eST Treatment Research Articles

Abstract 13453: Estrogen Regulates Expression of Fibrotic Markers in Ips-derived Cardiac Fibroblasts and Cardiomyocytes

Introduction: Cardiovascular disease (CVD) is the leading cause of death among women. Epidemiologic studies indicate that pre-menopausal women are protected against the development of CVD when compared to age-matched men. Women show less cardiac remodeling, with downregulation in fibrosis and inflammation pathways. Remodeling is manifested by morphology and electrophysiological changes including hypertrophy, apoptosis, fibroblast proliferation, fibrosis and accumulation of fibrillar collagens. Hypothesis: We hypothesize that female protection against CVD is associated with estrogen levels as the incidence and severity of CVD increases after menopause. Methods: Induced pluripotent stem cells (IPSC) from male (M) and female (F) controls were differentiated to cardiomyocytes (IPSC-CMs) and cardiac fibroblasts (IPSC-CFs). Samples were cultured for 30 days and collected for analysis before and after 48 hs incubation with 1uM of 17-b-estradiol (EST). In IPSC-CMs we analyzed gene expression, morphology and beating frequency. Gene expression, morphology and wound healing were analyzed in IPSC-CFs. Results: Expression of ESR1 , and MYH7 was higher in female IPSC-CMs with upregulation after EST treatment in females but not males. Male IPSC-CMs showed higher expression of COL1A1 , CX43 , MYH6 and RYR2 ; MYH6 , RYR2 and COL1A1 were downregulated after EST treatment. No differences were observed in morphology or beating frequency before or after EST treatment (F:27.5±4.5 bpm vs M:22±2.5 bpm). In IPSC-CFs, males showed higher expression of the fibrotic markers COL1A1 , POSTN, SMAD2 and CCL2 ; EST treatment decreased the expression of POSTN and SMAD2 . Wound healing assays showed a decrease of the wound healing % in women after EST treatment (CT F:86.4±19.7% vs EST F:57.43±10.1%, at 8 hs, *p<0.05). Male wound healing rate was slower than women in control and treated groups, with no changes after EST treatment (CT F: 86.4±19.7% vs CT M: 51.65±18.4%, at 8 hs, *p<0.05). Conclusions: Estrogen treatment exerts an effect on gene expression in female and male IPSC-CMs and IPSC-CFs with men showing higher expression of fibrotic markers, which are down-regulated by EST. Further functional studies will shed light on the phenotypic effects of gene expression changes.

Endothelial Relaxation Mechanisms and Oxidative Stress Are Restored by Atorvastatin Therapy in Ovariectomized Rats

The studies on hormone replacement therapy (HRT) in females with estrogen deficiency are not conclusive. Thus, non-estrogen therapies, such as atorvastatin (ATO), could be new strategies to substitute or complement HRT. This study evaluated the effects of ATO on mesenteric vascular bed (MVB) function from ovariectomized (OVX) female rats. Female rats were divided into control SHAM, OVX, and OVX treated with 17β-estradiol (EST) or ATO groups. The MVB reactivity was determined in organ chambers, vascular oxidative stress by dihydroethidine staining, and the expression of target proteins by western blot. The reduction in acetylcholine-induced relaxation in OVX rats was restored by ATO or EST treatment. The endothelium-dependent nitric oxide (NO) component was reduced in OVX rats, whereas the endothelium-derived hyperpolarizing factor (EDHF) component or prostanoids were not altered in the MVBs. Endothelial dysfunction in OVX rats was associated with oxidative stress, an up-regulation of iNOS and NADPH oxidase expression and a down-regulation of eNOS expression. Treatment with ATO or EST improved the NO component of the relaxation and normalized oxidative stress and the expression of those signaling pathways enzymes. Thus, the protective effect of ATO on endothelial dysfunction caused by estrogen deficiency highlights a significant therapeutic benefit for statins independent of its effects on cholesterol, thus providing evidence that non-estrogen therapy could be used for cardiovascular benefit in an estrogen-deficient state, such as menopause.

MiR-7-1 potentiated estrogen receptor agonists for functional neuroprotection in VSC4.1 motoneurons

Protection of motoneurons is an important goal in the treatment of spinal cord injury (SCI). We tested whether neuroprotective microRNAs (miRs) like miR-206, miR-17, miR-21, miR-7-1, and miR-106a could enhance efficacy of estrogen receptor (ER) agonists such as 1,3,5-tris (4-hydroxyphenyl)-4-propyl-1H-pyrazole (PPT, ERα agonist), Way200070 (WAY, ERβ agonist), and estrogen (EST, ERα and ERβ agonist) in preventing apoptosis in the calcium ionophore (CI)-insulted ventral spinal cord 4.1 (VSC4.1) motoneurons. We determined that 200nM CI induced 70% cell death. Treatment with 50nM PPT, 100nM WAY, and 150nM EST induced overexpression of ERα, ERβ, and both receptors, respectively, at mRNA and protein levels. Treatment with ER agonists significantly upregulated miR-206, miR-17, and miR-7-1 in the CI-insulted VSC4.1 motoneurons. Transfection with miR-206, miR-17, or miR-7-1 mimic potentiated WAY or EST to inhibit apoptosis in the CI-insulted VSC4.1 motoneurons. Overexpression of miR-7-1 maximally increased efficacy of WAY and EST for down regulation of pro-apoptotic Bax and upregulation of anti-apoptotic Bcl-2. A search using microRNA database (miRDB) indicated that miR-7-1 could inhibit the expression of L-type Ca2+ channel protein alpha 1C (CPα1C). miR-7-1 overexpression and WAY or EST treatment down regulated CPα1C but upregulated p-Akt to trigger cell survival signaling. The same therapeutic strategy increased expression of the Ca2+/calmodulin-dependent protein kinase II beta (CaMKIIβ) and the phosphorylated cAMP response element binding protein (p-CREB) so as to promote Bcl-2 transcription. Whole cell membrane potential and mitochondrial membrane potential studies indicated that miR-7-1 highly potentiated EST to preserve functionality in the CI-insulted VSC4.1 motoneurons. In conclusion, our data indicated that miR-7-1 most significantly potentiated efficacy of EST for functional neuroprotection and this therapeutic strategy could be used in the future to attenuate apoptosis of motoneurons in SCI.

