Microangiographic studies on vascularization of the human fetal corpus of the mandible.

Investigations were carried out on 40 human fetuses aged 4 to 8 months. It was found that an increase in number of blood vessels and their diameter are related to mineralization of teeth germs. The blood vessels influence the development of the mandible and teeth.


Variability in the course of the superficial temporal artery in man.

The temporal artery was investigated in 120 human cadavers using injection method. Division of the superficial temporal artery take place below the external acoustic meatus. The frontal ramus of the artery is larger than the parietal one.


Histological and quantitative studies of the muscle spindles in human fetal infrahyoid muscles.

Muscle spindles were investigated in 19 human fetuses, aged 11 to 17 weeks. First spindles were observed in 11th week. The number of muscle spindles increases with advancement of the development being highest in the omohyoid muscle.


Pulmonary arteries dimensions and their dependences on the age, the height and body weight.

The studies were carried out on 100 right and 100 left lungs taken from human corpses of both sexes whose age varied from 1 month to 80 years. The material was divided into 3 age groups. The pulmonary arteries were injected 65% duracryl and then digested in sulfuric acid. The specimens obtained were examined to measurement the length and the diameter of the interlobar portion of the pulmonary arteries and their correlation indexes to the age, the length and the body weight. It was certain, that the pulmonary arteries dimensions still grows during man's extrafetal life; the diameter of the right pulmonary artery is usual greater than the left artery. The length of the interlobar portion of the right pulmonary artery varied in both sexes from 5 to 43 mm, and from 6 to 43 mm on the left pulmonary artery. The diameter of the some part of pulmonary arteries varied from 3 to 21 mm on the right side, and from 4.5 to 20 mm on the left side.


Metric features of the human biceps brachii, brachialis and brachioradialis muscles.

Analysis of metric features of the biceps brachii, brachialis and brachioradialis muscles in adult humans showed distinct correlation between metric features of particular muscles and statistically significant dimorphic differences.


Variation of the median nerve in the forearm and the wrist.

Rare type of division of the median nerve in proximal 1/3 of the forearm was described. The finding was made during autopsy of 53 years old white man. A nerve branch that run separately from the main trunk of the median nerve appeared to be the common digital nerve to the 3rd and 4th finger. Between this branch and the main trunk the radial head of the flexor digitorum superficialis muscle was situated, that eventually converged into the tendon of the superficial flexor of the middle finger.


Innervation of the testis in human fetuses from 16th to 22nd week.

In fetuses of 16th week the tunica albuginea is composed of fibrous and vascular layers. In the vascular layer the nerve fibers are observed. Beginning from the 20th week on the nerve fibers appear between spermatic cords.


Influence of kinetin (6-furfurylo-amino-purine) on human fibroblasts in the cell culture.

Influence of kinetin on human fibroblasts cultured in vitro was studied. Investigations were carried out on I passage fibroblasts, taken from the human skin. Feulgen's method and radioautography were used. Quantitative estimations of DNA in nuclei of fibroblasts were done on cytophotometer. An increase in amount of DNA in nuclei of fibroblasts cytophotometer. An increase in amount of DNA in nuclei of fibroblasts cultured in the presence of kinetin was found. Stronger 3H uridine incorporation was recorded in cultures after administration of the lowest dosis of kinetin. Kinetin, derivative of adenine (6-furfurylo-amino-purine) belongs to cytokinins and it regulates the growth of plants.


Development of the muscle spindles in the human fetal oblique muscles of the eyeball.

The development of muscle spindles in human oblique eye muscles was investigated in 22 fetuses aged 12 to 24 weeks. The muscle spindles appear at 12th week as 1-2 intrafusal fibers surrounded by capsule. With advancement of the development the number of intrafusal myofibers increases. In 14th week the capsule is composed of 2 layers and in 17th week the external capsule appears.


Fine structure of the glial cells in the inferior ganglion of the vagus nerve in human embryos and fetuses.

Basing on electron microscopy the glial cells within the inferior ganglion of the vagus nerve may be recognized during 7th and 8th week. The satellite cells and lemmocytes are of similar structure and they differ as to location within the ganglion. During development the ratio of satellite cells to nerve cells changes and in 23rd week 4-8 glial cells surrounding single neuron are observed. The cytoplasm of glial cells is more electron dense than that of nerve cells and the nucleus is always oval or triangular. Cellular organelles are less numerous in comparison with nerve cells.


Anatomical basis for selective block of the sympathetic fibers in otolaryngological diseases.

In 40 human adults and 10 fetuses during anatomopathological autopsies, the method of blocking of the sympathetic fibers reaching the internal ear was described. The fibers pass around the vertebral artery. Needle was introduced in the place of decussation of horizontal line passing through superior (females) or inferior (males) margin of the cricoid cartilage with the medial margin of the sternocleidomastoid muscle.


Arterial vascularization of the vermiform appendix in human fetus.

In 50 human fetuses of the crown-rump length ranging from 88 mm to 185 mm (12 to 20 weeks), the variability of the arterial vascularization of the vermiform appendix was investigated using the injection method. 3 vascularization types were distinguished on the basis of the size and number of the supplying arteries. The obtained results were compared with those reported by other authors who studied vermiform appendix vascularization in adults. In most cases the vermiform appendix is supplied by single ramus arising from ileocolic artery.


The growth and topography of the human fetal uterus.

Investigations were performed on 12 embryos and 76 fetuses aged 6 to 40 weeks. The uterine cavity is formed in 12th week. During fetal period the uterus grows most intensively between 16th and 24th week. Up to 16th week the fundus of the uterus is at the level of 1st sacral vertebra. From this period the fundus is located at the level of 2nd sacral vertebra.


The development of the uterine ligaments in human fetuses.

In 12 human embryos and 76 fetuses the uterine ligaments were investigated. The round ligament appears at 8th week and the broad ligament is observed in 19th week. Up to 17th week the round ligament is connected with the uterine tube. It arises separately from the border of the uterus beginning from the 18th week. In fetuses of 11 and 12 weeks the broad ligament forms a thick plate enclosing vessels and nerves. The cardinal ligament appears at 10th week.


Architectonics of blood vessels in the wall of the human uterus in microangiographic studies.

Blood vessels of the human uterus were investigated in 50 uteri of women aged 35 to 65 years. The vessels are arranged into layers in the uterine corpus and isthmus. They pass irregularly in the fundus and cervix. With advancement of age the blood vessels in the uterine wall lose their regularity.


Influence of bilateral ovariectomy on the morphology and ultrastructure of the pineal gland in the pig (Sus scrofa)--quantitative and qualitative study.

Ovariectomy effects ultrastructure of pig pineal gland. It caused changes in both structure and number of cytoplasmic components and volume of nuclei. The most significant ones were observed two weeks after ovariectomy. The reported changes point to relationship between the ovaries and pineal in pig.


Investigation of muscle imbalance in the leg in symptomatic forefoot pes cavus: a multidisciplinary study.

The cross-sectional areas of the peroneal and anterior muscle compartments at the same level in the upper leg were measured using magnetic resonance imaging in 41 cases of forefoot pes cavus. The pes cavus group included idiopathic cases and pes cavus associated with Charcot-Marie-Tooth disease, Friedreich's ataxia, cerebral palsy, status postpoliomyelitis, nerve trauma, and spinal cord tethering. Thirty-nine of these cases were symptomatic. The results were compared with studies of 11 normal controls. It was found that in the majority of cases of forefoot cavus, the peroneal compartment was enlarged relative to the anterior compartment when compared with the normal controls. Biopsies of the tibialis anterior and peroneus longus muscles in 18 patients with forefoot pes cavus showed that any relative expansion of the peroneus longus was not due to pseudohypertrophy. Overaction of the peroneus longus in comparison to its antagonist the tibialis anterior is proposed as an important factor in the pathogenesis of the majority of symptomatic cases of forefoot pes cavus.


The validity of measurements made on standard foot orthoroentgenograms.

In certain clinical settings, standard x-rays may be misleading since they only provide two-dimensional representations of three-dimensional structures. After metatarsal osteotomy, the actual first metatarsal shortening may appear magnified on standard x-rays. We investigated variations in metatarsal and intermetatarsal angle (IMA) in a cadaver model and variations of first metatarsal length following in vivo Chevron osteotomies. For skeletal models, changing the foot arch from pes planus to pes cavus changed the first metatarsal length by as much as 19%, whereas the measured IMA remained relatively unchanged. Pronation of the first metatarsal was found to alter the IMA (decreased for planus, increased for cavus). In vivo studies revealed a 3.2-fold difference when actual metatarsal length change following chevron osteotomy was correlated to corrected x-ray change in length (9.5% versus 2.9% P < or = .005). Previous investigators have evaluated hallux valgus deformities with emphasis on metatarsal lengths and IMAs to determine choices of procedure. Based on the results of this study, treatment plans and further evaluation for forefoot surgery may be inaccurate if based solely on x-ray measurements.


Strength of fixation constructs for basilar osteotomies of the first metatarsal.

Twenty-four pairs of fresh-frozen human feet had a proximal osteotomy of the first metatarsal that was fixed using either screws, staples, or K wires. Each metatarsal was excised and the specimen was loaded to failure in a cantilever beam configuration by applying a superiorly directed force to the metatarsal head using an MTS servohydraulic test machine. Specimens with a crescentic osteotomy that were fixed using a single screw demonstrated higher mean failure moments than pairs that were fixed with four staples or two K wires; staples were the weakest construct. All specimens fixed with staples failed by bending of the staples without bony fracture; all K wire constructs but one failed by wire bending. Chevron and crescentic osteotomies fixed with a single screw demonstrated equal bending strengths; the bending strength of an oblique osteotomy fixed with two screws was 82% greater than for a crescentic osteotomy fixed with a single screw. Basilar osteotomies of the first metatarsal are useful in correcting metatarsus primus varus often associated with hallux valgus pathology. Fixation strength is an important consideration since weightbearing forces on the head of the first metatarsal acting at a distance from the osteotomy site subject the construct to a dorsiflexion bending moment, as simulated in our tests. Our results show that screw fixation is the strongest method for stabilizing a basilar osteotomy. Based upon the relatively low bending strengths of the staple and K wire constructs, we would not recommend these forms of fixation.(ABSTRACT TRUNCATED AT 250 WORDS)


Medial deviation of the first metatarsal head as a result of flexion forces in hallux valgus.

