MISOPROSTOL

EFFECTS OF DAYTIME 8.5

DEVELOPMENT

WHITE MICE (Mus musculus )

Authors

Andrs Fernndez Snchez1 B.Sc.M.Sc - Carolina Isaza of Lourido2 MD.MSc . Mauricio Palacios
Gmez3 MD , PhD .
1Researcher 1Professor Autonomous University Foundation of the Americas , Medicine Program
2Research Professor of Medicine Program Universidad del Valle, Department of Morphology
3 Research Professor of Medicine Program Universidad del Valle, Department of Physiology

Keywords:

Misoprostol - Prostaglandin E - Malformations - Animal Model Gestation.

ABSTRAC:

Exposure in utero to misoprostol has been associated with the presence of congenital malformations
(MFC) As tales "Moebius Syndrome", this is a clinical entity described since 1543 by Paul P. Moebius,
and was characterized by agenesis or hypoplasia nuclei VI and VII cranial nerve and sometimes
malformations in Upper and Lower Associated Members.
Recently published article shows increased frequency of visits to the clinic Dysmorphology Hospital
Universitary del Valle "Evaristo Garcia" (HUV) in Cali (Colombia), where children were found with
multiple malformations and Suspicion Moebius Syndrome, Associated heavily use employee failed in
the quarter fines misoprostol abortion primer scammers.
Despite the large number of clinical and basic studies on the subject, they have not yet been clarified
what triggers this clinical entity, and is considered today as heterogeneous. : In addition,
experimental models in laboratory animals have been unsuccessful para show teratogenic effects of
misoprostol, while accepting a clear association of C Use during pregnancy and Moebius syndrome in
humans.
When assessing the teratogenic effect of misoprostol in fetuses exposed to doses of 25 mg orally and
vaginally, the presence of major malformations in limbs found, as agenesis: Senior Fellow, meromelias
And Sirenomelia addition: malformations in Hall handset and microtia and anotia. Regarding the
revision of the seventh cranial nerve nuclei, in this mouse model of white (Mus musculus), was not
found agenesis of the VII cranial pair bathroom all individuals Litters.

INTRODUCTION
Misoprostol (11-16 dihydroxy -16 methyl methyl 9 -oxoprost 13 - E 1 oate ) is a synthetic prostaglandin
structurally related to prostaglandin E1 ; being approved and licensed by the Food Drug
Administration (FDA) to be used orally in the prevention of ulcer disease gastric and duodenal being
in turn is effective in treating hemorrhagic and erosive lesions caused by the administration of NSAIDs
(NSAIDS) . 1, 2 , 3, 4 , 5, 6 ,
O

H3C

OH
CH3
HO

OH

CH3

CH3

HO

OH

Figure No. 1. Structural formulas of Misoprostol and prostaglandin E1.

Being a prostaglandin causes uterine smooth muscle contraction also causes separation of the fibers
existing collagen in the cervix, so that in some countries where abortion is legal, misoprostol is used
as an abortifacient. However, the FDA recognizes that in certain circumstances, the use of some drugs
in conditions outside the usual indications is rational, appropriate and accepted in medical practice."
7.8, 9.10.
Pharmaceutically misoprostol is manufactured in preparation of oral tablets 100 mg and 200 mg of
hexagonally and grooved , carrying on one side the number ( 1461 ), these contain as inactive
products hydrogenated castor oil , hydroxypropyl methylcellulose , microcrystalline cellulose and
starch sodium glycolate , being this cellulose matrix which stabilizes the compound at room
temperature. 11, 12,13, 14, 15,16
Once the misoprostol is absorbed from the gastrointestinal tract becomes its pharmacologically
active metabolite called misoprostol acid. The plasma concentration of misoprostol acid reaches its
peak in about 30 minutes, and then descend quickly to 15 or 20 minutos.1,2,3,17,18,19,20 Its
bioavailability decreases with concomitant food intake or antacids. It is metabolized primarily in the
liver and 64% to 73% of its active metabolite is excreted in the urine, and the remaining by feces. 6 21,
22, 23
Pharmacodynamic this drug inhibits gastric acid secretion induces healing ulcers and mucosal
protection. The antisecretory activity is mediated by the action exerted directly on specific receptors
of prostaglandins oxyntic cells or gastric parietal inhibiting basal secretion of gastric acid during the
day and night, as well as inhibition of acid secretion stimulated by histamine, pentagastrin, coffee and
generated by the consumption of other foods also reduces the volume of gastric acid secretion and
pepsin at baseline. For such circumstances this medicine is a cytoprotective mucous gstrica.7
Misoprostol has no known interactions with other medications and does not induce the development
of enzyme cytochrome P-450 system heptico.6 Adverse effects include nausea, vomiting, diarrhea,
pain abdominal pain, chills and fever, all dose - dependent. 6.7
Concerning their abortive properties, these are well known by health professionals and often also for
the general population. As the drug is available at low prices, and our environment is marketed
without restriction or medical formula, many women use it as a drug abortive maneuver to terminate
their pregnancies, 13against it the pharmaceutical industry and public health experts have expressed
with great concern that if the attempt is unsuccessful abortion and pregnancy continues to term, fetal
exposure in utero to misoprostol, can increase the risk for birth defects occur. Most documented
casuistry of self-administration of misoprostol to induce abortion comes from Brazil; as a result, this
nation provides a unique opportunity to study the potential teratogenicity of misoprostol. 8,
14.24,25,26

