Relationship among serum levels of luteinizing hormone,estradiol, and progesterone during follicle stimulation and results of in vitro fertilization and embryo transfer (IVF-ET)

Eighty-eight IVF-ET cycles were classified into four groups according to the results of IVF-ET (Group A—conceptional cycles, 10 cycles; Group B—cycles with cleaved oocytes, 58 cycles; Group C—cycles with fertilized oocytes, 9 cycles; Group D—cycles without fertilization, 11 cycles). Serum luteinizing hormone (LH), estradiol (E ₂ ), and progesterone (P) levels during follicle stimulation were studied in these groups. Patients participated in our IVF-ET program due to irreparable tubal damage. Follicle development was stimulated with a clomiphene—human menopausal gonadotropin (hMG)—human chorionic gonadotropin (hCG) regimen. Group C showed a low E ₂ response to follicle stimulation. Groups B and D showed significantly higher serum P levels on day 0 (the day of hCG injection) than Group A (Group A, 0.73 ± 0.11, vs Groups B and D, 1.43 ± 0.15 and 2.17 ± 0.42 ng/ml; P <0.01). The effects of serum P and LH levels on the fertilization and pregnancy rates were studied. The pregnancy rate was not affected by the serum LH level but was only 2.7% in cycles in which serum P was 1.2 ng/ml on day 0, which was significantly lower than that in cycles in which serum P was <1.2 ng/ml on day 0 (19.1%) (P <0.05). The fertilization rate was significantly lower in the cycles with higher levels of serum P and/or LH than in cycles in which serum P was <1.2 ng/ml and serum LH was normal (50.5 vs 78.8%; P <0.01). These findings suggest that the serum P level, but not the LH level, during follicle stimulation is closely related to the achievement of pregnancy.     

Neurochemical correlates of selective neuronal loss following cerebral ischemia: role of decreased Na, K-ATPase activity

In order to investigate the role of Na+,K(+)-ATPase in the development of neuronal necrosis following cerebral ischemia, ischemia was induced in gerbils by occluding the common carotid artery unilaterally for 10 min. A time-course analysis revealed that significant reductions of the Na+,K(+)-ATPase activity in the cerebral cortex and hippocampus were manifested at 15 min, 30 min, and 1 h, and returned to the control level one day following recirculation. No apparent alterations of the Mg(2+)-ATPase activity, on the other hand, were obtained throughout the experimental period. Furthermore, Scatchard analyses of [3H]ouabain binding to the cerebral cortex membranes disclosed that the Bmax values invariably decreased without any change of Kd values following ischemia. It has also been shown that treatment of the animals with an agent known to mitigate ischemic neuronal necrosis, i.e. BY-1949, significantly reversed such derangements. These results suggest that the recovery of decreased Na+,K(+)-ATPase activity shortly after ischemia exerts a protective effect against ischemic brain damage.     

Lipid Peroxidation of Rat Erythrocyte Membrane Induced by Adriamycin-Fe3+

Adriamycin-Fe³⁺ caused lipid peroxidation of erythrocyte membrane in relation to its concentration. Adriamycin-Fe³⁺ had a high affinity for membrane and the adriamycin-Fe³⁺-binding membranes was also found to cause lipid peroxidation. Under aerobic conditions, adriamycin-Fe³⁺ caused a reduction of cytochrome c and ferrous iron formed spontaneously. Superoxide dismutase (EC 1.15.1.1) (SOD) strongly inhibited the reduction of cytochrome c; however, the enzyme promoted formation of ferrous iron independent of enzymatic action. These results suggest that cytochrome c was reduced by superoxide radical (O₂ ^?) or an adriamycin-iron-O₂ complex such as adriamycin-Fe³⁺–O₂ ^?, but not by adriamycin-Fe²⁺. The ferrous iron chelator bathophenanthroline sulfonate (BPS) completely inhibited oxygen consumption caused by adriamycin-Fe³⁺, indicating that ferrous iron is absolutely required for the lipid peroxidation. SOD and hydroxyl radical scanvengers did not inhibit the lipid peroxidation, indicating that O₂ ^? and hydroxyl radical were not involved in membrane peroxidation. The peroxidation reaction was dramatically inhibited by Tris buffer (2-amino-2-hydroxymethyl-1,3-propanediol). However, hydroxyl radical generation and lipid peroxidation in Tris buffer were not related obviously, indicating that Tris did not act as a hydroxyl radical scavenger. The initial rate of TBARS (thiobarbituric acid reactive substances) formation induced by a mixture of adriamycin-Fe³⁺ and adriamycin-Fe²⁺ was much faster than that indcued by adriamycin-Fe²⁺ or adriamycin-Fe³⁺ alone. These results made it became possible to speculate that the lipid peroxidation might be initiated by an adriamycin-Fe³⁺-oxygen-adriamycin-Fe²⁺ complex.     