Several questions with regard to the hallux valgus complex, which includes metatarsus primus varus, give rise to discussion. How do bunions develop? Is disturbed muscle balance at the first metatarsophalangeal joint important in the pathogenesis of the hallux valgus complex? What is the relation between dynamic plantar load distribution and pain in the ball of the foot? What is the cause of recurrences of deformity after surgery? To answer these questions, we started with the biomechanical model of Snijders et al., which states that contraction of flexor muscles of the hallux worsens its valgus angle and causes medial deviation of the first metatarsal head. The present study was designed to validate the model on patients. When pressing the hallux downward, simultaneously the force under the toe and the medial deviation of the first metatarsal head were measured on preoperative patients and on controls (35 subjects in all). We could demonstrate with statistical significance that (1) when the subjects with hallux valgus push the great toe on the ground, the first metatarsal head moved in medial direction; in other words the foot widened. In the controls, as an average, the foot became narrower. (2) The greater the valgus deviation of the hallux, the greater the effect of the toe flexors, and (3) the greater the valgus deviation of the hallux, the less maximal flexion force it can apply. Implications of these findings on conservative and surgical therapy are discussed. The recurrences of deformity after first metatarsal osteotomies are explained by the action of the hallux flexors.(ABSTRACT TRUNCATED AT 250 WORDS)


Calcaneal fracture after cortical bone removal.

Four patients developed calcaneal fractures while walking, soon after bone removal from the undersurface of the calcaneus. Heel spur or osteomyelitic bone removal can lead to this complication.


Transfer versus lengthening of the posterior tibial tendon in Duchenne's muscular dystrophy.

Transfer of the posterior tibialis tendon to the dorsum of the foot was compared with lengthening of this tendon in 15 patients with Duchenne's muscular dystrophy. Patients undergoing tendon transfer (N = 9) had a longer operating time and two perioperative complications, but retained plantigrade posture of their feet even after walking ceased. By contrast, lengthening of the posterior tibialis tendon (N = 6) was associated with a recurrent equinovarus foot deformity that frequently interfered with shoe wear. Although transfer of the posterior tibialis is out of phase and technically more demanding, the unique prolongation of posterior tibialis strength in Duchenne's muscular dystrophy makes this operation a better option for these patients.


[Characteristics of obese patients in intensive care].

The frequency of overweight, together with the associated increase in morbidity--in particular coronary heart disease--means that such patients often require intensive care. The particular features of intensive care of obese patients are the increased risk of aspiration pneumonia (in the presence of lowered pH and elevated gastric juice volume), respiratory insufficiency caused by a reduction in expiratory reserve volume, and a greater incidence of thrombosis and embolism. Ventricular hypertrophy and dilatation results in a reduced cardiac reserve. A clinically unrecognized insulin resistance may become manifest during intensive care. Digoxin, theophylline and aminoglycosides, commonly with a low therapeutic spectrum, when administered on the basis of weight, may lead to dangerously high serum concentrations in the overweight. At a weight of more than about 100 kg, it must generally be expected that the compensatory mechanisms of the body are exhausted and that there will be an increased susceptibility to additional disorders of homeostasis.


Quantitative determination of intermetatarsal pressure.

Pain, plantar to the metatarsophalangeal joints, is a common location for the presentation of pain in the forefoot. In the absence of fractures or specific inflammatory conditions common causes of pain in this area include: (1) nonspecific metatarsalgia, (2) plantar fat pad atrophy, and (3) interdigital (intermetatarsal) neuroma. The aggravation and possibly the etiology of these forms of forefoot pain may be related to acute or chronic extrinsic pressure to the forefoot. The role of nonweightbearing, compression of the metatarsal heads, weightbearing, and toe-stance (on the metatarsal heads) on the intermetatarsal pressure of the third interspace was measured in eleven asymptomatic volunteers. Intermetatarsal pressures were recorded using the Stryker Miniaturized Digital Fluid Pressure Monitor. Intermetatarsal pressures for nonweightbearing, nonweightbearing with medial-lateral compression of the metatarsal heads, weightbearing, and toe stance averaged 21 mm Hg, 22 mm Hg, 29 mm Hg, and 36 mm Hg, respectively. In comparison to nonweightbearing, significant increases in intermetatarsal pressures were measured with weightbearing (P = .0027) and toe stance (P = .0002). The change noted from weightbearing to toe-stance was also significant (P = .0005). These findings support the proposition that increases in forefoot plantar pressures convey greater pressures to the intermetatarsal space and metatarsal heads. Increased pressures to the intermetatarsal space may likewise lead to or exacerbate the symptoms of patients with fat pad atrophy, nonspecific metatarsalgia and Morton's neuroma.


Metatarsal lengthening by distraction osteogenesis: a report of two cases.

Two cases of lengthening of metatarsals by distraction osteogenesis are reported. One of these cases is an acquired deformity of the first metatarsal; the other is a congenital short fourth metatarsal. By following the principles set forth by Ilizarov, it was possible to lengthen the metatarsal bones and surrounding soft tissues without open lengthening of tendons or secondary bone grafting of the distraction gap. We believe this method is an improvement over previously described methods of metatarsal lengthening.


[Obesity and surgery].

BASICS AND AIMS: Obesity is generally considered to be a surgical risk factor, although its influence cannot be accurately defined. The risks of surgery in the obese patient are discussed on the basis of a review of the literature. In itself, obesity does not increase the surgical risk, but it is often associated with cardio-respiratory, metabolic or coagulation disorders that increase morbidity and mortality in obese patients. The only statistically significant difference between obese and non-obese patients is the higher rate of secondary healing in the former. Guidelines aimed at reducing perioperative risk in these patients are suggested.


A new type of dislocation of the first metatarsophalangeal joint: a case report.

We are reporting a case of traumatic dislocation of the first metatarsophalangeal joint that does not correspond to any previously reported type. This new type of dislocation was associated with a rupture of the lateral short sesamophalangeal ligament/plantar plate, partial rupture of the intersesamoid ligament, and separation of a previously bipartite tibial sesamoid. Closed reduction was easily performed; however, the distal portion of the tibial sesamoid remained trapped within the joint. The rationale for and result of surgical treatment are presented.


Transverse proximal diaphysial fracture of the fifth metatarsal: a review of 12 cases.

A review of 12 cases of transverse proximal diaphysial fractures 3.0 cm distal from the fifth metatarsal tuberosity with a 2-year follow-up is presented. Six cases treated with percutaneous pinning, cast, and partial weightbearing with crutches and six cases treated with cast, partial weightbearing, and crutches are analyzed by classification, age, and average time to union. The healing times were the same in both groups.


[Possible uses for callus distraction. Bridging bone defects and bone lengthening in leg length inequality].

Representation of the possibilities of callus distraction for bridging bone defects on the basis of four typical cases successfully treated with this procedure. The filling of large diaphyseal bone defects through the use of callus distraction is reliable, relatively simple to carry out, and in most cases can be done on an outpatient basis. After prior osteotomy, a segment of the remaining bone is forced, millimeterwise, into the defect zone. As a result of continuous distraction of the callus forning in the osteotomy, tubular bone with weight-bearing capability is formed, and the patient is spared the defect resulting at the donor site when an autologous bone graft is taken, or the implantation of exogenous material. The same principle is employed for leg lengthening, and also permits simultaneous compensation of axis malalignment.


[Parkinson therapy yesterday, today, tomorrow. Neuroprotection gains in importance].

Owing to a lack of knowledge of the pathophysiology and pathochemistry of Parkinson's disease, conservative treatment was long restricted to the treatment of symptoms. In recent decades, as the role of dopamine became better known, progressive improvements in therapy were achieved, which initially meant the administration of the precursor, L-Dopa, of the primarily non-replaceable neurotransmitter, and later augmentation of the activity of dopamine in addition. Amantadine, a highly effective drug with a wide spectrum of action and a high level of tolerability, was successfully introduced in 1969. The recently discovered NMDA antagonism, also in conjunction with a description of the mechanism of action of amantadine, which makes it possible to inhibit the effect of excitatory amino acids--in particular glutamate--in the CNS, led to the principle of neuro-protection, which is now considered the key to the treatment of Parkinson's disease.


[Obstructive respiratory tract diseases--new standards in therapy].

Presentation of modern-day therapeutic procedures in various stages of obstructive airway diseases. Subacute bronchitis and hypersensitivity of the bronchial system are early symptoms of obstructive diseases of the airways. Highly effective prophylactic treatment includes adrenocortical hormones (ACH) (corticosteroid) inhalation and antibiotics used individually or in combination for a variety of indications, are now available. In cases of manifest airway obstruction (Stage B), bronchodilators are additionally required. Of those presently available, the beta 2 sympathicomimetics head the list. For anticholinergics and theophylline there are additional indications. If the resulting response is unsatisfactory, oral ACH must be employed in addition to ACH inhalation and bronchodilators. Cromoglycate administered for a period of four to six weeks should always be tried. In the stage of severe exacerbation (C), intravenous water-soluble ACH in combination with antibiotics, are necessary. The entire spectrum of bronchodilators should also be utilized. Functional analysis during treatment prevents under- or overtreatment. In particular in the early stages, the aim of treatment of obstructive airway diseases is prophylactic.


[Cholecystectomy and colorectal cancer. Does cholecystectomy increase the risk of developing colon cancer?].

Within the course of the last ten years, a possible association between carcinoma of the colon and prior cholecystectomy has been discussed. In 342 patients with colorectal carcinoma, and in a control group of 182 patients with gastric carcinoma, the rate of prior cholecystectomy was investigated. The cholecystectomy rate in the case of colon carcinoma patients was 7.9, and thus differed non-significantly from that of the control group (5.5%). At least in our case material, no relationship was found between cholecystectomy and carcinoma of the colon. The results reported in other studies are compared and discussed.


[ACE inhibition with cilazapril. Major therapeutic aspects: hypertension and metabolic syndrome].

Today, essential hypertension is considered to be genetically closely related to disordered peripheral glucose metabolism, and this situation is described by the term metabolic syndrome. Both diseases--hypertension and type II diabetes--submit the heart and arterial vessels to an unphysiological, chronic stress, which they can compensate only for a certain time. Today, when antihypertensive treatment is indicated, drugs capable of preventing late vascular injury while at the same time having the potency to reverse already existing organic changes, are employed. ACE-inhibitors are presently considered to be the most potent substances that are capable of exerting a positive effect on hypertension-associated changes, while not increasing the individual risk profile in the development of arteriosclerosis. The present paper discusses the new ACE-inhibitor, cilazapril, which can be administered in a practical single dose and develops a profile of action typical of ACE-inhibitors in hypertensives with and without an accompanying metabolic syndrome.


