Ischemic carotid endothelium. Transmission electron microscopic studies

The endothelium of monkey and rabbit common carotid arteries subjected to ischemia was examined by transmission electron microscopy (TEM). The right carotid artery of 24 rhesus monkeys was occluded by proximal and distal placement of removable surgical clips for periods ranging from five minutes to four hours. A single clip was used to occlude the right carotid artery of 15 rabbits for periods ranging from 5 to 30 minutes. With TEM, numerous blebs, intracytoplasmic vacuoles, membranous whorls, and pseudopodia were found in the endothelium of arterial segments subjected to ischemia by double or single clipping for as little as five minutes. Following occlusion of one hour or longer, disruption of interendothelial junctions was also noted. These TEM findings were compared with earlier TEM studies of the response of endothelium to other injurious stimuli and with previous scanning electron microscopic studies in which the same ischemic models were utilized.     

Carotid artery balloon test occlusion: combined clinical evaluation and xenon-enhanced computed tomographic cerebral blood flow evaluation without patient transfer or balloon reinflation: technical note

OBJECTIVE: Clinical evaluation was combined with xenon-enhanced computed tomographic (CT) cerebral blood flow (CBF) evaluation during carotid artery balloon test occlusion (BTO), without patient transfer from the angiography suite to the CT scanner or balloon reinflation. TECHNIQUE: Thirteen patients underwent carotid artery BTO. Placement of temporary occlusion balloons was performed with patients positioned on the CT scanner table. If neurological testing revealed no changes within 10 minutes after balloon inflation, patients were positioned within the CT scanner gantry for xenon-enhanced CT CBF evaluation. CBF evaluations were begun 12 to 15 minutes after balloon inflation and required 8 minutes for completion. After completion of CBF evaluation, neurological testing continued during 30 minutes of arterial occlusion. RESULTS: One patient did not tolerate BTO, with the development of reversible hemiparesis. Reliable CBF data were not obtained because of patient motion in one case. Eleven patients clinically tolerated BTO and completed CBF evaluation. For five patients, xenon-enhanced CT scanning revealed no regions with CBF of less than 30 ml/100 g/min. For four patients, xenon-enhanced CT scanning revealed small regions with CBF of less than 30 ml/100 g/min within the anterior frontal lobe on the occluded side. For two patients, ipsilateral CBF decreased dramatically during BTO, with CBF in many regions of less than 30 ml/100 g/min and in some of less than 20 ml/100 g/min. CONCLUSION: Xenon-enhanced CT CBF evaluation can be combined with clinical testing during BTO without patient transfer, balloon reinflation, or increases in the duration of the procedure. We recognize that the value of CBF evaluation during BTO remains to be proven; our technique does, however, eliminate abbreviated clinical neurological evaluation, patient transfer, and balloon reinflation, which were previously associated with the use of xenon-enhanced CT CBF evaluation during carotid artery BTO.     

SEM evaluation of endothelial changes following end-to-side microvascular anastomoses in rats

A scanning electron microscopic (SEM) study was carried out to evaluate the endothelial changes in carotid end-to-side microsurgical anastomoses. This procedure, associated with occlusion of the brachiocephalic trunk, was performed in 24 Sprague-Dawley rats. The animals were sacrificed at time intervals ranging between 15 minutes and 14 days after surgery. The specimens were collected after perfusion-fixation in vivo and treated for SEM observations. The findings show that the first 45 minutes are the most critical time for thrombus formation. Reendothelialization could be detected at 3 days after the microsurgical procedure. Endothelial regeneration occurred earlier on the suture line than on the stitches, documented by the migration and morphological changes of the endothelial cells advancing from the normal endothelium to cover the defect and the suture material. The repair was usually completed within 14 days. Damage of the inner vascular wall, related to minor failures of the microsurgical technique, appears to be a cause of delayed reendothelialization.     

