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The Panel of Economists and Bankers, after the completion of its work on the introduction of Zakah and Ushr, was asked to give priority to the consideration of ways and means to eliminate interest from the financial system of the country. Already the Panel has held a number of meetings to consider this matter. It has been mainly engaged in the task of exploring alternative financial arrangements whereby, on the one hand, all traces of interest may be eliminated while, on the other hand, the multifarious financial requirements of the economy may continue to be met without any dislocation. The ground covered so far by the Panel has surely set in motion the process of crystalisation of potentially fruitful ideas. The Banker's Sub-Committee of the Pane£. has presented a detailed scheme for the elimination of interest from the commercial banking system. This has provided to the Panel a valuable basis for its discussion on the subject. Work has proceeded on ways and means of eliminating interest from other components of the financial system. On the basis of its deliberations so far, the Panel has come to the conclusion that though it is not yet in a position to submit the full set of its recommendations in regard to elimination of interest, it should make an Interim Report to the Council of Islamic Ideology containing its first recommendations on the subject.
The Panel is in complete agreement that interest has no place in an Islamic economy. However, it is mindful of the fact that interest is so deeply entrenched in the existing economic system that its elimination is sure to raise problems of great complexity. According to the economic thought and practice developed in the Western world interest is the kingpin of the modern financial system. It is unfortunate that though Islam prohibits interest, the economies of all Muslim countries at present operate on the basis of interest. Muslim thought also has remained nearly dormant in the field of money and banking for centuries so that there is very little in the available literature on which the Panel could draw in its work related to an interest-free economic system. A few experiments with interest-free banking that are being made in certain Muslim countries are too limited in their scope to be of any significant help in shaping a completely interest-free economic and financial system. The Panel therefore feels that measures to eliminate interest from the economic system must follow a deep and thorough study of all the problems involved, and precipitate action should be avoided in this highly delicate field.
The Panel would like to emphasise that prohibition of interest is part of the overall value system of Islam. The most important ingredients of the Islamic socio-economic order are honesty, hard work, accountability to God, respect of each other's rights and 'Adi The Islamisation process, of which the gradual and orderly elimination of interest will be an essential part, will have to be fed by reformatory efforts at moral building and eradication of false values of life acquired by a prolonged contact with an un-Islamic economic and social system.
In view of the complexity of the task and the present moral texture of the society the Panel recommends a cautious approach in the matter of elimination of interest. The Panel gave careful consideration to various selected areas in which a phased programme for the elimination of interest could be initiated, and it was agreed that as a first step interest may be eliminated from the operations of the National Investment (Unit) Trust and the Investment Corporation of Pakistan as well as from housing finance provided by the House Building Finance Corporation and Pakistani banks. The details of how this may be done are gjven in the later part of this Report.
The specification of the areas from which interest may be eliminated to begin with has been covered by a number of considerations. In recommending the elimination of interest from the operations of N.I.T. the Panel has been mainly guided by the consideration that in the very first phase of gradual elimination of interest, people should be provided with an investment medium in which they can invest their savings without any religious inhibition. Among other financial institutions, I.C.P. has been selected because a good part of its operations is even now free of interest so that complete elimination of interest from its operations can be effected with the least possible delay. Moreover, by the very nature of its functions, I.C.P. is an institution which would be called upon to shoulder greater responsibilities in an interest-free system, and it is therefore appropriate that it should begin to be equipped for the purpose with utmost expedition. In recommending abolition of interest from housing finance the Panel seeks to meet 'a basic need of the people on interest-free basis and to rid a major sector of the economy from interest.
The Panel would like to underscore the point that transition to an interest-free system would have been a less complicated problem if the economy was more buoyant. Due to reasons which are well known the profitability of industry has been adversely affected in recent years. The actual yield on equities has been rather low. On the other hand, on account of the inflationary situation and low rate of national savings, interest rates in the economy have gone on rising. Hence institutions like N.I.T. and I.C.P. which should normally deal mostly in equities had to take in interest-bearing securities in their portfolios to keep up a sufficiently attractive yield for their investors. Despite this Government had to guarantee a minimum return on N.I.T. Units to sustain investors' interest. Elimination of interest-bearing instruments from the portfolio of N.I.T. would involve a reduction in its income which would need to be compensated by enhanced subsidy by Government so long as the interest system continues to operate in the rest of the economy.
While making the transition to interest-free system it would be necessary to ensure that saving and investment processes are not hampered and allocation of investment resources is optimal, keeping in full view the scarcity of capital. The Panel feels that as the interest- free system evolves it would necessitate the creation of new financial instruments to replace the present interest-bearing securities. Immediate attention needs to be given in this context to the replacement of debentures by a financial instrument which is in conformity with Shari'ah. Companies need finance for varying periods and hence, in addition to equity, they have to be enabled to raise capital for specific time duration. The Panel is of the view that fixed period Profit/Loss Participation Certificates can take the place of debentures. For the sake of brevity, such certificates have been titled as Participation Term Certificates in the following section of the Report.
Measures that need to be taken to eliminate interest from the operations of N.I.T. and I.C.P. and from housing finance are indicated below:
National Investment (Unit) Trust
The National Investment (Unit) Trust was established in 1962 mainly to mobilise and pool small household savings through the sale of Units and to deploy these for productive investment in the corporate sector. The N.I.T. sells two types of Units viz., Registered Units and Bearer Units. Both types of Units are encashable at any time. However, while the former are encashable only through the Trust's authorised agents the latter are freely transferable. The income of N.I.T. consists partly of dividends on its holdings of corporate shares and partly of interest on its holdings of Government securities and corporate debentures. It also earns interest on balances which it holds with commercial banks in the form of fixed deposits. Another source of its income is capital gains realised from sales of the shares on the stock exchange from time to time. The aggregate income from all these sources, adjusted for operational costs, is distributed among the Unit holders annually. It will be seen from the above that basically the rate of return on investment in N.I.T. Units is not fixed but variable depending on the level of the net income earned during the year. However, for the past several years, the Federal Government has been guaranteeing a minimum rate of return to the Unit holders, which involves granting of some subsidy by the Government to the N.I.T. The Government also guarantees the minimum repurchase price. Moreover, investment in Registered Units up to 30 per cent of the Unit holder's annual income (subject to a maximum of Rs. 20,000), qualifies for tax relief as investment allowance under Section 15 AA of the Income Tax Act. Income of N.I.T. is exempt from income tax while the dividend income of Unit holders is also exempt from income tax up to a maximum of Rs. 10,000/00*.
Interest is involved in N.I.T.'s operations in the case of its investment in Government securities, debentures, shares of commercial banks and fixed deposits held by it with banks. For eliminating interest from the transactions of the N.I.T. the Panel recommends the following measures:
N.I.T.'s holdings of Government securities, which amount to about Rs. 20 million and represent a small proportion (less than 3%) of its investment portfolio, may be sold to commercial banks/State Bank and the proceeds may be utilised for purchase of ordinary shares.
Debentures holdings constitute a significant proportion of N.I.T.'s total investments and amounted to Rs. 135.74 million or 19.52 per cent of N.I.T.'s investment portfolio as on 30th June, 1977. These may be sold to the commercial banks and/or converted into Participation Term Certificates with the consent of the issuing companies. These Certificates should be given the same treatment as the pre-tax deductable interest income in the profit and loss accounts of the companies con cerned. This will not cause any revenue loss to the Government and will at the same time provide an incentive to the issuing companies to convert the debentures into Partici- pation Term Certificates. In the hands of the recipient, the treatment of the income from these Certificates for tax purposes should be the same as in the case of dividend income. This measure would increase the marketability of the Certificates. N.I.T. also holds preference shares but the amount involved is not significant. It is recommended that these may also be disinvested in the same manner as debentures.
Holdings of banks' shares which constitute a small proportion (about 3%) of the N.I.T.'s investment portfolio and amounted to Rs. 20.5 million as on 30th June, 1977 may be sold to Government-sponsored financial institutions or to the Federal Government.
N.I.T. also holds substantial funds in the form of fixed deposits with banks. As on 30th June, 1977 these amounted to Rs. 47 million. Fixed deposits provide an avenue for deploying its surplus funds for earning income and also help in ensuring requisite liquidity as their maturity is so arranged that the cash balance requirements of N.I.T. are easily met. The Panel recommends that the fixed deposits may be liquidated and the proceeds used for purchase of shares. However, since the stock market at present lacks sufficient depth and the amount involved is quite sizeable the Panel suggests that the proceeds of fixed deposits may be deployed for selective purchase of ordinary shares from banks and from Government holdings of shares of nationalised industries.