[Healthy aging? Longevity with reference to interdisciplinary gerontologic research].

The percentage of over-60-year-olds, 80- and even 100-year-olds in the general population is increasing rapidly in most countries. What can be done to ensure healthy ageing and assure a good quality of life in old age? What are the correlates of longevity associated with psycho-physical wellbeing? Longitudinal studies underscore the multi-faceted nature of the ageing process: genetic and biological factors influence longevity, as also do social and ecological factors, which in turn help determine the life style that promotes longevity.


[Effect of diuretics on serum lipoproteins].

BASICS: Drug treatment of the risk factor hypertension has not led to the expected reduction in morbidity and mortality of coronary arterial disease. A negative effect of antihypertensive drugs on the lipid metabolism is under discussion as a possible cause. Both the commonly employed thiazide diuretics and chlorthalidone and the loop diuretics furosemide, piretanide and xipamide lead, to varying extents, to an increase in total and LDL cholesterol as well as triglycerides in subjects with normal metabolism. In addition, some diuretics lower the levels of protective HDL cholesterol. These side effects can be avoided--at least in part--by a combination with such drugs as prazosin, pindolol or captopril, which have a favorable effect on metabolism. It is recommended to carefully monitor serum Lipoproteins during diuretic treatment so that possible adverse changes may be counteracted by appropriate measures.


[Therapy with H2 receptor antagonists in childhood. Use of ranitidine in duodenal and gastric ulcer and reflux esophagitis].

Representation of common characteristics and peculiarities of the treatment with H2-receptor antagonists of duodenal and gastric ulcer and gastro-esophageal reflux in childhood in comparison with adults, taking ranitidine as an example. In the treatment of primary and chronic duodenal and gastric ulcers, ranitidine at doses of up to 10 mg/kg/day, given in two fractions (up to 300 mg daily) is a proven form of therapy. The endoscopically confirmed healing rates after 6 to 8 weeks of acute treatment vary between 80 and 100%. The recurrence rate in children with duodenal ulcer is high, so that long-term treatment with 4 to 5 mg/kg/day (evening dose) is added. Following a recurrent ulcer, treatment should be continued for one to two years. Ranitidine has been used with success for the treatment of gastroesophageal reflux. Shorter intervals between administrations (3 times a day) with a total dose of up to 10 mg/kg/day are to be recommended. More experience is required in this area. At therapeutic doses, ranitidine shows hardly any side effects in children, even with long-term treatment, and can be considered to be very well tolerated by this age group, including babies.


Pancreatic and extrapancreatic sulfonylurea receptors.

The hypoglycemic effect of sulfonylureas and their analogues results from their binding to a high affinity site in the B-cell plasma membrane. This site seems to be a structural component of the ATP-sensitive K(+)-channel and represents the pancreatic sulfonylurea receptor. Binding of sulfonylureas causes closure of the ATP-sensitive K(+)-channel and thereby initiates a chain of events eventually leading to the release of insulin. Diazoxide inhibits insulin secretion via opening of the ATP-sensitive K(+)-channel. Sulfonylurea receptors resembling the pancreatic receptor occur in nerve cells, cardiac muscle, skeletal muscle and smooth muscle. Neither these extrapancreatic receptors nor low affinity receptors for sulfonylureas in myocytes and adipocytes contribute to the therapeutic benefit of sulfonylureas.


In vitro effects of glycosylated insulin and glucagon in perfused liver of the rat.

Using perfused liver of the rat, the hepatic uptake of glycosylated insulin (GI) and glucagon (GG) and its effects on hepatic glucose output were investigated. Insulin and glucagon were glycosylated in ambient high glucose concentration, and GI80 or GG80 (insulin or glucagon incubated with 0.08% glucose), GI350 or GG350 (incubated with 0.35% glucose), and GI1000 or GG1000 (incubated with 1% glucose) were prepared. The liver was perfused with the medium containing 1000 microU/ml insulin and 200 pg/ml glucagon or 200 microU/ml insulin and 1000 pg/ml glucagon. The fractional uptake of insulin or glucagon by perfused liver was not significantly altered by the glycosylation. In the liver perfused with 1000 microU/ml insulin and 200 pg/ml glucagon, glucose output was not changed by the glycosylation of the hormones, while in the liver perfused with 200 microU/ml insulin and 1000 pg/ml glucagon, GI1000 reduced its biological activity, as reflected by insulin-mediated decrease in glucose output. These results suggest that in the liver insulin incubated with markedly high concentration of glucose reduces its biological activity at a physiological concentration in the presence of high concentration of glucagon.


Effect of iodoarachidonates on thyroid FRTL-5 cells growth.

Excess iodide inhibits several thyroid parameters, by a putative organic iodocompound. Different iodolipids, including iodinated derivatives of arachidonic acid (IAs), are produced by rat, calf and pig thyroid. The action of two iodolactones, one bearing the iodine atom at the position 6 (IL-d) and the other at position 14 (IL-w) on growth of FRTL-5 cells was studied. KI, IL-w and IL-d exert a dose-related inhibition on FRTL-5 cell proliferation. The first two compounds caused inhibition at 1 microM while IL-d was effective at 10 microM. This inhibitory action of iodolactones (ILs) was not altered by 1 mM methyl-mercaptoimidazol (MMI), indicating that they exert their effect per se. The action of ILw on cell growth was reversible. The growth-stimulating effect of 10 microM forskolin was inhibited by IAs, showing that one possible site of action lies at the cAMP pathway. The present results give further support to our hypothesis about the role of IAs in thyroid growth autoregulation.


Saponin effects of prolactin-like stimulation of ornithine decarboxylase activity in mouse mammary gland explants.

Saponin, a naturally occurring plant glycoside, was found to elicit a prolactin-like stimulation of ornithine decarboxylase (ODC) activity in mouse mammary gland explants. A dose-response activation of ODC was observed with saponin at concentrations between 2 and 10 micrograms/ml. At concentrations of 10 and 15 micrograms/ml, saponin effected a response similar to that of PRL; when tested in concert, PRL and saponin caused a nonadditive response. The time-course of the saponin and PRL effects on ODC activation were not different; a maximum response occurred 2-4 hours after addition of saponin. The saponin and PRL responses were abolished by antibiotics (puromycin and cyclohexamide) that inhibit protein synthesis, but not by actinomycin D which inhibits RNA synthesis. Finally, saponin, by itself, did not affect the rate of milk product formation, but at higher concentrations (above 0.5 microgram/ml) impaired the PRL stimulation of lipid and casein synthesis.


Specific stimulation by estradiol of tissue-type plasminogen activator production in 7,12-dimethylbenz[a]anthracene-induced rat mammary tumor cells.

The hormonal regulation of two plasminogen activators, tissue-type plasminogen activator (t-PA) and urokinase (u-PA), was studied both in 7,12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary carcinoma and in DMBA-induced rat mammary dysplasia. t-PA activity in DMBA-mammary carcinoma was decreased markedly by oophorectomy and recovered upon estradiol administration to reach the maximum level at 12 hr. In contrast to its effect on DMBA-mammary carcinoma, estradiol had no effect on t-PA activity in DMBA-mammary dysplasia. Furthermore, DMBA-mammary carcinoma cells in primary culture displayed similar estrogen-dependency in production of t-PA, while t-PA production in DMBA-mammary dysplasia cells was not under the control of estradiol in vitro. Moreover, estrogen-stimulated production of u-PA activity was not observed in DMBA-mammary carcinoma cells or DMBA-mammary dysplasia cells both in vivo and in vitro. Taken together, these results suggest that estrogen stimulates the production of t-PA but not u-PA and that this estrogen dependency of t-PA is limited to malignant DMBA-mammary tumor cells.


Identification of müllerian inhibiting substance specific binding in human cell lines.

The receptor for Müllerian Inhibiting Substance (MIS), a gonadal glycoprotein hormone, has not been previously identified. Plasma membranes from MIS-sensitive human tumor cell lines (HTB-111, endometrial carcinoma; and A-431, vulvar squamous carcinoma) were detergent extracted and incubated with 125I-labeled MIS anti-idiotypic antibody, or radioiodinated human recombinant MIS (125I rhMIS), with and without unlabeled competitors. 125I anti-idiotypic MIS antibody bound to HTB-111 membrane extracts was displaceable by unlabeled anti-idiotypic antibody, but not by anti-isotypic antibody prior to cross-linking. Specific binding of the anti-idiotypic MIS antibody to endometrial carcinoma cells was verified using fluorescence activated cell analysis and fluoresceinated antibody. Furthermore, unlabeled anti-idiotypic MIS antibody competed for 125I rhMIS binding to A-431 vulvar carcinoma membranes. The labeled anti-idiotypic MIS antibody binding complex could be separated from 32P labeled EGF receptor in the A-431 membranes, indicating that EGF, a natural inhibitor of MIS activity, and MIS itself bind to different receptors. These studies demonstrate a specific, displaceable binder for MIS in the plasmalemmae of two human tumor lines. Purification of this cell surface receptor protein will be greatly aided by using the MIS anti-idiotypic antibody.


Venous versus arterialised venous blood for assessment of blood glucose levels during glucose clamping: comparison in healthy men.

The aim of this study was to investigate the influence of the arteriovenous (A-V) gradient in blood glucose concentrations at low and high insulin levels on the determination of glucose requirements during glucose clamping in 9 healthy, insulin sensitive, male volunteers. In a random order two clamps were performed, once using arterialised venous blood (A Clamp, mean pO2 = 11.5 +/- 0.36 kPa, 86 +/- 2.7 mmHg), and once using venous blood (V clamp, mean pO2 = 7.9 +/- 0.21 kPa, 59 +/- 1.6 mmHg). Insulin levels were maintained at 48 +/- 2.4 mU/l from 0-180 min and at 1054 +/- 114 mU/l from 180-360 min. Elevation of insulin levels caused a significant rise of the A-V gradient: from 0.3 +/- 0.1 to 0.5 +/- 0.1 mmol/l (p < 0.05) and from 0.2 +/- 0.1 to 0.3 +/- 0.1 mmol/l (p < 0.05) during the A and V clamps, respectively. Despite these A-V glucose gradients no significant differences were found for the glucose requirements during the last 30 min of each period of insulin infusion between the A and V clamps: 43.70 +/- 3.4 vs 44.8 +/- 2.8 mumol.kg-1.min-1 during the low insulin level and 77.3 +/- 5.0 vs 76.2 +/- 3.4 mumol.kg-1.min-1 during the high insulin level. We conclude that the A-V glucose gradient, even at high insulin levels, does not influence the assessment of glucose requirements to a measurable extent, allowing the use of the simpler technique of taking venous rather than arterialised venous blood for the measurements of glucose levels during glucose clamping.