Among the defects associated with the use of misoprostol that are consistent with this second criterion, the
presence of Moebius syndrome is, this being a rare clinical entity characterized by the loss of function of
cranial nerve engines (especially VI and VII. 12,14,30,31,32,33,34
This paper aims to identify the teratogenic effects of misoprostol on the intrauterine development of
rodent (Mus musculus) white mouse during the day 8.5 of gestational period within an experimental model
for them an experimental animal model was developed in mice white (Mus musculus ) to assess potential
teratogenic effects of misoprostol , which allowed us to establish the possibility of embryotoxicity and
teratogenicity of misoprostol and describe the external major malformations in fetuses of white mouse
(Mus musculus ) , exposed during the day 8.5 of the gestation at varying doses of misoprostol as evaluating
the presence histomorfologicamente core VII cranial nerve in fetuses white mice (Mus musculus) exposed
during the day 8.5 of gestation at varying doses of misoprostol .
MATERIALS AND METHODS
Experiments were performed with white mice (Mus musculus.) 8 weeks of age selected parental litters
strain reference CFW, acquired by the Vivarium of the Universidad del Valle, which were contained in metal
mesh cages with dimensions standards and stainless steel cover, according to management protocols
biolgicos54 reagents. They were fed a standard diet (Rodentina - Colombia) and distilled water ad libitum,
keeping in controlled environments of temperature, humidity and light cycles of 12 x 12 hours.
Since its birth took them control weight and height to verify your standard average at the 8th week of age,
which is 55 g for the male and 49.5 g - 50.0 g for females.
Design experimental animal model To evaluate the teratogenic effect on the fetuses of white mice (Mus
musculus) exposed to misoprostol, 2 doses (25 ug and 50 ug) and 4 different administration routes,
supplied in a single critical period, were considered the 8.5 day embryonic period.
As misoprostol LD50 for white mice is 25 mg to 75 mg / kg, 27.28,35,36,45,25 ug dose were used and 50 mg
that are within that range.
The experimental and control groups were established on two factors: Factor A (dose of medication) and
Factor B (Form drug administration); the observed variables were: (gestational weight, number of offspring,
the offspring size and number of malformed fetuses). These data were processed in the Prism 4 statistical
package (Graph - Pad) 46.
The results of the presence or absence of the nucleus of cranial nerve VII were evaluated with the regression
model Logstica47 (W.W. Daniel, 2002 - SS statistical package Pluss, 2002)48

b1= control

FACTOR A
(Dose Medication)

25g
50 g

FACTOR B

b2= oral

(Form
management
Medicine)