Thomas G. Orr Memorial Lectureship. The general, general surgeon

BACKGROUND: The obvious advantages of rapid arterial anastomoses have prompted the continuing search for more rapid anastomotic techniques to complement the standard sutured anastomosis. Nonpenetrating, arcuate-legged titanium vascular closure staple (VCS) clips were initially developed for microvascular anastomoses. The purpose of this study was to compare VCS clips with sutured arterial end-to-end anastomosis in larger vessels. METHODS: In 6 pigs, transacted iliac arteries were reanastomosed with VCS clips on one side and continuous 6-0 polypropylene suture on the other. RESULTS: The reconstruction time was 8.4+/-5.2 minutes for clip closure and 12.0+/-6.6 minutes for suture closure (P = 0.033). All vessels were patent half an hour after completing the anastomoses with no signs of early thrombosis. CONCLUSIONS: Arterial end-to-end anastomosis can be performed more rapidly with VCS clips than continuous sutures, and are potentially useful in situations where the clamp time of the vessel is critical.     

A combined microsurgical skull-base and endovascular approach to giant and large paraclinoid aneurysms

BACKGROUND: The treatment of giant and large paraclinoid aneurysms remains challenging. To improve exposure, facilitate the dissection of aneurysms, assure vascular control, reduce brain retraction and temporary occlusion time, enable simultaneous treatment of associated lesions, and achieve more successful treatment of "difficult" (atherosclerotic and calcified) aneurysms, we combined the skull-base approach with endovascular balloon occlusion of the internal carotid artery (ICA) and suction decompression of the aneurysm. METHODS: Sixteen female patients were treated, eight with giant aneurysms and eight with large aneurysms. Eight aneurysms occurred on the right side and eight on the left. Eight patients had an additional aneurysm; five were clipped during the same procedure. Three patients had infundibular arterial dilation. One patient had an associated hemangioma of the ipsilateral cavernous sinus. The cranio-orbital-zygomatic approach was used for all patients. The anterior clinoid was drilled, and the optic nerve was decompressed, dissected, and mobilized. Transfemoral temporary balloon occlusion of the ICA in the neck was followed by placement of a temporary clip proximal to the posterior communicating artery. Suction decompression was then applied. All aneurysms were then successfully clipped, except one that had a calcified neck and wall that could not be collapsed. Intraoperative angiography performed in 13 of 15 patients with clipped aneurysms confirmed obliteration of the aneurysm and patency of the blood vessels. RESULTS: Postoperative results were good in 14 patients. One patient had right-sided hemiplegia and expressive aphasia, which improved after rehabilitation. One patient with an additional basilar tip aneurysm clipped simultaneously died on the fifth postoperative day because of intraventricular hemorrhage. The origin of bleeding could not be determined on autopsy. Surgical difficulties and morbidity stemmed mainly from a severely calcified or atherosclerotic aneurysmal neck. CONCLUSION: The combination of skull-base approaches and endovascular balloon occlusion coupled with suction decompression is a successful option for the treatment of these challenging aneurysms.     

Chemical synthesis of D-ribo-phytosphingosine-1-phosphate, a potential modulator of cellular processes

OBJECTIVE: The nonpenetrating, arcuate-legged clip has proved its ability to provide a high-quality microvascular anastomosis. This study assessed the feasibility of constructing a coronary end-to-side anastomosis on the beating heart with a novel mechanical, sutureless anastomotic device that applies 12 circumferential clips simultaneously. METHODS: In 14 consecutive pigs (70-90 kg), the left internal thoracic artery (diameter, 3 mm) was grafted to the left anterior descending coronary artery (diameter, 3 mm) by means of a one-shot anastomotic stapler prototype. Endothelial denudation, medial necrosis, and intimal hyperplasia were analyzed quantitatively and compared with those seen in conventionally sutured anastomoses (n = 4). RESULTS: In 8 of 14 anastomoses, the one-shot anastomotic stapler successfully applied all 12 clips circumferentially across the everted arteriotomy edges. In the remaining, either 1 (n = 4) or 3 and 4 adjoining malaligned clips had to be replaced manually with a single-clip applicator. Coronary occlusion was limited to approximately 3 minutes. At follow-up, all anastomoses were patent angiographically. At 2 days, in 2 of 7 cases, a local coronary dissection was observed, and there was a considerable loss of endothelial cells and medial damage. At 28 days, however, minimal intimal hyperplasia was seen at the anastomotic lining, although more pronounced when compared with conventionally sutured anastomoses. CONCLUSIONS: The one-shot anastomotic stapler prototype enabled short-occlusive (3 minutes), sutureless end-to-side grafting on the beating porcine heart. In spite of early endothelial and medial damage and 2 local dissections, all anastomoses remained patent with minimal intimal hyperplasia at 4 weeks.     

