[Presence of microfilariae in the anterior chamber of the eye onchocerciasis]. Observation of a patient presenting 34 microfilariae in the anterior chamber of one eye, with negative biopsies, two small nodules and punctiform onchocercosis keratitis elementary microlesions. The patient has been observed five times during more than one hour each time. The author reviews the results of different surveys he has carried out in America and Africa, and which showed that the presence of microfilariae in the anterior chamber of the eye did not seem to forebode severe ocular lesions.


Can recurrent cholesteatoma be avoided?

Can recurrent cholesteatoma be avoided? After closed operation techniques recurrences of cholesteatoma are often caused by retraction of squamous epithelium under the bony annulus, under which it spreads into the attic or the mastoid cavity. Consequently, retractions under the bony annulus can be avoided, if the annulus is removed. An endaural operation technique is described in which the superior and posterior bony ear canal wall is completely removed and the posterior wall is reconstructed by means of a meatal skin flap. As the oval window is often partly hidden by the frame of the tympanic membrane it is more favourable for the restoration of the sound conducting mechanism if the frame and surface of the tympanic membrane are extended backwards. This can be done by repositioning the posterior cutaneous ear canal wall and grafting the tympanic membrane with fascia. By doing this, the attic is incorporated into the ear canal. Should the factors which originally induced the development of the cholesteatoma remain effective and cause a new retraction, the full breadth of the posterior ear canal skin will retract into the exenterated mastoid cavity. An easily accesible open cavity and not a recurrence will be the result. In a follow-up study of 112 patients it was found that 67 of them had developed such an open cavity, which caused no serious problems.


[Pseudo-tumoral manifestations of Schistosoma-mansoni bilharziasis observed on Madagascar. 93 cases]. Concerning respectively 93 observations got in the laboratory of "Pathology" of the Institut Pasteur (from 1954 to 1976) and the laboratory of "Pathology" of the Hospital of Befelatanana, Tananarive (from 1960 to 1975) the authors would like to point out to the medical doctors the pseudo-tumoral aspects of the bilharziasis due to Schistosoma mansoni. 1(0) The pseudo-tumoral aspects of the intestinal bilharziasis are relatively seldom complications of this parasitosis. 2(0) The malignant macroscopical aspect of these pseudo-tumors added to an often alarming symptomatology, if not possible association of cancer and bilharziosis, often brings mistakes in diagnosis. 3(0) Therefore and especially in endemic areas, biopsy is required or at least a medical proof treatment, before any surgical act at least with children or young people, in case of malignant pseudo-tumor which is actually but a bilharziasis.


Eradication of cholesteatoma.

Eradication of cholesteatoma. Cholesteatoma is, among the lesions appearing in chronic otitis media, the one creating most problems. We have selected 8 clinical situations, the most significant in our practice. After a short description of each, we expose our attitudes, past and present, in regard to each. Whether to use open or close techniques seems to be today's dilemma.


[Supplementary report on the study of resistance to coumafene in Rattus ratus observed at Marseilles]. A resistance to coumafene has previously been observed among Rattus rattus in a foodstuffs warehouse in the port of Marseille. New tests are here reported. They concern rats captured in two warehouses situated at a distance of 1,000 m. from one another, and separated by a distance approximately similar to that of the first warehouse explored. The experiments which were carried out, although limited in number, have clearly shown a resistance to coumafene administered in high doses in a rat of the first group and in three rats of the second group. There is every reason to fear a resistance extended to all the territories where Rattus rattus are found in the harbour and in the neighbouring regions.


Some considerations on middle ear cholesteatoma in 'foreign workers'. The treatment of cholesteatoma in patients without a permanent residence or in medically underdeveloped countries presents a serious problem in middle ear surgery. Methods which erradicate disease safely and reduce the risk of recurrence to a minimum are necessary. We differentiate 3 clinical types of cholesteatoma: 1 Retraction of Shrapnell's membrane or in the postero-superior quadrant with very little cholesteatoma lateral to the ossicles. A transcanal technique is used to evert the cholesteatomatous pocket into the meatus intact, followed by reconstruction of the outer attic wall. 2 Circumscribed cholestatoma lateral and often also medial to the ossicles. Combined approach tympanoplasty is used in such cases. 3 Marginal perforations with large cholesteatomas, situated medial to the ossicular chain. For these ears, classical radical mastoidectomy is the method of choice. (Radical mastoidectomy also has to be performed in all cases of type II, where luxation of the matrix in toto has failed.) No recurrence has been detected in cases where follow-up was possible.


Management of the labyrinthine fistula.

Management of the labyrinthine fistula. Preoperative, operative and postoperative findings in 97 cases of labyrinthine fistula are presented. The majority of patients had had symptoms of chronic otitis media for 20 years or more and manifested some degree of sensorineural hearing impairment. Two-thirds had experienced dizziness. The fistula was limited to the lateral semicircular canal in 83 cases and involved the labyrinth more extensively in 14 instances. The intact canal wall technique was used in less than 60% and an open cavity technique in a quarter of the cases. Severe or total sensorineural hearing impairment developed postoperatively in 8% of the lateral canal cases and in over half of the extensive fistula cases. Five per cent had incapacitating dizziness for up to 6 months postoperatively. When a labyrinthine fistula is encountered in an only hearing ear, a classified modified radical mastoidectomy is usually recommended. In other instances, the procedure performed will vary with the status of the opposite ear, the extent of the fistula, the sensorineural function of the involved ear and the size of the mastoid.


Tympanomeatoplasty with preservation of the bony bridge for cholesteatoma surgery. A technique of tympanoplasty which combines preservation of the medial part of the osseous canal wall with reconstruction of its lateral aspects is described. The results in terms of functional improvement and complications are discussed. It is concluded that better management of chronic upper respiratory disease may lead to more successful tympanomastoid surgery.


Tympanoplasty and eustachian tube function. In an attempt to assess the value of preoperative tubal testing in the selection of cases most likely to respond favourably to surgery, 100 ears with a central perforation following chronic otitis media were investigated. Tubal function was tested manometrically with different known tests. Postoperatively the healing and hearing results were correlated with the preoperative test results. No positive correlation was found between healing or hearing and tubal function. Postoperatively 50 ears were re-tested in a pressure chamber with a flow volume technique. It was found that the tubal function improves postoperatively in spite of no visible pathological changes in the aural end of the tube. Possible explanations are offered.


Tympanoplasty: the fundamentals of the concept. The development of tympanoplasty based on a fundamental understanding of the mechanisms of the middle ear in health and disease is described as it evolved. As a consequence of problems arising from the creation of an open cavity further developments led to the concept of a 'closed' operation which in turn has its own disadvantages. These are principally concerned with aeration of the tubotympanic cleft. The most recent phase of tympanoplasty is concerned with limiting the extent of the procedure to what is required to expose the disease process and re-establish function.


Importance of en bloc homograft in intact canal wall tympanoplasty. The incidence of cholesteatoma recurrence using intact canal tympanoplasty has been compared in 2 series of operations. The first group represents cases treated with the conventional conservative technique for attic and middle ear surgery. The second group represents cases on which the concept of 'radical attic and middle ear surgery' has been applied and an en bloc homograft has been used for reconstruction. A preliminary survey has shown a much lower incidence of cholesteatoma recurrence in ears belonging to the second group. The advantages of this surgical technique are discussed.


Auricular cartilage palisade tympano-, epitympano-, antrum- and mastoid-plasties. Autogenous auricular cartilage and perichondrium have been used to reconstruct the tympanic membrane and ossicular chain. The variations of technique are described.


TORPs and PORPs in tympanoplasty.

TORPs and PORPs in tympanoplasty. In an attempt to improve results in ossicular reconstruction, a newly introduced material, PlastiPore, has been evaluated over a short term. The techniques employed are described in detail and the use of a lateral covering material such as tragal cartilage is advocated as a means of preventing extension of this foreign material. The results in over 100 ears are better than those obtained with previous methods.


Ear disease in relation to age in the cleft palate child and adolescent. Ninety-eight cleft palate subjects were studied to ascertain the eventual adolescent and adult hearing. There were 72 complete clefts of lip and palate and 26 clefts of the palate alone. The incidence or side of ear disease did not appear to be influenced by this type of cleft. Other systemic anomalies were present in 11 cases with congenital middle ear abnormalities in 3 and sensory neural hearing loss in 8 subjects. Progressive improvement in incidence of hearing loss from 94% at 4 years to 42% at 20 years was noted with the strict definition of hearing loss as 10 dB conductive loss at any one frequency. 10% of ears were left with permanent structural changes in the middle ear. In only 2 patients was there a bilateral hearing loss of 20 dB or more through the speech range.


Research vistas in chronic otitis media. In spite of recent major advances in our knowledge of the basic factors in diseases of bone and mucosa the mechanisms responsible for the development of cholesteatoma are still not known for certain. The role of connective tissue in determining the histologic character of the overlying epithelium deserves more attention. It is possible that more radical removal of connective tissue from the mastoid would result in a lower incidence of post operative cholesteatoma. A study of human temporal bones provides little evidence that retraction pockets are caused by underlying scar tissue. Although a vast weight of suspicion indicts malfunction of the Eustachian tube as the primary cause of chronic middle ear disease, the proof is lacking providing a supreme challenge for otologists of the future.


Ventilation of the atelectatic ear.

Ventilation of the atelectatic ear. The absorption of gas from the middle ear--mastoid air cell system causes an average pressure decrease of 5 cm water per hour. Interference with the normal opening of the Eustachian tube causes increased negative pressure build-up, which stops gas absorption from the middle ear. High carbon dioxide tension is associated with metaplastic changes of mucosal stem cells into mucus producing cells. Ventilation of the middle ear is necessary for removal of the negative pressure as well as of the excess carbon dioxide. The problems associated with longterm ventilation tubes are reviewed, and some future prospects of permanent ventilation tubes are discussed.


A review of 50 cases of intracranial complications from otogenic infection between 1961 and 1977. The records of 50 patients with intracranial complications of middle ear disease treated over the past 16 years have been reviewed. Virtually all the known intracranial consequences of middle ear infection have been encountered, and include cerebral abscess, cerebellar abscess, meningitis, lateral sinus thrombosis, otitic hydrocephalus and cortical thrombophlebitis. The clinical features, neurological signs and investigative data for each group have been compared and contrasted. Treatment in the past has consisted of radical and modified radical mastoidectomy. It is proposed that these techniques will continue to be preferred for the management of chronic otitis media in the presence of a condition as serious as an intracranial complication. It is concluded that with a quoted incidence of postoperative cholesteatoma as high as 40% intact canal wall techniques are not justified in these patients at present.