b3= vaginal
b4= oral vaginal
b5= vaginal y oral

Sexing and Registration of estrus

Sexing was performed at 40 days of age. To differentiate between a male and a female was taken into
account the distance between the urinary meatus (anorectal triangle) and the anal region , distance is lower
in females than in machos.29,30,31,49 vaginal washing was performed with saline (0.9 % NaCl), using a
dropper , obtaining a sample of vaginal epithelial cells to microscopically determine the relationship with
the phase of the menstrual cycle (C.E.C.S.A,1968)50
Mating and drug Administration
Misoprostol (Cytotec, Searle) was employed - in chewable tablets of 100 mg, which were weighed, then
macerated until uniform solutions with 25ug or 50 ug required for each pregnant animal. The drug was
dissolved in distilled water to a volume of 1 microliter supplied by gavage or by dropper adapted for vaginal administration.
These females were taken estrus test and placed to cycle together to favor the effect of Whitten (1965) 51
with the aim that are at the same time prepared for gestational period, under similar environmental conditions. When their cycles were regulated, they were tested again took estrus and placed in other cages as
required experimental groups, where he lived one parental male. In addition they were given a time range
of 15 hours (time spent for courtship and fertilization). From this moment and the appearance of the vaginal plug it was determined as the 01 day experimental protocol.
During the period 8.5 (stage Theiler 13, equivalent to 8 days and 12 hours gestation)52, 53 were supplied
with misoprostol to pregnant women in order to verify their offspring if prostaglandin interferes with the
development of the VII cranial nerve, the route of administration of misoprostol group was established by
experience, the following experimental groups were used: (Control, oral, vaginal, oral - vaginal and
vaginal - oral), the dosage was 25 mg employee and 50 mg as stipulated in the experimental model.14, 18,
20.
OBSERVATION MACRO AND STEREOMICROSCOPIC OF SPECIMENS.
On day 15 of gestation under general anesthesia (Ketamine / Xylazine Intraperitoneal; 100/10 mg / kg) Ketamine: (Imalgene, 500 - Laboratory Merial, France - Colombia S. A. - Xylazine.
After the above procedure performed proceeded to perform intracardiac perfusion and subsequently
caesarean postmortem54. The specimens were examined with stereomicroscope to observe whether or not
external phenotypic characters dysmorphic.
General parameters and biometric standards for litters were taken; including the number of embryos per
litter and size, standard length, the for experimental and control groups, which were compared with the
tables development Kauffman (1999) 53 for the rodent (Mus musculus) white mouse.
EVALUATION OF THE CORE OF THE SEVENTH CRANIAL NERVE.
The embryonic and / or fetal specimens were fixed in picric acid and then be recorded photographically.
Three days after fixation, sections were performed by stereomicroscopy brain bulboprotuberancial level
sulcus area where the facial nucleus is. Finally we cut to 8 microns which were carried technique paraffin
(Paraplast) and colorations hematoxylin and eosin (H and E) were performed.

RESULTS
Following administration of misoprostol on day (9.0) died eight females belonging to the following
experimental groups: oral 25 mg - 50 mg vaginal (4/4) and vaginal group 50 mg - 25 mg oral (4/4),
remaining as total sample experimental 32 pregnant females. Postmortem pathological studies performed
at 8 females revealed no evidence of toxicity in target organs such as brain, liver, kidney, lung, spleen,
stomach, also not specimens, or fetal resorptions was found in uterine bodies.
Pregnant females who completed the experiment at day 15 were discriminated into the following groups:
25 mg oral (3/4), 25 mg vaginal - oral 50 mg (3/4), 50 mg vaginal - oral 50 mg (2/4), 25 mg vaginal (1/4) and
the control group (8/8). (Figure 2).

Figure No. 2. Number of females with pregnancy at the end of the experiment. The experimental groups
began with four females each.
After practicing cesarean section and perfusion with formalin buferad specimens in normal (healthy),
malformed and resorptions were found. (Figure No.3).

Figure No. 3. SPECIMENS BY TYPE OF TREATMENT DOSE : The bar chart shows the malformed fetuses that
were found in vaginal group of 25 mg (50%) and oral group of 25 micrograms
(5.2%) , fetuses They were
found in normal controls (100%), 50 vaginal mg (100%), and the group of 25 vaginal ug - 50 ug oral (100%);
fetal resorptions were presented at the 50 ug oral group (100%).
No malformations in the control group were found, systemic and vaginal administration of misoprostol is
related to a decrease in the number of specimens, an increase in the proportion of resorptions and
malformations in the different groups (Figure No.3), without being related to the Total administered dose or
SIZE OF THE FETUS AT DAY 15 OF GESTATION

