Indirect Fluorescent Antibody Test for Malaria Antibody

13 Pages·2007·2.99 MB·English

Indirect Fluorescent Antibody Test
for Malaria Antibody
WILLIAM A. SODEMAN, Jr., M.D., and GEOFFREY M. JEFFERY, Sc.D.
Smearseither used fresh preserved
THEINDIRECTfluorescentantibody(FA) are orare
test for malaria antibody, which has been m. a *.. ^ tune for treatment and are
used extensively in few research centers, has then bi?u«^Jto roomt^peratare in a dessica-
a
demonstrated utility in clinical and research tor- The slides are treated at room tempera-
ianvestigationsofmalarialinfectionsin and ture' the filmf ,not beinf P*.*!** *> &y be-
man
experimental hosts. Saliou (1) and Tobie {2) tweenstepsoftheprocedure. Afterthesmears
havepreviouslyreviewedtheapplicationofthis have been mounted.' *ey are <*amine<l for
mtheethtoesdtstohasstubdeieesn odinscmuaslsaerdiai.n tShpeecliiftiecriattyuroef, ^oFroesrctebneceA^osncaensucietabmliecrmoisccroopscyoipnethe pres-
^"^T
but far know there is published f* studv a L*ltz .cros°°Pe
evalusaotionoafsthweetest forsensitivitny,oreproduci- ^> w*h an,0sr5T.1HB_2?)^Jt source 2?)
bility,and ofapplication. Ourstudyseeks wa? used with a UG-2 or B0r-12 exciter filter
ease and UV absorbmg secondary fflter. The
tliomiptoaitnitonosutinainndterdpirsectuastsisonourrecseusltoifngerfrroormatnhde Parasiatef .re located bv usinS the B^~12
filter5 the degrees of fluorescence deter-
ewvaoylvtehdeatnedchunsiequdewiisllapbpeliperde.senTthedeimnetdehtoaidl.we "^Wlth theJ?0"2 filt(f fluowreesrceence is
graded on an arbitrary scale of 0 (no fluores-
Methodscence) to 4+ (maximum fluorescence). Titers
usTImngatjgZheeneFt-r.Aaal, tseesntjs.ei2,oarl.lmai.lnivaersi.tAa.il?gaatAnot_r.i-sibpord1eysehtnatxvlley mareTIwndh,eia.rcdi-,,hvaepnfd,lt.ufmorgroetsm,,hcteehncmeeeftic,n,haaonld-d,,bieolfu»dt-,eTKirmuoonvnios.fntr,taaestnt,ed-,ds-.,aesrsuom-
adioptedi xtihe met_.ihodi diescri-ibedi iby Kttuv.m andi
xta.siszoejcdi.aibt*,leosod(i/3«)\a.re Auais.re-d,dirai.sediant__tih,.ii.gnefn/_i/lpmrsepoaf*ra*pt\ai.roanssi.-, ttc,,,ih.eaemt_pestp_,eet_rd,of*.to,aor.msa_apdjne,_hcceie.rfiecotf,-Atoes^srteercpoorl,gootngxoii..cczioeldt-,,,e'sptwr,sie..ncihi.S.<ap_pfvle,eecsifa._iitx-n--
and, txlhese s-l.i,des are treated as folloxws:i. j. ncally our concerns ihave bifeen wi-t^h reagent_.
Stepsofprocedure Minutes standardization, quantitation of antigen, and
0.1 N HC1_ 5
Distilled water (dip)_
-
PBS pH 7-7.2 (first wash)- 5 #r# Sodeman is chief, gastroenterology section, Vet-
PBS pH 7-7.2 (second wash)- 5 Administration Hospital, Ann Arbor, Mich.,
Test serum-20-30
PBS pH 7-72 wash 7 .10 a mstructor i.n medi,c.i.ne, U»rniv.ersi.ty of/ M*i*-ch»i.gan
Specific conjugated antiglobulin_20-30 Medical School, Ann Arbor. Dr. Jeffery is acting
PBS pH 7-7.2 wash__-__ 10 chief, Laboratory of Parasite Chemotherapy, Na-
Mount in buffered glycerin pH 7-7.2-_- tional Institute of Allergy and Infectious Diseases,
Note: PBS.phosphate-buffered saline. PublicHealthService*
VoL 81, No. 11, November 1966 1037
with total volume of reactants, all of which re¬ globulin specific for the type of serum to be
quire particular consideration within the me- testedisaddedto each well; 0.05 ml.ofaglobu¬
chanics of the test as described. We have lin solution is used for each well. Slides are
evolved the following protocol: again placed in amoistchamber for20minutes
Air-dried thin films of parasitized blood are andrinsed, then washedinPBS for 10minutes.