Combined Human Chorionic Gonadotropin (HCG) and Human Menopausal Gonadotropin (HMG) Treatment in Gonadotropin-Deficient Males with Pituitary Dwarfism

The effects of hCG-hMG treatment in 13 boys with pituitary dwarfism associated with gonadotropin deficiency, were assessed.
No patients except one showed signs of puberty at a bone age of 13 years or above. The one patient with some signs of puberty did not become fully mature. The hCG-hMG was started at a mean age of 20.4 years. The hCG at a dose of 5,000 IU was injected intramuscularly twice a week and the hMG at a dose of 75 IU was given once a week at first. During treatment, the frequency of hMG injections was increased to twice a week in six patients who still had not produced normal sperm counts. After a mean duration of 19.23 months, spermatozoa appeared in eight patients, of whom four showed more than 20 × 10⁶ sperm/ml. Among six patients who did not have normal sperm counts and had increased hMG injections, one produced a pregnancy and four achieved sperm counts of more than 35 × 10⁶/ml. One patient had refractory azoospermia. In 13 boys with growth hormone and gonadotropin deficiency, hCG-hMG treatment produced normal spermatogenesis in nine patients, one of whom fathered a girl. Thus, hCG-hMG treatment, especially twice-a-week injections of both hCG and hMG, appears to be effective for gonadotropin deficiency in males.     

Transformation of NIH3T3 Cells with Synthetic c-Ha-ras Genes

Synthetic human c-Ha- ras genes in which amino acid codons were altered to those which are frequently used in highly expressed Escherichia coli genes were ligated to the 3'-end of Rous sarcoma virus long terminal repeat. When NIH3T3 cells were transfected with the plasmids having those genes with valine at codon 12, leucine at codon 61 or arginine at codon 61, transformants were efficiently produced. These results indicated that the synthetic c-Ha- ras genes are expressed in a mammalian system even though their codon usage is altered to correspond with that of E. colt. This expression vector system should he useful for studies on the structure-function relationships of c-Ha- ras , since the synthetic gene can be easily modified to have multiple base alterations, and can also be used simultaneously for the production of large amounts of p21 in E. coli for biochemical and biophysical studies.     

Leukocytosis in Mice Following Therapy with a Novel Antitumor Agent, RA-700

Nine daily intravenous (iv) injections of RA-700 (an antitumor cyclic hexapeptide) at doses of 2 to 6 mgAg/day caused increases of WBC counts at 4–6 days after treatment in normal C57BL/6 × DBA/ 2 (BDF1) mice. The percentages of neutrophils and lymphocytes were modified. There was a decrease of colony-forming units in culture (CFUc) in bone marrow to 40% of the control value on day 1 but CFUc rapidly returned to normal values on day 3. The colony-forming units in spleen (CFUs): in bone marrow decreased during treatment. On the other hand, CFUc and CFUs in spleen were increased from the initiation of treatment to the time prior to the increase of WBC count. Spleen weight increased after treatment, and histologically, increases of immature and also mature granulocytes and megakaryocytes were observed. However, RA-700 did not stimulate the progress of hematoprogenitors in vitro. The results indicated that RA-700 stimulates the progress of hematoprogenitors in the spleen, but this effect is probably indirect.     

The Syndrome of Self-Induced Water Intoxication in Psychiatric Patients

Abstract: This is a report on six psychiatric patients who indulged in excessive ingestion of water and subsequently developed tonic-clonic seizures in the course of the underlying mental disorders. On the basis of the DSM-III criteria, they were diagnosed as follows: schizophrenic disorder, 4; schizoaffective disorder, 1; borderline personality disorder, 1. The levels of serum electrolytes were estimated during five episodes of seizures in three patients. Hyponatremia was a consistent finding (serum sodium: mean = 120.6 mEq/liter). Plasma osmolality and plasma levels of arginine vasopressin (AVP) were determined during two episodes in two patients. The inappropriately high circulating levels of AVP relative to plasma hypoosmolality were documented. However, the response to the overnight fluid deprivation and acute water load during the period of no seizures in two patients revealed no evidence of the persistent SIADH, suggesting the temporal association of hyponatremic encephalopathy with inappropriate AVP secretion. It is not conclusive whether the transient SIADH is the cause or the consequence of hyponatremic encephalopathy, although a delusion or an auditory hallucination could play a critical role in drinking water excessively in three patients.     