Figure No. 4 . SPECIMENS BY SIZE OF TREATMENT OF PREGNANT FEMALES. The diagram shows the size of
each specimen by groups. Each symbol represents a fetus and the results are presented as the mean SD.
Differences were considered statistically significant for all experimental groups (p = 0.001) compared to the
control group (White 1 and 2). No resorptions were considered in this analysis.
Tracking maternal weight as variable before misoprostol administration and after administration of this, it
not determined significant when evaluated by the statistical package Graph Pad46 under nonparametric
regression analysis and multiple correlation differences.
Morphological description.
Group vaginal administration of 25ug:
Four females, 8 pups was obtained in a single pregnancy, fetal resorptions not found in any of the females. It
was found microtia right in an individual (Photo No. 1), left microtia and forelimb left thicker than the right
in another (Photo No.2), umbilical hernia in another individual (Photo No.3) and a fetus tetraphocomelia and
caudal regression - sirenomelia (photo No. 4). The nuclei of the seventh pair had normal histological
appearance in all individuals, applying statistical logistic regression test for this treatment, no significant
differences were found because 100% of the sample population of this group qualifies as valid (presence of
the core the seventh cranial nerve)48

Figure No. 5. Right side view, specimen with right microtia, compared to the location of the right ear of a
normal individual of the same litter.

Figure No. 6. (A). Specimen left microtia and also with (B). The thicker than the right (C) left upper
extremity compared with an individual of the same normal gestational age ventral disposition of the
upper limbs.

Figure No. 7. On the left herniation of viscera ( 1) is observed , a situation that is not characteristic of
this period of development, presence of redoubt of the tail (2) and the lateral margin of the
anterolateral abdominal wall defect denoting closing the cavity (3). The photo on the right shows a
normal fetus of the same litter with integrated abdominal wall (1) and omphalomesenteric normally
implanted conduit (2).

Figure No. 8. The photograph on the left shows a fetus of 15 days gestational age (TH = 24), which is
estimated physical characteristics corresponding to a specimen tetraphocomelia and caudal
regression (sirenomelia) - fixed in formaldehyde . The photograph on the right corresponds to the
same specimen after fixation in picric acid, which allows better detail (1) forebrain area, (2) maxillary
area, (3) mandibular area , (4) abdominal region (5) Region flow (6) Shoulder.

Figure No. 9. Histological sections of brain stem level bulboprotuberancial groove for specimens
subjected to 25 mcg of vaginal misoprostol. (A) Specimen with right microtia. (B) Right upper
extremity specimen thicker than the left and left microtia. (C). Specimen with umbilical hernia. (D).
Control specimen. - The numbers represent the following structures: (1) Kernels of facial, (2) Fourth
ventricle, (3) Cerebellum. (40X)
Group oral administration of 25 ug
Three females, 38 calves in total fetal resorptions not found in any of the females was obtained. 2
fetuses with malformations was found in the right upper limb , one of the individuals has focomelia of
right upper limb with mild and severe mesomelic commitment acromlico segment; and the other ,
focomelia with severe compromise of mesomelic and acromlico segments (photo No. 6) . The nuclei
of the seventh pair had normal histological appearance in all individuals, applying statistical logistic
regression test for this treatment , no significant differences were found because 100 % of the sample
population of this group qualifies as valid ( presence of the core the seventh cranial nerve)48

Figure No. 10 (A) shows a fetus that has focomelia of right upper limb with mild and severe mesomelic
commitment acromlico segment. (B) Fetus Focomelico with severe compromise of mesomelic and
acromlico segments (C) normal fetus of the same gestational age (Estereoscope at 40X).

Figure No. 11. Cortes brainstem level bulboprotuberancial groove for specimens exposed to 25 mg of
vaginal administration of misoprostol. (TO). Meromelia specimen. (B). Focomelia specimen - a small
pyknotic focus is observed near the nucleus of the facial. (C). Control- specimen (1). Facial nuclei. (2) Fourth
ventricle . (3) Cerebellum. (40X). All are the core of the seventh cranial nerve.
Group vaginal administration of 50 ug
2 females, 15 calves in total was obtained, no fetal resorptions were found in any of the females , as
malformations were not found . The nuclei of the seventh pair had normal histological appearance in all
individuals. They also exhibited a pattern of normal surface vascularization and exhibited characteristics of
a fetus at 15 days of gestation morphological characteristics , such as prosencephalic eminence , defined
upper and lower limbs , primordia visible nails on toes , five rows of visible vibrisas , presence of hair follicles
in the head and jaw region 53 (Photo No.12) .