prepared for testing by delineating circular After the slides removed from PBS, drop
are a
areas of uniform size, hereafter called wells of 0.02 percent Evans blue is placed on each
Our practice is to prepare each slidewiththree well as a counterstain. Keaction is permitted
wells 8mm. in diameter. Thewellsareformed in a moist chamber for 15 minutes. A PBS
by printingthe pattern with a spongelikeplas¬ rinse follows, then a 10-minute PBS wash on a
tic foam templet saturated with a 4 percent slide rotator. Smears are mounted in buffered
water-soluble silicone compound, Siliclad glycerin (pH 7.2) for viewing. Titers de¬
are
(S)(C). The only areas of the slidenot covered rived from the final dilution of test serum in
by a silicone film are the wells. The slides are whichfluorescenceof2+ orgreatercanbedem¬
dried on their sides in racks at 37° C. for 30 onstrated. We found this endpoint, which
minutes, theposition of the wells beingmarked was suggested by Voller and Bray (5), to be
with a diamond scribe. After the drying pe¬ themostreproducibleandeasilyobservableone.
riod, the slides are passed through 0.1 N HC1
for 5 minutes, through distilled water (a dip), Results and Discussion
and finally, through two phosphate-buffered In the titration of test serums for the end¬
saline pH 7.2 washes of 5 minutes each. After point of fluorescence, most investigators have
removal fromthefinal wash, theslideis tapped used serialtwofolddilutions. Achangeoftwo
on its side on an absorbent surface. Aqueous or more tube dilutions has generally been con¬
solutions will normally adhere only to the area sidered to represent a significant change in
ofthe wells; occasional dropletspersistingelse¬ antibody titer. In hands, replicate titra¬
our
where on the slide may be removed by touching tions of a single serum diluted in this fashion
them with absorbent paper. In this manner, did not, however, result in consistent endpoints
wells of a uniform size containinguniform vol¬ within the range of a single ± tube dilution;
umes of PBS may be quickly formed. about 1 in 20 titrations varied from the mode
Serums to be tested are initially diluted 1 to by 2 dilutions (table 1). We believe that this
5 with PBS and then are prepared in serial inconsistency reflects the cumulative error in¬
threefold tubedilutions with PBS as a diluent. herent both in the multiple manipulations and
Dilutions arepreparedby a loop microtitration in the assumptions associated with the test
technique commonly used in virus titrations performed in laboratory.an
as our error
(4). This method yields 0.05 ml. of each dilu¬ which efforts at standardization of reagents
tion of test serum with an error of ±1 percent. and techniques failed to eliminate. Because
The test is quickly performed, speed being thetest apparently could notbe dependedupon
increased without sacrifice of accuracy. To to distinguish antibody concentrations differ-
maintain a uniform volume of reactant, the ing by as much as three twofold tube dilutions,
total volume of each dilution is aspirated from adopted protocol using threefold dilu¬
we a
the diluting cup and transferred to the antigen tions. When the test was performed as
well. A melting point capillary fitted with a outlined previously, but using this protocol,
perforated rubber bulb provides a convenient antibody titers on replicate samples remained
disposable transfer pipette. Slides are placed within a single ± threefoldtubedilution ofthe
in a moist chamber at room temperature for modal titer. We tested this protocol by repli¬
20 minutes. After the reaction period, slides cate observations on individual serums (table
are rinsed with PBS and washed in PBS on a 1) andconsiderthatbythismethod adifference
slide rotator for 10 minutes. After the wash¬ of two or more threefold dilutions represents
ing, slides are removed from the PBS and a significant change in antibody titer. This
tapped on their sides to remove excess PBS as protocol decreases the sensitivity ofthe testbut
previously described. Fluorescein-conjugated makes interpretation easier.