Increase of glial fibrillary acidic protein fragments in the spinal cord of motor neuron degeneration mutant mouse

We analyzed protein fractions extracted from the spinal cord of the motor neuron degeneration (Mnd) mouse, a mutant that exhibits progressive degeneration of lower spinal motor neurons, by one- and two-dimensional polyacrylamide gel electrophoresis (PAGE) after solubilization of the tissue with medium containing sodium dodecyl sulfate (SDS)-urea during growth of the animal, in comparison with those of age-matched controls (C57BL/6). Several protein spots were detected around a region of pI 5.6–6.0 and molecular mass of 35–50 kDa in Mnd spinal cord tissue on the two-dimensional PAGE separation profile with Coomassie brilliant blue staining, while only a few spots around the same region were found in the control spinal cord. These spots were all immunoreactive with an antibody against glial fibrillary acidic protein (GFAP), a cytoskeleton filamentous protein specific to astroglial cells. The protein spot with molecular mass of 50 kDa showed immunoreactivity with anti-GFAP antibody, had a blocked amino-terminus, and is assumed to be intact GFAP. Several protein spots with slightly smaller molecular masses of 35 to 48 kDa lacked the head domain of the GFAP molecule as a result of cleavage at the 29th and 56th residues from the amino terminus. In Mnd spinal cord tissue, the densities of the immunoreactive GFAP bands with smaller molecular masses increased with development, and became dominant at the time of the appearance of behavioral paralytic gait around 6 to 7 months of age. These results suggest that the increased GFAPs devoid of head domains are related to the degenerative loss of motor neurons in the Mnd spinal cord. Histopathological and GFAP immunohistochemical examination of Mnd spinal cord preparation demonstrated progressive degenerative loss of motor neurons, and considerable increases in number of GFAP-stained astrocytes in the ventral horn at 7 to 9 months of age. These processes of degenerative loss of motor neurons and proliferation of reactive astrocytes with increased levels of fragmented GFAP in the Mnd spinal cord during development seem to be characteristic and preceded the deterioration of motor activities in this animal model of amyotrophic lateral sclerosis.     

Neural correlates of compensation after hemilabyrinthectomy

Vestibular brain stem neurons responsive to angular acceleration in the plane of the horizontal canal were examined in cats 30 to 52 days after contralateral labyrinthectomy and compared with similar units in cats with intact labyrinths and in other cats immediately after transection of the contralateral eighth nerve. In the compensated state, the mean spontaneous firing rate of type I neurons was 24 spikes/s, in contrast to the mean of 45/s observed immediately after contralateral labyrinthectomy. In intact cats, mean firing rate was 19 spikes/s. Sensitivity, as measured in spikes/s/deg/s², was significantly lower immediately after labyrinthectomy than in intact controls and remained so in compensated cats. On the other hand, time constants and the ratio of adapting:nonadapting units was unchanged. Ablation of the midline cerebellum including vermis and fastigial nuclei did not materially affect these results. We concluded that (i) the main defects in static posture and nystagmus in the uncompensated state were due to the striking difference in resting firing rates between the ipsi- and contralateral vestibular nuclei; (ii) compensation in the static posture was the result of a tendency to equalize the resting firing rates in the two vestibular nuclei; and (iii) recovery in the dynamic, head-turning situation was due to partial recovery of sensitivity on the ipsilateral side, bringing it to the relatively constant, unchanging depressed sensitivity on the contralateral side.     

Direct projections from the globus pallidus to the paraventricular nucleus of the thalamus in the rat

After injecting horseradish peroxidase into the thalamic regions around the paraventricular thalamic nucleus (Pv) in the rat, small neurons in the globus pallidus (GP) were labeled retrogradely with the ezyme. After injecting [³H]leucine into the GP, terminal labeling was autoradiographically observed in the Pv bilaterally with an ipsilateral dominance. These terminals in the Pv were shown by electron microscopic autoradiography to make asymmetrical synaptic contacts upon small dendrites of Pv neurons.