Complement activation occurs on subendothelial extracellular matrix in vitro and is initiated by retraction or removal of overlying endothelial cells

Vascular endothelium is continuously exposed to plasma complement, which could generate a potent proinflammatory signal if activated on the vascular wall. Normal endothelium, however, expresses an anti-inflammatory phenotype, which includes resistance to complement fixation. As activated endothelium converts to a proinflammatory phenotype, we investigated the effect of cytokines on endothelial susceptibility to complement fixation. Cytokine-treated HUVEC were exposed to human serum as a source of complement, and C3 deposition was quantified. IL-1beta and TNF-alpha in combination with IFN-gamma markedly increased endothelial C3 deposition; however, immunofluorescence microscopy revealed that the endothelial cells had retracted, and that bound C3 was concentrated not on cells but in areas of exposed subendothelial extracellular matrix (ECM). Studies with cell-free ECM indicated that complement activation required only ECM exposure and was independent of cellular activation. C3 deposition on ECM was reproduced by reconstituting the alternative pathway, which generated a stable C3 convertase on ECM, but not on endothelial cells. C3b and iC3b were identified on ECM exposed to purified alternative pathway components and serum, respectively. In conditions associated with endothelial disruption, exposure of subendothelial ECM could induce complement fixation and contribute to inflammation and vascular damage.     

Risks and benefits of diagnostic angiography after aneurysm surgery: a retrospective analysis of 597 studies

INTRODUCTION: Cerebral angiography performed after aneurysm surgery can identify causes of morbidity and mortality that may be corrected. The risks and benefits of angiography that is performed after aneurysm surgery, however, have not been clearly defined. We therefore reviewed our experience with postoperative angiography to determine its dangers and benefits. METHODS: During 10 years, 543 consecutive patients received treatment for cerebral aneurysms. A retrospective analysis of 597 diagnostic angiograms obtained after aneurysm surgery for 494 of these patients was performed. RESULTS: Catheter-induced vessel spasm and dissection, occurring most frequently in the internal carotid artery, were observed in seven (1.2%) and six (1%) studies, respectively. No angiography-associated strokes were identified. No association between age, smoking, hypertension, blood pressure, atherosclerosis, or severe vasospasm and angiographic complications was observed. Aneurysm remnants were identified in 36 (5.7%) of the 637 aneurysms that were surgically treated. Atherosclerosis (P < 0.01) or multiple clip applications (P < 0.01) were significantly associated with aneurysm remnants. Angiographic vessel occlusion was observed in 28 (5.7%) patients and resulted in stroke in 14 of these patients. Vessel occlusion was significantly associated with increasing aneurysm size (P < 0.001), atherosclerosis (P < 0.001), temporary clips (P < 0.001), multiple clips (P=0.03), multiple clip applications (P=0.001), and a new postoperative neurological deficit (P=0.002). Severe vasospasm and newly identified aneurysms were observed in 51 and 16 patients, respectively. CONCLUSION: Angiography after aneurysm surgery is safe and can be routinely performed. Angiography after aneurysm surgery should be particularly considered for patients with large aneurysms or cerebrovascular atherosclerosis and for those who develop new postoperative neurological deficits.     

Early detection of colonic cancer by colonoscopy

Experimental arterial occlusion was performed on eight dogs by means of inflatable and releasable balloons, in five of which a reliable and long lasting arterial occlusion was obtained. Three accidents occurred. One dog died when the liquid latex was emptied spontaneously from the balloon and the territory was embolized causing a severe general reaction. In two other dogs the emptying of the balloon, inflated with iodine, was followed by distal embolization of the empty latex balloon. The possibility of such accidents would be too great a risk for human application, but it now seems possible with the new glue, isobutyl-2-cyanoacrylate, to embolize vascular territories more accurately and more safely by means of inflatable and releasable balloons.     