1038 Public Health Reports
Table 1. Comparison of twofold and three¬ which lack uniformity discarded. Within
are
fold dilutionof serumsforindirect fluores¬ the recognized limitations, among a selected set
cent antibody determinations of slides prepared from a single souroe ofpara¬
sitized blood, the total antigen content of each
well can be considered relatively uniform as to
parasite size and number.
Whilethiscrudequantitation of antigenmay
reasonable consistency of the total
assure
amount ofantigen in eachtest preparation and
thusthenumberofantibodybindingsites,there
are other problemsrelatedtothe antigen. The
amount of fluorescence within the test well is
discrete fluorescent units
seen as or masses
(parasites) rather than diff fluorescence of
use
thetotal quantity of antigen in the well. Fur-
thermore, in this case, fluorescence represents
light emitted by fluorescein-tagged antibodies
boundtoantibodieswhichare,inturn,boundto
antigen. This situation is further complicated
because fluorescence is measured in subjective
units of brightness rather than precise
as a
physical unit. The comparative size of the
parasites on the antigen slide apparently mate¬
rially affectsthebrightnessoffluorescence;that
is, large parasites are brighterthan small para¬
sites. In titrations of FA endpoints, fluo¬
of the small parasites (rings)
rescence on a
bloodfilmwillfalltolessthan2+ twodilutions
before that of the larger parasites (schizonts).
Thus,testsensitivitywillvarybytwotubedilu¬
21 DDiilluuttiioonnss ssttaarrtteedd aatt 11::51.0. tions dependingonthesizeoftheparasites used
indicator. When the staining titers of
as an a
Evidence that the mechanics of the test may serum are compared among a number of differ¬
affect its sensitivity prompted us to examine ent antigen preparations, a slide containing
carefully each of the materials and methods many small parasitesmaynotbepositive,while
used in the test, including the antigen, the test one of the same species with a few large para¬
serum, the specific antiglobulin conjugate, and sites may show strong fluorescence. The only
the method of counterstaining. solutionseemstobetouseasinglelotofantigen
Antigen. The use of thin films of parasitized slides for a single experiment and to recognize
blood as the antigen preparation raises several that significantchanges intitermay occurwith
questions about uniformity of this reactant. changein lotofthe antigen slides.
a
We are not aware of an infallible method to Another difficulty that we have encountered
prepare large numbers of thin blood films of a with antigen is in its in vivo binding of anti¬
uniformcell density andcell distribution. For body. SuchbindinghasbeenreportedinPlas¬
this purpose we prepare as many films as prac¬ modium berghei infection in mice (6), and we
ticablefrom asinglesourceofparasitizedblood, have demonstratedthis binding phenomenon in
attempting in each film to obtain approximate¬ P. cynomolgi infection in rhesusmonkeys when
ly the same thickness (cell density) and an slides were obtained late in the course of infec¬
even distribution of cells in the central areas tion. To insure the desired reproducibility of
which are to be occupied by the wells. Slides the test, antigen should be obtained before sig¬
which are obviously too thick or too thin or nificant specific antibody -appears.
Vol. 81, No. 11, November 1966 1039
Tvst serum. We have already discussed the the conjugate. Conjugated globulin obtained
of threefold dilutions rather than twofold by diethylaminoethyl (DEAE) cellulose frac¬
use
dilutions. An equal volume of test serum tionation of tagged serum is our reagent of
should be placed on each well. This volume choice. This reagent proved superior toconju¬
helpsdeterminethesensitivity ofthetest, forif gates absorbed with tissue powder, for it re¬
thevolumevariesfromonetitrationtothenext, sulted in less background staining. Suitable
replicate determinations cannot be made. The commercial products are available. By using
simplest illustration of the effect of serum single lots of serum for each experiment, we
volumethatwehavebeenabletoperformwasas have avoided the necessity for more sophisti¬
follows: A slide prepared with three wells. cated reagent standardization. When indi¬
was
Thefirstreceived0.05ml.ofthetiter (endpoint) cated, determination of fluorescein-protein
dilutionofaserumthathadbeenpreviouslyrun. ratios and use of quantitative precipitin tests
The second well received 0.1 ml. of the next would permitbetterstandardization.
