
Thanking you
with regards
mallika pandey
Question) Consider a continuous culture provided with a sterile feed containing 10 mM glucose. The steady
state cell density and substrate concentration at three different dilution rates are given in the table
below.
Dilution rate (h-1) Cell density (g L-1) Substrate concentration (mM)
0.05 0.248 0.067
0.5 0.208 1.667
5 0 10
The maximum specific growth rate µm (in h−1), will be _____.
Thanking you
with regards
mallika pandey
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Bacterial input to wound-associated skin cancer
Binding of a bacterial protein by a white blood cell receptor at the site of wounds associated with chronic skin inflammation can tip the balance from wound healing to tumour formation. That is a major finding of a study published today in the journal Nature Communications from researchers in King's College London. The results of this mouse study may have important ramifications for therapies for patients suffering from chronic ulcers or skin blistering diseases.
It has been previously established that there is a link between chronic inflammation, tissue damage and tumour formation, however underlying mechanisms are not well understood. The current study is the first to show that bacteria on the skin could contribute to the formation of skin tumours. In the study, the researchers examined wounds in mice with chronic skin inflammation. They observed that tumours formed at the wound site, which depended on the presence of immune cells. The underlying signalling mechanism involved a pattern recognition receptor called Toll-like receptor 5 (TLR-5) on the surface of immune cells. TLR-5 recognises and binds to the bacterial flagellin protein, a protein monomer containing highly conserved regions which makes up the bacterial flagella, found on most motile bacteria.
The direct relevance of the findings to human skin cancer has yet to be investigated, however in an inherited chronic skin condition in humans called Epidermolysis Bullosa (EB), a protein called High mobility group box 1 (HMGB1), which is a nuclear protein involved in transcriptional regulation, is increased. HMGB1 is also increased in mice with chronic skin inflammation and in the current study, removal of TLR-5 from immune cells caused a reduction in HMGB1 levels in mice. These results suggest that therapies involving reduction of bacterial flagellin protein on the skin surface or targeting of TLR-5 could be effective in prevention of tumours in patients with chronic skin conditions.
Lead author Professor Fiona Watt, Director of the Centre for Stem Cells and Regenerative Medicine at King's College London, said: 'These findings have broad implications for various types of cancers and in particular for the treatment of tumours that arise in patients suffering from chronic ulcers or skin blistering diseases. In the context of chronic skin inflammation, the activity of a particular receptor in white blood cells, TLR-5, could tip the balance between normal wound repair and tumour formation…Our findings raise the possibility that the use of specific antibiotics targeting bacteria in wound-induced malignancies might present an interesting clinical avenue.'
Reference: Hoste E et al. Innate sensing of microbial products promotes wound-induced skin cancer. Nature Communications 6, Article number: 5932; doi:10.1038/ncomms6932
It has been previously established that there is a link between chronic inflammation, tissue damage and tumour formation, however underlying mechanisms are not well understood. The current study is the first to show that bacteria on the skin could contribute to the formation of skin tumours. In the study, the researchers examined wounds in mice with chronic skin inflammation. They observed that tumours formed at the wound site, which depended on the presence of immune cells. The underlying signalling mechanism involved a pattern recognition receptor called Toll-like receptor 5 (TLR-5) on the surface of immune cells. TLR-5 recognises and binds to the bacterial flagellin protein, a protein monomer containing highly conserved regions which makes up the bacterial flagella, found on most motile bacteria.
The direct relevance of the findings to human skin cancer has yet to be investigated, however in an inherited chronic skin condition in humans called Epidermolysis Bullosa (EB), a protein called High mobility group box 1 (HMGB1), which is a nuclear protein involved in transcriptional regulation, is increased. HMGB1 is also increased in mice with chronic skin inflammation and in the current study, removal of TLR-5 from immune cells caused a reduction in HMGB1 levels in mice. These results suggest that therapies involving reduction of bacterial flagellin protein on the skin surface or targeting of TLR-5 could be effective in prevention of tumours in patients with chronic skin conditions.
Lead author Professor Fiona Watt, Director of the Centre for Stem Cells and Regenerative Medicine at King's College London, said: 'These findings have broad implications for various types of cancers and in particular for the treatment of tumours that arise in patients suffering from chronic ulcers or skin blistering diseases. In the context of chronic skin inflammation, the activity of a particular receptor in white blood cells, TLR-5, could tip the balance between normal wound repair and tumour formation…Our findings raise the possibility that the use of specific antibiotics targeting bacteria in wound-induced malignancies might present an interesting clinical avenue.'
Reference: Hoste E et al. Innate sensing of microbial products promotes wound-induced skin cancer. Nature Communications 6, Article number: 5932; doi:10.1038/ncomms6932
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BCL11A: a novel overactive triple-negative breast cancer gene
Exciting results from a new study suggest that an overactive gene called BCL11A drives the development and progression of the aggressive triple-negative breast cancer. The study, carried out in both human cells and in mice, is published in the journal Nature Communications.