Just as the other groups, the nuclei of the seventh pair for 15 specimens of this group had normal
histological appearance. Therefore by applying statistical regression test logstica48 for this treatment, no
significant differences and dichotomy in this group were found, because 100% of the sample population
qualified as valid (core presence cranial nerve VII).
2 females, 32 fetal resorptions was obtained, which are characterized by the cylinder egg and fetal
membranes encapsulating the gastrular structure. 3 females, 45 fetuses were obtained, no fetal resorptions
were found in any of the females, as malformations were not found, Nuclei seventh pair had normal
histological appearance in all individuals. Therefore, applying statistical regression test logstica48 for this
treatment, 100% of the sample population qualified as valid (core presence cranial nerve VII) with no
significant differences and dichotomy in this group.
8 females, 108 fetuses were obtained, no fetal resorptions were found in any of the females, as
malformations were not found, Nuclei seventh pair had normal histological appearance in all individuals.
Therefore, applying statistical regression test logstica48 for this treatment, 100% of the sample population
qualified as valid (core presence cranial nerve VII) with no significant differences and dichotomy in this
group.
DISCUSSION OF RESULTS
Animal models are widely used for screening and evaluating new drugs in various phases (pre-clinical and
pharmacovigilance) 56 and its availability has made possible many advances in pharmacology and embriotoxicologia 56, 57. It is clear that the greater the degree of similarity between the animal experimental
model with a human disease, the greater the possibility of understanding the pathophysiology and
developing new drugs; and in this case, the greater the possibility of identifying the aspects that trigger a
disease or a pattern of birth defects that occur as a result of the administration of a developmental toxicant.
The 8.5 day gestation in white mouse (Mus musculus), is a time of prenatal development that belongs to
the embryonic period and could be equivalent to 23 to 28 gestational period humano53. During this stage
are already established a total of 14 to 16 pairs of somites, the presence of the first and second branchial
arch evidence, began the development of the limb buds and has already completed the closure of the
abdominal wall. Also in this period rombmeras 4 and 5, which are important elements in the development
of the nuclei of the seventh cranial (facial) 52, 53 are formed.

Concerning the percentage of pregnancies was achieved during the experiments, it was found that this was
low, amounting to 7.2% compared Reports Paumgarten, 24 who found in a series of 57 pregnant females
80% broods his Sample population present at Time of practicing them cesarean section on day 18 (Theiler
Stage = 24) 49 of gestational period a dose of 20 to 30 mg / kg of misoprostol, Reason why might consider
that misoprostol affects the number of pups of the Agreement dose used.
Fetal resorptions were found in litters significantly with intraperitoneal administration of prostaglandin
E263; prostaglandin E1 59, 60, 61, 62, but no air the exhibition A misoprostol, and This finding relates to the
dose used, regardless of the route of administration, since alone were observed in the experimental group
of 50 mg of oral administration and not in the other litters analyzed in our experimental model.
Malformations in fetuses found were similar to those reported in Paumgarten24 series of misoprostol and
prostaglandin E2 Persaud with TVN63. In this work, a different phenotype as meromelias that had not been
reported before with these medications was presented, but that literature if documents when a pregnant
woman uses the drug misoprostol as an abortifacient maneuver during the first quarter of gestacin.1,8,9,14,18,20,21,22,23,35,36,37; the sirenomelias not been associated with the use of misoprostol to
date, despite the possible etiology of vascular disruption may be associated with misoprostol in our estudio
38,39,40,41,42,43.
During the development work, the size was considered as a possible indicator of developmental toxicity,
data that has not been taken into account by any other author regarding the issue, because this is a
morphometric and meristic variable that is not specific protocols experimental work done with
prostaglandins, which to date have focused on acute and chronic toxicity, subacute and not the effect of
these segment II studies on reproduction. Due to the homogeneity of this parameter in normal fetuses
(with very little variation from the average) and considering that none of the experimental groups was able
to reproduce this feature of the control group, we can infer that this variable is very susceptible to alter and
a downward trend size related to high doses and is appropriate to consider it as a new indicator of
embryotoxicity for further study found.
The present study was motivated by the association between central lesion of cranial nerve VII described in
infants exposed to misoprostol in its embryonic stage and this finding a situation that had not been evaluated in studies with experimental animal models from administration other prostaglandins, which under the
criterion of temporality of association were engaged to be tested doses at various critical moments in the
development of rats and mice, but not for the purpose of associating the development of the VII pair,
however, our investigation found no histological alterations or absence of this structure at doses that fetuses exposed.
The use of misoprostol as a pharmacological method to cause abortion is a public health problem, that not
only should be discussed with legal, ethical and moral approaches, but as a possible source of an epidemic
of birth defects high social cost, if part the fact that these children are fruits of pregnancies in which there
was a motivation to be rejected, and that malformations at birth are an additional risk for future affective in
most cases. Therefore, the findings of this research, along with other studies in animal models and reports in
humans are sufficient evidence to consider misoprostol as a cause of preventable birth defects, so you can
identify the drug misoprostol is teratogenic and embryotoxic when a patient is administered during the first
trimester of pregnancy.