Intractable pelvic pain following Filshie clip application

Presented is the case of a 37-year-old South Australian woman who experienced intractable pelvic pain following laparoscopic sterilization with Filshie clips. The pelvic anatomy was normal and one Filshie clip was applied to each Fallopian tube. The patient stated she had experienced right-sided lower abdominal pain that radiated down the anterior part of her right thigh since regaining consciousness after general anesthesia. The pain had failed to resolve seven days after the procedure and the patient was unable to perform even simple tasks. Analgesics provided only temporary, partial relief. There were no signs of infection or any other exacerbating condition. At diagnostic laparoscopy, instillation of bupivacaine around the clip provided transient relief, but the pain returned the next day at the same level of severity. After one month of intractable pain, laparoscopic bilateral salpingectomy was performed to remove the clips and the pain disappeared. Although back pain has been reported in up to 14% of women undergoing laparoscopic sterilization, this is the first published case of long-term abdominal pain associated with the Filshie clip.     

Embolization with temporary balloon occlusion of the internal carotid artery and in vivo proton spectroscopy improves radical removal of petrous-tentorial meningioma

A highly vascular petroclival meningioma supplied by tentorial branches of the internal carotid artery was embolized by temporary balloon occlusion of the parent vessel distal to the tumor, followed by obliteration of the tumor vascularity with polyvinyl alcohol particles. Subsequently, in vivo proton spectroscopy showed necrosis of a large portion of the tumor and helped determine the timing of surgery. Both innovative techniques considerably facilitated the subsequent radical excision of the tumor with no neurological morbidity.     

Brain lipid peroxidation and antioxidant levels in fetal lambs 72 hours after asphyxia by partial umbilical cord occlusion

Giant middle cerebral artery (MCA) trifurcation aneurysms that cannot be excluded directly can be treated by flow inversion achieved by creation of an extracranial-intracranial bypass distal to the aneurysm, followed by occlusion of the parent vessel proximal to the aneurysm. As opposed to surgical occlusion, endovascular occlusion avoids dissection of the aneurysm area, and the site of occlusion can be chosen according to the flow distribution demonstrated on angiography performed during test occlusions. Two patients with giant aneurysms of the MCA trifurcation benefited from flow inversion treatment. Forty-eight hours after an MCA-superficial temporal artery bypass had been created, the M1 segment was occluded by inserting a coil in the first patient and the internal carotid artery was occluded with balloons in the second patient (there was no communicating artery in the latter case). Both occlusions were performed immediately after a clinical test of occlusion tolerance. The patients were clinically intact during the postoperative course. Follow-up angiography performed 11 and 4 months, respectively, after vessel occlusion showed that the aneurysm occlusion was stable.     

Novel technique and instrumentation for laparoscopic application of hemostatic clips

A novel device for applying hemostatic clips in laparoscopic surgery incorporates a distal hook into a multiple-firing titanium clip applier. The hook may be used for blunt dissection of tissue, and to displace and control ducts and vessels during clip application. A single instrument may be used to achieve hemostasis in areas that are difficult to reach, and past pointing problems encountered with straight on clip appliers are alleviated. Comparative testing of the holding force of the curved clips used with this device versus the straight clips used in conventional multiple-clip appliers demonstrated a higher mean pull-off force of 0.473 lbs versus 0.33 lbs. Clinical application of the device in laparoscopic procedures including cholecystectomy, vaginal hysterectomy, Nissen fundoplication, vagotomy, varicocelectomy, and lymphadenectomy show the utility of the hook clip applier.     

Facilitation of Chemoreceptor-induced reflex vasoconstriction by intravertebral arterial administration of clonidine

The influence of clonidine on the reflex vascular responses to stimulation of carotid body chemoreceptors and bilateral carotid occlusion was studied in morphine, chloralose-urethane anesthetized dogs. Bilateral carotid occlusion and intracarotid injection of nicotine (30 and 100 microgram) or sodium cyanide (200 and 500 microgram) elicited reflex vasoconstriction in the perfused gracilis muscle vascular bed. Infusion of clonidine (2-4 microgram/kg) into the vertebral artery significantly lowered blood pressure. Reflex vasoconstrictor responses to chemoreceptor stimulation were significantly enhanced after clonidine administration whereas reflex vasoconstrictor responses to carotid occlusion were markedly reduced. The facilitation of chemoreceptor reflex responses by clonidine was observed in dogs with intact or sectioned vagi and in animals in which the carotid arteries were perfused at constant blood flow. Inhibition of carotid occlusion responses by clonidine was observed in dogs with intact or sectioned vagi. Infusion of clonidine directly into the carotid arteries did not significantly alter responses to chemoreceptor stimulation. These experiments demonstrate that clonidine antagonizes the reflex vasoconstriction caused by carotid occlusion while potentiating the vasoconstriction elicited by chemoreceptor stimulation. The data suggest that clonidine exerts central actions which result in a facilitation of the chemoreceptor reflex and a simultaneously occuring hypotension which is probably due to an action on baroreceptor pathways.     

