Wednesday, April 25, 2012

South Africa in the 50s


Since I am dying I thought I might remember some of my experiences in South Africa in the Fifties. I am a Yank, and my mother married a South African doctor who was a general practitioner.  We moved to Johannesburg and after completing a matriculation, I entered the Bachelor of Science Medical science class at the Witwatersrand University.
 Wits was an active place in those days.
My first assignment as a science student was under the tutelage of Professors Strydom and Windham. Both had trained at the University of Pennsylvania in environmental health. The SA Chamber of Mines was having a severe problem sustaining a viable labor crew.  The Chamber of Mines group recruited Africans from small villages. The recruits were between the ages of 14 to 18. They were brought to the mines in Johannesburg and sent down to depths of 6 miles where they dug gold from the stopes. A stope is a shaft between two tunnels. There is little air and the work is very hard. The boys typically collapsed from heat stroke and the then therapy was to take them to the surface  and throw them into cold water. Their body temperature was about 104 to 105 degrees fahrenheit. The mortality was 50%.  Strydom developed a method where before going to the  stopes the Africans were acclimatized. He worked out the method on 6 of us science students. He placed us in a hot room with a red green blinking metronome and had us shovel ore for a week. He observed our temperatures and noted that it took about 3 days for our temperature to stabilize at about 99 degrees. He then set a red/green light metronome in one of the passage ways of the East Rand Proprietary Mine (6 miles down) and tested this on new African recruits.  The method was scientifically successful and the mortality decreased to under 10%. In retrospect how humanitarian was the approach?

I was lucky to have  Raymond Arthur Dart as my lecturer in Anatomy. He, of course, along with Phillip Tobias, was the discover of Australapithecus Africanus. They both detailed for us the anatomy and physiology of this  human “missing link,” which was discovered at Sterfontein, and explained their  thoughts on why it was a missing link.  These also detailed the differences  between the present-day indigenous  African  population and the European population. For example, Africans had no platysma, their endocrine glands were different and the size and shape of their skulls was different. I have often wondered why world class scientists had to discuss issues of apartheid.

The  University of Witswatersrand, like most of the other South African universities, had a great hall. I was privileged to see the first public performance of “King Kong,” with music composed by Todd Matshikia, one of the original journalists on Drum magazine. Miriam Makeba sang a leading role.  This show was a jazz opera with an all black South African cast. The first such theatrical effort in South Africa, it was a fantastic experience.   The ushers tried but failed to segregate the audience.

One other event at the great hall that I remember distinctly was when my  Professor of Botany Eddie Roux, author of a history of South Africa titled Time  Longer then Rope, and actively against the governments policies of apartheid, debated  the question whether Christ existed as history or myth. You must  remember the prevailing religious philosophy that pervaded the society was that  Africans were “ hewers of wood  and drawers of water.”  The Dutch Reform Church to which most Afrikaners belonged believed in a literal reading of the Bible. Of course this took a lot of bravado on Professor Roux’s part to take on this establishment by claiming that Christ’s life story was myth rather than fact. Roux debated with a theology school professor at a noon lecture and won the debate. Almost immediately  afterwards, he was house-arrested and his academic carrier ended. I believe that he died a  lonely man.

Another memory concerns the way some students tried to resist the apartheid system.  I was a member of a U.S. student organization, which I joined while enrolled at Reed College, prior to entering the University of Witwatersrand. I believe this is why I was contacted by Allard Lowenstein, later a Congressman from New York State.  Lowenstein ostensibly had come to SA to study the flora and fauna.  Somehow he learned that my step dad’s car  a Ford prefect was permitted free access to  native townships.  My step dad was a general practitioner whose practice included many Chinese patients who were forced to live and work in these townships. The Chinese were the shopkeepers to the Africans. Lowenstein asked me and some  SA friends to drive him around the townships. I agreed to Lowenstein’s request, despite the potential peril this might have caused my step dad and his family in Johannesburg, and we drove through many townships taking movies of the people and their living conditions. At about the same time, Hans Beukes  a Namibian student activist from South West Africa, had been awarded a scholarship to a Scandinavian university. The SA government refused to let him travel, so Lowenstein and friends from Wits placed him in the boot of a car and drove him to Swaziland.  Beukes flew to    Norway and then on to NYC, where he detailed the need to make SWA an independent state and thedire situation of non-whites in SA in a speech at the United Nations.  The United Nations then censured SA.

The politics of apartheid even entered the medical school curriculum at Wits; but sometimes we were able to outwit the authorities. At school, 6 students were assigned to dissect one cadaver.  When we wanted a break, my group of six would go to the movies at the “bioscope” (the South African name for a movie theatre).  One of the students in my dissection group was a Chinese student; by law,  Chinese people were forbidden from going to white movies. We informed the ticket agent that our friend was Japanese  and he was permitted to attend the films with us.  This was because South Africa, which did not recognize China diplomatically, had a vigorous trading relationship with Japan and did not discriminate against Japanese persons in the country.
I left South Africa after the Sharpesville massacre in 1959.  At that time, most of the professionals I knew felt there was going to be a blood bath in South Africa.

Michael T. Makler, MD
www malariaantibodies.com

Tuesday, June 3, 2008

Rapid Diagnosis of Plasmodium Knowlesi

Singh et al have recently described over 70 cases of human malaria utilizing polymerase chain reaction (PCR ) that were caused by P.knowlesi thought to originate from a crab eating Macaque (Macaca fascicularis) of Borneo, and originally mis-diagnosed as P.malariae by microscopy (1). White reviewed these finding and suggested that P.knowlesi malaria be considered the Fifth Human Malaria Parasite (2).