BIBLIOGRAPHY
1. Pastuszack Anne L, et al. Use of misoprostol and Moebius Syndrome During Pregnancy in infants. NEJM.
1998. Vol 338: 1881- 1885.
2. House Committee on Government Reform and Oversight Off - Label drug use and FDA review of
supplemental drug applications: hearing before the Subbcomitte on Human Resources and
Intergovernmental Relations. 10th Congress, 21nd sessions, September 12, 1996: 53-94.
3. Isaza, Carolina, Wilmar Saldarriaga, Reggie Garcia, Maria Cristina Lopez, Igncio Zarate. Misoprostol risk
when it fails as an abortifacient in the first quarter. Health- UIS magazine. N.1 38. 2006 Vol. P: 66- 70.
4. Zucker, Ronald, theories explaining Moebius Syndrome. Hospital for Sick Children of Toronto, Canada.
1986. In press.
5. Van Der Zwaag, HTFM Verijl, D - Beltran - Valero De Barnabas, et al. Mutation analysis in the candidate
genes Mobius syndrome PGT and GATA on cromosome 3 and EGR on cromosome 10. J. Med. Genetic. 2003.
p: 1-32
6. - Refaey H, Templeton A. Induction of abortion in the second trimester by combination of Misoprostol and
mifeprestone: a randomized comparison Between two misoprostol regimens. Hum.Reprod.1995; 10:
475-478.
7. Akdamar K Agrawal N, Ertan A. Inhibition of nocturnal gastric secretion in the normal human volunteers
by misoprostol, a synthetic prostaglandin E1 methyl ester analog. Am. J. Gastroenterol. 1982; 77 (12): 902
-904.
8. Alan Guttmacher Institute. Issues in brief. An overview of clandestine abortion in Latin America, http:
//www.guttmacher. Accessed rglpubslibl2.htmi.May 2002.
9. Costa SH, Vessey MP. Misoprostol and illegal abortion in Rio de Janeiro, Brazil. 1993 Lancet. 341 (8855):
1258-1261.
10. Faundes A, Santos LC et al .. Post-abortion complications after interruption of pregnancy with Misoprostol. 1996. Advances in Contraception.12 (1): 1-9.
11. Bugalho A, Bique C, Almeida L. Application of vaginal Misoprostol before cervical dilatation to provide
first trimester pregnancy interruption. Obste Gynecol. 1994; 83: 729-731.
12. Gonzlez Garamendi, Landa Tabuyo. Misoprostol as an abortive in Spain. On the subject of a court case.
Journal of Forensic Medicine No. 38. October 2004.
13. Teratology Society Public Affairs Committee. FDA classification of drugs for teratogenic risk. Teratology
1994, 49 (6): 446-447
14. Gonzalez CH, FR Vargas, Perez ABA, Kim Ca, Brunoni D, Marques - Dias MJ, et al. With or without limb
deficiency mobius sequence in seven Brazilian children Associated with Misoprostol use in first trimester of
pregnancy. Am. J. Med. Genet. 1993; 47:54 - 64.
15. Genest DR, Richardson A. et al. Limb defects, and omphalocele in a 17 week fetus first trimester misoprostol following exposure. Teratology.1994. 49 (5): 418.
16. Hofmeyr GJ. AM Gulmezoglu. Vaginal Misoprostol for cervical ripening and induction of labor. The
Cocharane Library, Issue 1. Oxford: Update Software; 2002