Menstrual cycle exhibits no effect on postprandial lipemia.

Menstrual cycle effects on postprandial lipemia were investigated in 13 healthy women. They were monitored (dietary intake, basal body temperature) for a period of one cycle. Statistically significant follicular-luteal differences were neither observed between fasting triglyceride serum levels nor between areas under triglyceride response curves after a standardized oral fat load.


Epidermal growth factor plasma concentrations in healthy control persons, acute and chronic stress and during pregnancy.

Plasma concentrations of Epidermal Growth Factor (EGF) in healthy control persons do not change significantly during adult lifetime and no diurnal changes could be detected. In addition, no alterations of plasma EGF concentrations occur during acute and chronic stress and in all three trimesters of pregnancy. Thus, the previously suggested theories that EGF might play a causative role in the activation of the hypothalamic-pituitary adrenal axis during the stress reaction and in the prevention of peptic ulcers in pregnancy could not be confirmed by the present data.


Urinary excretion of glycosaminoglycans in patients with postmenopausal osteoporosis.

The organic bone matrix contains glycosaminoglycans (GAG) of which the precise function and importance in bone mineralisation are still unclear. We examined 85 persons--35 healthy women (25 premenopausal [preMP] mean aged 40.7 years; 10 menopausal [MP] mean aged 59.3 years) and 50 patients with postmenopausal osteoporosis [PMOP] at a mean age 60.4 years. The dynamic of urinary excretion of GAG was measured in 24-hour collected urine by precipitation with cetylpyridinum chloride and spectrophotometry at 560 nm, corrected for the level of excretion of creatinine. There was a significant increase in GAG excretion in patients with PMOP compared with healthy persons (8.25 mg/g and 9.53 mg/g vs 24.11 mg/g; p < 0.0001). A significant positive correlation was established between GAG and calcium urinary excretion and a negative one between GAG and serum estradiol levels. During the treatment with calcitonin the excretion of GAG was decreased which can be used for monitoring the changes of bone metabolism.


Increased internal calcium mobilization in platelets of patients with chronic renal failure.

Platelet free calcium concentrations ([Ca2+]i) were measured with Fura-2 to elucidate the intracellular calcium kinetics in patients with renal disease. There were no significant differences of the resting [Ca2+]i among the control subjects (C) (n = 12), patients with chronic glomerulonephritis (CGN) (n = 8), and patients with chronic renal failure (CRF) (n = 12). In all groups, platelets [Ca2+]i were significantly increased by agonists (thrombin, adenosine diphosphate) compared with their respective basal level. Thrombin-induced [Ca2+]i rise was significantly higher in CRF (840 +/- 265 nM) than in C (600 +/- 163) and CGN (562 +/- 137). Also adenosine diphosphate elicited similar responses. In the presence of calcium chelator in the incubation buffer, the elevation of [Ca2+]i after thrombin stimulation was statistically higher in CRF (469 +/- 85 nM) than in C (275 +/- 60) and CGN (301 +/- 41). These findings suggest that platelets of CRF were capable of increasing [Ca2+]i in response to agonists, through further mobilization of calcium from the intracellular pool rather than the elevation of transmembrane calcium influx.


Molecular analysis of the sex-determining region from the Y chromosome in two patients with Frasier syndrome.

In the Frasier syndrome there is an association between XY gonadal dysgenesis and chronic renal failure. Owing to an observed sex reversal, the Y chromosomes of two girls with this syndrome have been analyzed. Using molecular-biology techniques, no major alterations of the known sex-determining area of the Y chromosome were found. Furthermore, the sequence did not reveal impairment of the recently described testis-determining factor SRY. These data suggest that in the Frasier syndrome, XY sex reversal and renal failure could be the result of either faulty gene(s) located downstream in the sex differentiation pathway during embryogenesis, or impaired SRY regulation. Preliminary results on the Wilms' tumor suppressor gene WT1, a candidate for acting downstream to SRY, are also provided.


Effects of estrogen, pituitary gonadotropins and prolactin on immunohistochemical localization of inhibin subunits in the ovary of hypophysectomized female rats.

Effects of estrogen, pituitary gonadotropins and prolactin on immunohistochemical localization of alpha- and beta A-subunits in the ovaries of hypophysectomized female rats were investigated. Hypophysectomy resulted in disappearance of immunoreactive inhibin subunits in the ovary. Administration of DES, FSH and LH to hypophysectomized rats provoked growth of follicles, and resulted in positive immunostaining for inhibin subunits in the granulosa cells. In contrast to follicle-stimulating hormone (FSH) and luteinizing hormone (LH), prolactin administration failed to demonstrate positive staining for inhibin subunits in the ovary. The present in vivo results suggest that several hormones which are known to stimulate granulosa cell growth and maturation, such as estrogen, FSH and LH, enhance inhibin subunit production by the ovary. The morphologic aspect of inhibin subunit production by the ovary in response to several hormones has been demonstrated in the present study.


Effects of endogenous and exogenous calcitonin on inflammation-mediated osteopenia in the rat.

Inflammation-mediated osteopenia (IMO) in the rat is characterized by loss of bone mass within 3 weeks after induction of nonspecific inflammation (s.c. talcum injections) in growing rats. Histologically, this shows as marked inhibition of osteoblasts 3 days after the initiation of IMO. The role of calcitonin (CT) was investigated in the present study. A reversible increase of serum CT levels was found after intraperitoneal calcium challenge in rats on day 4 after induction of IMO, which was thought to be a result from calcium efflux from bone. No difference in stimulated serum CT levels between the rats with and without IMO was seen on any other day during 4 weeks after initiation of IMO. Bone loss after IMO was more pronounced in normocalcemic and euthyroid rats with deficiency of endogenous CT (thyroidectomy with parathyroid gland reimplanted) (-12.9%) compared with sham operated controls with IMO (-3.25%). Daily subcutaneous injections of 100 mIU salmon CT in rats with and without IMO did not prevent the development of bone loss. This might have been due to the growing state of rats of this age group. Our results support the hypothesis that endogenous CT physiologically has a bone protective role. They furthermore are consistent with the view that endogenous CT itself is not pathogenetically involved in the development of osteoporosis.


Prediction of advanced puberty by height velocity in children with acute lymphoblastic leukemia.

Pubertal development was retrospectively evaluated in 58 children with cancer, mostly acute lymphoblastic leukemia (ALL), who are in complete remission and off chemotherapy. Six girls [5 patients with ALL, and 1 with malignant lymphoma (NHL)] showed advanced puberty (25.0%, 6 of 24 female patients with ALL and NHL) through the evaluation of their growth velocity. No evidence for advanced puberty was seen in the males. All 6 girls had received cranial irradiation for central nervous system prophylaxis and systemic chemotherapy including glucocorticoid. The mean age at onset of the pubertal growth spurt in these 6 girls was significantly lower than for girls with a solid tumor [6.90 +/- 0.10 and 9.00 +/- 0.77 (mean +/- SD) years, respectively (p < 0.01)]. By simply evaluating the height velocity, we could predict advanced puberty which was ultimately associated with short stature.


Final height attainment and gonadal function in girls with precocious puberty treated with cyproterone acetate.

Thirty-four girls with precocious puberty (27 idiopathic, 6 cerebral, 1 McCune-Albright syndrome) were treated with cyproterone acetate (CPA) for 1.2-8.4 years (3.71 +/- 0.31; mean +/- SEM) at a daily dosage of 66-150 mg/m2 (103.7 +/- 6.2). The mean chronological age (CA) and bone age at the beginning of treatment were 5.99 +/- 0.31 and 8.6 +/- 0.39 years, respectively, and 9.78 +/- 0.19 and 12.44 +/- 0.22 years, respectively, at the end of therapy. At the last evaluation, mean CA was 14.23 +/- 0.4 years, and 32 girls had reached final height. The control group consisted of 10 girls with idiopathic precocious puberty who, at their parents' request, were not treated. Mean CA at the onset of pubertal signs was 6.05 +/- 0.25 years. All patients had reached final height at the time of the last observation. There was no significant difference between final height of treated (152.43 +/- 1.36 cm) and untreated (149.55 +/- 1.99 cm) girls. Final height was significantly lower than target height in both treated (155.08 +/- 0.92 cm; p < 0.025) and untreated (156.45 +/- 1.29 cm; p < 0.0005) patients, but the mean height of treated patients is nearer to target height than that of untreated ones. A positive correlation was found between final height and target height both in treated (p < 0.005) and untreated (p < 0.05) patients. After the discontinuation of CPA treatment all girls resumed the progressive course of puberty.(ABSTRACT TRUNCATED AT 250 WORDS)


Growth after radiotherapy and chemotherapy in children with leukemia or lymphoma.

The effect of radio- and chemotherapy on auxological parameters was investigated in 30 children treated for acute lymphatic leukemia (ALL) or non-Hodgkins lymphoma (NHL). Growth velocity was decreased during the first year of treatment. Catch-up growth was insufficient during the following years. Thus, the whole group experienced a loss of height of 0.49 +/- 1.1 SD at 6.8 +/- 2.6 years after diagnosis. Height and growth velocity were not different between children who received 18 or 24 Gy cranial irradiation; however, growth velocity was significantly lower in children who were treated for more than 2 years or who had the more intensive chemotherapeutic protocol. Evaluation of the growth hormone (GH) response to pharmacological stimulation revealed reduced GH peaks in 47% of the patients, but there was no correlation of GH peak with growth or treatment parameters. In conclusion, the impairment of growth in children after treatment for ALL or NHL might be related to the intensity and duration of chemotherapy.


Familial adrenal feminization probably due to increased steroid aromatization.

5/10 members of a North African family (father, 2 male and 2 female siblings) had gynaecomastia, early growth and short final stature. The 8-year-old propositus had advanced bone age, facial acne, gynaecomastia, pubic hair and prepubertal testicular volume. Basal oestrone (E1) was elevated (670 pmol/l) and increased with adrenocorticotropic hormone (ACTH; 826 pmol/l). After human chorionic gonadotropin stimulation testosterone (T) responded normally whereas E1 and oestradiol (E2) remained unchanged. ACTH-dependent adrenal feminization was confirmed by a transient reduction of breast tissue following dexamethasone or cypropterone acetate treatment. Testolactone increased T/E2 (from 5.6 to 20.3) and A/E1 (from 3.4 to 31.4) ratios and temporarily reduced the breast tissue. In conclusion, this is a familial type of adrenal feminization with increased adrenal androgen aromatization. This is the first time that male-to-male and male-to-female transmission has been reported.