After the dismantling of fixed deposits with banks, the N.I.T. would need to have a mechanism whereby it can invest its funds on short-term basis in shares and acquire necessary cash, whenever needed, by selling shares. Since in the present state of the stock exchange it cannot be taken for granted that N.I.T. will be able to acquire shares or replenish its cash resources by selling shares at short notice, the Panel recommends the adoption of the following two measures:
(a) An over-the-counter market may be opened by the I.C JP. for N.I.T. on the same lines as is provided by the institution for its investment account holders. Under this arrangement, I.C.P. should be provided with the list of shares held by the banks, other financial institutions and Federal and Provincial Governments which can be sold to N.I.T. as and when the N.I.T. has surplus funds for investment.
(b) Funds may be placed at the disposal of I.C.P. in the nature of a revolving fund for the sale and purchase of the shares. Whenever the need arises the N.I.T. may replenish its cash resources by selling shares from its portfolio to the I.C.P.
These measures would ensure that the N.I.T. is in a position to invest its surplus funds in non-interest income earning assets at any time and is also able to liquidate them whenever the need arises.
The Panel further recommends that in order to meet situations when N.I.T. may not be able to adequately meet its liquidity requirements through disinvestment from its portfolio on account of the risk of capital loss or undesirable repercussions on the stock market, it may be provided borrowing facilities by the State Bank free of interest. It may be mentioned that provision of this facility by the State Bank to the N.I.T. would not require any amendment in the State Bank of Pakistan Act.
With the elimination of the interest-bearing investments, N.I.T. is likely to suffer a fall in its income. In case the profitability of the companies whose shares are held by the N.I.T. and the conditions in the stock market do not improve markedly, the amount of subsidy which the Government has been providing to the N.I.T. to support the minimum repurchase price and the minimum return on the Units would need to be raised considerably. The Panel feels that continuation of Government guarantee of minimum repurchase price and minimum return on N.I.T. Units is essential for sustaining the interest of the investors for so long as the interest system continues to operate in the rest of the economy. The question of permissibility of a guarantee of this type is the transitional period from the viewpoint of Shari'ah may be decided upon by the Council of Islamic Ideology.
Investment Corporation of Pakistan
The I.C.P. was established in 1966 with the objectives of broadening the base of equity investments and encouraging the growth of the capital market in Pakistan. Its major functions consist of underwriting new issues of shares and debentures; maintaining investors' accounts and providing margin loans to the account holders; floating closed-end mutual funds; buying and selling shares on the stock exchange to help maintain stability in share values; managing investment portfolios on behalf of individual investors; and providing professional counsel for investments in the corporate sector.
Interest receipts and payments constitute substantial proportions of C.P.'s earnings and expenditure. For instance, during the year 1976- 77 interest accounted for Rs. 32.4 million or 57 per cent of total income and for Rs. 27.99 million or 63 per cent of total expenditure of I.C.P.* I.C.P's transactions involving interest consist of loans from Government of Pakistan and from the State Bank of Pakistan; issue of debentures; provision of bridging loans; loans under the Investors' Scheme; Mutual Funds operations; and interest on other investments and loans.
The Panel recommends the following measures to eliminate interest from the I.C.P.'s operations:
Debenture: The existing debentures held by I.C.P. may either be disinvested to the commercial banks or converted into Participation Term Certificates with the consent of the issuing companies. The terms and conditions on these certificates would be the same as suggested in the section on N.I.T. The Panel feels that the second alternative would be preferable keeping in view the fact that in the financial plans of companies there is continuing need for term finance.
The Debenture Investment Scheme introduced recently by I.CJP. should be replaced by the Participation Term Certificate scheme.
Underwriting: There are three types of underwriting techniques that are usually adopted by investment banks viz., 'best effort', 'standby' and 'firm commitment'. The 'best effort' and the 'firm commitment' techniques are not currently in use in Pakistan. Normally the 'best effort' technique is followed in highly sophisticated capital markets which have a wide net work of brokers and investment salesmen. The 'firm commitment' technique has not been applied primarily because it presupposes freedom in negotiating the price of the new issues at a discount which is not permissible under the Companies Act at present. The only technique that is in vogue in Pakistan is the 'stand-by' technique. Owing to a variety of reasons this technique has given birth to bridge financing arrangements whereby the underwriters give an interim loan at fixed rate of interest equivalent to their underwriting commitment for meeting the financial needs of the sponsor during the gestation period of the projects. Although underwriting business as such is not repugnant to Shari'ah, tije arrangement of bridge financing makes such type of underwriting incompatible with SharVah. The Panel feels that the present 'stand-by' technique of underwriting should be substituted by 'firm commitment' underwriting and neces- sary changes made in the Companies Act to provide freedom to the underwriter to negotiate the price of the new issues at a level below par value.
In case the present underwriting arrangements are substituted as recommended, the need for bridge financing, involving the element of interest, would be eliminated.
Mutual Funds: Mutual Funds are largely free of interest element. Interest payments are, however, involved in some mutual funds which have been advanced loans by the I.C.P. in order to compensate for the loss of income on investment in the shares of companies located in former East Pakistan. This interest element can be eliminated if Government takes over such shares.
On the income side, Mutual Funds receive fixed interest income on their deposits and debenture investments. The interest receipts on deposits are small in amount and represent interest payments by I.CP. pending investment. Interest payment by I.C.P. to the Mutual Funds in this case may be substituted by an appropriate profit/loss-sharing arrangement. As regards debentures they should either be substituted by shares or replaced by Participation Term Certificates.
Loans Under the Investors' Scheme: Under the Investors' Scheme, the I.C.P., apart from rendering advisory services, provides margin loans to investors for investment in shares of public companies listed on the stock exchange and charges interest on such loans. The interest element involved in this scheme can be eliminated by replacing the present loaning arrangements by a joint ownership scheme, under which funds would be provided by I.C.P. to investors on the basis of profit/loss-sharing. The existing investors' accounts involving interest should be converted to the new system if investors so agree; otherwise they may be tapered off in course of time.
Loan from Government of Pakistan: These comprise loans received from the Government of Pakistan during the period 1966—71. The outstanding amount of these loans as on 30th June, 1977 was Rs. 276.55 million or 64.6 per cent of the I.C.P.'s total borrowings. During 1976-77 interest payments on these loans by the I.CP. constituted about 38 per cent of its overall interest payments. Thus a substantial part of the interest element on I.C.P.'s expenditure side would be eliminated if the outstanding interest-bearing loans of the Government are converted into Participation Term Certificates. Under this arrangement the Government will share in the profit or loss of the I.C.P. on a basis to be mutually agreed between the Government and the I.C.P. Any future assistance by the Government to the I.CP. may also be on the same basis.
Loans from the State Bank: These loans are secured by the guarantee of the Government of Pakistan and carry interest at the bank rate. The outstanding amount of these loans as on 30th June, 1977 was Rs. 90 million and constituted 21 per cent of I.C.P.'s total borrowings. State Bank of Pakistan loans may also be converted into Participation Term Certificates. Any future assistance by the State Bank to the I.C.P. may be on the same basis.
Amounts Received in Respect of Debentures Issued: These represent the amounts received in respect of non-convertible Government guaranteed debentures issued by the Corporation. The debentures are redeemable at par in six annual instalments and bear interest at 2xh per cent per annum above the bank rate. The Corporation receives thereagainst a subsidy from the Government at the rate of 4 per cent per annum. The amount under this head stood at Rs. 61.8 million constituting 14.4 per cent of I.C.P.'s total borrowings. Interest payable on these debentures amounted to Rs. 10.5 million during 1976-77. These debentures may be converted into Participation Term Certificates to eliminate interest.
Investment in Government Securities: I.C.P.'s investments in Government Treasury Deposit Receipts, income tax bonds and compensation bonds are rather small and amounted to only Rs. 12.64 million or 2.8 per cent of its total investment as on 30th June, 1977. To eliminate interest income pertaining to this item, these securities may be sold to other financial institutions.
At present loans for construction of houses or pruchase of built- up houses are being provided by the House Building Finance Corporation and commercial banks to the general public and by the Federal and Provincial Governments, banks and certain other institutions to their own employees. All housing loans provided to the general public by House Building Finance Corporation and commercial banks are interest-bearing. Loans provided to employees by Government and other employing institutions also carry interest except in a few cases where loans up to certain maximum amounts are free of interest.