Osteoplastic approach in chronic otitis media by means of a microsurgical reciprocating saw. We have devised a reciprocating saw for microsurgery, which performs 100 oscillations per second. The amplitude of the oscillations is adjustable within the range of 3-3 mm, by pressing a pedal. The width of the saw cuts is 0.2-0.3 mm. In the osteoplastic approach the superior and posterior segment of the ear canal wall is temporarily removed. Starting from a mastoidectomy 4 cuts are performed: cut 1 from the superior anterior angle of the mastoidectomy in a horizontal direction parallel to the dural plate through the whole thickness of the lateral epitympanic wall. Cut 2 from the ear canal above the short process of the malleus in a superior direction so as to meet cut 1. Cuts 3 and 4 start from the facial recess, one from the inside of the ear canal, the other from the mastoid cavity, both meeting in a groove along the facial ridge. This exposes all middle ear spaces as in a radical mastoidectomy. After elimination of disease and reconstruction of ossicles the canal firmly into its bed, needing no further fixation. Healing of the re-implanted canal segment presents no problem.


Excision arthroplasty with delayed wound closure for the infected total hip replacement. Methods of management of infections in total hip replacement vary from the use of antibiotics with reimplantation to a complete removal of the prosthetic unit. This is a series of 10 consecutive patients having documented sepsis following total hip replacement characterized by gram-negative organisms which was managed with the method of excision (Girdlestone) arthroplasty with delayed wound closure. Parenteral antibiotics were used while the wound remained open. Once wound healing had been accomplished, the patients were placed on appropriate oral antibiotics and continued on same for approximately 6 months. The duration of follow-up was 3-5 years. All patients have continued to remain free of clinical sepsis. One patient has chronic pain. All patients are fully ambulatory and are fully satisfied with their functional capacities. Recognizing a serious infection following total hip replacement, especially characterized by the presence of gram-negative organisms, requires drastic treatment methods. Excision arthroplasty with delayed wound closure is a reasonable alternative, especially in view of the functional results obtained.


Subcapital prosthesis fracture in total hip arthroplasty: case report. There have recently been multiple reports of late complications of femoral stem fractures occurring after total hip replacement arthroplasty of the hip. The case presented is the first case reported in the English literature of a subcapital prosthesis fracture involving a Charnley-Mueller design femoral component. Revision surgery was carried out with satisfactory replacement of the failed component.


Bilateral total hip and knee replacement in adults with rheumatoid arthritis: an evaluation of function. Sixteen patients with advanced rheumatoid arthritis who underwent total joint replacements of both hips and both knees were followed for an average of 22.8 months postoperatively. Their functional status before and after surgery was assessed by use of a special rating system for function. Fourteen of the 16 patients demonstrated higher total function scores at follow-up. Improvement was most marked in walking endurance, need for walking aids, and ability to climb stairs. Factors which were believed to predispose to less functional improvement included older age at initial surgery, longer duration of disease, more severe upper extremity involvement, and more frequent medical illnesses. There were 8 patients in whom combined hip and knee flexion in one or both lower extremities did not exceed 190 degrees after surgery. When this deficiency was combined with severe involvement of the upper extremities, difficulty with activities such as climbing stairs and arising from a chair was more common. Therefore, the goal of surgical treatment is to provide combined hip and knee flexion in excess of 190 degrees in these patients. Total joint arthroplasty has increased the likelihood of functional improvement in patients with severe rheumatoid arthritis involving the hips and knees, and encouraged the surgeon to operate earlier in the course of the disease as well as on patients with severely deformed joints.


Total hip arthroplasty for failed endoprostheses. Ninety total hip Charnley arthroplasties were performed in 80 patients with failed endoprostheses between July, 1970, and November, 1975. Pain was completely alleviated or minimal in 97.7% of cases. The ability to ambulate improved in all cases with 93.2% good or excellent results. Low grade infection must be considered in treating failed endoprostheses. Appropriate surgical debridement and antibiotic therapy offer a reasonably good chance for success.


Stability of the Giliberty bipolar hip: report of three cases. The Giliberty Bipolar Hip Prosthesis consists of an unbonded acetabular cup which snap-fits over the head of the femoral prosthesis. Because of the free movement permitted at the cup-pelvic junction as well as at the snap-fit interface, there has been a great deal of concern over the potential for dislocation. Three patients in whom this unit was implanted sustained significant skeletal trauma subsequent to surgery. None of them dislocated. In each case the prosthetic head was known to be in a relatively vertical orientation with respect to the transverse axis of the pelvis prior to the injury. In 2 of the 3 cases radiographically identifiable movement of the cup was present before and after the injury. These observations suggest that the persistently mobile, vertically positioned unbonded cup remain stable despite the stress of significant trauma. It is probable that the potential for cup movement, even where on the X-ray it appears to have stopped, acts as a safety valve in absorbing force that might otherwise dislocate the hip or cause fractures of the femur or the pelvis.


Perosseous venography in the diagnosis of viability in subcapital fractures of the femur. Although perosseous venography of the hip is simple, variations in the circulation of the femoral head and difficulties in prediction of viability account for its limited acceptance. The venogram accurately demonstrates viability in 77% of 50 routinely fixed subcapital fractures. Factors causing variations in the circulation of the fractured head are: method of surgical fixation; efficiency of the fixation, and the clinical course of healing. These and other less obvious factors make it impossible to correctly interpret a venogram. Because of a 3 week delay before cell death becomes apparent, a histological examination of a further 20 femoral heads excised after venography is equally unsatisfactory. In 50 fractures labelled with tetracycline preoperatively, subjected to venography, and then treated by hemiarthroplasty, the level of tetracycline fluorescence by the osteocytes can be correlated with the venogram predictions. There is a 92% correlation of venography with tetracycline evaluations of head viability. Venography can distinguish the dead femoral heads from those with potential or adequate vascularity and thereby provide essential clinical information for the operating surgeon.


The results of medial displacement osteotomy for unstable intertrochanteric fractures of the femur. The results of medial displacement osteotomy for unstable intertrochanteric fractures demonstrate that instability exists whenever there is lack of continuity of bone cortex on opposing surfaces of the 2 main fragments after reduction of the fracture. In 88 patients, 5 were lost to follow-up; 83 were reviewed personally by the authors at an average follow-up period of 27 months; the complication rate (both local and general) was high. Eight per cent showed non-union of the fracture; 13% developed deep infection (2 suffered from septicemia). Fifteen of 83 patients required a second operation. The mortality rate at 6 months was 13%, owing to heart failure, bronchopneumonia or thromboembolism. Of the 56 living, only 24 (43%) demonstrated a good functional result. Medial displacement osteotomy should be combined with a sliding screw-plate device. Without a sliding screw plate, medial displacement osteotomy for unstable intertrochanteric fractures of the femur may not be as successful as would appear from reports in the literature.


A special guide for insertion of multiple pins for fracture of the hip: experience with 200 cases. Two hundred cases of intracapsular femoral neck fractures have been treated with a multiple parallel pin technique similar to that originated by Austin Moore. The technique employs a personally designed guide which facilitates insertion of multiple parallel pins. The fixation obtained satisfactorily resists shear stress and rotational stress and allows for continuous compression at the fracture site. The technique is most effective in treating intracapsular fractures of the femoral neck.


Internal fixation of slipped femoral capital epiphyses. The problems related to internal fixation for the treatment of slipped femoral capital epiphyses were reviewed in 164 cases and by a questionnaire of 59 orthopedic surgeons. Significant problems were reported with all fixation devices. With Trifin nails, the main difficulty was getting them to stay in in place. With threaded pins, there was difficulty removing them. The best fixation was obtained with the cast Vitallium Knowles pin, but it requires additional modification and improvement.


Changes in the upper femur after low friction arthroplasty. In 169 low friction arthroplasties, an extension to 10 years of the radiological appearances, noted at 4 years after acrylic cement had been in the medullary cavity of the femur, there was no grossly visible evidence of deterioration. An increased number of femora showed hypertrophy with normal bone texture. The number showing atrophy remained the same at about 4.8%. Absorption of the medial neck of the femur averaged 5.5 mm over the whole study and was often preceded by small cystic areas. Evidence of demarcation of cement from the endosteal surface of the femur was absent in 66%. When demarcation did occur, in most cases it was confined to the tip or below the midlevel of the stem. In only 2.4% did it reach slightly above the midlevel. No tendency could be found for the radiological appearances of medial femoral neck absorption or cement demarcation to predominate in patients with lower grade levels of function. Improved techniques, by which more perfect fixation between cement and bone in the upper level of the prosthetic stem is achieved, may eliminate instances of loosening and localized bone erosion encountered elsewhere.


The natural history of congenital dislocation of the hip: a critical review. Studies of the natural history of congenital dislocation of the hip do not distinguish which particular features indicate a poor prognosis. In complete dislocation we have found that the development of a false acetabulum with thick sclerotic iliac bone more frequently leads to pain than those with little or no bony reaction of the ilium. Congenital dislocation leads to functional disability in all patients but only half have significant pain. Congenital subluxation and dysplasia of the hip are responsible for a high proportion of coxarthrosis. The lack of symptoms, and the subtle physical findings until irreversible degenerative changes occur, makes early diagnosis difficult. When recognized early, correction of the deranged anatomy must be done if coxarthrosis is to be prevented. Concentric hip motion and normal muscular function are essential for proper development of the joint. A reciprocal relation exists between the development of the proximal femur and the acetabulum. Structural abnormality of either eventually leads to joint dysfunction.


The effect of the inverted limbus on closed management of congenital hip dislocation. A comparative study of 20 congenital dislocations of the hip with inverted limbus treated by closed reduction were compared with a group of 20 controls in which the limbus was non-obstructive as demonstrated by arthrography. In each group 3 hips developed signs of avascular necrosis. Seventeen of the 20 hips with inverted limbus and 11 of the 20 controls required subsequent femoral or acetabular procedures. Half of the hips in both groups at follow-up (mean 9 years) were dysplastic with a CE-angle of less than 20 degrees. Thus, the incidence of avascular necrosis and late clinical and radiographic results were comparable in both groups.