Triple-negative breast cancer has a particularly poor prognosis, being associated with high incidence of recurrence and metastasis to sites including the brain and lung. Treatment is complicated by its lack of oestrogen receptor, progesterone receptor and the HER2 protein, all of which are targets for therapy in other forms of breast cancer, and very few genomic aberrations have been identified for triple-negative breast cancer. This lack of effective therapies for triple-negative breast cancer makes identification of underlying mechanisms and tailored therapies all the more important. Approximately 20% of patients are affected by triple-negative breast cancer; the majority of triple-negative tumours are of the basal-like subtype.
In the current study, breast cancers from nearly 3000 patients were examined in order to identify genetic changes associated with behaviour of stem cells and developing tissues. Previous work by the same research team had suggested that mutations to such genes often drives cancer development. One of these genes was BCL11A. Senior author Dr Pentao Liu of the Wellcome Trust Sanger Institute explains further: "Our understanding of genes that drive stem cell development led us to search for consequences when these genes go wrong. BCL11A activity stood out because it is so active in triple-negative cancers. It had all the hallmarks of a novel breast cancer gene."
The results indicated that increased BCL11A activity occurred in approximately 8 out of every 10 patients with basal-like breast cancer and was associated with a more advanced grade of tumour. If the copy number of the BCL11A gene was increased, survival prospects for the patient were reduced. In a mouse cancer model, inactivation of BCL11A resulted in no tumour development in the mammary glands, while all mice in which the gene was not inactivated developed mammary tumours.
Manipulation of BCL11A in human and mouse cells impacted on their cancer-like properties. Joint first author Dr Walid Khaled of the Wellcome Trust Sanger Institute and University of Cambridge says: "Our gene studies in human cells clearly marked BCL11A as a novel driver for triple-negative breast cancers. We also showed that adding an active human BCL11A gene to human or mouse breast cells in the lab drove them to behave as cancer cells. As important, when we reduced the activity of BCL11A in three samples of human triple-negative breast cancer cells, they lost some characteristics of cancer cells and became less tumorigenic when tested in mice. So by increasing BCL11A activity we increase cancer-like behaviour; by reducing it, we reduce cancer-like behaviour." The research team also established that BCL11A is needed for normal development of breast stem cells and their progenitor. It is thought that these are the cells which give rise to basal-like breast cancer upon mutation.
The research team have proposed that their results indicate BCL11A as a strong candidate for development of a possible targeted treatment of triple-negative breast cancer.
Reference: Khaled, WT, Lee SC et al. (2015) BLC11A is a triple-negative breast cancer gene with critical functions in stem and progenitor cells. Nature Communications, published online in advance of print publication; doi: 10.1038/ncomms6987
Triple-negative breast cancer has a particularly poor prognosis, being associated with high incidence of recurrence and metastasis to sites including the brain and lung. Treatment is complicated by its lack of oestrogen receptor, progesterone receptor and the HER2 protein, all of which are targets for therapy in other forms of breast cancer, and very few genomic aberrations have been identified for triple-negative breast cancer. This lack of effective therapies for triple-negative breast cancer makes identification of underlying mechanisms and tailored therapies all the more important. Approximately 20% of patients are affected by triple-negative breast cancer; the majority of triple-negative tumours are of the basal-like subtype.
In the current study, breast cancers from nearly 3000 patients were examined in order to identify genetic changes associated with behaviour of stem cells and developing tissues. Previous work by the same research team had suggested that mutations to such genes often drives cancer development. One of these genes was BCL11A. Senior author Dr Pentao Liu of the Wellcome Trust Sanger Institute explains further: "Our understanding of genes that drive stem cell development led us to search for consequences when these genes go wrong. BCL11A activity stood out because it is so active in triple-negative cancers. It had all the hallmarks of a novel breast cancer gene."
The results indicated that increased BCL11A activity occurred in approximately 8 out of every 10 patients with basal-like breast cancer and was associated with a more advanced grade of tumour. If the copy number of the BCL11A gene was increased, survival prospects for the patient were reduced. In a mouse cancer model, inactivation of BCL11A resulted in no tumour development in the mammary glands, while all mice in which the gene was not inactivated developed mammary tumours.
Manipulation of BCL11A in human and mouse cells impacted on their cancer-like properties. Joint first author Dr Walid Khaled of the Wellcome Trust Sanger Institute and University of Cambridge says: "Our gene studies in human cells clearly marked BCL11A as a novel driver for triple-negative breast cancers. We also showed that adding an active human BCL11A gene to human or mouse breast cells in the lab drove them to behave as cancer cells. As important, when we reduced the activity of BCL11A in three samples of human triple-negative breast cancer cells, they lost some characteristics of cancer cells and became less tumorigenic when tested in mice. So by increasing BCL11A activity we increase cancer-like behaviour; by reducing it, we reduce cancer-like behaviour." The research team also established that BCL11A is needed for normal development of breast stem cells and their progenitor. It is thought that these are the cells which give rise to basal-like breast cancer upon mutation.
The research team have proposed that their results indicate BCL11A as a strong candidate for development of a possible targeted treatment of triple-negative breast cancer.