The Panel recommends that the present system of loaning by House Building Finance Corporation and commercial banks which is based on interest may be substituted by a system whereby these institutions may provide funds to prospective builders of houses or purchasers of houses on joint ownership basis with rent-sharing arrangements. The procedure that may be followed is outlined below:
The prospective builder of the house will submit an application to the financing institution for financing the construction or purchase of house on a joint ownership basis that would subsist until the entire amount provided by the financing institution is repaid. The application would provide all the relevant information such as monthly income of the applicant, the amount he would invest from his own resources, whether he already owns a plot or wishes to complete a house thereon which cannot be completed for lack of own funds etc. or wishes to pruchase a built-up house.
The financing institution, if satisfied with bonafides of the applicant and the feasibility of his plan, would enter into an agreement for provision of housing finance on joint ownership basis. The term of the agreement may be roughly as follows:
The respective financial contributions of either party would be specified. The value of land on which house is to be constructed and any construction costs already incurred will be taken into account in determining a party's financial contribution.
The period of joint ownership will be specified in the agreement.
The schedule of instalment payments, after a mutually agreed grace period covering the actual construction of the house, will be set up. With the payment of successive instalments the share of the financing institution in the ownership of property would go on declining and will finally cease on payment of the last instalment.
The initial rental value of the house will be determined at the time of agreement (with the help of expert appraisers, if necessary) on the basis of quality of construction, accommodation, location and prevalent rents in the locality. The rental value will be reviewed and refixed after every three years.
The share of the financing institution in the rent would be on a pro rata basis and the amount payable to the financing institution by way of rent will go on declining with the successive payment of instalments. The calculated share in rent will allow for the usual rate of depreciation and payment of taxes and property dues.
In case the builder/purchaser of the house wishes to sell the house before the expiry of the agreement, he would be free to do so and any capital gain/loss would be shared between the two parties proportionately according to the respective outstanding shares.
The builder/purchaser of the house would also be free to terminate the joint ownership agreement by paying off the outstanding amount of investment of the financing institution at any time during the period of agreements
The financing institution will have the right to terminate the agreement in case of fraud and to auction the property in the event of default in the payment of instalments and rent.
Apart from eliminating interest, the system suggested by the Panel above would provide substantial relief to prospective house builders/ purchasers. A person borrowing Rs. 1,00,000 for 20 years from House Building Finance Corporation has to pay at present a total of Rs. 1,64,000 by way of interest. An example worked out by the Panel on the basis of existing property tax structure in Karachi shows that under the proposed scheme if a person contributes Rs. 50,000 from his own resources and obtains Rs. 1,50,000 from House Building Finance Corporation and the initial rent fixed is Rs. 1,000 with a ten per cent escalation every three years, he will have to pay a total of only Rs. 71,000 by way of rent over a twenty year joint ownership agreement.
The Panel recognised the possibility of a notable rise in the demand for funds for housing following the introduction of the system proposed by it. The Panel, therefore, recommends that if such a situation develops and prices of construction materials and the wages of construction labour come under pressure, the financing institutions may be asked to appropriately reduce the maximum ceilings on their participations so as to protect the interests of the builders of small houses and bring pressure on builders of larger houses to use more of their own resources.
In the case of construction companies, funds would be provided by the financing institutions under a joint ownership agreement on profit- sharing basis.
As regards loans granted by the Federal and Provincial Governments, and other institutions to their employees, the Panel recommends that these should be provided free of any charge up to the maximum entitlements of the employees concerned and treated as a fringe benefit to them.
At present, the House Building Finance Corporation finances its loaning programme to a largd extent from borrowings from the State Bank of Pakistan against issue of debentures bearing an interest rate at 2 per cent below the bank rate. The Panel recommends that after the House Building Finance Corporation switches over to the new system, the State Bank may provide assistance to it on profit/loss-sharing basis for specific periods.
Source: Money and Banking in Islam, Ziauddin Ahmed; Munawar Iqabal; M. Fahim Khan. Republished with permission.


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Drew Paul
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Who is Drew?
Dr. Drew Paul, DD, CEA was born to an American father and a Palestinian mother. Noted technologist, artist and entrepreneur, Drew Paul currently resides in New York City where he operates several technology, marketing, and multimedia production based businesses.
Drew relocated to Florida, USA at an early age. Educated in the public school system, Drew Paul supplemented his education with autonomous extra-curricular studies throughout his early childhood including studies in art, history, music, science, astronomy, and medicine. It is rumored that Drew Paul began his first successful business at the age of 15. At the age of 17, Drew Paul graduated high school early and with honors.
He then studied International Business with a minor in Art and Design at St. Petersburg College and graduated with an Associate in Arts Degree in 2003. Afterwards, Drew attained several high profile internships with companies including Rainbow Art and Design, which led to a position as lead designer and IPP, inc. which led to positions in upper management. Drew Paul's education continued studying International Business at the University of South Florida in 2006, while operating the first of his several marketing companies.
After beginning his professional career, he pursued graduate program studies at the Massachusetts Institute of Technology. Drew Paul also trained through the University of California's Berkeley School of Music to attain his certification as Audio Engineer.
For a period of ten years whilst living in Miami, Florida, Drew Paul gained insight from an enlightening mentorship with Former US Deputy Assistant Secretary of State Dr. Steve Pieczenik, MD, PhD whose body of work includes acting as State Department official under Henry Kissinger, membership to the Council on Foreign Relations, over two dozen New York Times Bestsellers and membership to the Academy of Motion Picture Arts and Sciences.
Following in the footsteps of his grandfather, Drew Paul was ordained as a minister in 2016 and was granted his Doctorate in Divinity shortly thereafter based on his dissertational research.
Drew Paul currently holds the U.S. patent rights for four inventions in the fields of Alternative Energy, Marketing and Software Development. Drew Paul is the author of two books containing over seven scientific papers pertaining to the field of Applied Physics.
In 2008, Drew Paul is credited for the development of the first Website Builder of its kind.
Drew Paul was awarded the 2012 Citizen Life Saving Award by Tampa Police Chief, Jane Castor, the OSHA Excellence Award, and was recognized by the National Safety Council with the award of the Distinguished Service to Safety Award.
He was awarded the Dade County Young Entrepreneur Award in 2013 and was rated #1 Graphic Designer in Miami for two successive years in 2013 and 2014.
In 2015, Drew Paul founded the INUNI Organization, the Initiative for National United Nations Intervention.
Drew Paul was awarded as the Grand Prize Winner of the 2015 Make Energy Contest- International Challenge sponsored by Arizona State University, Apple Computers, Arduino, The Rep Rap Organization, Laboratorio para la Ciudad, New America, Green Momentum and Technologico de Monterrey for his contributions to the development of alternative energy solutions.
In 2016, Drew contributed to the Grand Prize winning submission to the 27th Annual Carnegie Institute of Science's Fundamental Physics Competition.
Drew Paul has worked as Audio Engineer and Producer alongside Grammy nominated and Rock & Roll Hall of Fame artists. He was the founder of The Studio 305 and Lead Audio Engineer at the historical Blue Note.
Drew Paul is the owner and founder of nine businesses ranging from marketing and media production to industrial science and alternative energy.
Blabor.com
The INUNI Organization
Drew Paul is a free thinker. Once described as a "free radical," Drew is determined to push the boundaries of what is possible. Guided by the belief that "All is One," Drew Paul is driven to contribute more to this world than he takes.
Drew keeps a hand in multiple public projects and a hand in several personal passions.
A revolution is upon us.
"The only difference between a scientist and a magician is that a scientist tells you how it's done."
-Drew Paul
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129 S. 8th St., Salina, KS 67401
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CM&L Now Offering Services in Lincoln, Kansas
Five Selected for Inclusion in Best Lawyers in America©
Four Selected for Inclusion in The Best Lawyers in America©
Defense Verdict Obtained in Surgery Case
Clark, Mize & Linville Named a Tier 1 Law Firm by U.S. News and Best Lawyers®
Defense Verdict Obtained in Bowel Perforation Case
Dustin J. Denning and Jacob E. Peterson successfully defended our client, an obstetrician/gynecologist (OBGYN), in a medical malpractice jury trial in which the plaintiff alleged that our client was negligent when performing a laparoscopic assisted vaginal hysterectomy ("LAVH") during which her small bowel was perforated through and through with a 5 mm optical trocar. The perforation occurred during the initial introduction of the optical trocar into the abdomen, but was not observed intraoperatively during the nearly 2-hour surgery. As compared to traditional trocars that involve the "blind" insertion of a Veress needle and bladed trocar, the plaintiff argued that the optical method is arguably a safer method of obtaining access into the abdominal cavity because the surgeon should be able to visualize each layer of tissue as the tip of the trocar is advanced into the abdominal cavity. The plaintiff alleged at trial that our client deviated from the standard of care in failing to recognize the bowel perforation at the time it occurred and failing to recognize it during the surgery prior to closing up plaintiff.