Long-term results of closed reduction of complete congenital dislocation of the hip in children under one year of age. Eight to 29 year follow-up investigation of the results of closed reduction in 40 completely dislocated hips in children, treated under one year of age, show that 92.5% of the hips had good functional results and 77.5% had good anatomical results. The slant and depth of the acetabular roof rapidly improved during the first year after reduction and slowly during the following 8 years. Femoral head necrosis occurred in 12 hips (30%). The necrosis was mild in 4 hips and severe in 8. The 8 hips with severe femoral head necrosis had poor anatomical results. Surprisingly, the development of the acetabulum was not influenced by femoral head necrosis. Femoral head necrosis was frequently observed when the head was in a high position in relation with the acetabulum prior to reduction. Closed reduction should not be attempted unless the head can be brought down below the level of the acetabulum.


Rheumatoid synovial cyst of the hip joint: a case report. Rheumatoid synovial cyst of the hip joint in a 52-year-old man including iliopsoas bursal extension was demonstrated by arthrography and excision biopsy. Surgical excision of the cyst and femoral head replacement were carried out. There was no recurrence for 15 months postoperatively. The condition should be suspected in patients with degenerative changes in the hip joint and adjacent unexplained palpable mass.


Coxa vara in three patients with concommitant cerebral palsy. Many theories have been advanced to explain the final neck shaft angle in the developing proximal femur. Muscle spasticity and delayed weight-bearing are the postulated causes of the development of the valgus deformity in patients with neuromuscular disease. Three patients with coxa vara exhibited some degree of spasticity and delayed weight-bearing. In spite of this, each child developed a varus rather than a valgus hip deformity. Existing theories seem not to adequately explain the development of the final neck shaft angle.


Functional results of iliopsoas transfer in myelomeningocele hip dislocations. Paralytic hip dislocations in 37 children with myelomeningoceles have been treated by open reduction, iliopsoas tendon transfer and various other procedures about the hip. At follow-up, less than half the patients were community ambulators and had maintained their reductions. The level of the spinal lesion, alignment of the lower extremities in relation to the trunk and pelvis and the presence of scoliosis including pelvic obliquity were more important factors in determining the patient's degree of function than the status of the hip reduction.


Femoral osteotomy in Perthes disease.

Femoral osteotomy in Perthes disease. A subtrochanteric osteotomy was performed on 25 hips of 24 patients affected with Perthes disease. Twelve of these patients have been followed 4 or more years postoperatively. Coxa plana, or "mushroom" deformity, never occurred. Roundness of the head was always achieved. Internal fixation was first secured with a 4 screw plate and later advantageously substituted by Hoffmann external fixation. Post-operative shortening was always present but gradually resolved to less than 1 cm in most cases. A Trendelenburg positive sign was always present postoperatively and usually disappeared within 2 years. Internal rotation of 40-60 degrees was obtained at surgery. The resulting external rotation deformity of the leg corrected spontaneously. Varus deformity, on the other hand, changed little within this follow-up. Varisation of the hip is nevertheless an essential part of the osteotomy. The overall results were directly related to the amount of epiphysis involved. The fate of the epiphyseal plate seems to play a major role in the neck deformities and the resulting coxa vara. The absence of metaphyseal changes in the face of significant epiphyseal involvement is an ominous sign.


Autologous transfusion following frozen storage at -85 degrees. Two cases are presented in which frozen autologous blood stored at -85 degrees was used in surgical autologous transfusions. No complications were encountered in either patient intraoperatively or postoperatively. At the time of discharge hemoglobin and hematocrit were normal. Frozen autologous blood is recommended when more than 4 units of blood are required; it is particularly useful when crossmatching is not possible owing to presence of antibodies in the blood.


The orthopedic aspects of Gaucher disease. Orthopedic problems are an important feature of the adult, chronic or non-neuropathic form of Gaucher disease. Thirty-five South African patients have been investigated and of these 29 had significant skeletal complications. Twenty-one had nonspecific bone pains, 14 had episodes of pseudoosteomyelitis and 2 had acute hematogenous osteomyelitis. Twenty experienced collapse of femoral heads and in 9, prosthetic joint replacement had been successfully undertaken. Four had kyphoscoliosis due to wedging of vertebral bodies and three had pathological fractures of tubular bones. Twenty-eight of the patients were of Askhenazi Jewish stock and the minimum prevalence of the disorder in this group in South Africa is one in 4,300. If skeletal problems and splenomegally coexist in a Jewish patient, the diagnosis of Gaucher disease warrants serious consideration.


Revascularization and replantation after upper extremity trauma: experience with interposition artery and vein grafts. Improved techniques and experience in microvascular surgery make it possible to repair vessels and replant digits and extremities with increasing success. Patients with sharp or localized crush injuries are excellent candidates, and the expected survival of the extremities is greater than 80%. A good functional result can be anticipated. Generalized crush and avulsion injuries, which were common in our clinical experience, are difficult problems requiring careful selection criteria, appreciation of diffuse vessel damage that may require bypass vein or artery grafts, and the appropriate use of anticoagulants to prevent vessel thrombosis. Severe crush or mangled hand injuries, brachial plexus nerve avulsions, multilevel crush or power takeoff injuries, and excessive bacterial wound contamination are unfavorable situations for revascularization efforts, and the expected results are poor. To warrant attempts at revascularization, it must be anticipated that nerve regeneration and muscle function will be achieved and that functional use will be better than that possible with a prosthesis.


Hydroxyproline centiles for normal adolescent boys and girls. Centiles for 24 hour urinary total hydroxyproline are useful in monitoring growth and the effect of treatment in children with orthopedic diseases such as scoliosis, where growth is difficult to measure by anthropometric means. The centiles have been calculated from the data of a 5 year longitudinal study on 100 normal adolescent schoolchildren. Hydroxyproline estimations and assessments of growth were done 3 times a year. The rate of growth (height velocity) can be correlated with hydroxyproline levels found in adolescents.


Eight year results of Charnley arthroplasties of the hip with special reference to the behavior of cement. Five hundred and forty-seven total hip replacements by the Charnley technique during 1967 and 1968 have been studied for the quality of the late results. The average follow-up was 8.3 years. Mechanical failure at the cement-bone interface occurred in only 2.2% of cases. A search was specially directed towards evidence of incipient pathology at the bone-cement interface in the absence of any suspicion of clinical defect. This was found in 12% of cases but it is emphasized that the criteria were exceptionally rigorous and entirely radiological. Because the cement technology used at that time is now considered unsophisticated, mechanical failure of cement is a preventable condition. A previous report of socket migration in 9.5% of cases is not confirmed by the present survey of a more recent series of cases.


Dense mineralized perivascular tubules in osteopetrotic bone. Samples of osteopetrotic bone were deorganified and examined with the scanning electron microscope. Densely mineralized tubules with walls measuring from 1-10mu in width and with a luminal diameter of 70-90mu were observed scattered throughout the processed surface of the bone. Adjacent sections of bone prepared by routine histological methods for light microscopy showed that the tubules occur in areas of bone where "hyaline" structures are seen surrounding blood vessels. The tubules are not present in normal bone. The nature of these tubules and their possible significance in the pathological physiology of osteopetrosis requires further investigation.


Bone remodeling as an expression of altered phenotype: studies of fracture healing in untreated and cured osteopetrotic rats. The healing of tibial fractures was studied in 3 groups of young littermate rats; untreated osteopetrotic (ia/ia), normal littermates (ia/+) and osteopetrotic (ia/ia) cured of the disease 3 days earlier by transplanted normal spleen cells. The fracture site was completely remodeled in 4 weeks in normal rats, 6 weeks in transplanted (cured) osteopetrotic rats and 9 weeks in untreated osteopetrotic rats. The delayed healing in untreated osteopetrotic rats resulted from reduced resorption present in this mutation. Increased resorption brought about by transplantation of normal spleen cells accelerated healing in the transplanted osteopetrotic rats. Fracture repair took longer in the latter group than in normal littermates presumably because tibial fracture occurred before the 2 week interval required for skeletal transformation from osteopetrotic to normal phenotype following transplantation of normal spleen cells. These data show that alteration of the phenotype of osteopetrotic rats is reflected in fracture healing.


Adjustable intramedullary replacement of the knee: evolution of surgical technique and prosthesis. The total knee replacement developed at the New York Orthopaedic Hospital is a nonhinged, nonconstrained variety in which the principle fixation mechanism is via the intramedullary stems of both components. The plastic segment (high density polyethylene) is press fitted into the metallic component of the tibial segment to minimize "plastic flow" and deformity. The operative technique calls for conservation of the bone of the proximal tibia by a minimum of bone resection following complete soft tissue release including removal of the cruciates. Accuracy of replacement becomes possible when the cruciate ligaments are excised and the mediolateral soft tissue structures are balanced in tension following maximum spacing of the joint to provide stability. Provisions are made for routine replacement of the patellofemoral joint.


The mechanical testing of a sliding meniscus knee prosthesis. A 3-component sliding meniscus knee prosthesis was tested mechanically to evaluate the direct loads and the anteroposterior and mediolateral bending moments on the prosthetic components during the loading phase on a pendulum prosthesis testing machine. The bending moments changed with variation of the component positions. Tilting the tibial plateau produced the greatest change in lateral bending moment on the femoral component but alteration in plateau rotation or femoral tilting produced little change in the low values of the 2 bending moments. It is necessary to ensure that the tibial component is positioned horizontally on the tibial plateau, but it is not necessary to be concerned about the angle of the femoral component to the vertical or the misalignment rotationally of the tibial component. The load ideally passes through the condylar component to the tibia much more centrally than if it were a constrained prosthesis system during the loading range, reducing the likelihood of displacement of the 2 fixed components.


The use of polyethylene mesh (Marlex) as an adjunct in reconstructive surgery of the extremities. When Marlex mesh is inserted into the body it serves as a scaffold into which migrates fibroblastic tissue. Throughout the process of maturation of fibrous connective tissue, the mesh remains inert. Experience covering 15 years demonstrates that Marlex mesh is useful for repair of ligament, muscle, and bone defects.


The result of replacement of partial or total collateral ligaments with Marlex mesh in the knees of dogs. Marlex mesh is a synthetic material which has been used clinically to replace ligamentous structures. It serves as a scaffold for infiltration of the fibroblasts eventually becoming mature fibrocytes. In dogs, almost totally mature fibrocytes appear within the framework of the Marlex mesh prosthesis by the end of 3 months of healing.