higher dilution, and the third well received 0.2 Gounterstain. Diggs and Sadun (9) suggest
ml. of the next higher dilution. Reaction was the use of Evansblue as acounterstain, and we
carried out per protocol. All three wells have found this reagent of singular help in de¬
stained with the brightness of 2+ as confirmed terminingtheendpoint ofthetitration. With¬
bythree observers. Thisresultsuggeststhatto out a counterstain, the fluorescence falls
maintainauniformsensitivity,constantquanti¬ gradually from4+ tothe0 levelofbackground
ties ofeach dilutionmust be applied. fluorescence. The color change is from bright
Specific antiglobulin conjugate. Quantita¬ apple-green to faint apple-green. With the
tive considerations with regard to antigen counterstain, the color changes from bright
and test serum are less important with specific apple-green to red. The change is distinct,
antiglobulin conjugate than with other sharp, and less questionable from thesubjective
re¬
agents provided rule is observed. Enough point of view than without counterstain.
one a
conjugated antiglobulin must be addedto satu- Properly applied, the counterstain appears to
ratethebinding sites of all globulin inthe anti¬ cause little, if any, artifact from the quenching
gen preparations. For economy, we use the of specific staining.
maximum effective dilution of conjugate. We Thequestionhasbeenraisedofcomparability
determine this for each lot of conjugated anti¬ of results from the various centers currently
serum,using aknown positivetestserumtofind performingthe test. Wesentquestionnairesto
the highest dilution of conjugate that will still most of the investigators using this test, and a
give maximum fluorescence. summary of the methodsthey reported appears
A detailed discussion of the techniques of in table 2. There is wide variation in the spe¬
conjugation is beyond the scope of this paper. cific method. For example,there are wide dif¬
We recommend the methods of Marshall and ferences in the of the slidestained (from
areas
associates (7) for conjugation and those of 0.2 to 9.35 sq. cm.), and in the volumes of the
Kiggs and associates (8) for fractionation of test serum appliedto theslide (from 0.05to 0.5
Table2. Variabilityof methodsappliedby numberofinvestigators
a
1040 Public Health Reports
ml.). In the absence of careful comparison of Immunodiagnosis. Amer J Trop Med & Hyg
the results obtained by various laboratories on 13: 185-203 (1964).
the same serums, there can be little meaningful (3) Kuvin, S., et al.: Antibody production in human
comparison at this time between titers obtained malaria as determined by the fluorescent anti-
body technique. Science135: 1130-1131 (1962).
by thevariouscenters.
(4) Sever, J. L.: Application of a microtechnique to
viral serologictitrations. J Immun 88: 320-329
Summary (1962).
The indirect fluorescent antibody test for (5) Voller, A., and Bray, R. S.: Fluorescent antibody
malaria antibody can provide auseful measure- staining as a measure of malarial autibody.
Proc Soc Exp Biol Med 110: 907-910 (1962).
ment of the immunological status during ma-
(6) Kreier, J. P., and Ristic, M.: Detection of a
larial infection. Variations in the method of Plasmodiwm berghei-antibody complex formed
performance of the test, however, may alter its in vivo. Amer J Trop Med & Hyg 13: 6-10
sensitivity and reproducibility. In antigen (1964).
preparation, special attention should be di- (7) Marshall, J. D., Eveland, W. C., and Smith, 0. W.:
Superiority of fluorescein isothiocynate (Riggs)
rected toward obtaining smears with a-uniform
for fluorescent-antibody technicwith a modifica-
distribution of parasites and toward maintain-
tion of its application. Proc Soc Exp Biol Med
ing uniformity of size in the area tobe stained. 98: 898-900 (1958).