The plaintiff began showing signs consistent with a post-operative ileus the day after surgery (Post-Op Day 1), and her condition continued to worsen until she was returned to the operating room the morning of Post-Op Day 2 for exploratory surgery that revealed the through and through small bowel perforation. The plaintiff developed sepsis and other significant complications following the second surgery but recovered and was discharged from the hospital 35 days later. The plaintiff sought approximately $870,000 at trial.
We defended the case by showing that while our client preferred the optical port because he felt it was a safer alternative to the "blind" approach, the optical port is still not without risks, including injury to the bowel, and that such a complication occurred absent negligence on the part of our client. We demonstrated that the layers of tissue through which the optical trocar passes can be distorted for various reasons, including the patient's body habitus, and that failing to observe the perforation intraoperatively is not a deviation from the standard of care. Medical literature suggests that perforations go unrecognized at the time of surgery in 30-50% of all cases.
After a four-day jury trial, a Riley County, Kansas, jury agreed and returned a verdict in favor of our client after deliberating about 45 minutes.
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Threatened the University Hospital in Northern Norway
TOPICS:FinesNorthern NorwayNorway TodayUniversity Hospital
Tor Ingebrigtsen, CEO of UNN . Photo: UNN
Posted By: Pieter Wijnen 12. August 2017
The Health Authority threatened the University Hospital in Northern Norway with daily fines
The Norwegian Health Authority has on several occasions threatened to impose fines on the University Hospital in Northern Norway (UNN) of NOK 10,000 a day in order to get answers to inquiries.
– It is very rare that hospitals do not respond to our and other supervisory authorities' inquiries, says Director of the Norwegian Public Health Authority, Jan Fredrik Andresen, to Dagens Medisin (Today's Medicine).
The hospital shall have failed to respond despite reminders from the health inspectorate. On three occasions the Health Inspectorate has notified of coercive, meaning that the hospital must pay an amount daily, if they did not respond to inquiries.
– It has proved incredibly difficult to receive responses from UNN to inquiries we demand answers to, says Andresen.
Today's Medicine has been in contact with the CEO of University Hospital in Northern Norway, Tor Ingebrigtsen, who does not wish to comment on the matter at the moment.
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Mix With The Masters
Início/Educacional, Estúdios de Gravação/Mix With The Masters
Mix With The MastersDavid Molho2021-11-17T23:28:51+00:00
Mix with the Masters, founded by Grammy-nominated producer Maxime Le Guil and Victor Lévy-Lasne, is a global leader in music production education, curating an ongoing series of in-person and online seminars with award-winning producers and engineers on the art of record making for a global audience of aspiring hit-makers. When the Mix with the Masters team decided to build a new flagship location in the heart of Paris to meet all of their remote and in-person production needs, they retained the services of WSDG to design the ultimate teaching studio that would make their students and artist partners feel equally at home.
The Mix with the Masters Studio is located in a two story building in a residential area within the inner ring of Paris. The recording studio is located on the bottom floor and the top floors consist of a series of production suites and office space used by the Mix with the Masters staff for day to day needs. The WSDG design team was tasked with ensuring acoustic excellence across the complex, as well as ensuring technological and ergonomic flexibility between rooms to allow for multiple configurations of the space for both educational and recording needs.
The studio consists of two main rooms, the 48m2 Studio A which serves as the primary teaching area but can also be converted into a live room, and the 19.5m2 Studio B. Studio A is designed to seat up to 20 students, and its console is located on an elevator which allows it to disappear into the floor and easily convert the room into a live room where needed, with Studio B serving as the control room. Studio A is equipped with acoustic dampening curtains in order to provide maximum flexibility for its various uses and is also set up for Dolby ATMOS and video projection.
The studio also features a 15m2 Drum Room, side room, a pair of ISO Booths and a lounge. All the rooms have flexible connectivity to allow for multiple useful configurations. The lounge area of the studio was formerly a garden, and as such now features a glass ceiling and windows in order to preserve the aesthetics and feel of a calm, outdoor space.
Visit the Mix With The Masters Official Website
Read Resolution Magazine Article
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Read Recording Mag Article
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by Nicole Carbon on April 9, 2018
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The highly-anticipated opening of Downtown Sarasota's Art Ovation Hotel is about to make its debut and paint the town red. They are partnering with the Ringling College of Art + Design to celebrate and embrace the vibrant arts and culture scene Sarasota is built upon and thrives on. This new addition to downtown will host a private opening of its inaugural exhibit on Thursday, April 12, "LEGACY: An Exhibition by Ringling College of Art and Design Faculty and Alumni." The works will be on display through June 27.
During the event, those in attendance can experience all aspects of the hotel including the lobby's Gallery Lounge and the Perspective Rooftop Pool Bar to appreciate collections by contemporary artists from the Ringling College of Art + Design.
At the opening, works on display will be by artists who include Jason Aponte, Joe Fig, Thomas Carabasi, Cynthia Mason, Jill Taffet, and Jeff Schwartz.
Proceeds from the sales at the opening go directly to the College's scholarship fund.
The downtown Sarasota hotel has been designed to "fully immerse guests in the creative and artistic experience." It will offer an Artists-in-Residence program and will engage its guests in the area's abundant events centered around its cultural scene. It will continue this exclusive collaboration with the Ringling College with opportunities for students to enrich their education and experience and guests will gain access to the College's lectures, openings, and opportunities to join in special events and galas.
Even at the close of season, it's an exciting time for the city and especially the year-round residents who have an appreciation for art and design. This isn't something being done for tourists and seasonal residents only, but rather for the city as a whole and an enriching offering to visiting guests.
Most importantly, this is an initiative to recognize and support what the students of the College are doing to enhance their education and grow their artistic talents. For more information visit, https://artovationhotel.com/.
Nicole Carbon
Nicole Carbon is the Editor-In-Chief of VODA Magazine. She is based in beautiful Sarasota, FL. For over a decade, she has been writing for both print and online media outlets about food, wine, travel, and other lifestyle-related finds. She received her Level 2 Award in Wines certification from the Wine and Spirits Education Trust (WSET) and writes a monthly wine column in the Venice Gondolier Sun. She contributes to Edible Sarasota, writes a health and wellness blog, and various feature articles and content for other media outlets. Previously, she wrote two magazine columns titled "A Girl Walks Into a Bar," and "Inside the Kitchen." Her work has appeared in the Austin American Statesman, Austin MD, Austin Monthly, Austin Woman, Austin Man, Citysearch Austin and Los Angeles, CultureMap.com, JetBlue's blog, SRQ Magazine, and the list goes on. She has also made TV appearances on local NBC and Fox news affiliates as well as Cooking Channel's Eat Street.
#art exhibit art #art opening #Art Ovation Hotel #artists #downtown hotel #downtown Sarasota #Sarasota
by Nicole Carbon
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text:
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Let's amplify unspeakable things
Amplification of female voices
(workshops)
This is a documentation of a series of meetings/workshops I am organising together with Christina Karagianni. Our intentions come from personal experiences and we seek to explore the silencing of excluded voices, and go beyond their barriers. We combine our practices -her practice lies on choreography and dance and mine on social interaction, voice and sound- in moments of Leeszaal, an open local library, a public space of diversity. How can we engage politically with the exclusion of specific voices from the public sphere? We are borrowing methods from feminist groups and protest movements and vocal warming up exercises by Pauline Oliveros. For example, protesters would amplify the speaker's voice by repeating collectively their speech to make their presence visible. Feminists would create safe spaces where women could speak about domestic violence and make a dialogue based on listening.
during the workshops we've realised that we should reconsider the terms female/male and how to refer to those voices when inviting people
people from Leeszaal and our environment, who find themselves related and interested to this topic. The produced material is being included in an audio archive,
player.
this is how female/high-pitched voices have been described since acient times as Anne Carson observes
"Blocking our Voices"
(podcast)
The podcast "Blocking our Voices" includes a series of episodes about the ways we learn to self-control and sensor our speech. All the podcasts are made from parts of the "player", an online archive, with my voice as a narrator. The voices and sounds, you will listen to, come from workshops I have co-organised, soundwalks and media sources from a body of research.
"Voice's Transition"
The podcast "Voice's Transition" includes a series of episodes about the electronical transformation of one's voice into another voice in terms of gender. All the podcasts are made from parts of the "player", an online archive, with my voice as a narrator. The voices and sounds, you will listen to, come from workshops I have co-organised, soundwalks and media sources from a body of research.