Socket wear in Charnley low friction arthroplasty of the hip. Five hundred forty-seven Charnley hip arthroplasties were measured for wear radiographically after an average period of 8.3 years. The operations were performed in 1967 and 1968 and the average rate of wear was 0.07 mm per year. This is only half that recorded for the operations in 1963, 1964, and 1965. Clinical radiographic methods of measuring socket wear are valid provided that the socket is inserted with the wear marker not more than 10 degrees from the coronal plane. This can be estimated provided that radiopaque cement is used.


Total hip arthroplasty: a review of four years experience. Forty-nine total hip replacements are reviewed after an average follow-up of 32 months. There is an 8.2% dislocation rate and an infection rate of 4.0%. There is a 39% incidence formation of "significant" amounts of heterotopic bone. From a review of the hip rating forms it is apparent that in most patients marked improvement is brought about by the operation.


Massive ankylosis following total hip anthroplasty. A total hip arthroplasty was undertaken in a 49-year-old man who had previously spontaneously ankylosed both hips. In addition, the patient possessed hyperostosis of the spine and calf, not characteristic of any previously described spondylitic or hyperostotic syndrome. Nine months postoperatively the patient reankylosed the total hip arthroplasty. This case provides further evidence that total hip arthroplasty must be undertaken with caution in patients with hyperostotic tendencies.


Primary Thompson prosthesis for acute femoral neck fractures. Two hundred fifty cases of F.R. Thompson endoprosthesis replacement without methylmethacrylate in acute femoral neck fractures between 1969 and 1973 were reviewed. Follow-up evaluation was obtained in 94 patients averaging 4.8 years postoperative utilizing the Harris hip questionnaire. None of the patients were personally examined, but each was interviewed and rated for pain, function, and satisfaction with the hip surgery. The average Harris score was 63 points giving 56% overall satisfactory results. Pain relief was satisfactory in 85% of cases. Functional results were 71% fair or better, and 90% were satisfied with the overall results. Mortality at one year postoperative was 23%. Incidence of deep wound infection was 2% and prosthesis dislocation was 1%. Both infection rate and dislocation rate were higher in posterior exposure compared to anterior lateral surgical approach. The lack of methylmethacrylate cement apparently did not delay postoperative ambulation, as 60% of all patients were discharged within 4 weeks of fracture. The indications for methylmethacrylate with the Thompson endoprosthesis is an unanswered question and requires further investigation.


Zickel nail fixation of bilateral subtrochanteric fractures. The introduction of the Zickel nail has provided a significant advance in the treatment of subtrochanteric fractures. The device allows early mobilization and a high rate of union. In a 24-year-old man, the device was used bilaterally for subtrochanteric fractures, thus obviating the need for prolonged hospitalization and promoting early ambulation, and return to work. It must be emphasized that a bilateral operation for subtrochanteric fracture is a formidable procedure but can be accomplished in an otherwise healthy individual.


Hematogenous infection in total joint replacement. Hematogenous infections may appear at any time after joint replacement operations. As distinguished from delayed infections where general symptoms of infection are absent, hematogenous infections cause malaise, chills and fever, usually of the septic type. Roentgenograms of the joint at an early stage of hematogenous infection do not show any sign of infection as these take time to develop. Nor is the scintigram positive at an early stage. The diagnosis is beyond doubt only when the same strain of bacteria is cultured from the joint, a primary focus and blood.


Intraoperative femur fractures during total hip replacement. Eleven intraoperative femur fractures occurred during 605 total hip replacements at Rancho Los Amigos Hospital, an incidence of 1.8%. Fractures occur more often in women, are usually located in the proximal femur, and mostly occur during femoral reaming. The main predisposing factors were osteoporosis and previous hip operation. The treatment may be operative or non-operative depending on the location and type of fracture. This complication is associated with a high morbidity and a significant delay in convalescence.


A noncemented total hip prosthesis.

A noncemented total hip prosthesis. Although total hip arthroplasty stabilized by cement has been a great step forward, loosening is becoming more and more frequent and constitutes a significant problem. This is a report of the design of a non-cemented prosthesis in which fixation of the prosthesis is secured by the in-growth of newly-formed bone into the irregularities of the metal. In more than 2000 cases with a follow-up of 5 1/2 years, this fixation design proves to be sound. The complications are relatively rare and the results are generally good.


Five to fourteen year interim results of uncemented total hip arthroplasty. One thousand eight hundred eight uncemented total hip replacements have been performed during the last 14 years. The prototype and early models, 11-14 years later have retained a satisfactory result in 56% of cases, and the valgus prostheses, 6-11 years after implantation retain a satisfactory result in 85% of cases. The models in current use, with a standard neck shaft angle, and a range of implants to fit the medullary canal securely, have given a satisfactory result in 94% of patients in a one to 6 year survey. Of the whole series 101 hips (5.5%) have required further surgical treatment, mainly for interface pain but only 9 patients (0.5%) have finally ended with a resection-pseudarthrosis, all for infection. The overall results of the operation appear comparable with those of cemented metal on metal implants, but the frequency of minor discomfort on weightbearing is probably greater than that which follows a cemented on plastic articulation. Against this must be balanced the low risk of infection, and the ease with which revisional surgery can be performed.


Nonfatal chloroquine poisoning.

Nonfatal chloroquine poisoning. Acute overdosage with chloroquine is a life-threatening emergency. Adults who have ingested greater than 3 gm of chloroquine phosphate have died within 2 hr after ingestion. We describe the rapid onset of typical gastrointestinal, central nervous system, respiratory, and cardiovascular symptomatology in a 49-year-old female who ingested 19.5 gm of chloroquine phosphate. Prompt emesis, lavage, administration of activated charcoal, and aggressive treatment of hypotension and respiratory distress were responsible for the patient's recovery within 6 hr.


Methyl ethyl ketone polyneuropathy in shoe factory workers. Three cases of polyneuropathy in shoe factory workers are described. The etiologic agent is felt to be methyl ketone, a substance previously thought to be a safe solvent. The concentration of this substance in the ambient air of the patients was below the threshold limit value. This suggests that there may be a cumulative effect and that we should reexamine the threshold limit values set for all the organic solvents with respect to inhalation and absorption from skin surfaces.


Effects of morphine and methadone on the nuclear uptake of estradiol by the brain. Rats were treated for ten days with morphine (50 microgram/kg) or methadone (10 mg/kg). On the tenth day the animals were injected with 3H-estradiol, and the nuclear uptake of the steroid by the central nervous system was evaluated by autoradiography. Neither of the opiates were found to have any significant effect on the nuclear uptake of 3H-estradiol by the central nervous system. The data suggest that the mechanism by which methadone and morphine inhibit normal reproductive function in addicts is probably not the inhibition of nuclear uptake of estradiol.


Blood cadmium level and hypertension in humans. Blood cadmium level was determined in 29 nontreated hypertensive male subjects and 29 controls. All were individually matched for sex, age, and smoking habits. No differences were found between hypertensives (3.3 +/- 0.4 ng/ml) and normotensives (2.6 +/- 0.4 ng/ml).


[The journey of the Guarayos to the land of the dead. A transcultural psychiatric study]. The investigation of the being of a culture has the power of revealing something about the existence of the human beings that form it. The ethnographic study of the Guarayos, in which we find the great 'mythologem' of the agricultural people, allows us to understand the collective and permanent drunkenness that characterizes it.


Phototoxicity of Dictamnus alba.

Phototoxicity of Dictamnus alba. Phytophotodermatitis from Dictamnus alba was observed in three patients. Thin-layer chromatography and spectrophotometry disclosed the occurrence of 5-methoxypsoralen and 8-methoxypsoralen in the plant extracts which was also demonstrated by their phototoxic activity in vitro.


Photocontact allergy to 6-methylcoumarin. A proprietary sunscreen-induced photosensitivity reactions in a small number of users. Laboratory study revealed that the reactions were of the photoallergic type and were due to the presence of a synthetic fragrance, 6-methylcoumarin. By photomaximization testing  6-methylcoumarin was found to be a potent photocontact allergen.


Triclocarban: evaluation of contact dermatitis potential in man. Triclocarban was subjected to a profiling of its dermatitis producing potential including irritancy (21-day cumulative irritancy potential and application to 213 normal controls), phototoxicity (method of Marzulli), predictive contact sensitization (modified Draize method), predictive phototesting and battery screening in 2200 dermatitis patients, in an effort to define its relative dermatitis potential. The allergic contact dermatitis potential of triclocarban following bar soap use appears minimal.


The fine structure of the vegetative cells of Erythrocystis montagnei, a symbiotic red alga. Erythrocystis montagnei lives as a symbiont on the thallus of another red alga, Laurencia paniculata. This electron microscope study carried out on its vegatative cells shows that the plastids have a fine structure adequate to perform photosynthesis, different from that observed in other parasitic red algae. Starch granules occur frequently in all the cells of the thallus except the large vacuolized basal cell, which joins together the symbionts. These features suggest that Erythrocystis lives on its host plant for reasons other than a lack of photosynthesis.


Plasma membrane of Phycomyces: sequential changes in the structure during spore formation. The plasma membrane of sporangiospores (spores) of Phycomyces blakeleeanus has distinct structural features that distinguish it from the plasma membrane of the sporangiophores. The plasma membrane of the spores is, however, derived from that of the sporangiophore. A study of sequential development of spores shows that the mass of cytoplasm in a sporangial initial is cleavaged by proliferation and subsequent anastomosis and membrane fusion of the clevage vesicles. The plasma membrane of the cleavage vesicles has a reversed curvature (inside out) to that of the sporangial plasma membrane. Otherwise, the distribution of intercalated membranous particles (Imp) on the fractured faces is similar to that of the plasma membrane of the sporangium. Lather, as a result of fusion of cleavage vesicles, curvature of the plasma membrane in the spore initials returns to their original state. In the processes of spore maturation, a new type of structural feature appears in the plasma membrane, which is not present either in the plasma membrane of spore initials or in the plasma membrane of sporangiophores or mycelia. This feature often is seen as large particles (30--35 nm) which are randomly distributed on the exoplasmic face (EF) with corresponding depressions on the plasmic half (PF) in freeze-fractured replicas. They increase in number with increasing spore maturity. These particles are, in fact, the inward protrusions of the plasma membrane of the spore. A change in the number of 5--8 nm intramembranous particles on the fractured faces is also noted during spor maturation, with an increase on the EF face and a corresponding decrease on the PF face. This alteration may result from vertical movement of Imp or from a change in the fracture plane as a consequence of alteration in the lipid content during maturation.