17. Schuler L, Pastuszak A. et al. Pregnancy outcome after exposure to Misoprostol in BRAZIL- A prospective, controlled study. Reproductive Toxicology. 1999. 13 (2): 147-151
18. Gonzalez CH, Marques-Dias M. J. et al. Congenital abnormalities in Brazilian children Associated with
Misoprostol misuse in first trimester of pregnancy. Lancet. 1998. 351 (9116): 1624-1627.
19. Coelho KE, MF Sarmento et al. Misoprostol embryotoxicity: Clinical evaluation of fifteen Patients With
arthrogryposis. American Journal of Medical Genetics. 2002. 95 (4): 2197-301.
20. Pastuszack AL, L Schuler et al. Use of misoprostol and Moebius syndrome During Pregnancy in
infants. New England Journal of Medicine. 1998. 338 (26): 1881-188.
21. Teratology Society Public Affairs Committee. Of Clasifications drugs for teratogenic risk. Teratology.
1994, 49 (6): 446-447.
22. I. M Orioli, Castilla EE. Epidemiological assessment of misoprostol teratogenicity. British Journal of
Obstetrics and Gynecology. 2000. 107 (4): 151-523 5.
23. Vargas FR, Schuler-Faccini L et al. Prenatal exposure to misoprostol and vascular disruption defects: A
case-control study. American Journal of Medical Genetics. 2002. 95(4): 302- 306.
24. Paurmgartten FJR, Magalhaes-de-Souza CA et al. Embryotoxic effects of Misoprostol in the mouse.
Brazilian Journal of Medical and Biological Research. 1995. 28(3): 355-361.
25. Shepard TH. Proof of human teratogenicity. Teratology. 1994. 50(2): 97-98.
26. Downie WW. Misuse of misoprostol. Lancet. 1991. 338(8761): 247.
27. Sifton DW, ed. Physicians Desk Reference, 57th ed. Montvale, NJ: Medical Economics Company. 2003.
28. MacKenzie WE. Misoprostol and the politics of fear. Lancet 2001; 357 (9264):1296.
29. Nutting, E. F. And P. F. Cammarata. Effects of Prostaglandins on fertility in female rats, 1999 Nature,
222: 287.
30. Fonseca W, Alencar AJ et al. 1993. Congenital malformation of the scalp and cranium after failed first
trimester abortion attempt with misoprostol. Clinical Dysmorphology. 2(1): 76- 80.
31. Palmer CA. Mebius syndrome. EMedicine Journal. 2(1), www.neuro/topic6l2.htm, accessed May
2002.
32. Fontelle Luca, Alexandra Prufer De Q.C, Araujo, Rosiane S. Fontana. Sndrome de Moebius, relato de
caso. Arquivos de Neuro psiquiatra. 2002. Vol.59 no 3B Sao Paulo.
33. Perez - Torrero E, Torrero C, Salas M. Neonatal nutricional rehabilitation of morphological features in
facial motoneurons altered by prenatally food deprivation in the rat. Nut. Neurosciences. 2002. 5 (4):
263-268.
34. Goodard, Judy, M, Mireille Rossel, Nancy R, Manley; Mario R.Capechi, Mice with targeted disruption of
Hoxb-1 fail to form the motor nucleus of the VII nerve. Development, 1996. 122, 3217 3228.
35. Barrow, R.Jeffrey; Mario R. Capecchi. Targeted Of disruption of the Hoxb-2 locus in mice interferes
with expression Hox B1 and Hox B4.Development. 1996. 122. 3817-3828.
36. Stevenson Re, Lyons Jk, Phelan Mc, Jones Mc, Barr M, Clericuzio C, Et Al. Vascular steal: the pathogenetic mechanism producing sirenomelia and associated defects of the viscera and soft tissues. Pediatrics