"Mediating Speech"
The podcast "Mediating Speech" includes a series of episodes about the amplification of one's speech through the voice of others. All the podcasts are made from parts of the "player", an online archive, with my voice as a narrator. The voices and sounds, you will listen to, come from workshops I have co-organised, soundwalks and media sources from a body of research.
(audio archive)
The player is an audio archive of voices and sounds from workshops I have co-organised, soundwalks and media sources from a body of research. It is a digital utterance that intends to be a reminder to the first forms of excluded female vocal expressions. I am using this material in my podcasts by mixing and overlayering them and creating a form of narrative with my own voice.
This work has been produced in the context of the graduation research of Angeliki Diakrousi from the Experimental Publishing (
XPUB
XPUB is a two year Master of Arts in Fine Art and Design that focuses on the intents, means and consequences of making things public and creating publics in the age of post-digital networks
) Master course at the Piet Zwart Institute, Willem de Kooning Academy, Rotterdam University of Applied Sciences.
https://xpub.nl
Special thanks to Christina Karagianni for organising together these inspiring workshops, Angelique, Fatima, Lidia, Dionne and other women from
Wereldvrouwen
Wereldvrouwen Rotterdam Foundation does various social, educational and recreational activities for and by women from various cultures, many of them with a low income and low education level. One of their meeting points is at Leeszaal
for participating in the workshops, and all the other participants who came in the beginning, but also the online participants who generously contributed with their voices to the website. More thanks to my tutors, XPUB comrades and all my friends with whom I had interesting discussions and tryouts that pushed my research further.
The last 6 workshops are part of
Women@Art
(free translation) Wereldvrouwen Rotterdam Foundation does various social, educational and recreational activities for and by women from various cultures, many of them with a low income and low education level. Most women hardly ever come into contact with the world of Art & Culture. Participants are encouraged to develop with inspiring examples from the art world by visiting exhibitions / studios in the city. A number of female (amateur) artists give workshops on their own work. The aim is to make the world of art more accessible, to help women to develop their own creativity and to transform it into strength
and funded by the
Municipality of Rotterdam
The municipality granted to us a structural subsidy called Pitcher Perfect 010
The studies of Angeliki Diakrousi were funded through a scholarship by the Onassis Foundation
Free Art Licence
[Angeliki Diakrousi, Let's Amplify Unspeakable Things, 2019. Rotterdam].
Copyleft: This is a free work, you can copy, distribute, and modify it under the terms of the Free Art License http://artlibre.org/licence/lal/en/
Source code of the website
'Let's Amplify Unspeakable Things' is an online platform of exploration and research, regarding
The term 'female voices' refers as well not only to the feminine, queer, non-binary voices but also to the primitive and non-rational ways of expression, that the contemporary democracy tends to exclude
in public and their amplification, developed by Angeliki Diakrousi. It creates a space for excluded voices to be explored (online and physical spaces), that questions the establishment of authoritative voices — which construct exclusive speech platforms and binaries, along the assumption that voices have to be rational in public dialogues. It includes a set of
Different frictions emerged, between the technical and non technical
that revisit and overlay an archive of audio recordings, produced in situated
and soundwalks, and media sources from a body of research. 'Let's Amplify Unspeakable Things' is based on the concept that amplification can provide presence through repetition and multiplication, as the structure of the workshops and the website reveals. The project itself is a process of feedback and iteration of actions, throughout a period of four months, situated around the space of Leeszaal and hub.xpub.nl. It exists in between the friction of institutions (a public library and an academy) and different public spheres (wereldvrouwen, academic), and it aims to go beyond their binaries.
"Let's Talk About Unspeakable Things"
This is a thesis of three essays which relate to female and collective voices, and their mediation. The essay address the voice as a feminist tool for communication; studying how the female voice creates conditions for forms of listening and making space. Historically, some modes of address have been marginalized and shut out of the public domain. Collective voices are marginalized under the realm of patriarchal individualistic society. Even though voice is a medium for collective practice it is situated in a context that tends towards social binary structures and oppositions that restrict its possibilities. These binaries have held influence on Western thinking since antiquity. Nevertheless, the nature of voice and its mediation overpass oppositions of gender, nationality, culture, space, technology and power relations. My research seeks to unravel these political capabilities of voices, in order to explore democratic ways of communication that embrace excluded forms of address.
Diary of West Rotterdam
(first experiments)
This is a diary from my first experiments. It includes mostly material from my walks. I recorded sounds from the West Rotterdam,
while walking around the neighborhood of
Leeszaal for several days. This action was part of my first experiments when I was in attempt to understand how gender binaries regarding voice are reflected in space, especially public space.


Try Me is a studio album by Rosie Gaines originally planned for release in 1994, but eventually released digitally in 2005; many of the tracks had since been worked on further and released on Closer Than Close, released in 1995. Try Me features one song with writing and musical input by Prince - In The Socket (previously released by the NPG Music Club in a remixed form as Hit U In The Socket).
Work on a solo Rosie Gaines album began while she was still a member of the New Power Generation band, in 1990 (I Want U was first recorded in late 1989, before work on the album commenced). Work continued until September 1991, when she and Prince fell out regarding her solo career. She was contracted to Paisley Park Records, however, and continued to work on the album without Prince following her departure from the band (after the Diamonds And Pearls Tour). The album became known as Concrete Jungle.
After Paisley Park Records closed, Rosie Gaines worked on the album further, now renamed Try Me. The album still remained unreleased, however, until she worked on it further, renamed it Closer Than Close and released it on Motown Records (see that album's entry for full details of its release).
In The Socket, as released on Concrete Jungle, is the original 1991 recording, which was later remixed for its release in 2001 through the NPG Music Club.
No singles were released to promote the album, and the album did not enter any charts, receiving a release only through RosieGaines.com.
No physical CD is known to exist, therefore the tracks below are listed from the RosieGaines.com download version.
1 Written by Rosie Gaines and Prince.
No other production credits available.
This page was last modified on 28 December 2018, at 07:04.


text:
class ProcessGallery < Struct.new( :gallery_id )

    def perform
      @gallery = Gallery.find(gallery_id)
      @zipfiles = @gallery.assets.where('asset_file_name like ?', '%zip').all

      @zipfiles.each do |zip|
        asset_filename = zip.asset.path;
        `(test -d tmp/gallery/#{@gallery.id} || mkdir -p tmp/gallery/#{@gallery.id}); unzip -j #{asset_filename} -d tmp/gallery/#{@gallery.id}/`

        Dir.glob(File.join("tmp/gallery/#{@gallery.id}/",'*')).each do |picture_path|
          if File.basename(picture_path)[0]!= '.' and !File.directory? picture_path
            asset = @gallery.assets.where(:asset_file_name => File.basename(picture_path)).first
            if !asset
              asset = @gallery.assets.create(:title=>File.basename(picture_path),:description=>'Extracted from'+asset_filename)
            end
            asset.asset = File.new(picture_path)
            asset.save
          end
        end
        @gallery.save
        `rm -rf tmp/gallery/#{@gallery.id}`
      end
    end

end

redpajama_set_name: RedPajamaGithub


ENEOS 5W-30 - Upgraded Fully Synthetic Formulation!
• ENEOS Motor Oil 5W30 meets the latest API SN standard for gasoline engine oils.
• ENEOS Motor Oil 5W30's excellent engine cleanliness and wear resistance have been proven in official laboratory tests in the United States. This performance has also been confirmed in a fill range of vehicle tests.
• The consumption of motor oils depends largely on the type of base oil. Because ENEOS Motor Oil 5w30 is blended from high-performance oils, it has little oil consumption, so users can look forward to adding less oil to their engines.


Works well on ceramic stove tops, electric stoves and gas stoves.
Stainless steel contains some amount of Nickel (Metallic).
which is known to the State of California to cause cancer.
ManPans believes in the quality of it's products and offers a 30 day satisfaction guarantee*. If this product does not perform to your satisfaction then it can be returned for a refund within 30 days from purchase.
* Under normal home use and following care & use instructions. This excludes damage from misuse or abuse. Minor imperfections and slight color variations are part of normal use.
ManPans warrants all products against defects in material or workmanship from date of purchase for a period of 5 years. We will repair or replace defective products, but will not be responsible for any consequential damages including any claim of lost profits.
Metal utensil safe means that metal whisks, spatulas and other non-sharp metal utensils may be used without damage. In fact, we recommend a metal spatula when using our fry pan and saute pans. Do not use sharp instruments or appliances such as electric mixers to cut or chop against the pan as this may damage the coating and will not be covered under this warranty. Immersion blenders can be used with bottom safety guards. Food sticking to the pan is not a covered by the Warranty.