Cytokinesis in Impatiens balsamina and the effect of caffeine. A description of cytokinesis in cells of roots of Impatiens balsamina is given, and the effect of caffeine on this process. The disposition of organelles, microtubules and vesicles which form the new cell plate is similar in normal and caffeine treated roots. In sections cut in the plane of the developing cell plate, membranous arms radiating from common centres appear to play an important role in promoting efficient fusion of the vesicles, and caffeine reduces their occurrence. The possible presence of callose in the newly formed cell plate, and the formation of plasmodesmata, are discussed. Mechanisms of membrane fusion and effects of caffeine on this event are also considered.


Chromosomes and meiosis in two species of Dermaptera. The chromosome cytology of two species of Dermaptera, namely Diplatys gladiator and Forficula sp., has been studied. The diploid number of chromosomes in male Diplatys gladiator is 18 (16AA + XY), whereas in male Forficula sp. it is 27 (24AA + X1X2Y). These are the two new chromosome numbers for Dermaptera. The evolutionary status of Forficula sp. in the family Forficulinae has been emphasized.


On the mechanism of prometaphase congression: chromosome velocity as a function of position on the spindle. Rates of movement of univalents at prometaphase and of half-bivalents at anaphase in living cricket and grasshopper spermatocytes were determined as a function of the distance from the pole toward which the movement was directed. In the artificially produced univalents of cricket cells, correlation coefficients for rate versus distance form the pole were widely disparate from movement to movement and there was no consistent relationship between velocity and distance from the pole. However, in the naturally occurring univalents of grasshopper cells, there was a significant positive correlation between velocity and distance from the pole. In both cricket and grasshopper cells, there was no consistent correlation between rate of movement and distance from the pole for half-bivalents at anaphase. The prometaphase data from grasshopper cells support the simple hypothesis of Ostergren (1950) that congression results from the application to chromosomes of forces which increase with increasing distance from the pole. Furthermore, these data are consistent with models of force production which suppose that the relationship between force (reflected as velocity) and distance from the pole is a linear one.


DNA sequence organisation in avian genomes. By means of renaturation kinetics of DNA of the three avian species Cairina domestica, Gallus domesticus and Columba livia domestica the following major DNA repetition classes were observed: a very fast reannealing fraction comprising about 15% of the DNA, a fast or intermediate reannealing fraction that makes up 10%, and a slow reannealing fraction of about 70%, which apparently renatures with single copy properties. --Comparing the reassociation behaviour of short (0.3 kb) and long (greater than 2 kb) DNA fragments of duck and chicken it becomes apparent that only 12% (duck) and 28% (chicken) of the single copy DNA are interspersed with repetitive elements on 2 to 3 kb long fragments. The lengths of the repetitive sequences were estimated by optical hyperchromicity measurements, by agarose A-50 chromatography of S1 nuclease resistant duplexes and by electron microscopic measurements of the S1 nuclease resistant duplexes. It was found that in the case of the chicken DNA the single copy sequences alternating with middle repetitive ones are at least 2.3 kb long; the interspersed moderate repeats have a length average of at least 1.5 kb. The sequence length of the moderate repeats in duck DNA is smaller. The results show that the duck and the chicken genomes do not follow the short period interspersion pattern of genome organisation, characteristic of the eucaryotic organisms studied so far.


Absence of satellite DNA synthesis during meiotic prophase in mouse and human spermatocytes. Mouse spermatocytes were labelled in situ with 3H-thymidine at successive stages of meiosis. Isolated mouse as well as human spermatocytes were similarly labelled under in vitro conditions. DNA synthesis was followed either by tracking radioactivities in Cs2SO4 gradients or by measuring reassociation kinetics. Mouse satellite DNA and the 3 satellites of human DNA are labelled during S-phase but not during pachytene. In the mouse genome, there is a preferential labelling of regions containing foldbacks (human spermatocytes were not analyzed in this respect). The absence of detectable pachytene synthesis in satellite DNA is consistent with genetic evidence on the absence of crossing-over in constitutive heterochromatin.


Responses of mammalian metaphase chromosomes to endonuclease digestion. Digestion of fixed metaphase chromosomes by endonucleases (micrococcal nuclease and DNase II) under optimal digestion conditions followed by Giemsa staining produces sharp banding patterns identical to G-bands. In 3H-thymidine labeled, synchronized metaphase cells of the chinese hamster (CHO line), the band induction is accompanied by the removal of DNA. The single strand specific nuclease S1 and DNase I do not produce such banding patterns.


Higher order structure in metaphase chromosomes. I. The 250 A fiber. Metaphase chromosomes released from cells in the presence of Joklik's suspension media by vortex-mixing with 0.5 mm glass beads have been analyzed by electron microscopy. In these preparations the chromosomes are composed of series of loops (200-300 A in diameter) which are, in turn, composed of closely-apposed arrays of nucleosomes. Negative-staining of these preparations has allowed the identification of several distinct patterns within the loop which appear to arise from variations in nucleosome packing. Analogous patterns are also observed in chromatin fragments generated by brief micrococcal nuclease digestion. From these data we have deduced certain features of nucleosome-nucleosome interactions in higher-ordered chromatin fibers.


Higher order structure in metaphase chromosomes. II. The relationship between the 250 A fiber, superbeads and beads-on-a-string. The morphology of metaphase chromosome-derived chromatin fibers released from cells by non-ionic detergent cell lysis in the presence of divalent cations has been studied by electron microscopy. In these preparations the euchromatic arms appear as a series of loops, 200-300 A in diameter, which are composed of closely-apposed nucleosome arrays. The higher order fiber in chromosomes derived from detergent-lysed cells appears to be less stable than chromatin fibers obtained by mechanical cell lysis. The fiber breaks down into a series of non-uniform nucleosome aggregates (superbeads) and finally to chromatin in a beads-on-a-string morphology upon incubation at 31 degrees for 20 min. These observations allow us to suggest a relationship between uniform thick fibers, superbead-containing fibers, and beads-on-a-string chromatin within metaphase chromosomes.


Meiotic chromosome pairing and synaptonemal complex transformation in Culex pipiens oocytes. The synaptonemal complexes of the oocytes of the mosquito Culex pipiens quinquefasciatus have been reconstructed from serial sections. A diffuse structure, probably a chromocenter composed of centromeric heterochromatin, was present during pachytene. As no synaptonemal complexes were visible inside the chromocenter the continuity of the 2 arms of a bivalent was lost. The telomeric ends were clustered on a small area of the nuclear membrane in a bouquet arrangement; they were associated in pairs, and sometimes joined through a special structure. One pair was composed of the 2 telomeres of the shortest bivalent and a ring configuration was thus formed. The other 2 chromosomes may form one or two rings. During a short transitional stage, after the disappearence of the synaptonemal complexes, several thousand annuli, 1200-1500 A in diameter, were present in the nuclei. The annuli disappeared as material originating mainly from the transverse filaments of the synaptonemal complexes formed a "capsule" around the chromosomes during diplotene.


[Schizophrenia and addiction].

[Schizophrenia and addiction]. The relationship between schizophrenia and drug abuse has been approached using physiopathological, psychopathological, and cybernetics communicational techniques. The hypotheses which have been proposed with regard to the physiological mechanism involved in schizophrenia and drug abuse all have in common, among other things, the idea that neurotransmitters play a role in the development of these behaviors. Likewise, the psychopathological symptoms presented by schizophrenia and drug abuse offen many similarities. The mechanism by which drug abuse may lead to schizophrenia is discussed. The analysis of the communicational system reveals the correlation between there two syndroms. None of the three techniques which have been described should be exclused from clinical practice.


Preparation, purification, and stability of high specific activity 125I-labeled thyronines. A modified chloramine-T method is described for the preparation of several radioiodothyronines by an exchange reaction, which results in low specific activity preparations, or by an addition reaction, which yields radioiodothyronines with specific activities up to 7 mCi/microgram. Purification by paper chromatography is shown to be more convenient than by LH-20 chromatography and provides better resolution among the various thyronines. Radioiodothyronines with only a single iodine atom in the outer (3,5,3'-triiodothyronine and 3,3'-diiodothyronine) are stable for several months when stored in organic solvents. The least stable radioiodothyronines are those with two 125I atoms in the outer ring (3,3',5'-triiodothyronine (rT3) produced from 3-iodothyronine and thyroxine (T4) from 3,5-diiodothyronine). The stability of rT3 and T4 stored in human plasma at 4 C is much greater than when stored in buffer at the same pH. The use of high specific activity [125I]rT3 has permitted the development of a radioimmunoassay with a sensitivity of 1 pg rT3/ml incubation volume.


Mechanisms for the testicular hypertrophy which follows hemicastration. The hormonal and testicular effects of hemicastration have been examined using rodent models. When rats were hemicastrated at 5 days of age, significant testicular hypertrophy was noted within 5 days. Hypertrophy decreased as the age at hemicastration approached 20 days and did not occur in rats 45 days of age or older. The changes in testicular weight were associated with significant (P less than 0.001) increases in serum FSH values from 10-20 days, but no significant alterations in the intratesticular concentration of testosterone occurred. Hemicastration also caused significant hypertrophy in testes depleted of germ cells. This was reflected by a substantial increase in testicular protein and DNA when compared to intact controls at 10, 15, and 20 days of age; however, the concentration of protein/microgram DNA was increased only at 15 days. These data indicate that 1) changes in serum FSH and not intratesticular testosterone are associated with the testicular hypertrophy which follows unilateral castration of immature animals; 2, a significant proportion of the hypertrophy can be attributed to non-germinal cells, the Sertoli cell being the prime candidate;3) the increase in testicular weight is primarily the result of an increase in cell number.


Acute effects of thyroid-stimulating hormone on cultured thyroid cell morphology. The acute effect of TSH on the ultrastructure of dog thyroid cells in monolayer tissue culture was examined by scanning electron microscopy. Although thyroid cells in monolayer culture remained responsive to TSH stimulation, the cell surface changes induced by TSH were different from those seen in intact thyroid tissue. Morphological changes were observed within 15 min after the addition of TSH, the earliest time point examined. These changes were fully developed within 4 h. Spontaneous morphological transformation was not observed in the absence of TSH. Morphological alterations induced by TSH were 1) the thickening and retraction of the cell border, 2) the formation of a network of cytoplasmic projections resulting from both cell retraction and active cytoplasmic extension, and 3) an increase in the number of surface microvilli. The relationship between this acute TSH action and thyroid hormone synthesis and secretion remains to be clarified.