Body positions and movement patterns in female patients with congenital adrenal hyperplasia.

Female patients with congenital adrenal hyperplasia (CAH; N = 33; 11-41 years), simple-virilizing (SV) patients (N = 19), salt-wasting (SW) patients (N = 13), and sister controls (N = 14) were compared with regard to their body positions and movement patterns. Data collection comprised both self assessments and mothers' assessments using 20 sex-dimorphic items with corresponding "more masculine" and "more feminine" versions for each variable, represented in photographs (forced-choice approach). Primarily based on mothers' assessments, single-item results suggested slightly more masculine positions and patterns for female CAH patients compared to sisters, for SW patients more distinct than for SV patients. Results from an 11-item scale ("motor behavior", alpha = 0.59) revealed differences between SW (more masculine) and SV patients for self assessments (P, one-tailed, < 0.09); sisters were in an intermediate position closer to the SV patients. According to mothers' assessments, the CAH patient group as a whole differed (more masculine) from sisters (P < 0.06); this finding was mainly accounted for by the SW group (P < 0.04). Complex analyses on the relationship of motor behavior and intervening variables (e.g., postnatal androgenization, onset of puberty, menarche, height, weight, sexual orientation) revealed very few significant results. Findings rather suggested organizational hormonal effects on body positions and movements prenatally; they are in line with main results from the interview section of the Hamburg CAH study (e.g., "Gender-related behavior"). An approach of this kind seems to be justified for investigating motor behavior in future psychoendocrine studies.


Effects of acute and repeated exposure to stress on the hypothalamo-pituitary-adrenocortical activity in mice during postnatal development.

Hypothalamo-pituitary-adrenocortical (HPA) response to a mild stressful procedure was investigated in mice at Days 8, 10, 12, and 14 of postnatal development. Pups that were removed from the dam and exposed to a novel odor (clean bedding) for 15 min showed higher plasma corticosterone levels than pups whose mother was removed from the cage for 15 min or unhandled pups at all ages, although statistically significant differences were only evident at Days 12 and 14. Lower HPA axis responding in younger mice was not due to immaturity since 8-day-old mice showed a significant and larger increase of plasma corticosterone levels when separated from the mother and isolated from littermates in the absence of bedding. Mice daily exposed to clean bedding (15 min) for the first 13 days of life did not show reduced plasma corticosterone response when reexposed to the stressor at 14 days of age. Conversely, increased plasma corticosterone levels in dams in response to removal of pups was not detectable after repeated exposure to this manipulation (14 days) regardless of the procedure their pups were submitted to, thus ruling out a role of maternal corticosterone passing through the milk on which the pups were fed. These results demonstrate that 15 min exposure to clean bedding is a noninvasive procedure able to elicit HPA axis response in developing mice over a wide age range without producing habituation.


Winning, losing, mood, and testosterone.

In two experiments, male college students either won or lost $5 on a task controlled entirely by chance. In both studies, winners reported a more positive mood change than did losers and, in Experiment 2, winners reported a more positive mood change than a neutral group that did not win or lose money. After the task was completed, winners exhibited significantly higher testosterone levels than losers. Levels of cortisol, a hormone associated with stress and arousal, did not differ among the groups, suggesting that a hormone-behavior response pattern for winning and losing is specific to testosterone. These data suggest that winning can alter testosterone levels in men and that mood may mediate such changes.


Investigation of familiar and novel chemosensory stimuli by golden hamsters: effects of castration and testosterone replacement.

Estrous hamsters secrete an odorous vaginal discharge that intact male hamsters investigate vigorously during copulatory behavior. Castrated animals are not attracted to this vaginal discharge. In this study we observed that repeated exposure of intact and castrated hamsters to this vaginal discharge reliably produced habituation of investigatory behavior. Presentation of the odor from a novel female to a habituated male caused an increase in investigation (dishabituation). Castration produced a decrease in investigation within 1 week of surgery. However, the surgery did not produce a decrease in dishabituation until 3 months afterward. Testosterone treatment increased chemosensory investigation in castrated animals. It seems likely that the dishabituation observed in this study may represent a sensory component of the Coolidge Effect.


Time course of androgenic modulation of odor preferences and odor cues in male meadow voles, Microtus pennsylvanicus.

During the breeding season, male meadow voles prefer female over male odors and females prefer male over female odors. Testosterone control of males' odor preferences and production of odors attractive to females differ. A male meadow vole's preference for female versus male odor was still evident 1 week after castration, but not 1 week later. This preference was reinstated in testosterone-treated male voles 2 weeks after the onset of hormone replacement. The attractiveness of male odors to females did not disappear until 3 weeks after castration. The attractiveness of male odors was reinstated 1 week after castrated males were treated with testosterone. The time course for the androgenic modulation of production of odors attractive to females may facilitate breeding. For example, at the end of the breeding season males may emit an odor that is still attractive to females. Similarly, at the beginning of the breeding season males may emit an odor that is attractive to females.


Sexual differences in hormonal control of release calls in bullfrogs.

Release calls in anuran amphibians are given when animals are inappropriately clasped by others. Since other call types, such as mate calls, are sexually dimorphic in frogs and toads, sonogram analysis was used to determine whether release call characteristics might also be sexually dimorphic in bullfrogs (Rana catesbeiana). Only intercall intervals differed significantly between males and females. Call duration, dominant frequency characteristics, and the display of secondary or tertiary harmonic frequencies were similar in both sexes. In the spring, but not the fall, calling rates were significantly lower in female bullfrogs, compared to males. Females also had significantly lower plasma androgen concentrations and higher plasma estrogen in the spring, compared to males. In both sexes, plasma androgen and estrogen were significantly higher in the spring, compared to the fall. The neuropeptide arginine vasotocin significantly decreased release call rates in females in the spring while it significantly increased rates in males. Vasotocin had no significant effect in the fall. Prostaglandin E2 significantly inhibited release calling in both males and females. On the other hand, prolactin significantly inhibited calling in female bullfrogs, but had no affect in males. Thus, although acoustic characteristics of release calls were similar in male and female bullfrogs, hormonal control of call rates was sexually dimorphic and seasonally variable.


An assessment of agonist/antagonist effects of tamoxifen in the female mouse brain.

Ovariectomized CFW mice were treated with tamoxifen (TAM) alone or in combination with estradiol benzoate (EB) to determine its ability to promote/block lordotic behavior and the induction of hypothalamic progestin receptors (PR). Across a range of doses, TAM plus progesterone treatment did not activate female sexual behavior. When given with EB, TAM suppressed lordotic behavior in a dose-dependent fashion. TAM did not induce PR when given alone and it completely blocked the ability of EB to induce PR. It therefore appears that for these responses TAM functioned as a pure antagonist in the female mouse brain, although the degree of its antiestrogenicity varied with the response under consideration. A potential mechanism mediating this differential effectiveness is discussed.


Regulation of sociosexual communication in female Long-Evans rats by ovarian hormones.

Two experiments examined the role of the steroid hormones, estradiol (E2), progesterone (P), and testosterone (T), in activating scent marking and 50-kHz ultrasonic vocalizations in ovariectomized Long-Evans rats in response to a devocalized male rat. In Experiment 1, females received, in a counterbalanced order, a series of six hormone treatments consisting of two injections (48-54 and 4 hr before behavioral tests). The six treatments were 8 micrograms E2 followed by 500 micrograms P or oil, 2 micrograms E2 followed by 500 micrograms P or oil, and oil followed by 500 micrograms P or oil. Injections of either the high or low dose of E2 followed by P resulted in high levels of vocalizations. Neither E2 by itself or P by itself were very effective. Surprisingly, none of the hormone treatments were effective in activating marking above the level seen when the females received control injections of oil. Four other hormone treatments were examined in Experiment 2: daily injections of 500 micrograms T, daily injections of 50 micrograms E2, implantation of silastic capsules of E2 (5% E2, 5 mm length) followed by 500-micrograms P injections before behavioral tests, and implantation of silastic capsules of E2 followed by oil injections. Animals receiving E2 by silastic capsule followed by P injection displayed the highest levels of marking and vocalizations across the five weekly tests. These results suggest that while E2 and P synergize for the display of female-typical behaviors similar to the hormonal regulation of lordosis, the mechanism of E2 action may be different for the two signaling behaviors. Scent marking appears to be responsive to the tonic levels of E2 released from silastic capsules.


The abnormal conduct of capercaillies Tetrao urogallus.

The frequency, behavior, and testosterone levels of deviant capercaillies were studied in southern Finland. Roughly 1% of the population was estimated to behave abnormally. Deviant males display and show threatening behavior toward not only humans, but also attacked stuffed capercaillie males and without hesitation copulated with a stuffed female. Deviant females appear tame and crouch in front of humans, but avoid males. The testosterone level in deviant males was about five times higher than that of normal displaying males. The origin of this striking phenomenon is still unclear, although it must be connected with display. Two, though not mutually exclusive, proximate mechanisms for the development of this abnormal behavior are presented: anomalous sex hormone concentrations and incorrect sexual imprinting. The ultimate reason for the apparent increase in numbers of deviant capercaillies is probably logging which diminishes populations and, thereby, favors the spreading of abnormal behavior.


Behavioral and hormonal correlates of alternative reproductive strategies in a polygynous lizard: tests of the relative plasticity and challenge hypotheses.

Species with alternative reproductive tacts are good models to investigate the poorly understood question of whether individual variation within sexes results from the same physiological mechanisms that control variation between sexes. We have shown previously that adult male tree lizards, Urosaurus ornatus, of different throat color morphs express different levels of aggression in the laboratory. Further field results support the suggestion that the two morphs practice alternative reproductive tactics because the two morphs express different levels of aggressive behavior under field conditions and exhibit dramatic and opposite responses to aggressive challenges. However, despite these behavioral differences, the two morphs do not differ in levels of testosterone or corticosterone either in undisturbed situations or following aggressive challenge. These results are consistent with the relative plasticity hypothesis which proposes that organizational, rather than activational, actions of steroid hormones will be more important in morph differentiation when morphs are fixed in adult life, as they are in tree lizards. These results also support the hypothesis that steroid hormonal levels are insensitive to social modulation in males of species such as U. ornatus without paternal care.