Copyright 2019 LloydPans Kitchenware. All Rights Reserved.


Predictive analytics first gained massive attention in the 2011 baseball film "Moneyball," adjusted from the popular business book "Moneyball: The Art of Winning an Unfair Game." Both film and book inform the story of how Oakland Sports manager Billy Beane changed the whole professional sports industry by using mathematics and stats to pick gamers and create strategies. The transformation of present analytics before predictive analytics extends well beyond sports into applications as differed as Netflix's film suggestions and Google's self-driving cars and trucks. In its first rule of leadership, it is now also about reinventing the software testing market by helping services benefit from DevOps continuous deployment pipelines to minimize the expense, time and danger of software delivery.
Making use of predictive analytics helps companies comprehend ways to benefit from customer-facing feedback, in addition to ways to forecast and avoid flaw patterns in future software application releases. Such mobile data capture in a business management system can help managers determine what problems need to be prioritized and how they can improve their services.
Shift left testing, screening previously in the software application advancement lifecycle while an application is being developed.
Shift right testing, screening in production after an application is launched.
The two screening methods aren't equally special. Depending upon the risk-level your company wants to shoulder, moving right and testing in production might be the fastest and most efficient method to record end-user feedback on a specific application. If the released software application has problems, shift left testing permits you to release your bug repairs that much quicker.
In DevOps shift left testing, screening is carried out earlier in the software application advancement procedure, with the objective of increasing quality, reducing long test cycles and lowering the possibility of software application flaws when developing their production code. This is frequently done by moving combination screening to the left of its normal position in the lifecycle so that it happens as close as possible to the development procedure. Considering that combination testing is where lots of disruptive, substantial flaws are typically found, this enables groups to obtain feedback on code quality much faster, which is even quicker when the feedback is inputted into a cloud management platform or system, with more precise outcomes.
Constant testing is a huge part of shift left testing, which includes automating manual tests and running those tests as early and as typically as possible, sometimes utilizing virtualized elements and environments. Agile automated testing is necessary in a DevOps pipeline because manual screening is a lengthy and labor-intensive procedure. Predictive analytics help groups figure out which of the hundreds and even countless tests that have to be run manually versus an application in the pipeline that can be automated.
In shift right testing, the quality level of an application in production is constantly kept an eye on and customized as required. Adjustments are done proactively instead of reactively, based upon a predictive analysis of production information sets, that include application efficiency, user interaction information, consumer feedback, resource use and other functional metrics. Analytics is likewise beneficial here in picking and automating proper test cases, that is, the conditions or variables that identify whether an application being evaluated satisfies requirements or works properly.
Predictive analytics is the practice of drawing out beneficial info from information sets using analytical algorithms and artificial intelligence in order to anticipate patterns and habits. When applied to software application screening, predictive analytics makes it much easier to recognize exactly what to evaluate and what quality problems to anticipate prior to and after they take place in production. This is especially helpful because of the brief advancement cycles of modern-day DevOps tasks. Predictive analysis and constant feedback from end-users make it much easier for QA groups to anticipate the danger levels of various applications in their DevOps pipeline.
Identifying the danger level of various applications is essential on DevOps tasks considering that nimble software application groups might effectively choose to take more risk on specifically released applications in order to beat their competitors to market or to obtain end-user feedback to confirm a hypothesis. In risk-based software application screening utilizing predictive analytics, concern for repairing flaws is based upon the threat capacity of the flaws. This is because, on hectic nimble jobs, bug repairs for low intensity bugs will get low priority and are normally just arranged when time is free.
There are 2 primary factors to consider in this kind of risk-based software application screening: the likelihood of the software application problem taking place and the effect of the problem when it happens. High effect or potential bug repairs should be set up first. For instance, a bug in a specific module of code for an online shopping cart algorithm that keeps a service from processing deals need to be arranged for repair first. On the other hand, a bug that presents a really small rounding mistake in that same transaction is a lower priority.
Shift right testing includes more than simply bug repairs, nevertheless. Predictive analytics permits nimble groups to engage with end-users far more proactively, such as in a case of the shopping cart abandonment, when consumers put products in their online shopping carts, but then leave in the before finishing the purchase. In a case like this, the nimble group and business leaders can use analytics and information mining methods to shine info from large transaction datasets in order to increase conversions, potentially by either simplifying the checkout procedure or by retargeting buyers with e-mails after they have actually left a site.


text:
Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA
Aline S Ferraz1,
Elza FT Belo1,
Ligia MCC Coutinho1,
Ana P Oliveira1,
Andréia MS Carmo,
Daniele L Franco1,
Tatiane Ferreira1,
André Y Yto1,
Marta SF Machado1,
Monica CG Scola2 &
Elizabeth De Gaspari1
BMC Infectious Diseases volume 8, Article number: 30 (2008) Cite this article
10k Accesses
A simple filter paper method was developed for, the transport and storage of monoclonal antibodies (Mabs) at room temperature or -20°C after spotting on filter paper, for subsequent serotyping of outer membrane antigens of N.meningitidis by dot-blot ELISA.
Monoclonal antibodies (Mabs) were spotted within a 0.5–1 cm diameter area of Whatman grade 903 paper, which were stored individually at room temperature or at -20°C. These MAbs were stored and analyzed after periods of one week, 4 weeks, 12 months, or 13 years in the case of frozen Mab aliquots, or after 4 weeks at -20°C or at room temperature (RT) in the case of Mabs dried on filter paper strips. Assays were performed in parallel using dot-blot ELISA. In addition to the MAbs specific for serotyping class 1, 2 or 3, we used a larger number of Mabs for polysaccharides, lipooligosaccharides (LOS), class 5 and cross-reactive antigens for native outer membrane of N.meningitidis. The Mabs dried on filter paper were eluted with phosphate-buffered saline (PBS) containing 0.2% gelatin.
Mabs of the isotypes IgG and IgM dried on filter papers were not affected by duration of storage. The detection by serotyping Mabs was generally consistent for dried filter paper MAb samples stored frozen for over 1 year at -20°C, and although decreased reactive antibody titers were found after storage, this did not interfere with the specificity of the Mabs used after 13 years as dry spots on filter paper.
The use of filter paper is an inexpensive and convenient method for collecting, storing, and transporting Mab samples for serotyping studies. In addition, the samples occupy little space and can be readily transported without freezing. The efficiency of using immunoglobulin G (IgG) or M (IgM) eluted was found to be consistent with measurement of IgG or IgM titers in most corresponding, ascites Mabs stored frozen for over 1 year. The application of meningococcal typing methods and designations depend on the question being asked.
Meningococcal disease (MD) is a significant cause of mortality and morbidity throughout the world [1, 2]. The incidence of MD in Brazil has been monitored since the occurrence of serogroup A and C epidemics between 1971 and 1974. In 1974, the incidence was greater than 179 cases per 100,000 inhabitants. From 1980 to 1992, the annual incidence of MD ranged from 1.0 to 1.4 per 100,000 inhabitants in different states of Brazil. During the period between 1981 and 1987, the mean proportion of serogroup B isolates identified was about 83%, while serogroup C strains represented only 6% of isolates. In 1988, the incidence of MD in the greater Sao Paulo area exceeded 4.06 per 100,000 inhabitants, suggesting a new epidemic in that region. This epidemic differed from previous ones because it was caused by serogroup B strains in 1988 and 1989 and serogroup B and C strains in 1990. The incidence of MD caused by Neisseria meningitidis serogroup C in greater São Paulo has been low since the end of the epidemic situation in 1971 and 1972. In that region, the prevalence of serogroup C strains increased from 4 to 14% and 8 to 32% during the years 1989 and 1990, respectively. Serotype 2b isolates were responsible for most of this increase, representing approximately 22 and 74% of the serogroup C strains isolated in 1989 and 1990, respectively [3, 4]. In greater São Paulo, there has been a constant increase in the incidence of serogroup C meningococcal disease since the late 1980s [3, 4].
The current serotyping system for meningococci is based on a battery of Mabs [5, 6] which recognize antigenic differences in the outer membrane proteins of class 2 or 3 and 1, respectively [7]. The monoclonal antibody (Mab)-based typing system was developed because of the difficulties encountered with the use of absorbed hyperimmune polyclonal sera for typing.