Effect of maternal hypothyroidism and triiodothyronine on the fetus and newborn in rats. Of 36 adult female rats studied 19 were radiothyroidectomized; 3 weeks later all rats were bred. Radiothyroidectomized rats were divided into Groups I-III. Group I was untreated. Maternal weight gain was not noted until day 9 and was steady thereafter. Group III rats were treated with triiodothyronine up to the first day of pregnancy, but were hypothyroid during gestation (elevated TSH and low T3). Group II rats were treated with T3 up to the day of delivery. The pregnancy, its outcome, and TSH and T3 levels were similar to the normal controls (Group VI). Pregnancy, its outcome, and TSH and T3 levels were not different from controls when T3 was given to normal rats up to day 1 of pregnancy (Group IV). T3 concentrations were elevated in the normal rats which were given T3 continuously up to the day of delivery (Group V). However, the animals did not appear to be thyrotoxic and TSH was not suppressed. The size of the litters of Group I mothers was significantly less than in other groups and this difference may be attributed to an increased tendency to resorb implanted fetuses in untreated hypothyroid mothers. However, activity, day of eye opening weight gain, and TSH and T3 levels of the pups of all the groups were similar up to age 21 days. The data indicate the independence of the fetal pituitary-thyroid axis from maternal thyroidal status in rats.


Estrogen and progesterone interactions in the rabbit uterus in vivo after steroid administration. After a single injection of Estradurin (polyestradiolphosphate), rabbit plasma 17 beta-estradiol (E2) levels rose to a maximum in 3 days and were maintained for at least a 13-day period. The uterine E2 concentration rose slowly so that at day 12 the tissue/plasma E2 ratio was similar to that in untreated controls. The gain in tissue weight after the Estradurin injection correlated well with the increase in tissue E2 concentration. In a group of rabbits to which progesterone (P) was administered 7 days after Estradurin injection, the E2 concentration in plasma did not change significantly. However, the tissue E2 concentration showed a significant decrease after four daily injections of P. When P injections were halted (progesterone withdrawal) for 2 days, the tissue E2 levels rose and were greater than those found in the group not receiving P. These levels were significantly higher than those found in animals which had received daily injections of P for 2 or 4 days. These results indicate that P is able to suppress the accumulation of E2 by uterine tissue, in agreement with published evidence that P is able to reduce the levels of cytoplasmic E2 receptors.


Intracellular transport and packaging of prolactin: a quantitative electron microscope autoradiographic study of mammotrophs dissociated from rat pituitaries. Dispersed pituitary cells prepared from estrogen-treated female rats were subjected to pulse labeling with [3H]leucine (5 min) followed by a chase incubation (up to 3 h) in order to study intracellular transport of PRL in mammotrophs. Sites of synthesis, rates of transport, and sites of packaging and storage of PRL were determined by quantitative electron microscopic autoradiography. Results of grain counts show that label is initially (end of pulse) distributed randomly over the rough endoplasmic reticulum (ER), but rapidly (5--15 min of chase) moves to the stacked Golgi cisternae where concentration into secretion granules takes place. The label moves successively from small (Type I) immature granules (15--55 min of chase) to large (Types II and III) polymorphic granules (55--115 min) in the Golgi region, to rounded or ovoid mature (Type IV) granules (55--185 min) usually found in the peripheral cytoplasm, indicating that these types of granules represent successive stages in granule concentration and assembly. Analysis of the relative grain density (percentage of total grains/percentage of total area) confirmed that there was progressive concentration (up to 20--150 times) along the transport route with the concentration lowest in the ER, higher in the Golgi, and highest in immature and mature secretion granules. These data indicate that synthesis of PRL occurs randomly in the ER, transport to the Golgi occurs rapidly (within 5--10 min), and is completed rapidly (90% within 15--20 min), and concentration into granules and aggregation of small granules into larger forms also occurs rapidly (by 15--20 min), but goes on over a prolonged period of time (up to 3 h). Use of dispersed cells has allowed a more precise determination of the location and kinetics of steps in the intracellular processing of PRL than has been possible previously using other systems.


Methylation of estradiol-17 beta by a partially purified preparation of bovine pineal hydroxy-indole-O-methyltransferase. A partially purified preparation of hydroxy-indole-O-methyltransferase (HIOMT) from bovine pineal was shown to O-methylate estradiol-17beta (E2). The HIOMT preparation was incubated with E2 and [3H]-S-adenosylmethionine (SAM). The [3H]-3 methyl-ether or estradiol [MeO-E2] produced was identified by thin layer chromatography (TLC) and crystallization to constant 3H/14C ratio in the presence of [14C]-MeO-E2 and unlabeled MeO-E2 standards. The kinetics of the enzyme reaction for melatonin formation from N-acetyl-serotonin (NAS) and for MeO-E2 were compared.


Aromatase activity in the developing rabbit brain. The formation of 17beta-[3H]estradiol from [1,2,6,7-3H]testosterone was assessed in placenta and central nervous system tissues from rabbit embryos that varied in age from 13-28 days of gestation. In the fetal brain, significant rates of aromatase activity were limited exclusively to the forebrain, and the highest rates of activity (approximately 0.5 pmol/h/mg protein) were found in the diencephalon both male and female embryos between days 19 and 25 of gestation. These rates of aromatase activity are second only to the fetal ovary when expressed per mg protein; moreover, forebrain is the only tissue in the male embryo capable of synthesizing significant amounts of estrogens in vitro. When projected to the whole organ, the capacity of the diencephalon for aromatization exceeds the capacity of the fetal ovary approximately 9-fold. Placental aromatase activity was high (2.1 pmol/h/mg protein) on day 13 but fell to a level approximately 20-fold lower by day 19 of gestation. These findings indicate the potential importance of the forebrain as a source of estrogens during embryogenesis.


Intracellular prolactin in rat corpus luteum and adrenal cortex. This study was done to examine whether PRL, which appears in both active milk secretory cells (MSC) (1) and milk (1,2) during normal lactation, could also be detected in PRL target cells which do not transfer this hormone into an exocrine secretory product. Ovaries, adrenals, and mammary and pituitary glands were collected from actively nursing rats decapitated on day 15 post-partum. All four tissues were processed for light microscopic immunohistochemical identification of PRL. Immunoreactive PRL was again found in pituitary PRL cells and in MSC. It was also detected within both lutein and adrenal cortical cells (zona fasciculata). In all three target tissues, PRL was present in target cell cytoplasm, but, in MSC and adrenal cells, it was also found occasionally in nuclei. These findings, which seem to indicate that transfer into an exocrine secretory product cannot be the only possible explanation for the appearance of this protein hormone inside its target cells, extend the evidence suggesting that PRL may have intracellular sites of action.


Electrophoretically separable forms of rat prolactin with different bioassay and radioimmunoassay activities. The possible existence of forms of rat PRL with different bioassay (BA)/RIA activity ratios was investigated. Anterior pituitary tissue and medium in which explants of rat adenohypophyses had been incubated were processed by polyacrylamide disc gel electrophoresis using a Tris-glycine buffer. The gel columns were cut into five segments and eluates from these were tested for PRL by BA and by RIA. Most of the BA- and RIA-detectable PRL in tissue and medium was found in the same gel region to which purified rat PRL (RP-1) migrated. The BA/RIA ratio of this major form of PRL was 2.2-4.0. Other more slowly migrating forms of PRL were found in the regions where albumin and growth hormone migrate. The BA/RIA activity ratios of these forms varied depending on the sample processed (e.g., tissue vs. medium; sex and/or age of the adenohypophyseal donor; duration of incubation) but were generally considerably higher than those of the respective major forms. PRL that migrated faster than the major form was also detected by the RIA in several cases, but in only one experiment (with 4-day culture medium) was bioassayable PRL present in this more anodal region. In this case the BA/RIA ratio was very high (78.6). These results indicate that forms of rat PRL with different BA/RIA activity ratios exist within and are secreted by rat adenohypophyses.


Estrogen receptor in rat liver: translocation to the nucleus in vivo. Following in vivo ethinyl estradiol (EE2) administration (100 microgram sc) to adult female rats, the estradiol-specific binding sites (ESBS) of liver cytosol were markedly reduced at 30 and 60 min. The reduction at 30 min was to one-quarter of that found in rats treated with vehicle alone. Coincident with this reduction, nuclear ESBS were increased. The ESBS of partially purified cytosol and of dense sucrose-purified nuclei were determined by gel filtration after incubations with tritiated estradiol using exchange assay conditions. An elevated temperature during the exchange assay incubations was necessary to demonstrate most of the ESBS in purified nuclei of EE2-treated rats and suggested that estrogens are attached at these sites. Following administration of 5 microgram EE2, the decrease in cytosol ESBS and the increase in nuclear ESBS were smaller. In contrast, 5 microgram EE2 was as effective as 100 microgram EE2 in substantially increasing the ESBS observed in uterine nuclear fractions. The low level of ESBS found in whole brain purified nuclei was unchanged by 100 microgram EE2 administration. The steroid specificity and proteolytic enzyme sensitivity of the purified nuclear ESBS of treated rats were similar to that of the partially purified cytosol ESBS of rats treated with vehicle alone. The data are consistent with the ESBS being estrogen receptor proteins which translocate from the liver cytosol to the nucleus after estrogen administration in vivo.


Angiotensin II/aldosterone dose-response curves in the dog: effect of changes in sodium balance. The possibility that the responsiveness of plasma aldosterone concentration to angiotensin II alters with changes in sodium balance was investigated in male beagle dogs under conditions of controlled sodium and potassium intake. Angiotensin II was infused at four different rates (usually 3, 6, 12, and 24 ng/kg/min), each for 1 h, 1) after periods of normal sodium diet (32 mEq/day), 2) after moderate sodium depletion (negative cumulative sodium balance 25-58 mEq), 3) after severe sodium depletion (65-116 mEq negative cumulative sodium balance), and 4) after sodium loading (150-212 mEq positive sodium balance), daily potassium intake remaining constant (26 mEq/day) throughout. Angiotensin II/aldosterone dose-response curves after moderate sodium depletion were both elevated and steepened in comparison with those found during normal sodium intake. Severe sodium depletion was associated with even greater elevation of dose-response curves, but individual aldosterone responses to angiotensin II were irregular and unpredictable. Sodium loading significantly diminished aldosterone responsiveness to angiotensin II. Blood pressure increments during angiotensin II infusion were attenuated by sodium depletion.


