Application of molecular orbital calculations to interpret the chlorophyll spectral forms in pea photosystem II.

The energy and oscillator strength of electronic transitions of chlorophyll (Chl)-amino acid complexes were calculated by using molecular orbital methods. The energies varied widely with coordinated amino acids and the difference between the maximum and minimum energy was about 830 cm-1. This energy difference was comparable with the spreading of absorption bands for light-harvesting Chl-protein complexes of photosystem II (LHC II) of green plants. The feature of the Qy band for pea LHC II was interpreted with the aid of the calculated energies and oscillator strengths. Four spectral components of the band were assigned to individual Chl-amino acid complexes.

Characterization of three forms of light-harvesting chlorophyll a/b-protein complexes of photosystem II isolated from the green alga, Dunaliella salina.

Three forms of light-harvesting chlorophyll a/b-protein complexes of photosystem II (LHC II) were isolated from the thylakoid membranes of Dunaliella salina grown under different irradiance conditions. Cells grown under a low intensity light condition (80 micromol quanta m(-2) s(-1)) contained one form of LHC II, LHC-L. Two other forms of LHC II, LHC-H1 and LHC-H2, were separated from the cells grown under a high intensity light condition (1,500 micromol quanta m(-2) s(-1)). LHC-L and LHC-H1 showed an apparent particle size of 310 kDa and contained four polypeptides of 31, 30, 29 and 28 kDa. LHC-H2, with a particle size of 110 kDa, consisted of 30 and 28 kDa polypeptides. LHC-L contained 7.5 molecules of Chl a, 3.2 of Chl b and 2.1 of lutein per polypeptide, analogous to the content in higher plants. LHC-H1, with 5.6 molecules of Chl a, 2.5 of Chl b and 1.8 of lutein per polypeptide was similar to that in the green alga Bryopsis maxima. LHC-L and LHC-H1 maintained high efficiency energy transfer from Chl b and lutein to Chl a molecules. LHC-H2 showed a high Chl a/b ratio of 7.5 and contained 3.4 molecules of Chl a, 0.5 of Chl b and 1.4 of lutein per polypeptide. Chl b and lutein could not completely transfer the excitation energy to Chl a in LHC-H2.

Peripheral catecholamine metabolites and menstrual irregularity in patients with polycystic ovaries.

OBJECTIVE: To investigate the catecholamine status of patients with polycystic ovary syndrome. DESIGN: Three parallel groups with polycystic ovary were diagnosed by ultrasound: (a) 5 patients with regularly ovulatory menstruation; (b) 10 with anovulatory menstruation; (c) 13 with secondary amenorrhea who responded to progestagen with withdrawal bleeding. Blood samples for measurement of LH, testosterone, and catecholamine metabolites were drawn during cycle days 4-7. RESULTS: (1) Serum LH and testosterone of the patient groups (b) and (c) were significantly higher than those of controls. (2) Plasma 3,4-dihydroxyphenylglycol (DOPEG) and the DOPEG/DOPAC ratio were elevated in patients, and DOPAC levels were reduced. However, there was no significant difference of catecholamine metabolites among the three patient groups. CONCLUSIONS: The androgen status in polycystic ovary diagnosed by ultrasound is correlated, but catecholamine status is not correlated, with the menstrual irregularity of polycystic ovary syndrome.

On the relationship between endocrine and ovulatory abnormalities, and polycystic ovaries as diagnosed by ultrasonography.

The present study was undertaken to investigate the relationship between serum testosterone levels, the menstrual cycle, and polycystic ovaries as shown by ultrasonographic methods. The patients were divided into four groups: (1) cases with normal-appearing ovaries and with regular menstruation (control); (2) cases with normal ovaries on ultrasound, but with abnormalities of the menstrual cycle (MA); (3) cases with polycystic ovaries, but normal menstrual cycle (PCO); (4) cases with both polycystic ovaries and menstrual abnormalities (PCOMA). The results were as follows. (1) The ovarian area was significantly larger in the PCOMA group than in the control group. (2) The number of cases with elevated LH (greater than 30 mIU/mL) was significantly higher in the PCOMA group than in the other groups. (3) In comparison with controls, the percentage of cases with LH/FSH ratios greater than 3.0 was significantly higher in all three groups. (4) The estrone/17 beta-estradiol ratio for the PCOMA group was significantly greater than those of the other groups. (5) The testosterone levels of the PCO and PCOMA group were significantly greater than the control value. (6) The percentage of endocrine criteria (LH greater than 30 mIU/mL or LH/FSH ratio greater than 3.0, estrone/17 beta estradiol ratio greater than 1.0 testosterone greater than 75, greater than 100, and greater than 125 ng/dL) for PCOMA was significantly higher than that of the MA group and the control group, but not significantly greater than the PCO group. It is suggested that morphological changes in the ovaries show a closer correlation with abnormalities of endocrine function than does the presence or absence of abnormalities of the menstrual cycle.