Gonadal hormones determine sex differences in timing of incubation by doves.

Male and female ring doves express a sexually dimorphic pattern of incubation. The dimorphism is temporal rather than motoric. The male incubates for a block of time in the middle of the day and the female incubates the rest of the time. The present study explored the role of gonadal hormones in the control of the temporal dimorphism. Female-female pairs incubated their eggs, but it could not be predicted which of the partners would be sitting on the nest at any given time. Male-male pairs did not incubate and instead destroyed the nests that were provided and displayed aggressive behavior. Some intact males incubated when paired with gonadectomized males, although the castrates tended to ignore the nest. In contrast, when castrated birds were given heterotypical hormonal replacement therapy, they and their same-sex partners incubated the eggs that had been provided, with the gonadectomized birds sitting at a time appropriate to the hormonal state. The results indicate that gonadal hormones influence not only the expression of incubation behavior, but also its phase and duration.


Structure and linkage of the D2 dopamine receptor and neural cell adhesion molecule genes on human chromosome 11q23.

The gene encoding the D2 dopamine receptor (DRD2) is located on human chromosome 11q23 and has been circumstantially associated with a number of human disorders including Parkinson's disease, schizophrenia, and susceptibility to alcoholism. To determine the physical structure of the DRD2 gene, we utilized cosmid cloning, isolation of yeast artificial chromosomes (YACs), and pulsed-field gel electrophoresis to construct a long-range physical map of human chromosome 11q23 linking the genes for the DRD2 and neural cell adhesion molecule (NCAM). The D2 dopamine receptor gene extends over 270 kb and includes an intron of approximately 250 kb separating the putative first exon from the exons encoding the receptor protein. The resulting physical map spans more than 1.5 mb of chromosome band 11q23 and links the DRD2 gene with the gene encoding the NCAM located 150 kb 3' of the DRD2 gene and transcribed from the same DNA strand. We additionally located the sites of at least four hypomethylated HTF islands within the physical map, which potentially indicate the sites of additional genes. High-resolution fluorescent in situ suppression hybridization using cosmid and YAC clones localized this gene cluster between the ApoAI and STMY loci at the interface of bands 11q22.3 and 11q23.1.


Multiple variants in subtelomeric regions of normal karyotypes.

We describe a human genomic cosmid clone, 56.1.1, that contains subtelomeric sequences present on multiple human chromosomes. In particular, using fluorescence in situ hybridization, we have identified 16 sites of hybridization on 12 chromosomes. In a sample of 8 unrelated individuals, 10 of these sites showed interindividual variation. Co-hybridization with other polymorphic probes allowed us to demonstrate cytologically heterozygosity at three sites in six individuals. The chromosomal distribution of hybridization sites in a family strongly suggests that these variants are inherited in a Mendelian fashion. These data show that subtelomeric repeats are a rich source of genetic variability. Possible mechanisms of generation of such variants are discussed.


Application of automated DNA sizing technology for genotyping microsatellite loci.

Highly polymorphic microsatellite loci offer great promise for gene mapping studies, but fulfillment of this potential will require substantial improvements in methods for accurate and efficient genotyping. Here, we report a genotyping method based on fluorescently labeled PCR primers and size characterization of PCR products using an automated DNA fragment analyzer. We capitalize on the availability of three distinct fluorescent dyes to label uniquely loci that overlap in size, and this innovation increases by threefold the number of loci that can be analyzed simultaneously. We label size standards with a fourth dye and combine these with the microsatellite PCR products in each gel lane. Computer programs provide very rapid and accurate sizing of microsatellite alleles and efficient data management. In addition, fluorescence signals are linear over a much greater range of intensity than conventional autoradiography. This facilitates multiplexing of loci (since signal intensities often vary greatly) and helps distinguish major peaks from artifacts, thereby improving genotyping accuracy.


Evolution of DNA sequence homologies between the sex chromosomes in primate species.

Cloned DNA sequences from 18 X-Y homologous loci have been used to examine the evolution of regions of homology between the human X and Y chromosomes. The pattern of X-Y linkage in different primate species has enabled the charting of the chronology of their appearance and removal from the sex chromosomes during evolution. Examination of the pattern of differences in restriction enzyme sites at different loci has been used to estimate the degree of divergence in three different regions of homology. These studies have indicated that (1) blocks of homology have arisen at different points in evolution, (2) different regions of homology are heterogeneous in composition in that they contain X-Y homologous sequences of different age, and (3) the combination of X and Y locations together with the point of evolutionary origin has defined five new patterns of homology.


Closure of a genetic linkage map of human chromosome 7q with centromere and telomere polymorphisms.

We have constructed a 2.4-cM resolution genetic linkage map for chromosome 7q that is bounded by centromere and telomere polymorphisms and contains 66 loci (88 polymorphic systems), 38 of which are uniquely placed with odds for order of at least 1000:1. Ten genes are included in the map and 11 markers have heterozygosities of at least 70%. This map is the first to incorporate several highly informative markers derived from a telomere YAC clone HTY146 (locus D7S427), including HTY146c3 (HET 92%). The telomere locus markers span at least 200 kb of the 7q terminus and no crossovers within the physical confines of the locus were observed in approximately 240 jointly informative meioses. The sex-equal map length is 158 cM and the largest genetic interval between uniquely localized markers in this map is 11 cM. The female and male map lengths are 181 and 133 cM, respectively. The map is based on the CEPH reference pedigrees and includes over 4000 new genotypes, our previously reported data plus 29 allele systems from the published CEPH version 5 database, and was constructed using the program package CRI-MAP. This genetic linkage map can be considered a baseline map for 7q, and will be useful for defining the extent of chromosome deletions previously reported for breast and prostate cancers, for developing additional genetic maps such as index marker and 1-cM maps, and ultimately for developing a fully integrated genetic and physical map for this chromosome.


The CEPH consortium linkage map of human chromosome 2.

This paper describes the Centre d'Etude du Polymorphisme Humain (CEPH) consortium linkage map of chromosome 2. The map contains 36 loci defined by genotyping generated from the CEPH family DNAs. A total of 73 different markers were typed by 14 contributing laboratories; of these, 36 loci are ordered on the map with likelihood support of at least 1000:1. Markers are placed along the length of the chromosome but no markers were available to anchor the map at either telomere or the centromere. Multilocus linkage analysis has produced male, female, and sex-averaged maps extending for 261, 430, and 328 cM, respectively. The sex-averaged map contains five intervals greater than 15 cM and the mean genetic distance between the 36 uniquely placed loci is 9.1 cM.


Cloning and characterization of the human sperm receptor ligand ZP3: evidence for a second polymorphic allele with a different frequency in the Caucasian and Japanese populations.

The human ZP3 gene, encoding the glycoprotein responsible for sperm-egg recognition, has been cloned using mouse ZP3 DNA as a probe. Genomic and cDNA cloning revealed eight exons, spread over 18 kb, encoding a protein of 424 amino acids with a 67% homology to mouse and hamster ZP3. Southern blotting, gene cloning, and sequence analysis were used to show that ZP3 is not a single-copy gene and that the human genome contains a second polymorphic locus which, due to an extra G residue in exon 8, has the potential to encode a truncated protein of 372 amino acids. Direct sequence analysis of polymerase chain reaction-amplified exon 8 DNA of 56 individuals of various human populations revealed three different sequence patterns: one containing only ZP3-424-coding sequences and two containing ZP3-424- and ZP3-372-coding DNA. The distribution of these three sequence patterns is significantly different between the Caucasian and Japanese populations, as indicated by ZP3-372 allele frequencies of 69 and 21%, respectively. Isolation of ZP3-424 and ZP3-372 cDNAs suggests that both loci represent functional transcription units. Therefore, it is hypothesized that throughout the human population during oogenesis ZP3 is translated from mRNAs derived from two to four transcription units. Provided that ZP3-372 mRNA is translated in vivo, corresponding differences in ZP3-372 protein levels might have an impact on human zona pellucida composition.


Cloning and linkage mapping of three polymorphic tetranucleotide (TAAA)n repeats on human chromosome 21.

We report the cloning, sequencing, and mapping of three short sequence repeat polymorphisms due to tetranucleotide (TAAA)n repeats from human chromosome 21. These DNA markers (D21S221, D21S225, D21S226) have been cloned from the chromosome 21-specific plasmid library of J. C. Fuscoe, C. C. Collins, D. Pinkel, and J. W. Gray (1989, Genomics 5: 100-109) and were shown to be polymorphic by polymerase chain reaction amplification and polyacrylamide gel electrophoresis. Genotypes were determined in informative CEPH pedigrees and used in linkage analysis relative to other mapped markers on human chromosome 21. One of these markers, D21S221, is closely linked to the amyloid precursor protein gene (APP), which has been implicated in the etiology of familial Alzheimer disease in some families.


Characterization of the mouse apolipoprotein Apoa-1/Apoc-3 gene locus: genomic, mRNA, and protein sequences with comparisons to other species.

In this report we present the genomic, cDNA, and predicted protein sequences for mouse apolipoproteins A-I and CIII, as well as sequence comparisons with other species. The genes for these apolipoproteins are within 2.5 kb of each other and convergently transcribed. The almost 9 kb of genomic sequence presented extends from 1298 bp 5' to the apolipoprotein A-I (Apoa-1) gene to 1249 bp 5' to the apolipoprotein CIII (Apoc-3) gene. The mouse Apoa-1 gene is 1.76 kb in length with four exons and three introns. The 5' flanking region contains TATA and CCAAT box sequences, an interferon responsive element homology, and potential binding sites for transcription factors CTF/NF1 and HNF4. Translation of the cDNA predicts that the mouse Apoa-1 primary transcript is 264 amino acids. The mouse Apoc-3 gene is 2.2 kb in length and also consists of four exons and three introns. The 5' flanking region contains TATA and CCAAT box sequences, RXR-1 and ARP-1 binding sites, and potential binding sites for transcription factors HNF4, NFkB, AP-1, and CTF/NF1. Translation of the cDNA predicts that the mouse Apoc-3 primary transcript is 99 amino acids. The clustering and genomic organization of the mouse Apoa-1 and Apoc-3 genes are similar to those of the rat and human genes. Significant sequence homologies between species exist for the proximal promoter and exonic regions of each gene, but not for the intronic or intergenic regions.(ABSTRACT TRUNCATED AT 250 WORDS)


Identification and localization of microsatellite markers covering human chromosome 18.