The size or stage of the parasites to be stained (8) Riggs, J. S., Loh, P. C., and Eveland, W. C.:
will also affect the interpretation of results. A simple fractionation method for preparation
Uniform quantities of the serum dilutions offluorescein-labeledgammaglobulin. ProeSoc
Exp Biol Med 105: 655-658 (1960).
should beused for staining. The sensitivity of
(9) Diggs, C.-L., and Sadun, E. H.: Serological cross-
the test must be adjusted to the precision of reactivity between Plasmodium vivaxc and Pla8-
performance. Use of Evans blue as a counter- modium falciparum as determined by a modified
stain simplifies the reading of these tests for fluorescent antibodytest. ExpParasit16: 217-
223 (1965).
malarial antibodies.
EQUIPMENT REFERENCES
REFERENCES (A) Osram HBO-200lamp, Unex Products Corp., New
(1) Saliou, P.: Diagnostic serologique du paludisme York, N.Y.
humainparl'immuno-fluorescence. BoscFrkres, (B) Leitz model SM fluorescence microscope, E. Leitz,
Lyon, France, 1964. Inc.,NewYork,N.Y.
(2) Tobie, J. E.: Detection of malaria antibodies- (C) Siliclad®, Clay Adams Co., New York, N.Y.
Selective Typhoid Immunization
The Public Health Service Advisory Committee on Immunization
Practices advises against routine typhoid immunization in the United
States. Instead, it recommends selective immunization for people
intimately exposed to a known typhoid carrier as would occur with
continuedhouseholdcontact,livingincommunitiesorinstitutionswith
outbreaksof-typhoidfever,ortravelingtoforeignareaswheretyphoid
fever is endemic.
The committee also recommends discontinuing the practice of vac-
cinating people in summer camps and in flooded areas. It holds that
paratyphoid A and B antigens when combined with typhoid vaccine
may increase the occurrence of vaccine reactions and consequently
discourages the use of paratyphoid A and B vaccines.
The committee states that less than 500 cases of typhoid fever are
reported in the United States annually and a continuing downward
trend can be expected.
Vol. 81, No. 11, November 1966 1041
Single Unit Cardiac Monitor-Defibrillator-Pacemaker
Rapidlyaccumulatinginforma¬ ties for reducing mortality by prompt observa¬
tion concerning cardiac arrest tion, diagnosis, andtreatmentofcardiac arrest.
points to the need for a single The comprehensive units are, however, still too
unit device for monitoring, d.c. cumbersome, heavy, and complicated for port-
defibrillation, and pacemaking ability. A compact, inexpensive, but highly
that is readily available, simpleto operate, rea¬ efficient combined unit could be placed inevery
sonable in cost, and of maximum efficiency. surgical operating room, emergency suite, car¬
Time, the crucial factor in such cases, must be diac patient's hospital room, and ambulance.
minimized. The simpler the combined unit, The unit illustrated combines the three ef¬
the greaterthe chance of successful application fective modalities. It has one power cord.
in an extreme emergency; the cheaper the cost Combining the three units in a single small
ofsuchaunit,themorelikely will onebe avail¬ cabinet eliminates duplication of many parts,
able near a stricken person. thus reducing weight to approximately 38
Recently developed, elaborate cardiac units pounds.a nurse could carry the apparatus if
have demonstrated convincingly the opportuni¬ necessary. In addition, it has only one cable
Circuit diagram of the single unit
PACEMAKER D.C.DEFIBRILLATOR UTILIZING SEMICONDUCTOR OEVICES
Employing Z-oxitondpovrarsupplyofoscilloscop*
SEMICONDUCTOR
(Otcilloscopt)
J~U_7=«
470K £k>K i»70K '
ifV '* it'°' ^®^f -?POAUCTEPMUATKER
I7VACJ U
POWER SUPPLY
* +75V
OSCILLO- -7h 6ND
SCOPE-i ..-I2V
3°00oooo£||7VAC
-»-IOOV
Note: Six 1,600-volt silicon-controlled rectifiers make possible rapid charge and discharge of thecondensers.
Amaximum200-volttransistormakespossibleapacemakerwithoutvacuum tubes.
1042 Public Health Reports
to the patient. The contour-fitted defibrilla- Electrodes incorporated in a "vest" are con¬
ting-typemonitorelectrodesproduceanelectro¬ venient and reliable for use with patients
cardiogram ofthehighestfidelity, although the prepared for surgery
electrodes placed in the customary position
are
for defibrillation. The electrodes used for
monitoring can be instantly converted to de-
fibrillating pacemaking Thus, cardiac
or use.
disaster betreatedtheinstantitisindicated
can
by the monitor.