37. Lipson AH, Gillerot Y, Tannenberg AEG. Two cases of maternal antenatal splenic rupture and
hypotension associated with Moebius Syndrome and cerebral palsy in offspring; further evidence for a
utero placental vascular aetiology for Mobius Syndrome and some cases of cerebral plasy. Eur J.
pediatr. 1996; 155-800-4
38. Shepard TH. Personal correspondence. Article to be published in Teratology 2002. With coauthors
Robert Brent, Jan Friedman, Ken Jones, Richard Miller, Janine Polifka, and Cynthia Moore.
39. Fonseca W, Alencar AJ et al. Misoprostol and congenital malformations. Lancet. 1991.
338(8758):56.
40. Graf WD, Shepard TH. Uterine contraction in the development of Mbius syndrome. Journal of
Child Neurology. 1997. 12(3): 225-227.
41. Persaud, T.V.N. Pregnancy and progeny in rats Treated With prostaglandin E1. Rev. Prostaglandins.
March 1973, V3, N3.
42. C. W. Jackson and T.V.N. Persaud. Pregnancy and progeny in rats Treated With prostaglandin A1.
Teratology, Research Laboratory, Department of Anatomy, University of Manitoba, Winnipeg, Canada.
Thesis: 1976. 95: 40-49
43. Suzanski. T. W. & T. V .N. Persaud. early embryonic development in rats after maternal treatment
with PGE2. Department of Anatomy, University of Manitoba, Faculty of Medicine and Dentistry, Winnipeg, Manitoba, R3E OW3, 1982.Canad. P401 -410.
44. Esaki, K, Sasa H, Umemura T. Effect of intragastric of Misoprostol dispersions on reproduction in
rats and Mouse II. Experiment on drug administration in the pre and early gestation periods. Jitchuken
Zenrinsho Kenkyuho. 1985. 11: 167-187.
Evoy 45.Mc G.K. American Hospital for service, information and drug formulation. 1996. Bethesta, MD,
217.
46. Graph Pad Software. Prism. References Windows or Macintosh, Parametric and Parametric Statistical No. Version GPW4- 12345-12356. 2002.
47. Daniel, Wayne W. Biostatistics: basis for the analysis of health sciences. 2002. 3rd Edition. Mexico:
Editorial Limusa Wiley.
48. Statistical Package SS. Pluss. 6.1. Innsightful Corporation W.DC. Seattle Washintong, 1998-2003
Logistic Regression. Dicotomic variably.
49. Jonathan B. L. Bard and Mathew H. Kaufman. The Development life of table Mouse. Department of
Biomedical Sciences, University Edinburgh, United Kingdom. Academic Press.1999. 256P
50. CECSA. Biology practices - keeping model white mice Mus musculus environments Vivarium, 1986.
1st ed. Barcelona, Spain.
51. Whitten, W. K. Occurrence of anestrous in caged mice in group. J. Endocrinology.1959; 18: 102-107.
52. http://genex.hgu.mrc.ac.uk/Databases/Anatomy/new/ - Theiler Stage, to map mouse development. Univ. Edinburgh 2001, U.K.
53. Kauffman, H. Mathew, Jonathan B. L. Bard. The Anatomical Basis of Mouse. Development. Academic Press.1999. San Diego California. pp: 298

54. Zuiga Jesus, Mary Joseph Tur, Silvana N, Miloco. Science and Animal Experimentation. 1. Edition.
Editorial Mc Graw Hill - Interamericana. 2001. Experimental Surgical Procedures. PP: 361 -513.
55. Schambra Uta, Jean M. Lauder, J. Silver. Atlas of the Mouse Brain Prenatal. 1992. Academic Press, Inc.
San Diego California. P: 510
56.Ramrez, Jorge H, et al, Lack of correlation between pressure values, the target organ damage and the
presence of retinopathy in the animal model of hypertension with L- name, Medical Colombia, OctoberDecember, 2006, Vol 37, N.004, Universidad del Valle, Cali, Colombia.
57.Sun ZJ, Zhang Ze. Historic perspectives and recent major advances in the animal models of hypertension. Acta Pharmacol. Without. 2005. 26: 295-301.
58. OECD. 415 - Repeated Dose 28-day Oral Toxicity Study in Rodents. OECD, Paris, France. Guidelines for
Testing of Chemicals; 1999. Report No .: 407.
Fonseca 59. W; Alencar A. J. C; Motta Coelho F. S. B. and H .L. L. Misoprostol and Congenital Abnormalities.
1991. Lancet, 338-356.
60. Kotsonis F.N. Dood D.C., F. E. Regnier B. Kohn Preclinical Toxicology profile of Misoprostol. Digestive
Diseases Sciences, 1985. Supply 30. 142S - 146S
61. Theil R, Chung M, Pastor C, G & Chahoud Bochert I. (1993). Evaluation of historical monitoring data:
correlation and fetal skeletal ossification Between weight in Han: NMRI - mice and Wistar Rats. Teratology,
48: 35A, _____ quoted by Paumgartten, F. J. R; Magalhaes- De Souza; Carvalho, R.R. (nineteen ninety five).
Embriyotoxic effects of Misoprostol in the Mouse. Brazilian Journal of Medical and Biological Research .. 28:
355-361.
62. Persaud T.V.N. Reproductive and teratological studies With prostaglandins. In Persaud T.V.N. (Editor),
Advances in the study of Birth Defects. Vol. 2 (teratological Testing). MTP Press Limited, Lancaster, 1979.
161 - 177.
63. Persaud T.V.N. The Efects Of Prostaglandin E2 Embrionic On Pregnancy And Development In Mice.
Toxicology, 1975. (5): 97-101.