To generate microsatellite markers from chromosome 18, we have cytogenetically localized a large number of lambda phage using a deletion mapping panel of somatic cell hybrids. Here we describe the identification of 65 new CA-repeat-containing phage and the localization of five markers developed in other laboratories. This approach allows the selection of a subset of markers that are well spaced across the chromosome and can be developed as genetic markers. The use of PCR-based markers should allow for the rapid genomic screening of disease genes on chromosome 18.


A panel of human chromosome 22-specific sequence tagged sites.

A panel of 29 sequence tagged sites (STSs) covering the long arm of chromosome 22 has been assembled. STS primer pairs were synthesized using available chromosome 22 sequence derived from the GenBank and EMBL DNA sequence databases, as well as published cDNA and genomic sequence, or from previously published and communicated primer pairs. Each STS was optimized for the polymerase chain reaction using a chromosome 22-only hybrid and human genomic DNA. Further STS content analysis on a panel of somatic cell hybrids that incorporated two chromosome 22 translocations resulted in the mapping of the X-box binding protein (XBP), D22S156, and transcobalamin II (TCN2) genes to 22q11-q13.1. The panel of STSs was used for the rapid determination of the STS content and thus the chromosomal DNA content of a new irradiation hybrid.


Human dopamine transporter gene (DAT1) maps to chromosome 5p15.3 and displays a VNTR.

The human dopamine transporter (DAT1) gene is localized to chromosome 5p15.3 by in situ hybridization and PCR amplification of rodent somatic cell hybrid DNA. Analysis of a 40-bp repeat in the 3' untranslated region of the message revealed variable numbers of the repeat ranging from 3 to 11 copies. These results will aid in the investigation of a role for this gene in genetic disorders of the dopaminergic system in humans.


Localization of the human CYP17 gene (cytochrome P450(17 alpha)) to 10q24.3 by fluorescence in situ hybridization and simultaneous chromosome banding.

The gene for human P450(17 alpha) (CYP17) was previously mapped to chromosome 10 through analysis of somatic cell hybrids. Using a modified procedure of fluorescence in situ hybridization, this gene has now been visualized on simultaneously banded chromosomes and localized to a specific subband of chromosome 10 at q24.3. This precise assignment may facilitate the understanding of the molecular basis of 17 alpha-hydroxylase/17,20-lyase deficiency and the evolution of the CYP superfamily of genes.


The CEPH consortium linkage map of human chromosome 15q.

The CEPH consortium map of chromosome 15q is presented. The map contains 41 loci defined by genotypes generated from CEPH family DNAs with 45 different probe and restriction enzyme combinations contributed by 10 laboratories. A total of 29 loci have been placed on the map with likelihood support of at least 1000:1. The map extends from 15q13 to 15q25-qter. Multipoint linkage analyses provided estimates that the male, female, and sex-averaged maps extend for 127, 190, and 158 cM, respectively. The largest interval is 21 cM and is between D15S37 and D15S74. The on-average locus spacing is 5.6 cM and the mean genetic distance between the 21 uniquely placed loci is 8 cM.


Localization of the gene for branchiootorenal syndrome to chromosome 8q.

Branchiootorenal syndrome is an autosomal dominant disorder that affects an estimated 2% of profoundly deaf children. In addition to hearing impairment, it is characterized by a lop-ear deformity, preauricular pits, branchial cleft sinus tracts, and renal anomalies. The pathogenesis of the disease remains unknown; however, the defective gene has been localized to chromosome 8q by family linkage studies.


Mapping of the versican proteoglycan gene (CSPG2) to the long arm of human chromosome 5 (5q12-5q14).

Versican is a major chondroitin sulfate proteoglycan of vascularized connective tissues whose eponym reflects its functional versatility in macromolecular affinity and interactions. In this report we have localized the versican gene (CSPG2) to the long arm of human chromosome 5 by utilizing a combination of somatic cell hybrids, Southern blotting, polymerase chain reaction, and chromosomal in situ hybridization. The proteoglycan gene segregated concordantly with hybrid cell lines containing the long arm of chromosome 5, comprising the 5q12-q14 band regions. To refine this locus further, we screened a chromosome 5-specific library and isolated several genomic clones encoding a portion of the 5' end of versican. One of these genomic clones was used as a probe for in situ hybridization of human chromosome metaphases. The results corroborated the data obtained using somatic cell hybrids and further refined the assignment of the versican gene to the narrow band region of 5q12-5q14, with the primary site likely to be 5q13.2. The availability of novel genomic clones and the mapping data presented here will make possible the identification of any defect genetically linked to this proteoglycan gene.


The L-isoaspartyl/D-aspartyl protein methyltransferase gene (PCMT1) maps to human chromosome 6q22.3-6q24 and the syntenic region of mouse chromosome 10.

We have mapped the genes for the human and mouse L-isoaspartyl/D-aspartyl protein carboxyl methyltransferase (EC 2.1.1.77) using cDNA probes. We determined that the human gene is present in chromosome 6 by Southern blot analysis of DNA from a panel of mouse-human somatic cell hybrids. In situ hybridization studies allowed us to confirm this identification and further localize the human gene (PCMT1) to the 6q22.3-6q24 region. By analyzing the presence of an EcoRI polymorphism in DNA from backcrosses of C57BL/6J and Mus spretus strains of mice, we localized the mouse gene (Pcmt-1) to chromosome 10, at a position 8.2 +/- 3.5 cM proximal to the Myb locus. This region of the mouse chromosome is homologous to the human 6q24 region.


YAC-assisted cloning of a putative G-protein mapping to the MHC class I region.

We report the successful use of whole yeast artificial chromosomes (YACs) as probes for direct positional cloning of novel expressed sequences in a given genomic fragment. The class I region of the human major histocompatibility complex, in particular the chromosomal fragment spanning the HLA-E locus, was investigated. The screening of a cDNA library with a 210-kb-long YAC clone led to the identification of a new gene, positionally conserved in the major histocompatibility complex of the mouse genome and encoding a putative GTP binding protein. Although its precise function remains unknown, the interspecies conservation of both sequence and map position suggests a regulatory or functional link with the histocompatibility cluster.


Syntenic conservation of HSP70 genes in cattle and humans.

A phage library of bovine genomic DNA was screened for hybridization with a human HSP70 cDNA probe, and 21 positive plaques were identified and isolated. Restriction mapping and blot hybridization analysis of DNA from the recombinant plaques demonstrated that the cloned DNAs were derived from three different regions of the bovine genome. One region contains two tandemly arrayed HSP70 sequences, designated HSP70-1 and HSP70-2, separated by approximately 8 kb of DNA. Single HSP70 sequences, designated HSP70-3 and HSP70-4, were found in two other genomic regions. Locus-specific probes of unique flanking sequences from representative HSP70 clones were hybridized to restriction endonuclease-digested DNA from bovine-hamster and bovine-mouse somatic cell hybrid panels to determine the chromosomal location of the HSP70 sequences. The probe for the tandemly arrayed HSP70-1 and HSP70-2 sequences mapped to bovine chromosome 23, syntenic with glyoxalase 1, 21 steroid hydroxylase, and major histocompatibility class I loci. HSP70-3 sequences mapped to bovine chromosome 10, syntenic with nucleoside phosphorylase and murine osteosarcoma viral oncogene (v-fos), and HSP70-4 mapped to bovine syntenic group U6, syntenic with amylase 1 and phosphoglucomutase 1. On the basis of these data, we propose that bovine HSP70-1,2 are homologous to human HSPA1 and HSPA1L on chromosome 6p21.3, bovine HSP70-3 is the homolog of an unnamed human HSP70 gene on chromosome 14q22-q24, and bovine HSP70-4 is homologous to one of the human HSPA-6,-7 genes on chromosome 1.


Conserved regulatory elements in the promoter region of the N-CAM gene.

Genomic clones containing 5'-flanking sequences, the first exon, and the entire first intron from the chicken N-CAM gene were characterized by restriction mapping and DNA sequencing. A > 600-bp segment that includes the first exon is very G + C-rich and contains a large proportion of CpG dinucleotides, suggesting that it represents a CpG island. SP-1 and AP-1 consensus elements are present, but no TATA- or CCAAT-like elements were found within 300 bp upstream of the first exon. Comparison of the chicken promoter region sequence with similar regions of the human, rat, and mouse N-CAM genes revealed that some potential regulatory elements including a "purine box" seen in mouse and rat N-CAM genes, one of two homeodomain binding regions seen in mammalian N-CAM genes, and several potential SP-1 sites are not conserved within this region. In contrast, high CpG content, a homeodomain binding sequence, an SP-1 element, an octomer element, and an AP-1 element are conserved in all four genes. The first intron of the chicken gene is 38 kb, substantially smaller than the corresponding intron from mammalian N-CAM genes. Together with previous studies, this work completes the cloning of the chicken N-CAM gene, which contains at least 26 exons distributed over 85 kb.


Conservation of coding and transcriptional control sequences within the snRNP E protein gene.

The snRNP E protein is one of several proteins associated with the U family of small nuclear RNAs that are involved in RNA processing. Isolation and characterization of the snRNP E protein cDNA sequences from mouse and chicken revealed a 100% conservation of the predicted amino acid sequence when compared to that of the human homologue. Further characterization of a genomic clone for the mouse snRNP E protein gene revealed that the 5' untranslated region and the immediate 5' upstream region have also been highly conserved: 72 and 70%, respectively. Conserved 5' regions include multiple copies of the CTTCCG hexamer sequence which are involved in regulating transcription of the human snRNP E protein gene. Mobility shift assays using corresponding DNA fragments from both human and mouse reveal that both fragments can compete for binding of at least one common transcription factor. These studies demonstrate that along with the amino acid sequence conservation between human and mouse, the snRNP E protein gene has also maintained a high DNA sequence conservation within its basal promoter structure.


Localization of three novel hybrid breakpoints and refinement of 18 marker assignments in the human 3cen-p21.1 region.

Using the human/hamster cell line UCTP2A-3, we have generated and isolated three hybrids, each containing a novel human chromosome 3p break. All chromosome 3 materials distal to the breaks were lost. Two of the breakpoints were produced using aphidicolin induction; the third breakpoint occurred spontaneously. The aphidicolin-induced breaks were localized to 3p21.1 in hybrid AR1 and to p14.1 in hybrid AR2. The spontaneous break was localized to 3p11 in hybrid 2A-3-1. These hybrids were used to sublocalize 18 chromosome 3 probes further to five regions within 3cen-p21.1. The new hybrids and sublocalized markers will be useful in the study and characterization of the 3p11, 3p14.1, and 3p21.1 segments of chromosome 3.