Refusion of anterior cerebral artery after aneurysm clipping--a case report

Herein, we are describing an unusual case suffering from a left anterior cerebral artery aneurysm (A1). Both the anterior cerebral arteries were supplied by the left internal carotid artery as was found in digital substraction angiography (DSA) preoperatively. The postoperative angiograms revealed that left anterior cerebral artery was supplied from the left internal carotid artery and the right anterior cerebral artery by the right internal carotid artery respectively. This finding of cerebral angiograms is interesting and rarely mentioned in the literature. Its hemodynamic change and pathogenesis were unclear and different to that of the coronary circulation, ischemic change of gut and skeletal muscle. The causes may include: 1) vasospasm at the anterior communicating artery after manipulation at surgery; 2) desiccation or shrivelling the adjacent artery by intraoperative electrocoagulation; 3) occlusion the anterior communicating artery by the wing of clip. 4) the deprivation of the blood flow from the left internal carotid artery after totally or partially narrowing left A1 by an aneurysm clip. The redistribution of blood volume in the previously hypoplastic right anterior cerebral artery and decreased caliber of the left anterior cerebral artery (A1) are likely playing a role in this case.     

Internal carotid artery occlusion related to seat belt shoulder strap: report of two cases

Two cases of traumatic internal carotid artery occlusion probably related to the seat belt shoulder strap are reported. Case 1. A 20-year-old woman was driving and was struck on the right front side of her car by another car. There were neither bruises, abrasions on her neck, nor weakness in her extremities. About 4 hours later, she developed left hemiplegia, and CT scan taken on the following day revealed low density areas in the capsulostriatal area on the right. The right carotid angiography revealed occlusion of the internal carotid artery about 3 cm distal to the bifurcation. Case 2. A 43-year-old man was driving and was struck on the front of his car by a hard iron railing. He sustained a sternum fracture, but there was no disturbance of consciousness or paresis of the extremities. His neck was unremarkable externally. About 50 days later, he developed left hemiplegia. CT scan and MRI revealed a massive infarction in the distribution of the right middle cerebral artery territories. The carotid angiography revealed occlusion of the right internal carotid artery about 3 cm distal to the bifurcation. In each cases, the driver was wearing a three-point shoulder seatbelt when the car was struck on the front or on the right front. Previous experimental studies have revealed in these situations the neck is flexed right anteriorly, and then quickly overextended left posteriorly. The overextension of the neck probably injured the intima of the internal carotid artery ipsilateral to the shoulder fixed in the seatbelt, resulting in the subsequent occlusion by a thrombus.     

Arteriovenous fistulae complicating central venous catheterization: value of endovascular treatment based on a series of seven cases

OBJECTIVES: To point out the usefulness and effication of endovascular treatment of arteriovenous fistulae complicating the insertion of central venous catheter. MATERIAL AND METHOD: Retrospective study of 7 cases treated between 1983 and 1993. The central venous catheterization had involved the right internal jugular vein in 5 cases and the right subclavian vein in 2 cases. RESULTS: Isolated endovascular therapy was performed in 5 cases. Surgery was necessary in 1 case to remove an hematoma. In another case, occlusion of a large AVF was performed during intraoperative endovascular temporary occlusion of the AVF. The choice of the occluding agent was primarily dictated by the hemodynamic and anatomical conditions (releasable balloons, coils). Vertebro-vertebral fistulae (3 cases) could be treated with patency of this vessel. No complications were encountered. CONCLUSION: Endovascular approach is a reliable and safe method to perform occlusion of post catheter AVF.     

A new balloon-controlled temporary vascular clip

We devised a temporary clip that is manipulated by inflation or deflation of a balloon. Prior application of this temporary clip to the proximal artery is convenient for controlling distal bleeding during microsurgery.     