Although large, cumbersome, complicated
units sell for approximately $4,000,the unit de¬
scribed canbeproducedto sell for about$1,200.
Theinstrumentpanelissosimplethatitsopera¬
tion can be learned instantly. The low cost,
compactness, and simplicity of operation of the
unitmake production and therefore wide¬
mass
spread distribution possible..J. Robert Close,
M.D.,OrthopaedicHospital,LosAngeles,Calif.
This invention developed under Public
was
HealthServicegrantNo.NB-2300.
cardiac monitor-defibrillator-pacemaker
PACEMAKER O.C.DEFIBRILLATOR UTILIZING SEMICONDUCTOR DEVICES
TRANSISTORIZATION(COMPLETE)BYINSERTIONOFONETRANSISTOR TRANSISTORIZATION COMPLETE
V-.MAX. 200VOLTS INPLACEOFTRANSISTOR2N657.
,
SEMICONDUCTOR
eo8p-oucniitotrs.bra*n.k SWITCH
2omp F K)m»Q -»»"?l>-
A wtj
(£*<&<&(5: +±h_
(OMillotcopt)
47+ok2II0V
iNmAjoS
-.:--rL|gIXNTT::00--I3B6OVV
470K ,7°* J .¦« . -?PACEMAKER
OUTPUT
m / _u*nr_r» FIBRILLATOR UVV7T +27V HWW.?+2I0V
CELHIAMNIGNEATOEFSTERXATNESIRSNTAOLR2N657FORAV.MAX.200VOLTS. V.A3cilpMlWMfJ*-if OUT-PU»iT. T.nKK1r| .*4W7WK, IO0K J£a:a.3.«U^TI-I40A-4I70IWK.li ,l_l
AMPLIFIER(SHADED PDnOuWitFWO «SsU.PdPdiLYv SEMICONDUCTOR
AREA) +75V TERMINATQR
,_._-._+3.00wV
OSCILLO 1-h. +765NVD IOK DELAY JIIlJ_1_
SCOPE
3«»oooo(J||7VAC
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(over)
Vol. 81, No. 11, November 1966 1043
Electrodes (physically separated) can be immediately applied in an emergency
<Gp*
Note: The electrodes are bipolar and ground isolated. The chassis is grounded. No ground wire is con¬
nected to the patient lest stray voltages occur. The bar separating the electrodes is a safety device for the
individual operating the unit.
1044 Public Health Reports
with better understanding of the
physical, mental, and social needs
ofthe patient as a basis for improv¬
ingnursingcare, patientmorale, and
job satisfaction of nursing staff.
The manual was prepared by the
public health nursing section of the
Colorado State Department of Pub¬
HHeepaalttihtisI.nfoPrHmSatPiuobnliSceartiieosnNNoo..48426,; umseedfiuclalinfiltmhseamneddifcialmlsttrriapisnitnhgatparroe¬ ltiiconHepraoljtehctastopdaertteromfinaedtehmeonesxttrean¬t
rpeevris1e0d0.19D6e6f;inelseafhleepta;ti5tisceantnsd, e$x2- gagreanmcsieosftohnaetocrommoprriesoeftthheemFeedmebrearl mofainrethaaibnielditatbiyonncuarrseingthatpercsaonnnbeel
plains the difference between infec¬ Advisory Council on Medical Train¬ and the rehabilitation nursing that
Dtieosucsrihbeepsatitsiysmpatnodmss,erurmechoepmamteitnidss. minegntAidosf.thMeemAbremrys, iDnecplaudretmDeenptarto¬f can be taught on an inservice basis.
treatment by a physician, and ad¬ the Navy, Department of the Air Depuration Plant Design. PHS
vises how toreduceriskofinfection. Force, VeteransAdministration, and PublicationNo.999-FP-7;1966; 119
thePublic Health Service. Provides pages; 75 cents. Outlines technical
Tapeworm. PHS Publication No. film descriptions arranged by sub¬ points in the design of shellfish de¬
158, Health Information Series No. jectwithatitleindexandacategory puration plants which safeguard
48; revised 1966; leaflet; 5 cents, listing. Lists materials currently oysters, clams, and mussels through
$2 per 100. Describes beef, dwarf, available for borrowing or renting; the purification process known as
and pork tapeworms that occur in nofilmslisted are forsaleonly. shellfish depuration.
humans. Warns against self-diag-
Radionuclide Analysis of Large
nosis and self-treatment. Explains Reported Tuberculosis Data, 1964.