After realizing the need for sensitive subtyping methods almost 20 years ago, an ambitious project to develop a Mab-based subtyping system was undertaken by researchers at The Netherlands National Institute of Public Health and Environmental Protection and by others. A panel of Mabs for serotyping and serosubtyping is now available at the website of (University of Oxford, UK). Before that dream was realized, an international interlaboratory comparisonof these reagents with 85 geographically and temporally diverse isolates of N.meningitidis serogroup B was was carried out in 1992 [8]. One of the problems with the Mab-based serotyping and subserotyping methods reported in that study was that a large proportion of isolates were nontypeable [8].
We described several years ago a simple method for the collection, preservation, shipment, and testing of minute amounts of dried monoclonal antibodies for typing N. meningitidis B [9]. The Mabs collected on filter paper were extracted with PBS and evaluated by dot-blot and immunoblot analysis employing whole cells of N. meningitidis B as antigen. The dried filter paper with Mabs could be stored at room temperature for as long as 30 days without detectable changes in antibody response when used for typing outer membrane antigens of N. meningitidis B in 1994 [9]. At that time, we used ascites and culture supernatant for two monoclonal antibodies of IgG isotypes specific for class 5 of N meningitidis B by dot-ELISA and immunoblot, and subsequently, we performed a better characterization of the monoclonal [10]. We did not analyze Mabs of IgM and IgG isotypes for other outer membrane antigens of N.meningitidis, nor was there a study of long-term cold storage of filter papers with dried Mabs.
This paper describes a simple filter paper procedure for collecting monoclonal antibodies of IgG and IgM isotypes on Whatman grade 903 paper, which are then easily air-dried and stored at room temperature, or at -20°C for serotyping for N.meningitidis. The papers can then be easily transported from the clinic or field to the laboratory. A similar procedure has been widely used for collecting spots of dried blood or saliva to screen for various infectious agents [11–21], and metabolic and genetic diseases, for the presence of specific genes, also for forensic purposes. For several years, blood spot specimens on blotting paper have been frequently used for the diagnosis and seroepidemiologic investigation of infectious diseases. This method has been applied to the diagnosis and seroepidemiologic survey of bacterial, viral and parasitic diseases [22, 23].
We also examined 135 N. meningitidis strains of serogroup A, 66 strains of serogroup C (from the 1972–1974 epidemics) from the culture collection at the Adolfo Lutz Institute, and 122 strains of serogroup B (from 1992) obtained from the reference center of N. meningitidis in Brazil. The National Reference Center for Meningitis (Adolfo Lutz Institute) serotypes all the Brazilian isolates of N.meningitidis. In our study, we also used reference strains for serogroups, serotypes, subtypes and immunotypes of N.meningitidis with a larger number of Mabs. The method described here permits the sending of Mabs to other Central Public Health Laboratory for serotying local isolates, thereby improving the epidemiologic monitoring of meningococcal infection.
We analyzed 135 N. meningitidis strains of serogroup A, 66 strains of serogroup C (from the 1972–1974 epidemics) from the culture collection at the Adolfo Lutz Institute, and 122 strains of serogroup B (from 1992) obtained from the reference center of N. meningitidis in Brazil, Adolfo Lutz Institute. In our study, we also used reference strains of N meningitidis in which cells were grown in (trypticase soy broth, Difco BRL products, Gaithersburg, MD) supplemented with 1% horse serum (Sigma, St.Louis, MO) in plates in a 5% CO2 atmosphere at 37°C [9]. The immunotype reference strains used were: 126E (L1), 35E (L2), 6275(L3), 89I(L4), 981(L5), M992 (L6),6155 (L7), M978 (L8), 120M (L9), 7880 (L10), 7889 (L11) and 7897 (L12).
The Mabs normally used for N.meningitidis typing belong to cell banks. Most of them are described at the website of (University of Oxford, UK). When new monoclonal antibodies are obtained, they must be compared with the reference strains [10] to determine whether they are new or have specificity comparable to that of existing antibodies. Tables 1, 2, 3, 4 describe the analysis of the Mabs studied here with the reference and case strains of N.meningitidis.
Table 1 Dot-blot ELISA using Monoclonal antibodies of serogroups for N. meningitidis.
Table 2 Dot-blot ELISA of serotypes and subtypes using Monoclonal antibodies for N. meningitidis.
Table 3 Dot- blot ELISA of class 5 using Monoclonal antibodies for N. meningitidis.
Table 4 Dot- blot ELISA of immunotypes using Monoclonal antibodies for N. meningitidis.
Monoclonal spot test
For the spot test, monoclonal antibodies for N meningitides, in the form of ascites or culture supernatant (10 μL), were applied on a 0.5–1 cm diameter area of Whatman grade 903 paper. The Mabs were dried for 18 h at RT before laboratory testing.
Collection, transport and storage
All paper strips with Mabs prepared were put into individual self-sealing (ziplock) polythene bags with a few grains of silica to keep out moisture. The Mabs were decanted into polypropylene vials and stored in a freezer at -20°C. The samples were collected once a month by the research staff. Half of these bags were stored at room temperature (25–30°C) and the rest at -20°C until analysis.
Storage of dried MAb spots on filter paper
Specimens were stored under two conditions. The first was in a freezer at -20°C, and the second was a non-air-conditioned room in the laboratory at a temperature ranging from 15 to 25°C. All Mabs were tested at 4 weeks. Mabs from ascites or culture supernatant, and dried Mab spots on filter paper were determined in parallel. Tables 1, 2, 3, 4 show the comparison between filter paper Mabs of different isotypes of IgG and IgM analyzed by dot-blot ELISA of filter paper eluate at different times.
Reconstitution of sample from filter papers
The disks containing 10 μL of ascites or culture supernatant were punched out of the filter paper [9]. The paper disks were soaked in 200 μL of phosphate-buffered saline (PBS) supplemented with 0.2% gelatin for 18 h at 4°C on a platform shaker (New Brunswick Scientific Model 2R) at 75 oscillations per min. Eluates contained the equivalent of a 1/20 dilution. The following day the supernatant was stirred, collected and analyzed, together with the corresponding Mab pairs stored frozen at -20°C. Each Mab dilution was previously determined using dot-blot ELISA for N.meningitidis reference strain [10].
Dot-blot ELISA
The method used for serotyping and serosubtyping was essentially that of Abdillahi and Poolman [6]. For preparation of samples (whole cells) for dot-blot, cells were suspended in PBS, pH 7.7, containing 0.02% sodium azide. The cells were heat-inactivated at 56°C for 30 min and the absorbance of the suspension was adjusted to 0.1 at 650 nm using a spectrophometer (model Spectronic 88). The bacterial cell suspension was stored at 4°C. For dot-blot ELISA, 1 μL of meningococcal cell suspension was spotted on 0.22-μm nitrocellulose strips (BioRad). After drying, the strips were incubated for 1 h in blotting buffer containing 5% gelatin (Sigma Chemical Co., St. Louis, Mo.), in PBS. Mabs (Table 1, 2, 3, 4) were pipetted directly into the blocking buffer, diluted as previously determined and incubated continuously overnight at 4°C. The strips were washed separately with PBS six times. Antibody binding was detected after 2 h incubation with a 1:2500 IgG or 1:5000 IgM of rabbit anti-mouse immunoglobulin conjugated to peroxidase (Kirkegaard & Perry Laboratories, Inc., Gaithersburg, MD.). The reaction was visualized by adding a freshly prepared solution containing H2O2 (30%) and as substrate 0.04% of 3-amino-9-ethyl carbazole(AEC) (Pierce).
The results of the serological tests performed with ascites or culture supernatant samples and dried Mab spots on paper kept at -20°C or at room temperature are presented in Tables 1, 2, 3, 4. The titers were mostly equal and the differences did not alter the final results when the dried Mab spots were stored at -20°C or at room temperature for a period of one year. After 13 years storage, the titers of IgG and IgM antibody isotypes did not interfere with the specificity (Figure 1) of the antibodies when analyzed by dot-blot ELISA. However, none of the differences resulted in a change in serotyping with epidemic strains when we used Mabs of serogroups A, B or C eluted after one year at -20°C (Figure 1). We also included in assays control culture supernatant lacking antibodies to N.meningitidis to look for false positives. In all cases where Mab samples were antibody-negative, the corresponding Mab samples dried on dot-Blot ELISA paper tested negative after one week, 4 weeks, 1 year or 13 years of storage at -20°C. Most of the Mabs used in the present investigation are included at the website of, (University of Oxford, UK) and were produced by Dr.W.D Zollinger and Dr. J.T Poolman. Comparison between filter paper Mab and freezer Mab can be seen in Tables 1, 2, 3, 4. The application of these MAbs for serogroup identification of meningococci was demonstrated by their abilities to correctly identify 323 clinical isolates in Brazil by dot blot-ELISA (Figure 1). Serogrouping of N.meningitidis (or meningococci) is important because the disease caused by some serogroups can be prevented by active immunization [24].