Postischemic hyperglycemia worsens neurologic outcome after spinal cord ischemia

PURPOSE: The neurologic effect of induced hyperglycemia in the postischemic period was investigated with a rat aortic occlusion model. METHODS: Sprague-Dawley rats weighing 200 to 350 gm were anesthetized, intubated, and ventilated with 1% to 1.5% halothane. Temperature was continuously monitored and maintained at 37 degrees +/- 0.5 degrees C. The chest was opened, the thymus excised, and the aortic arch exposed. Snares were placed around the aorta distal to the left subclavian artery and the right and left subclavian arteries. The three vessels thus isolated were occluded for 8 minutes. With snare release and withdrawal, the rats received an intraperitoneal injection of 5% dextrose in water (2 gm/kg) or an equivalent volume of 0.9% saline solution. In a second group of rats the administration of glucose or saline solution was delayed until 30 minutes after snare release. Blood samples for blood glucose determination were obtained before operation, before occlusion, immediately after occlusion, and 15, 30, 45, 60, and 240 minutes after occlusion. A neurologic deficit score was assigned at 1, 4, 18, and 24 hours after occlusion to quantify hindlimb neurologic deficit based on 15-point scale (0 = normal, 15 = severe deficit). Sham-operated rats received the same operation and injection, but the snares were only manipulated and not made occlusive. RESULTS: The rats that were administered glucose immediately after snare release showed a statistically significant exacerbation of lower extremity neurologic deficit at 24 hours after occlusion (p < or = 0.05, Mann-Whitney U test). The sham-operated rats were normal (0 score) at 24 hours. Significant elevation of blood glucose (321 +/- 33 mg/dl) was seen in the glucose-injected rats at 15 minutes and continued for up to 4 hours after occlusion (p = 0.040 and 0.014, respectively; Student's t test). CONCLUSION: Postischemic hyperglycemia immediately after a standard spinal cord ischemic stress worsens neurologic outcome.     

Ultrastructural alterations caused by immunological reactions after intracardiac injection of allogeneic antibodies against blood group antigens: an experimental study using the in vitro whole-rat embryo culture

The effects of intracardiac injection of 0.5 microliter allospecific hemolyzing rat-antirat antibodies, directed against the blood group antigens, on the endothelium of the dorsal aortae were studied in 9-14 somite-staged Wistar and RIV:Tax rat embryos, using both transmission electron microscopy (TEM) and immunoelectron microscopy (IEM). In a TEM study it was further investigated if either apoptosis or cell necrosis occurred as a result of the forementioned intracardiac injection. The results were compared to ultrastructural findings of the dorsal aortae in sham- and noninjected rat embryos of the same gestational age. In the control rat embryos, the aortic vascular wall consisted of a single continuous layer of endothelial cells. No clear basal lamina was present in TEM. Furthermore, no immunoreactivity against the endothelium or the intravascular blood cells was noted. Embryos injected with hemolyzing rat-antirat antibodies displayed clefts or pores, and diaphragmatic fenestrations of the endothelial lining of the dorsal aortae after 2 hr. Alterations resembled those induced by vasoactive mediators such as histamine, serotonin, bradykinin, and prostaglandins. The above changes had disappeared 4 and 6 hr after injection with complete restoration of the endothelial lining. Immunogold staining demonstrated Ig depositions along the luminal side of the endothelium, in the vicinity of the intercellular spaces, and in the subendothelial space of the dorsal aortae. Numerous particles were seen located inside intracytoplasmatic vesicles, indicating involvement of transcytoplasmatic transport as well as intracytoplasmatic phagocytosis. Similar depositions were observed in and around intravascular embryonic blood cells. Apoptosis, or programmed cell death, an important component in immunological reactions, occurred in rat embryos injected with hemolyzing rat-antirat antibodies. The excessive amount of apoptosis seen in this study is in accordance with the pathogenetic cell degeneration found in our earlier studies. Cell necrosis was not observed. The results from this study indicate that the endothelium of the dorsal aortae and intravascular blood cells only display a transient reaction following injection with hemolyzing rat-antirat (RAR) antibodies. The temporary reaction is presumably due to the release of vasoactive mediators. The smaller vessels and capillaries are still in an earlier stage of development, displaying fenestration, making them more susceptible for injury after immunological interaction. The results are indicative that the pathogenetic effect of the immunological reaction after intracardiac injection takes place at the level of the microcirculation by "switching on" apoptosis. Programmed cell death is essential in embryogenesis and development. Therefore excessive apoptosis, i.e., inappropriate apoptosis, will eventually induce congenital malformations.