Further evidence concerning catecholamine metabolism in polycystic ovary syndrome following Gn-RH administration.

The role of catecholamine activity in the GnRH-LH system among cases of polycystic ovary syndrome was investigated by high-performance liquid chromatography using an electrochemical detector. We measured plasma levels of catecholamine metabolites, 3-methoxy-4-hydroxyphenylglycol (MHPG) and 3,4-dihydroxyphenylacetic acid (DOPAC), in relation to LH-RH administration in 23 women with polycystic ovary syndrome. MHPG concentrations were significantly higher in polycystic ovary syndrome patients than in ten control subjects in early follicular phase. The peak values of LH after LH-RH administration were significantly correlated with plasma MHPG concentrations. These data suggest that an excess of norepinephrine is present in polycystic ovary syndrome and that the hypophyseal response to LH-RH administration is correlated with catecholamine, especially norepinephrine activity.

[Changes in serum androgen levels in patients with polycystic ovary syndrome].

In order to study the changes in androgen levels in patients with polycystic ovary syndrome (PCOS), serum androgen levels (testosterone [T], free-testosterone [free-T] and sex-hormone-binding globulin [SHBG]) were measured and related to the incidence of menstrual abnormalities, obesity and hirsutism. Tests were made in 61 cases of PCOS identified by transvaginal ultrasound and in normal controls. The following results were obtained: 1) The PCOS group with menstrual irregularities had significantly higher levels of T and free-T than the control group, but SHBG levels were not significantly altered. 2) The obese group had significantly higher free-T levels than the non-obese group, and SHBG was significantly lowered. 3) There were significant correlations between the levels of obesity and free-T and SHBG. 4) T and free-T were moderately high in the hirsutism group and SHBG was moderately low, but not significantly so. These findings indicate that, in PCOS patients, serum androgen levels, especially free-T and SHBG, are closely related to menstrual irregularities and the level of obesity.

Peripheral catecholamine metabolites and free testosterone in patients with polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is characterized by chronic anovulation, elevated serum estrogen and androgen levels, and inappropriate gonadotropin secretion. The present study compares the androgen levels (free testosterone) and catecholamine metabolites (DOPAC as a marker of central dopaminergic activity, DOPEG as one of adrenergic activity) in patients with PCOS and normal controls. The mean serum LH and free testosterone levels of polycystic ovary patients were significantly higher than those of normal controls. The mean DOPEG/DOPAC ratio in the polycystic ovary patients was significantly higher than those of the control group and there was a significant correlation between free testosterone and DOPEG or the DOPEG/DOPAC ratio. It is suggested that, in vivo, norepinephrine excess is present in women with PCOS and that a hyperandrogenemic status is correlated with catecholamine activity.

Plasma catecholamine metabolites and polycystic ovary syndrome.

It has been postulated that catecholamine metabolism may be altered in cases of polycystic ovary syndrome. To search for possible correlations between catecholamine metabolism and hormonal disturbances, we have studied the serum LH, LH:FSH ratio, testosterone, and plasma catecholamine metabolites in patients with polycystic ovary syndrome and in control subjects with normal ovulatory cycles. The metabolites studied were 3-methoxy-4-hydroxyphenylglycol (MHPG) and 3,4-dihydroxyphenylacetic acid (DOPAC) as markers of adrenergic activity and dopaminergic activity, respectively. The polycystic ovary was divided into 2 patterns [general cystic pattern (GCP) and peripheral cystic pattern (PCP)] as determined by ultrasound. The results were as follows: 1) Serum LH, LH:FSH ratios, and plasma MHPG levels in patients with polycystic ovary syndrome were significantly higher than in the controls. 2) In cases of polycystic ovary syndrome, serum LH, LH:FSH ratios, and testosterone showed no significant correlations with catecholamine metabolites. 3) Using the ultrasonographical classification, we found that plasma MHPG levels of the GCP group were significantly higher compared with the PCP group in patients with polycystic ovary syndrome. Thus, catecholamine metabolism is altered in patients with polycystic ovary syndrome, and ultrasonography revealed different patterns of catecholamine metabolism.

Transvaginal ultrasound is an effective method for screening in polycystic ovarian disease: preliminary study.