Dot- blot ELISA using monoclonal antibodies for serogroups A, B and C eluted from filter paper stored for one year at -20°C. Dot-blot ELISA results of analysis of 323 case strains of N meningitidis . The strains was dotted (1 μL) as whole cells suspension.A) Reactivity of the Mab WRAIR 2D7B5B5B2 at dilution 1:25,000(serogroup A); B) Reactivity of the Mab WRAIR 5C1-3H7 at dilution 1:50,000 (serogroup B) and C) Reactivity of the Mab WRAIR 7H94 at dilution 1:25,000 (serogroup C).
We also analyzed a series of murine monoclonal antibodies to serotypes and subtypes of N meningitidis that were specific for outer membrane proteins of classes 1, 2 or 3. In our laboratory, we prepared and used a Mab to P1.9 strains and others [9, 10, 25] (Table 2).
The use of monoclonal antibodies for serotyping of meningococci is feasible and easy to perform, and it appears to have significant advantages over the use of polyclonal typing sera [7, 26–29]. The analyses using several class 5 Mabs to opacity proteins (Opa), a family of antigenically variable outer membrane proteins of N. meningitidis [30], also did not show variability according to dot-blot ELISA after 1 year as dried spots on filter paper (Table 3)
Monoclonal antibodies (Mabs) have largely replaced rabbit antisera as the LOS typing reagent, but a set of Mabs that recognizes most of the important LOS antigens is not available, and many of the available Mabs lack a complete structural definition of their cognate epitopes [31–33]. The pathogenic Neisseria, N.gonorrhoeae and N.meningitidis, possess an outer membrane protein(OMP), designated H.8, with a conserved Mab binding epitope [34], which was also shown be stable in our studies using a new Mab that was produced earlier [35]. Other cross reactive Mabs also remained stable for one year on dry filter paper at -20°C (data not shown).
It is known that the titer of the Mabs used in dot-blot ELISA for serotyping depends on several factors, mainly on the quality of the anti-IgG or IgM employed as well as the choice of substrate. In our study, we used AEC, which shows a high sensitivity. Before choosing the titer of the Mabs to be used, we carried out a study with different dilutions using reference strains [10] (data not shown).
Another factor examined in this study was the type of filter paper utilized. In an earlier study, we used Whatman 1 MM and observed that the type of filter paper used did not result in an appreciable change in Mab elution [9]. However, in the present study, we used Whatman grade 903 paper.
Typing of N. meningitidis includes the determination of groups, serotypes, and subtypes by either whole-cell ELISA or dot-blot ELISA using Mabs. The serotypes, subtypes, LPS, class 5 and cross-reactive specificity of Mabs are defined by their reactivity with whole cells or native outer membrane proteins of the strains by immunoblot analysis.
Despite its limitations, the Mab-based serotyping and serosubtyping method is attractive because it is easy to perform, is relatively inexpensive, and does not require sophisticated equipment. This method is ideal for use in developing countries. The preparation and characterization of additional MAbs and further standardization of the method will increase the utility of this method.
In an ideal world, strains could be characterized using a wide selection of the available typing techniques to create the fullest picture possible of the infective organisms. In practice, this is rarely necessary, and time and financial constraints limit the choices available; therefore, one needs to consider which procedure is the most suitable for a specific situation [36, 37].
Thus this method, which is a simple alternative to complicated conventional methods of collection, transport and storage of biological specimens, may be applied to monoclonal antibodies for a host of pathogens. The method is being used routinely in this laboratory for diagnostic and screening purposes with regard to epidemic strains.
Public health specialists are especially interested in the early detection of outbreaks. Filter paper has been used to collect blood for public health purposes for more than 40 years. The paper is made from high-purity cotton linters and is manufactured to give accurate and reproducible absorption of blood specimens according to specifications of the National Committee for Clinical Laboratory Standards (NCCLS) [38]. In this study we used Mabs filter papers with different specifivities that were stored in a freezer for a long period of time. This may have an advantage because we have a large quantity of IgG or IgM antibody for one epitope. In addition, it simulates the reality of using filter papers, usually under conditions where electrical power is not available. We paid special attention to keeping the filter papers dry at room temperature and found no significant reduction in antibody titer after one month, in order to simulate the transport. The detection of IgG or IgM Mabs eluted from filter papers is a useful tool for epidemiologists [39–43].
In summary, MAb samples dried on filter paper and stored for periods of time at -20°C, and later eluted, give results concordant with those of corresponding to Mab samples tested by dot-ELISA. Here, we describe the use of a large number of Mabs of different isotypes and different specificities for antigens of N. meningitids. Our laboratory has been working for more than 10 years on the development of new monoclonal antibodies against this pathogen [9, 10, 25, 35, 44] and also with others pathogens [45–50]. When we need to send our monoclonals to confirm their utility, this becomes an efficient approach. However, here most of the Mabs used for serotyping this pathogen were used in the characterization of a new monoclonal antibody.
This study has created a mechanism for future validation of filter paper handling and storage of monoclonal antibodies.
The determination of the most appropriate screening methods and subtype analysis depends on the immediate requirements of an investigation. A panel of well-characterized subtype-specific MAbs proposed for use in screening a large number of isolates during an outbreak should be selected to identify the majority of strains. This method of epidemiologic screening has proven effective in assessing outbreaks during the last 17 years.
The agreement between filter paper eluates and Mabs of IgG or IgM isotypes was substantially high. The agreement was higher with frozen Mabs and the corresponding filter paper Mab eluates. Therefore, in outbreak situations as well as in routine surveillance of this disease, the results of serogroup determination may lead to decisions on the public health intervention measures to be taken. Serogroup determination may also help in understanding the changing epidemiology of meningococcal disease. Murine hybridoma monoclonal antibodies (MAbs) were produced against the capsular antigens of serogroups B, C, Y, W135, 29E, Y and Z meningococci. Each serogroup-specific MAb reacted with the extracted capsular polysaccharide from its homologous serogroup only and did not react with capsules from the three other serogroups.
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Pre-publication history
The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2334/8/30/prepub
We gratefully acknowledge Dr. W.D Zollinger and Dr. J.T Poolman of the Walter Reed Army Institute of Research, Washington, D.C. and NVI (Netherlands Vaccine Institute; previously the RIVM), respectively, for providing the most of the monoclonal antibodies used in the present investigation. Some of the monoclonals antibodies used in the present investigation are described at the website of (University of Oxford, UK). Other Mabs used in the present investigation were produced in the Immunology Section by E.N. De Gaspari. This research was supported by grants from FAPESP, São Paulo, Brazil, awarded to E.N. De Gaspari since 1996 for several projects using monoclonal antibodies for the Immunology Section of Adolfo Lutz Institute. To Dr. A. Leyva for English editing of the manuscript. We also thank 2 anonymous reviewers for their suggestions.
Immunology Section, Adolfo Lutz Institute, São Paulo, São Paulo, Brazil
Aline S Ferraz, Elza FT Belo, Ligia MCC Coutinho, Ana P Oliveira, Daniele L Franco, Tatiane Ferreira, André Y Yto, Marta SF Machado & Elizabeth De Gaspari
Bacterial Culture Collection, Adolfo Lutz Institute, São Paulo, São Paulo, Brazil
Monica CG Scola
Aline S Ferraz
Elza FT Belo
Ligia MCC Coutinho
Ana P Oliveira
Andréia MS Carmo
Daniele L Franco
Tatiane Ferreira
André Y Yto
Marta SF Machado
Elizabeth De Gaspari
Correspondence to Elizabeth De Gaspari.
The author(s) declare that they have no competing interests.
All authors contributed equally to the research. ASF, EFTB, LMCCC, APO, AMSC, AYY, MSFM are Master's students under the supervision of EDG who had used or are using the monoclonal antibodies from our laboratory. MCGS was a research technician that managed the growth and control of the N.meningitidis strains used in this study, DLF was a fellow of PAP/IAL/CCD/SES-SP (2005–2006), EDG was responsible for design of all the study.
Ferraz, A.S., Belo, E.F., Coutinho, L.M. et al. Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA. BMC Infect Dis 8, 30 (2008). https://doi.org/10.1186/1471-2334-8-30
DOI: https://doi.org/10.1186/1471-2334-8-30
Outer Membrane Protein
Neisseria Meningitidis
Whatman Grade
Central Public Health Laboratory