Numbers of Food and Water Sam¬
preventive methods. PHS Publication No. 688; 1966; 41
ples. PHSPublicationNo.999-RH-
pages; 80 cents. Presents data and
Ringworm. PHS Publication No. statistics on tuberculosis morbidity 17; 1966; by Esther Ferri, Paul J.
46, Health Information Series No. andmortalityandincludesrecentin¬ Mdgno, and Lloyd R. Setter; 28
6; revised 1966; leaflet; 5 cents, $2 formation about the status of the pages. Describes the processing of
per 100. Describes symptoms, pre¬ diseasein the United States. Points large numbers of food and water
ventivepractices, and warns against to changes that have influenced re¬ samples for gross and specific radio¬
self-diagnosis and self-treatment for centtrendsandpinpointsthevariety activity as conducted by a regional
ringworm of the feet (athlete's ofcomponents thatrequireconsider¬ laboratory of the Division of Radio¬
foot), nails, body, and scalp. ation in assessing the applicability logicalHealth. Describestechniques
Public Health ServiceFilm Catalog, and effectiveness of local, State, or ftoerresat saernidesfoofr rcaedritoaniunclisdteabsleofeilne¬¬
1966. PHS Publication No. 776; nitaiteis.onal tuberculosis control activ¬ ments in foods, grossalpha and beta
revised March 1966; 99 pages; $1. determinations,gammaspectralanal¬
Revised list of health and medical Grade "A" Pasteurized Milk. PHS ysis, and determination of radium
films available from the Public Publication No. 1011; 1966; leaflet; 226 and strontium 90 in water.
Health Service Audiovisual Facility, 5 cents. Gives the history of the Briefly discusses manpower and
Communicable Disease Center, At¬ Cooperative State-PHS Program for equipment requirements for various
lanta, Ga. Certification of Interstate Milk measurements.
Hospital Environment Makes the Shippers. Describestheoperationof
Difference. PHS Publication No. theprogram,itsgrowth,benefits,and This section carries announcements of
9S0-C-14; 1966; leaflet. Outlines accomplishments. new publications prepared by the Public
the significance of the hospital en¬ Elementary Rehabilitation Nursing pHreaelptahreSdervwiictehaFneddeorfalselseucptpeodrtp.ublications
vironment to patients, staff, and Care. PHS Publication No. 1486; Unlessotherwiseindicated, publications
community in terms of infection 1966;100pages,illustrated;55cents. for which prices are quoted are for sale
control, personnel safety, and hos¬ Presents a manual for teaching by the Superintendent of Documents, U.S.
pital utilization. Summarizes the the fundamentals of rehabilitation Government Printing Office, Washington
Division of Hospital and Medical nursingcaretonursingandancillary Dpa.nCi,ed20b4y02c.ash,Ordcehercsk,shoorulmdobneeyacocrodme¬r
Facilities' programs in research, ed¬ personnel in nursing homes, hos¬ and should fully indentifythe publication.
ucation, andconsultation specifically pitals, convalescent facilities, and Public Health Service publications which
directed to solving hospital environ¬ public health agencies. Employs do not carry price quotations, as well
mental problems. practical and effective rehabilitation aprsicseisngalreesasmhpolwen,cocpainesboeftohbotsaeinfeodrwwhiitchh¬
procedures based on the use of in- out charge from the Public Inquiries
Film Reference Guide for Medicine expensiveequipmentandaminimum Branch,PublicHealth Service,Washington,
and Allied Sciences. PHS Publica¬ of personnel. Provides the nursing D.C, 20201.
tionNo.487;revised1966;897pages; personnel,throughapplicationofthe The Public Health Service does notsup¬
$2.50. Includes entries for selected principles.and training techniques, ply publications other than its own.