A knowledge base for predicting protein localization sites in eukaryotic cells.

To automate examination of massive amounts of sequence data for biological function, it is important to computerize interpretation based on empirical knowledge of sequence-function relationships. For this purpose, we have been constructing a knowledge base by organizing various experimental and computational observations as a collection of if-then rules. Here we report an expert system, which utilizes this knowledge base, for predicting localization sites of proteins only from the information on the amino acid sequence and the source origin. We collected data for 401 eukaryotic proteins with known localization sites (subcellular and extracellular) and divided them into training data and testing data. Fourteen localization sites were distinguished for animal cells and 17 for plant cells. When sorting signals were not well characterized experimentally, various sequence features were computationally derived from the training data. It was found that 66% of the training data and 59% of the testing data were correctly predicted by our expert system. This artificial intelligence approach is powerful and flexible enough to be used in genome analyses.


Structure of DNA near long tandem arrays of alpha satellite DNA at the centromere of human chromosome 7.

The centromeric regions of human chromosomes contain long tracts of tandemly repeated DNA, of which the most extensively characterized is alpha satellite. In a screen for additional centromeric DNA sequences, four phage clones were obtained which contain alpha satellite as well as other sequences not usually found associated with tandemly repeated alpha satellite DNA, including L1 repetitive elements, an Alu element, and a novel AT-rich repeated sequence. The alpha satellite DNA contained within these clones does not demonstrate the higher-order repeat structure typical of tandemly repeated alpha satellite. Two of the clones contain inversions; instead of the usual head-to-tail arrangement of alpha satellite monomers, the direction of the monomers changes partway through each clone. The presence of both inversions was confirmed in human genomic DNA by polymerase chain reaction amplification of the inverted regions. One phage clone contains a junction between alpha satellite DNA and a novel low-copy repeated sequence. The junction between the two types of DNA is abrupt and the junction sequence is characterized by the presence of runs of A's and T's, yielding an overall base composition of 65% AT with local areas > 80% AT. The AT-rich sequence is found in multiple copies on chromosome 7 and homologous sequences are found in (peri)centromeric locations on other human chromosomes, including chromosomes 1, 2, and 16. As such, the AT-rich sequence adjacent to alpha satellite DNA provides a tool for the further study of the DNA from this region of the chromosome. The phage clones examined are located within the same 3.3-Mb SstII restriction fragment on chromosome 7 as the two previously described alpha satellite arrays, D7Z1 and D7Z2. These new clones demonstrate that centromeric repetitive DNA, at least on chromosome 7, may be more heterogeneous in composition and organization than had previously been thought.


Breakpoints in Robertsonian translocations are localized to satellite III DNA by fluorescence in situ hybridization.

We characterized 21 t(13;14) and 3 t(14;21) Robertsonian translocations for the presence of DNA derived from the short arms of the translocated acrocentric chromosomes and identified their centromeres. Nineteen of these 24 translocation carriers were unrelated. Using centromeric alpha-repeat DNA as chromosome-specific probe, we found by in situ hybridization that all 24 translocation chromosomes were dicentric. The chromatin between the two centomeres did not stain with silver, and no hybridization signal was detected with probes for rDNA or beta-satellite DNA that flank the distal and proximal ends of the rDNA region on the short arm of the acrocentrics. By contrast, all 24 translocation chromosomes gave a distinct hybridization signal when satellite III DNA was used as probe. This result strongly suggests that the chromosomal rearrangements leading to Robertsonian translocations occur preferentially in satellite III DNA. We hypothesize that guanine-rich satellite III repeats may promote chromosomal recombination by formation of tetraplex structures. The findings localize satellite III DNA to the short arm of the acrocentric chromosomes distal to centromeric alpha-repeat DNA and proximal to beta-satellite DNA.


Linkage mapping of melanoma (MLM) using 172 microsatellite markers.

The incidence of malignant melanoma is currently increasing faster than any other cancer and in 5-12% of cases occurs in a familial context in which the disease cosegregates as an autosomal dominant trait. To identify the location of genes that predipose individuals to familial melanoma (MLM), we have carried out linkage analysis in three large Australian melanoma pedigrees using 172 microsatellite markers spread across all autosomes. Three additional smaller families were typed for 70 of the same markers. In five of the six families we found lod scores between 1.0 and 2.3, which may provide evidence for the location of melanoma genes in proximity to some of these markers. If this turns out to be the case, these data potentially demonstrate that MLM is genetically heterogeneous since there was no marker for which all families gave significantly high LODs. These data provide the foundation for an exclusion map for melanoma and, more importantly, high-light areas of the genome for others to substantiate the potential positions of some of the genes that may be responsible for susceptibility to MLM.


Junctions between genes in the haptoglobin gene cluster of primates.

To investigate the nature of the recombination that generated the haptoglobin three-gene cluster in Old World primates, we sequenced the region between the second gene (HPR) and the third gene (HPP) in chimpanzees (15 kb), as well as the region 3' to the cluster in humans (14 kb). Comparison to the previously sequenced human haptoglobin (HP) and HPR genes showed that the junction point between HP and HPR in humans (junction 1) was not identical to the junction point between the HPR and HPP genes of the chimpanzee (junction 2). An Alu sequence was found at each junction, but both Alu sequences lacked short direct repeats of the flanking genomic DNA. The lack of direct repeats implies that both junction Alu sequences are the products of recombination between different Alu elements. In addition, other insertion and deletion events are clustered in the regions near the junction Alu sequences. The observation that Alu sequences define the junctions between genes in the haptoglobin gene cluster emphasizes the importance of Alu sequences in the evolution of multigene families.


A gene for Usher syndrome type I (USH1A) maps to chromosome 14q.

Usher syndrome (US) is an autosomal recessive disease characterized by congenital hearing impairment and retinitis pigmentosa. It is the most frequent cause of deaf-blindness in adults and accounts for 3 to 6% of deaf children. Here, we report the genetic mapping of a gene for US type I (USH1A), the most severe form of the disease, to the long arm of chromosome 14, by linkage to probe MLJ14 at the D14S13 locus in 10 families of Western France ancestry (Z = 4.13 at theta = 0). Among them, 8 families originated from a small area of the Poitou-Charentes region (Z = 3.78 at theta = 0), suggesting that a founder effect could be involved. However, since not all US type I families were found to be linked to this locus, the present study provides evidence for genetic heterogeneity of this condition (heterogeneity versus homogeneity test HOMOG, P < 0.05; heterogeneity versus no linkage, P < 0.01).


Linkage of Usher syndrome type I gene (USH1B) to the long arm of chromosome 11.

Usher syndrome is the most commonly recognized cause of combined visual and hearing loss in technologically developed countries. There are several different types and all are inherited in an autosomal recessive manner. There may be as many as five different genes responsible for at least two closely related phenotypes. The nature of the gene defects is unknown, and positional cloning strategies are being employed to identify the genes. This is a report of the localization of one gene for Usher syndrome type I to chromosome 11q, probably distal to marker D11S527. Another USH1 gene had been previously localized to chromosome 14q, and this second localization establishes the existence of a new and independent locus for Usher syndrome.


Localization of two genes for Usher syndrome type I to chromosome 11.

The Usher syndromes (USH) are autosomal recessive diseases characterized by congenital sensorineural hearing loss and progressive pigmentary retinopathy. While relatively rare in the general population, collectively they account for approximately 6% of the congenitally deaf population. Usher syndrome type II (USH2) has been mapped to chromosome 1q (W. J. Kimberling, M. D. Weston, C. Möller, et al., 1990, Genomics 7: 245-249; R. A. Lewis, B. Otterud, D. Stauffer, et al., 1990, Genomics 7: 250-256), and one form of Usher syndrome type I (USH1) has been mapped to chromosome 14q (J. Kaplan, S. Gerber, D. Bonneau, J. Rozet, M. Briord, J. Dufier, A. Munnich, and J. Frezal, 1990. Cytogenet. Cell Genet. 58: 1988). These loci have been excluded as regions of USH genes in our data set, which is composed of 8 French-Acadian USH1 families and 11 British USH1 families. Both of these sets of families show linkage to loci on chromosome 11. Linkage analysis demonstrates locus heterogeneity between these sets of families, with the French-Acadian families showing linkage to D11S419 (Z = 4.20, theta = 0) and the British families showing linkage to D11S527 (Z = 6.03, theta = 0). Genetic heterogeneity of the data set was confirmed using HOMOG and the M test (log likelihood ratio > 10(5)). These results confirm the presence of two distinct USH1 loci on chromosome 11.


Expression and generation of interleukin-8, IL-6 and granulocyte-macrophage colony-stimulating factor by bronchial epithelial cells and enhancement by IL-1 beta and tumour necrosis factor-alpha.

We have tested the hypothesis that the bronchial epithelium has the capacity to generate and release cytokines that could contribute to inflammatory events associated with inflammatory lung diseases. Messenger RNA (mRNA) for interleukin-6 (IL-6), IL-8 and granulocyte-macrophage colony-stimulating factor (GM-CSF) was identified in human bronchial epithelial cell primary cultures, characterized on the basis of staining for cytokeratin, using both in situ hybridization and Northern blotting. Using in situ hybridization we have shown that the majority of the cells expressed mRNA for IL-6 and IL-8, whereas fewer than 20% of cells expressed message for GM-CSF. The numbers of cells expressing message were increased by culture with tumour necrosis factor-alpha (TNF-alpha) (20 ng/ml, 24 hr). These observations were substantiated by Northern blotting, which showed that both TNF-alpha and IL-1 beta were able to induce a dose-dependent increase in IL-8-specific mRNA. Immunoreactive IL-6 and GM-CSF were detected and quantified in the culture supernatants by ELISA, and IL-8 by radioimmunoassay. The levels of immunoreactivity were increased by incubation of epithelial cells with either IL-1 beta or TNF-alpha for 24 hr. A transformed tracheal epithelial cell line (9HTEo-) expressed mRNA for IL-6, IL-8 and GM-CSF but, whereas levels of immunoreactive IL-6 in culture supernatants were comparable with those in primary cell cultures, levels of IL-8 were low and GM-CSF trivial. These observations indicate that the bronchial epithelium has the potential to be a major source of IL-8 and a number of other cytokines, and that production can be amplified substantially by IL-1 beta and TNF-alpha. The bronchial epithelium is ideally situated to modulate inflammatory and immunological events in and around the airways, and these observations suggest that it could contribute to promote and sustain inflammatory and immunological processes in inflammatory lung diseases such asthma.