Preprogramming mechanism of luteinizing hormone in the determination of the lifespan of the rat corpus luteum. Luteal regression was studied in female rats bearing an isografted pituitary under the renal capsule and the day of ovulation was designated as day 0. These females had pseudopregnancy-like cycles and ovulations at 16-day intervals. After each ovulation, corpora lutea were activated and progesterone concentrations increased in peripheral blood until day 5, after which concentrations gradually decreased. Removal of the in situ pituitary on day 0 or 1 resulted in maintenance of luteal function; progesterone concentrations were sustained. This effect of hypophysectomy on day 0 was prevented by daily administration of small amounts of LH from days 0-5; however, luteal regression occurred if hypophysectomy was performed on day 4 or later and progesterone concentrations decreased. Luteal regression was prevented if anti-LH serum was administered from days 1-5 to pituitary-isografted rats and was followed by hypophysectomy on day 5. These results indicate that the lifespan of corpora lutea in pseudopregnant rats is programmed between days 2-4 after ovulation through the hypothalamohypophysial axis via secretion of LH.


Excess production of free alpha subunits by mouse pituitary thyrotropic tumor cells in vitro. Primary cultures from three serially transplanted mouse pituitary thyrotropic tumors, Furth 97A, 97B, and 97C, and one newly generated tumor, NIH 101A, were shown to secrete free alpha subunits and a beta subunit (TSH-beta) in addition to intact TSH. The three glycopeptides were measured in specific, heterologous radioimmunoassays. Basal secretion of alpha subunit was found to be in excess in all four thyrotropic tumor cultures, ranging from 5.7- to 8.9-fold higher than that of intact TSH and 30- to 35-fold higher than that of free TSH-beta. In cultures from 97A, TRH (10(-7) M) stimulated the secretion of TSH, alpha subunit, and TSH-beta by 175-185% and (T4, 10(-5) M) inhibited TSH to 19%, alpha subunit to 68%, and TSH-beta to 58% of control. Secretion of TSH and its subunits in cultures from 97B was not consistently affected by TRH or T4. In cultures from 97C, TSH and alpha secretion was inhibited in a dose-dependent fashion by T3 from 2.5 x10(-9) to 1.6x10(-7) M. Gel chromatography of incubation media from tumor 97A revealed elution volumes of TSH and alpha similar to those of purified rat glycopeptides and confirmed the excess secretion of free alpha subunit. In contrast, the alpha to TSH ratios in media from cultures of pituitary cells from normal and thyroidectomized rats were 1.6 and 0.8, respectively. The total production of TSH and its subunits was measured in cultures from tumor 97A and confirmed that alpha subunit was synthesized excessively. The ratio of the plasma concentrations of alpha to TSH in tumor-bearing mice was 1.6 compared to 1.3 in thyroidectomized mice, and 0.36 in thyroidectomized rats. The alpha to TSH ratios in extracts of the corresponding thyrotropic tissues (tumor or pituitary) were 2.7, 1.7, and 0.40, respectively. These data are analogous to those reported in humans with thyrotropic and certain other pituitary tumors, and support hypothesis that the subunits of glycoprotein hormones are independently synthesized.


The effect of bombesin and related peptides on prolactin and growth hormone secretion in the rat. Bombesin and other related peptides isolated from the skin of anuran species and found in mammalian brain stimulate PRL and GH release in steroid-primed male rats. On a molar basis, bombesin and alytesin are the most active peptides in this assay system. Their PRL- and GH-releasing activity is not affected by the type of anesthetics used, but their minimal effective dose is lower after intravenous administration as compared with intracisternal administration. The in vivo stimulatory effect of bombesin on PRL secretion is not modified by the histamine antagonist diphenhydramine or the opiate antagonist naloxone. By contrast, naloxone reverses the GH-releasing activity of bombesin, suggesting an opiate-dependent mechanism of action of this peptide on GH secretion. The absence of an in vitro effect of bombesin and related peptides on PRL and GH secretion and a minimal effective dose of approximately 30 ng in bombesin and alytesin administered in steroid-primed rats, make them the most active peptides reported so far to act on the brain to modify PRL and GH secretion.


Testosterone secretion in the rat in response to chorionic gonadotrophin: alterations with age. It has been reported by others that both prevailing blood concentrations of testosterone and Leydig cell response to gonadotrophin (up to 1 h after injection) are reduced in the aged male rat. Although resting levels of plasma testosterone in our aged (24-26 months old) Sprague-Dawley rat are also depressed compared with young (3-4 months) or mature (12 months) animals of the same strain, subcutaneous injection with human chorionic gonadotrophin for 3 days restores secretory function, producing testosterone levels indistinguishable from those of similarly stimulated younger rats. In short term experiments, old rats did show a diminished testesterone secretory response to human chorionic gonadotrophin 1 h after a single intravenous injection, consistent with previous reports, but restoration of normal stimulated levels was observed by 2 h, and persisted up to 24 h. These findings differ from the demonstrated intrinsic testicular hyporesponsiveness to gonadotrophin of aged men, and probably represented a state of chronic understimulation of the aged rat Leydig cells, due to low prevailing levels of LH.


Evidence of a small molecule in mouse Leydig cell tumors which inhibits the conversion of estrogen receptor from 4S to 5S. The activation process of cytosol estrogen receptor (RE) from mouse Leydig cell tumors was investigated. When RE from tumors which were affected in their growth by estrogen was heated at 25 C for 30 min in high ionic strength buffer it was converted from native 4.0S to a transformed (5.3S) state. Dialysis of the cytosol from these tumors at 0-4 C in either the presence or absence of estradiol also resulted in conversion to 5.3S when estradiol was added before the sample was applied to the sucrose density gradient. When the dialyzed cytosol was applied to a sucrose density gradient without estradiol, the peak of estrogen binding capacity tested by adding E2 to the fractions from the gradient, occurred at approximately 4S. The dialysate of cytosol had an inhibitory effect on activation by warming. Dialysis of cytosol in the presence of estradiol enhanced the translocation of estradiol-receptor complex to nuclei. Molecular weight (approximately 128,000) and frictional coefficient (f/f0 1.62) of the estrogen-receptor complex dialyzed in the presence of high salt concentration were identical with those of warmed RE complex, but quite different from the native RE (approximately 73,000 daltons, frictional coefficient 1.48). This evidence indicates the presence in the cytosol of one or more small molecules which, when noncovalently bound to RE, inhibit the conversion of the native cytosol RE to the transformed state R'E2RE, having an additional protein unit.


Transport of maternal[3H]melatonin to suckling rats and the fate of [3H]melatonin in the neonatal rat. The question of whether maternal melatonin could be transported in milk to suckling rats was investigated because melatonin is probably not produced in these animals during the first 10 days of life. [3H]Melatonin was found to be rapidly transferred from the maternal circulation into lactating mammary tissue, and the stomach of each suckling rat was found to contain [3H]melatonin. To study the fate and tissue distribution of [3H]melatonin originating in the neonatal stomach, suckling rats were given [3H]melatonin by stomach tube; [3H]melatonin was recovered from plasma and seven tissues, including brain, 15 and 60 min later. The general tissue distribution of [3H]melatonin was similar to that found in adult rats. The major [3H]melatonin metabolites in the urine of suckling rats, as the adult rats, were the conjugates of 6-hydroxy-melatonin.


Serum somatomedin stimulation in thyroxine-treated hypophysectomized rats. Somatomedin activity in rat serum was measured by sulfate incorporation into embryonic chick cartilage. Hypophysectomized male rats were treated daily for 21 days with 20 microgram/kg L-thyroxine (T4) and/or 25 microgram bovine (b) GH. Serum from rats given T4 alone or with bGH had somatomedin potencies of 0.88 and 0.83, respectively, comparable to normal serum (1.00) and significantly greater than serum from bGH-treated hypophysectomized rats (0.62). Hypophysectomized rats had no measurable somatomedin activity. T4 treatment resulted in a return to normal of the basal metabolic rate and serum T4 and T3 concentrations. Addition of T4 or T3 to serum from normal rats to produce concentrations comparable to those found in serum of hypophysectomized rats after T4 and/or bGH treatment did not alter the somatomedin activity. Higher concentrations of T4 (10(-7) M) or T3 (10(-8) or 10(-7) M) did, however, enhance the sulfation potency of normal serum. T4 (10(-9) M to 10(-6) M) or T3 (10(-10) M to 10(-7) M) added to serum-free medium provided minimal sulfation activity that represented only a fraction of the activity of serum. T4 may stimulate production of somatomedin in the absence of adequate bGH or may be metabolized to an active sulfation factor.


Conversion of L-thyroxine to triiodothyronine in rat kidney homogenate. Rat kidney homogenates, in phosphate-EDTA buffer, consistently catalyzed the formation of T3 from added L-thyroxine (T4). The formation of T3 was assessed by both paper chromatography and RIA of T3. Conversion of T4 to T3 appeared to be enzymatic, showing pH and temperature optima (pH 7.0 and 37 C, respectively) and tissue and time dependence. Formation of T3 was unaffected by azide, cyanide, or catalase, nor was it dependent upon oxygen; indeed, under anaerobic conditions conversion of T4 to T3 was enhanced. Dialyzed homogenate retained full activity, and no cofactor requirement was demonstrated. A role of iron and thiol groups in the enzymatic formation of T3 from T4 was suggested by the inhibitory action of iron chelators and thiol-blocking reagents. The capacity of kidney for T3 formation was considerable and increased with increasing T4 concentrations, being approximately 2 nmol/g tissue/h at very high T4 levels. The apparent Km was estimated to be 3 x 10(-6) M. The conversion of T4 to T3 was inhibited by propylthiouracil at micromolar concentrations whereas methimazole, iodide, and lithium salts were without effect. The enzymatic activity of the homogenates was associated with its particulate components, the readily sedimenting fractions corresponding to plasma membranes and mitochondria being most active, and was absent from nuclei and cytosol.


Elevation of dopamine in fetal plasma and the amniotic fluid during gestation. Catecholamines were determined by a radioenzymatic assay in maternal and fetal rat plasma and in the amniotic fluid during the last five days of gestation. Dopamine was significantly higher in fetal than in maternal plasma whereas norepinephrine was the same. Epinephrine was sighificantly lower in fetal than in maternal plasma on days 18 and 19, but was the same during the remainder of gestation. Dopamine was the predominant catecholamine in the amniotic fluid on days 20-22 of gestation showing the largest percent increase prior to parturition. The data suggest that dopamine may have a peripheral hormonal role during fetal development and parturition.