Utilizing, a rapid malaria test (RMT) that detects plasmodium lactate dehydrogenase (pLDH) we were rapidly able to diagnose P. knowlesi. Depending upon the selection of monoclonal antibodies (mabs) to pLDH we could differentiate P. knowlesi from the other four forms of human malarias.

Figure 1 illustrates the principle of our pLDH assay using human blood from a finger prick that contains both P. vivax and P.falciparum.








Figure 2 demonstrates a series of actual immunochromatographic (dip) strips. These strips were stripped with 5 antibodies. The uppermost antibody is a control line containing goat anti mouse (G) antibody. It will react with all mouse mabs. The four other lines: 9C1, 10D, 11D, and 17E, contain mabs designed to be specific to: P.falciparum, P.vivax, P,malariae and P.ovale. Observe that : P.falciparum reacts only with mab 17; P.vivax with mab 11D; P.ovale with mab 10; P. malariae with mab 9 ; and P.knowlesi reacts with 13 and 17. P.knowlesi reacts with 11D and 17E. P. knowlesi reacts with no other mabs.





Figure 3 is an analysis of the the reaction of P. knowlesi first utilizing the Sequence of LDH from P. knowlesi deduced from genomic DNA fragments sequenced by the Sanger malaria genome project. LDH isoforms from Plasmodium vivax, malariae, ovale, berghei, yoelli, and falciparum were compared with that of knowlesi. Residues that were unique to both P. knowlesi and P. vivax are shown in blue, while residues unique to both P. knowlesi and P. falciparum are shown in red. The upper right side of this figure also demonstrates the binding specificity of many different anti-pLDH antibodies. Shown are the reactivities of the indicated monoclonal antibodies to the pLDH from 7 Plasmodium species. These include human, primate, avian, and murine malaria. P. vivax LDH is immobilized only by 11D9 (13H11) and P. falciparum LDH was only immobilized by 17E4(7G9). P. knowlesi LDH was immobilized by both 11D9(13H11) and 17E4(7G9).

The lower part of the figure is a model of P. knowlesi LDH and specific epitopes. A three dimensional model for P. knowlesi LDH was calculated using Modeller V9 and the P. falciparum and P. vivax crystal structures (PDB: 2A94 and 2AA3). Shown is the monomer as well as the assembled tetramer. The NAD co-factor analog 3-acetyl pyridine adenine dinucleotide is shown in black. Residues important for substrate binding and
catalysis are shown in yellow. P. knowlesi residues shared only with P. vivax are shown in blue and indicate where the 11D9/13H11 epitopes could be. P. knowlesi residues shared only with P. falciparum are shown in redand indicate a critical determinant of the 17E4/7G9 epitopes.


We conclude that the pLDH RMT is able to speciate all 5 forms of human malaria. This diagnostic assay takes 10-20 minutes to complete. The one limitation of the pLDH RMT is that it is currently unable to distinguish between a mixed infection with P.vivax and P.falciparum. However, in these infections, unlike the P. knowlesi infection there is often a much greater difference in the intensity of staining of the two bands, consequently together with light microscopy, this combination of tests should provide adequate diagnosis of this serious condition.


References
1.Cox-Singh J, Davis TM, Lee KS, Shamsul SS, Matusop A, Ratnam S, Rahman HA, Conway DJ, Singh B. Plasmodium knowlesi malaria in humans is widely distributed and potentially life threatening. Clin Infect Dis. 2008 Jan 15;46(2):165-71
2. White NJ. Plasmodium knowlesi: The Fifth Human Malaria Parasite Clin Infect Dis. 2008 Jan 15:46(2): 172-73
Authors
Michael T. Makler, MD Flow Inc. Portland Or 97239: 503 481 7994
Robert C. Piper, PhD , Phys, U of Iowa, Iowa City, IA 52242: 319 335 7842

Wednesday, April 4, 2007

Some pldh mabs reacting with animal- human malaria



ANIMALARIA™

Some pldh mabs reacting with animal- human malaria

A combination of 5 of our antibodies (from Flow Inc. Portland Oregon ,USA) directed to parasite lactate dehydrogenase enables a new rapid test we have called Animalaria™ to diagnose all Plasmodium spp. (malaria). That is , with this Animalaria™ test , we are so far able to detect an infection with any Plasmodium species. Initial evidence ,using combinations of our mabs, permits speciation.

Flow's test is unlike the rapid malaria tests which detect the HRP2 antigen. These HRP2 tests can only detect certain genetic variants of nonviable and viable P.falciparum. Consequently, they have no role in therapeutic monitoring , and are also quite limited in evaluation of the distribution of the multiple forms of animal malaria around the world. For example:

Honey creepers in Hawaii (P.relictum), black faced penquins in Southern Africa (P.gallinaceum), gorillas's in Africa (P.reichenowi) are ill and dying from malaria.
These parasites can be easily detected and their illness followed by a single line assay which uses our (Flow Inc. Portland Oregon USA) antibodies directed to parasite Lactate Dehydrogenase.
We can no longer expect to control malaria by just diagnosing and treating malaria in humans. We must also be aware of the types of malaria present in the animal populations where we live. (See Balbir Singh and colleagues in Lancet March 2004). Look at the chart above , and please note that P. brasilianum (in monkeys) is identical to P. malariae (in humans), and that P. reichenowi (in gorillas) is very similar to P.falciparum (in humans).