To assess the usefulness of transvaginal ultrasound scanning (TVS) for screening in polycystic ovarian (PCO) disease, we symptomatically and endocrinologically compared 9 cases (group A) with PCO noted by transabdominal ultrasound scanning (TAS) with 8 cases (group B) with PCO noted by TVS only. No difference in the mean value of LH/FSH ratio, E1/E2 ratio and testosterone was observed between the two groups. However, the frequency of endocrine abnormalities, menstrual disorders and hypertrichosis in group A was significantly more than in group B. Therefore, we suggest that the stage of PCO noted by TAS is more progressive than that noted by TVS only and TVS is useful for screening of PCO disease.

Changes in plasma catecholamines and pulsatile patterns of gonadotropin release in subjects with a normal ovulatory cycle and with polycystic ovary syndrome.

Plasma LH levels in women and female laboratory animals fluctuate. Since catecholamines play a role in gonadotropin secretion, we investigated the possible correlation between catecholamine metabolites and pulsatile patterns of gonadotropin release by measuring two catecholamine metabolites, MHPG and DOPAC. LH levels had a pulsatile pattern, and the levels of MHPG fluctuated. The average of LH and MHPG in polycystic ovary syndrome was significantly higher than that of a normal ovulatory cycle, and the average fluctuation of LH and MHPG in polycystic ovary syndrome was also significantly greater than that in a normal ovulatory cycle. There may be a correlation between the pulsatile pattern of gonadotropin and catecholamine activity, particularly adrenergic activity. It is suspected that adrenergic activity may correlate with the pulsatile pattern in polycystic ovary syndrome.

[Experimental studies of skin wound healing process by first intention in streptozotocin-induced diabetes mellitus rats].

Various parameters were used in observing the process of wound healing in rats with streptozotocin-induced diabetes (STZ diabetes). Sections stained according to the Hematoxylin-Eosin, Van-Gieson, and Azan methods were used in observing histological changes. At the same time, wound strength during the healing was measured as a parameter for evaluating the healing process. In addition, changes in leukocytes, plasma fibrinogen, activated factor XIII (aXIII), collagen content of the incised wound, and metabolic changes were determined. Results 1. Histological studies showed that, in STZ diabetes, the inflammatory response was minimal and occurred later than in normal cases. In the incised wound, cellular infiltration of polymorpho-nuclear leukocytes and fibrin nets accumulated poorly. The fibrin net was coarse and fragile. Furthermore, epithelialization of the wound was late: it did not occur until 5 days after the operation. In cases of STZ diabetes, patterns of hyperplasia and fibroblast arrangements were abnormal. Collagen regeneration and proliferation processes were remarkably retarded. 2. In normal, wound strength increased from the 5 postoperative day. After 10 days had passed, it increased remarkably until, after 30 days, it had returned to the preoperative level. In STZ diabetes, however, no increase in wound strength occurred for the first 14 days after the operation. There after strength increased slowly; but, 40 days after the operation, 80% of the preoperative level still had not been reached. 3. Changes in leukocytes were much later occurring in STZ diabetes than in normal. Recovery took longer than in normal. 4. In STZ diabetes, increases in plasma fibrinogen and decreases of the aXIII factor were slower than in normal. The a XIII factor decreased remarkably, and recovery was slow. 5. In terms of collagen content in the wound incision, in STZ diabetes, tropocollagen increase occurred later than in normal. But, from the 5 to the 14 postoperative days, its level was higher than that in normal. Maturation-process collagen and mature collagen increased still more slowly. In normal, mature collagen had reached preoperative level 20 days after the operation, in STZ diabetes, 80% of preoperative level still had not been reached 30 days after the operation. 6. These studies showed that the following factors hinder wound healing in cases of diabetes mellitus: minimal inflammatory response, incomplete formation of the fibrin nets, retardation of epithelialization, retarded action of plasma fibrinogen and the a XIII factor, reduced fibroblast activity, and slow increase in collagen content.

[The relations between doses of streptozotocin and pathosis in induced diabetes mellitus].