Reference: Khaled, WT, Lee SC et al. (2015) BLC11A is a triple-negative breast cancer gene with critical functions in stem and progenitor cells. Nature Communications, published online in advance of print publication; doi: 10.1038/ncomms6987
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Discovery of new antibiotic which bacteria cannot resist
The growth of resistance of bacteria to antibiotics has outstripped the development and introduction of new antibiotic compounds, leading to a public health crisis with the rise of multidrug resistant ‘superbugs’. However, a study published this week in the journal Nature describes a new antibiotic called teixobactin, derived from growth of uncultured bacteria. The results of this study, from researchers in Northeastern University in Boston, Massachusetts, the University of Bonn in Germany, NovoBiotic Pharmaceuticals in Cambridge, Massachusetts, and Selcia Limited in the United Kingdom, offers hope of a new source of resistance-free antibiotics, as uncultured bacteria comprise approximately 99% of all bacterial species in external environments.
Most antibiotics currently available were produced by screening of cultivable soil microorganisms. However, this is a finite resource which according to the authors of the current study was essentially already over-exploited by the 1960s. Researchers on this study have been involved in developing novel methods for growing uncultured bacteria by cultivating them in situ or using specific growth factors. The work in this field by Northeastern University researchers and study authors Professor Kim Lewis and Professor Slava Epstein resulted in the founding of NovoBiotic. Their methods involve a miniature device called the iChip, which Prof Epstein's team created in order to isolate and help grow single cells in their natural environment. This has greatly improved the access of researchers to uncultured bacteria. The work has resulted in NovoBiotic assembling approximately 50,000 strains of uncultured bacteria and discovery of 25 new antibiotics. Prof Lewis says that teixobactin is the latest and most interesting of these.
In the current study, the researchers discovered that teixobactin inhibits bacterial cell wall synthesis by binding to lipid II and lipid I, which are precursors of the bacterial cell wall components peptidoglycan and teichoic acid respectively. Testing of teixobactin on mutant Staphylococcus aureus or Mycobacterium tuberculosis strains did not reveal any mutants that were resistant to the antibiotic. Prof Lewis said: "Our impression is that nature produced a compound that evolved to be free of resistance. This challenges the dogma that we've operated under that bacteria will always develop resistance. Well, maybe not in this case." The team now hope to develop teixobactin into a drug.
According to Gerard Wright, a professor in the Department of Biochemistry and Biomedical Sciences at McMaster University who was not involved in this research, it may yet prove that as yet unidentified mechanisms of resistance to teixobactin exist in the environment. In a separate Nature article published alongside the research article Prof Wright comments that nevertheless, the team's work could lead to identifying "other 'resistance-light' antibiotics…(The researchers') work offers hope that innovation and creativity can combine to solve the antibiotics crisis."
References:
Ling LL et al. A new antibiotic kills pathogens without detectable resistance. Nature (2015); doi:10.1038/nature14098
Wright G. Antibiotics: An irresistible newcomer. Nature (2015); doi:10.1038/nature14193
Press release: Northeastern University; available at http://www.eurekalert.org/pub_releases/2015-01/nu-nda010815.php
Most antibiotics currently available were produced by screening of cultivable soil microorganisms. However, this is a finite resource which according to the authors of the current study was essentially already over-exploited by the 1960s. Researchers on this study have been involved in developing novel methods for growing uncultured bacteria by cultivating them in situ or using specific growth factors. The work in this field by Northeastern University researchers and study authors Professor Kim Lewis and Professor Slava Epstein resulted in the founding of NovoBiotic. Their methods involve a miniature device called the iChip, which Prof Epstein's team created in order to isolate and help grow single cells in their natural environment. This has greatly improved the access of researchers to uncultured bacteria. The work has resulted in NovoBiotic assembling approximately 50,000 strains of uncultured bacteria and discovery of 25 new antibiotics. Prof Lewis says that teixobactin is the latest and most interesting of these.
In the current study, the researchers discovered that teixobactin inhibits bacterial cell wall synthesis by binding to lipid II and lipid I, which are precursors of the bacterial cell wall components peptidoglycan and teichoic acid respectively. Testing of teixobactin on mutant Staphylococcus aureus or Mycobacterium tuberculosis strains did not reveal any mutants that were resistant to the antibiotic. Prof Lewis said: "Our impression is that nature produced a compound that evolved to be free of resistance. This challenges the dogma that we've operated under that bacteria will always develop resistance. Well, maybe not in this case." The team now hope to develop teixobactin into a drug.
According to Gerard Wright, a professor in the Department of Biochemistry and Biomedical Sciences at McMaster University who was not involved in this research, it may yet prove that as yet unidentified mechanisms of resistance to teixobactin exist in the environment. In a separate Nature article published alongside the research article Prof Wright comments that nevertheless, the team's work could lead to identifying "other 'resistance-light' antibiotics…(The researchers') work offers hope that innovation and creativity can combine to solve the antibiotics crisis."
References:
Ling LL et al. A new antibiotic kills pathogens without detectable resistance. Nature (2015); doi:10.1038/nature14098
Wright G. Antibiotics: An irresistible newcomer. Nature (2015); doi:10.1038/nature14193
Press release: Northeastern University; available at http://www.eurekalert.org/pub_releases/2015-01/nu-nda010815.php
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