Vol. 81, No. 11, November 1966 1045
STOCKWELL, EDWARD G. (University of Connecticut): Use of socioeconomic
status as a demographic variable. PublicHealth Reports, Vol. 81,November1966,
pp. 961-966.
The relations between the three proc¬ the basis of these observations. First,
esses of demographic change (fertility, socialandeconomicfactorsaremajorde-
mortality, and migration) and socioeco¬ terminants of demographic behavior, but
nomic status have been analyzed. The there is a need for more research to dis¬
data show that levelsoffertilityandmi¬ cover more precisely the nature and ex¬
gration tend to be positively associated tent of the relations between particular
withsocioeconomicstatus,whereaslevels demographic variables and particular
of mortality are negatively correlated indexesofsocioeconomicstatus. Second,
with all the various socioeconomic in¬ research inthisendeavormustclearlybe
dexes: occupation, education, income. cogniaantofthemultidimensionalnature
Thedataalsoshow thatthedemographic ofsocioeconomicstatus. Whatisneeded
variables are not necessarily related in is a research approach which recognizes
any consistent fashion to all the socio¬ that socioeconomic status is many
economic variables. Mortality, for ex¬ things: that it is composed of a number
ample, was more related (negatively) to of different variables, that each of these
income than to any other variable variables may act independently of one
whereas educationappeared (positively) another in specific situations, and that
to be the most significant factor in the each does not necessarily have to bear
migration socioeconomic differential. thesamerelationtoanygivenbehavioral
Two conclusions have been drawn on phenomena.
EISENTHAL, SHERMAN (Veterans Administration), FARREROW, NORMAN L.,
and SHNEIDMAN, EDWIN S.: Followup of neuropsychiatric patients in suicide
observation status. Public Health Reports, Vol. 81, November 1966, pp. 977-990.
Afollowupstudy wasconductedof912 formation, andneuropsychiatrichospital¬
patients in a Veterans Administration ization data did not reveal any single
neuropsychiatric hospital who had been pattern typifying the suicidal person.
placed in suicide observation status be¬ The success of the best predictors in
W00M tween1954and1958. Completefollowup forecastingsuicide wasfrom8toashigh
data were obtained for 90 percent of as 13 percent, while the best predictors
the patients. Forty percent of these pa¬ of attempted suicide forecast the event
tients subsequently manifested furtheT with success ranging from 23 to as high
suicidal behavior; 6 percent committed as29 percent. Inthisparticularpopula¬
suicide, 17 percent made nonlethal sui¬ tion, suicideismuch more probable than
cidalattempts, and17percentmanifested in the neuropsychiatric hospital popula¬
suicidal ideation. Analysis of the pa¬ tion or the general population.
tients' suicide histories, demographic in¬
BOURDILLON, JAQUES (New York State Department of Health), and VANDER-
LINDE, RAYMOND E.: An improvedscreeningprocedurefor bloodphenylalanine.
PublicHealthReports, Vol.81,November1966,pp.991-995.
An improved method is described for trolled conditions, this figure was
the automated screening ofblood phenyl¬ constant and independent of the concen¬
alanine. When dry blood spots on filter tration. The fluorescence ofnonphenyla¬
paper are exposed to hot steam, the pro¬ lanine substance was equivalent to only
teins are irreversibly bound to thepaper, 0.8mg. ofphenylalanineper100ml.
whereas the other solutes can be subse¬ With a smaller fluorometric cell, the
quently released by water elution. This method could be run at 60 specimens per
step takes the place of dialysis and per¬ hour without carryover. A modified
mits bypassing the dialyzer in the Tech¬ pump allowed the speed to be increased
nicon automated methods, resulting in a to 120 specimens per hour. The method,
fivefold increase in sensitivity. Dry which requires only a one-quarter-inch
blood disks containing added phenylala¬ paper disk of dried blood, is considered
nine and subjected to autoclaving for 3 accurate enough for screening purposes
minutes at 121° C. gave a phenylalanine and sensitive enough to detectborderline
recovery ofabout75percent. Undercon¬ cases.
1046 Public Health Reports