Tail-vein injections of streptozotocin (STZ) in various doses (20-60 mg/kg body weight) were used to induce diabetes mellitus in male rats of the Sprague-Dawley strain. Resulting pathosis was observed on the basis of various parameters (body weight, blood glucose, plasma insulin, glucose-tolerance tests, uroscopy, hemodiagnosis, and skin strength) Results 1. Diabetes mellitus was induced in all rats of the group in which the dosage was STZ 40 mg/kg body weight (S-40 group). Blood-glucose level in this group was about 340 mg (3 times the quantity in controls). Plasma-insulin level was about 8.9 microU/ml (about 1/3 the quantity in controls). Glucose-tolerance tests and observation of urino-glucose showed reductions in glucose tolerance and plasma-insulin response. 2. In the S-40 group, pathosis of induced diabetes mellitus had stabilized 30 days after and persisted for 90 days after STZ injection. 3. Skin strength decreased for 20 days after STZ injection. The level remained stable at half the strength of control skin from 30 to 90 days after the injection. 4. In groups in which doses were STZ 20 mg/kg body weight or STZ 30 mg/kg body weight, diabetes mellitus was induced in some of the rats. The animals tended to recover from the induced pathosis. 5. Diabetes mellitus was induced in all rats to which doses of STZ 50 mg/kg or STZ60 mg/kg body weight were induced. shortly after injection, the induced-diabetes pathosis changed for the worse; and grave complications (hypo-albuminosis, diabetic ketocacidosis, and diabetic proteinuria) were observed. A large number of the rats in these groups died. 6. The results of this study confirm the opinion that STZ 40 mg/kg body weight is the optimum dose for STZ induction of diabetes mellitus in rats for experimental studies. The suitable term for such studies is from 30 to 90 days after STZ injection.

[Study on hypoosmotic swelling test for infertile male patients].

The hypoosmotic swelling test was done on 74 infertile men, 7 patients who achieved pregnancy by AIH (AIH group) and 10 fertile men and the usefulness of its test was assessed. 1) The rate of total swollen sperm and that of g type swollen sperm from infertile men were significantly lower than rates in the patients in the AIH group and fertile men. 2) There were significant but weak positive correlations between semen analysis (total sperm concentration, motility, motile sperm concentration) and hypoosmotic swelling test results (total swollen sperm, g type swollen sperm). 3) When the cut off value of the rate of total swollen sperm and g type swollen sperm was set at 42% and 14%, respectively, rates were above the cut off value except for g type swollen sperm in patients in the AIH group.

Ultrasonic evaluation of the normal uterus in the neonate.

Using real-time ultrasound, a study was undertaken to evaluate the morphology of normal uterus in the neonate delivered at term. The uterus was visualized in 41 (89.1%) of the 46 neonates. The cervical volume (CE-V) was 3.65 +/- 1.36 cm3, the corporeal volume (CO-V) 1.18 +/- 0.42 cm3, and the CE-V was significantly larger than the CO-V (p less than 0.001). The uterine volume (U-V) was 4.83 +/- 1.57 cm3, and U-V showed a good correlation with birth weight (r = 0.42, p less than 0.01). The endometrial echo was constantly depicted as a mixed echo of a highly echogenic line and/or a myometrial halo. These normal values and the morphology of the neonatal uterus provide useful information for distinguishing a possible pathologic pelvic mass and screening of congenital genital disorders.

Effect of a traditional herbal medicine (shakuyaku-kanzo-to) on testosterone secretion in patients with polycystic ovary syndrome detected by ultrasound.

In 20 infertile Japanese with polycystic ovary syndrome Shakuyaku-Kanzo-To was used to lower plasma testosterone levels and hence to induce pregnancy. The polycystic ovary was classified into two types; general cystic and peripheral cystic patterns. Plasma testosterone was decreased in 18/20 (90%) and 5/20 (25%) became pregnant. The plasma testosterone concentration in the case of the general cystic pattern was significantly higher than that of peripheral cystic pattern, and the pregnancy rate in those with the general cystic pattern was lower. The efficacy of Shakuyaku-Kanzo-To therefore seems to vary according to the type of polycystic ovary syndrome treated.

Ultrasonographic evaluation of adrenal involution during antenatal and neonatal periods.

Using real-time ultrasonography, 25 adrenal glands from prenatal to neonatal periods were prospectively examined to assess the process of shrinkage. A similar sonographic structure was seen in both prenatal and neonatal adrenal glands. The area of adrenal gland (AGA) and the rate of decrease were calculated during antenatal and early neonatal periods. The values of AGA were 350 +/- 19 mm2 within a week of delivery, 304 +/- 17 mm2 (87 +/- 2%) just after delivery, 273 +/- 25 mm2 (77 +/- 5%) on the 1st day, 246 +/- 24 mm2 (70 +/- 5%) on the 2nd day, 215 +/- 23 mm2 (61 +/- 5%) on the 3rd day, 196 +/- 22 mm2 (56 +/- 5%) on the 4th day, 173 +/- 18 mm2 (50 +/- 4%) on the 5th day, 154 +/- 14 (44 +/- 4%) on the 6th day, 140 +/- 12 mm2 (40 +/- 2%) on the 7th day, respectively. In conclusion, the postnatal involution of the adrenals was documented by ultrasound.