The effect of feeding and starvation upon immunoreactive glucagon in the pancreatectomized pig. The response of 13 pigs to total pancreatectomy was compared with the response of 9 animals to sham operation. Approximately half of each group had free access to food while the remainder received oral water only. Completeness of pancreatectomy was confirmed at autopsy, and the disappearance of insulin and the rise in blood glucose to 14-19 mmol/l. Immunoreactive glucagon (IRG) increased in fed pancreatectomized pigs from the 3rd day postoperatively to a level of 350-400% of basal values by the 8th day. There was no rise in starved pancreatectomized animals, or in either group of sham-operated animals. The rises in glucose, free fatty acids, triglycerides and ketones were similar in the two groups of pigs.


In vitro insulin-stimulated conversion of [U-14C]glucose to 14CO2 by rat thymocytes. Addition of bovine insulin to thymocytes from adrenalectomized rats resulted in stimulation of [U-14C]glucose conversion to 14CO2. A significant enhancement of 14CO2 formation by insulin occurred by 30 min of incubation, and was consistently observed at an insulin concentration of 10(-8) M. The response to insulin was similar at 0.55 and 1.1 mM glucose, and was obtained at three cell concentrations (0.5, 1.0, 2.0 X 10(8) cells/ml). The incorporation of [3H]leucine into trichloroacetic acid-precipitable material was significantly increased by 10(-6) and 10(-8) M insulin. Cycloheximide, at a level of 2.5 X 10(-5) M, suppressed [3H]leucine incorporation by 93% and inhibited the stimulation of 14CO2 formation by insulin. We conclude that insulin can enhance the formation of 14CO2 from [U-14C]glucose by thymocytes in vitro, and that this response may require the synthesis of one or more proteins.


Non-parallel variations in the preferential secretion of 3,5,3'-triiodothyronine (T3) and 3,3',5'-triiodothyronine (rT3) from dog thyroid. rT3, T3, and T4 were measured in effluents from perfused dog thyroids and in hydrolysates of the thyroids. rT3 was determined by a newly developed sensitive radioimmunoassay (sensitivity approximately 0.1 pg rT3/tube). The rT3 and T3 contents of thyroid effluent were always much higher than would be expected from the relative amounts of T4, T3, and rT3 in thyroid hydrolysate. In four two-sided thyroid perfusions the T4/rT3 ration (wt/wt) in thyroid hydrolysates was 59.3 +/- 26.3 (mean +/- SD) and in effluents from unstimulated thyroids it amounted to 15.1 +/- 7.8. The T4/T3 ratio in thyroid hydrolysates was 10.3 +/- 1.9 and the ratio in effluents from unstimulated thyroids was 6.0 +/- 0.3. A continuous infusion of TSH, 100 micromicron/ml, enhanced the secretion of all three iodothyronines. During the stimulation a biphasic response was observed in the T4/T3 and T4/rT3 ratios in the effluent. At first the preferential secretion of T3 and rT3 was enhanced. Then the T4/T3 ratio returned to prestimulation values while there was a further increase in the T4/rT3 ratio to nearly twice the prestimulation values. Dog thyroids stimulated and examined in vivo showed a similar preferential secretion of rT3 and T3. Thus, both rT3 and T3 were secreted preferentially to T4 from the thyroid. The preferential secretions of rT3 and T3 did not vary in parallel. During a prolonged TSH stimulation the degree of preferential secretion of T3 was maintained despite a considerable increase in hormone secretion. On the contrary, the preferential secretion of rT3 decreased during prolonged TSH infusion.


Steroidogenesis in rat Leydig cells: effect of gonadotropins on the activity of 5-ane and 5-ene 3alpha- and 3beta-hydroxysteroid dehydrogenases during sexual maturation. The in vivo influence of gonadotropins on the activities of oxidoreductases of androst-5-ane and androst-5-ene steroids and pregnenolone was examined in testes from young rats. Animals were given daily injections of human CG for 5 days starting at 20 days of age and the testicular 12,000 X g supernatants were assayed for steroid oxidoreductase activities. Marked increases (up to 8-fold) were noted in the rate of oxidation of the 3beta-hydroxyl of 3beta-hydroxy-5beta-androstan-17-one, 3 beta-hydroxy-5alpha-androstan-17-one, 5alpha-androstane-3beta,17beta-diol, dehydroepiandrosterone, and pregnenolone, and in the 3-keto reduction of 17beta-hydroxy-5alpha-androstan-3-one, 17beta-hydroxy-5beta-androstan-3-one, 5beta-androstane-3,17-dione, and 5alpha-androstane-3,17-dione. The hormone response required a certain amount of time as no response was detected until 72 h after the first injection. As little as 1 IU hCG/injection resulted in significant increases in 3beta-oxidoreductase (3beta-HSD) activities. FSH and TSH gave no significant increases and 25 microgram NIH-LH-S18 resulted in increases only when the hormone was suspended in a sesame oil-beeswax mixture. Hormone treatments did not result in increased 5-ane-3alpha-HSD activities. Rats receiving chronic human CG treatment starting at 66 days of age showed less marked increases in 5-ane-3beta-HSD activities than the younger rats and no significant enhancement in 5-ene-3beta-HSDs. It is suggested that during sexual maturation the testicular biosynthesis of active 5-ane androgen(s) proceeds via 5-ane precursors with the help of age and gonadotropin-dependent 5-ane 3beta-oxidoreductase.


Data on the sites of stimulatory feedback action of gonadal steroids indispensable for luteinizing hormone release in the rat. The sites of the stimulatory feedback action of gonadal steroids on LH release were investigated in estrogen-primed ovariectomized rats. Either estradiol benzoate (E2) or estrone (E1) injections 72 h after E2 priming induced a significant increase in serum LH 30 h later, whereas injections of progesterone (P) did so 6 h later. Horizontal sections placed above the medial preoptic area prevented the increase after E2 injections but not after E1 and P injections. Retrochiasmatic sections or bilateral lesions placed in the medial-basal part of the suprachiasmatic area prevented any increases in serum LH induced by steroid injections. Intracerebral implantations of E2 and P in E2-primed ovariectomized rats induced a similar significant increase in serum LH when E2 was implanted into the bed nucleus of stria terminalis (BST), the lateral septum (l-SEPT) and the preoptic suprachiasmatic area (POSC), and when P was implanted in to the medial amygdala, the diagonal band of Broca (DBB), the POSC, the l-SEPT and the anterior hypothalamic area (AHA). These results suggest that the stimulatory feedback effect of E2 is exerted on the limbic structures, especially the BST and the l-SEPT, while the main sites of the stimulatory feedback action of P are located in the DBB, the POSC, and the AHA.


Pancreatic polypeptide responses to a meal and to intraduodenal amino acids and sodium oleate. Serum concentrations of immunoreactive pancreatic polypeptide (IR-PP) were measured in dogs in response to a meal and to intraduodenal infusions of amino acids, Na oleate, and HCl. In addition, the effects of somatostatin on meal-stimulated IR-PP concentrations were studied. In response to a meal, IR-PP rose from 151 +/- 19 pg/ml to 296 +/- 35 pg/ml (P less than 0.01) at 15 min, reached a plateau of 367 +/- 56 pg/ml at 45 min, and remained elevated for 4 h. In response to intraduodenal amino acids, IR-PP rose from 196 +/- 22 pg/ml to 342 +/- 19 pg/ml (P less than 0.01), and serum alanine rose from 251 +/- 18 mM to 361 +/- 30 mM. Peak concentrations of IR-PP, however, were reached before alanine rose above baseline. During administration of intraduodenal Na oleate, IR-PP rose from 181 +/- 21 pg/ml to 348 +/- 17 pg/ml (P less than 0.01). Intraduodenal HCl had no effect on IR-PP. Intravenous somatostatin (100 microgram/h) suppressed meal-stimulated IR-PP from 450 +/- 68 pg/ml to 133 +/- 23 pg/ml (P less than 0.01). The data indicate 1) IR-PP is increased after a meal and intraduodenal administration of amino acids and Na oleate; 2) the increases in serum IR-PP precede increases in serum levels of absorbed alanine; and 3) somatostatin suppresses the meal-stimulated increase in serum IR-PP.


Suppression of plasma luteinizing hormone by prolactin in the male rat. Although a direct effect of PRL on gonadotropins has been previously suggested, it has not been convincingly demonstrated. The secretion of LH and FSH was studied in response to the stimuli of castration and LH releasing hormone administration in adult male rats made hyperprolactinemic with ectopic pituitary glands. Although plasma LH and FSH levels were similar in non-castrate hyperprolactinemic rats vs. controls, LH concentrations 24 h postcastration were less in hyperprolactinemic animals as compared to controls (P less than 0.001). The level of LH achieved was inversely correlated with the PRL concentration generated (r = -0.71; P less than 0.01). LH suppression was evident in hyperprolactinemic rats at 1 and 3 days postcastration but was no longer observable at 7 days postcastration. After LH releasing hormone administration to non-castrate rats the rise in plasma LH was significantly less in the hyperprolactinemic animals (P less than 0.05). These experiments support the hypothesis that PRL directly inhibits LH secretion, presumably at the pituitary level.


Localization of delta4-5alpha-reductase in immature rat testes. Testes of 29-35-day-old rats were separated into seminiferous tubules and interstitial tissue by collagenase treatment and examined by incubation studies with radioactive substrates. The activity of testosterone 5alpha-reductase/g protein in the interstitial tissue was 37 times greater than that in the tubules. The site of formation of 5alpha-reduced C21- and C19-steroids from progesterone was found to be primarily in the interstitial tissue. In the interstitial tissue, testosterone 5alpha-reductase activity, which was stimulated 300-fold by the addition of NADPH, was localized in the microsomal fraction (8,000-105,000 X g precipitate). NADPH was five times as effective a hydrogen donor as NADH when the washed microsomal fraction was the source of the enzyme. The formation of 5alpha-reduced C21- and C19-steroids from pregnenolone and progesterone was demonstrated in the microsomal fraction of interstitial tissue. These results indicate that in 29-35-day-old rat testes, 5alpha-reduction of C19-delta4-3-ketosteroids and the formation of 5alpha-reduced C21- and C19-steroids from pregnenolone take place largely in the microsomes of interstitial tissue.