Role of MT1-MMP in Estrogen-Mediated Cellular Processes of Intimal Hyperplasia

Hormone replacement therapy increases intimal hyperplasia (IH) following vascular intervention. Matrix metalloproteinases (MMPs) play a role in IH development. We have shown estrogen up-regulates MT1-MMP expression, a transmembrane protein that activates MMP-2, and increases vascular smooth muscle cell (VSMC) collagen invasion via increased MMP-2 activity. Here we hypothesize inhibition of MT1-MMP will prevent hormonally-stimulated increased MMP-2 activation and the downstream cellular processes of IH pathogenesis. VSMCs from a postmenopausal donor were transfected with MT1-MMP or negative control siRNAs, treated with estrogen (Est), analyzed by q-PCR, Western blot, zymography, migration, invasion, and proliferation assays. Est treatment of MT1-MMP silenced cells still resulted in increased MT1-MMP expression (C = 41% ± 4%; Est = 52% ± 2%; P < 0.05). Silencing of MT1-MMP decreased basal MMP-2 activity (nonsilenced = 100%; MT1-silenced = 87% ± 3%; P < 0.05) but had no effect on basal invasion or proliferation. Est treatment of MT1-MMP silenced cells still resulted in increased MMP-2 activity (C = 87% ± 3%; Est = 101% ± 4%; P < 0.05) and invasion (C = 89% ± 6%; Est = 109% ± 3%; P < 0.05) compared with MT1-MMP silenced control cells. However, silencing of MT1-MMP did inhibit Est- and serum-stimulated proliferation (C = 106% ± 18%; Est = 104% ± 16%; FBS = 121% ± 24%; P = NS). Silencing of MT1-MMP in aged VSMCs results in impaired but not complete inhibition of basal and Est-stimulated increases in MMP-2 activity. Other mechanisms appear to be playing a role in hormonally-regulated cellular processes of IH pathogenesis. Future studies will target other signaling cascades, with the goal of identifying mechanisms responsible for hormonally-modulated unbalanced MMPs. In vivo manipulation of the expression patterns of MT1-MMP will be examined for the prevention of IH in animal models of vascular disease.

High eggshell temperatures during incubation decrease growth performance and increase the incidence of ascites in broiler chickens

High eggshell temperatures (EST; ≥38.9°C) during the second half of incubation are known to decrease the body and organ development of broiler hatchlings. In particular, relative heart weights are decreased by a high EST, and this may increase the incidence of metabolic disorders that are associated with cardiovascular development, such as ascites. The current study investigated the effects of a high EST on chick quality, subsequent performance, and the incidence of ascites later in life. Eggs were incubated at a normal (37.8°C) or high (38.9°C) EST from d 7 of incubation onward. After hatching, the chickens were housed per EST in pens, and a normal or cold temperature schedule was applied during the grow-out period. Hatchability, hatchling quality, BW, feed conversion ratio, total mortality, mortality associated with ascites, slaughter characteristics, and ascites susceptibility at 6 wk of age were evaluated. Except for total ventricle weight, no interaction was found between EST and the grow-out temperature. Hatchability was comparable between the EST treatments, but the percentage of second-grade chickens was 0.7% higher at the high EST. Yolk-free body mass was 3.0 g lower, and heart weights were 26% lower at hatch in the high compared with the normal EST treatment. Body weight continued to be less during the grow-out period after the high EST incubation. However, breast meat yield was 1.0% higher in the high than in the normal EST. Feed conversion ratio did not differ between EST treatments. Total mortality was 4.1% higher and mortality associated with ascites was 3.8% higher in the high compared with the normal EST treatment. The ratio between the right and total ventricle was 1.1% higher in the high compared with the normal EST treatment at slaughter age. In conclusion, a high EST from d 7 of incubation onward decreased hatchling quality and growth performance, but increased breast meat yield. Furthermore, high EST incubation increased the incidence of ascites, which may be related to the reduced heart development at hatch.

A deer antler extract prevents bone loss in postmenopausal osteoporosis model rats

This study was conducted to examine the effects of a deer antler extract in water (DAE) on bone metabolism. 8 week-old female rats (Sprague-Dawley) were subjected to establish the postmenopausal osteoporosis model by feeding a low calcium diet for 4 weeks after ovariectomy operation. The model rats were divided into 4 groups (n=32); Control, without DAE DAE 2.5, treated with 2.5% (wt/wt) DAE 5.0, treated with 5% (wt/wt) Est, treated with 17β-estradiol (10μg/kg BW). The rats were fed basal diet (AIN-93M) or experimental diets containing two levels of DAE for 6 weeks. Serum, femur and urine samples were collected. Weight gain was not affected by DAE supplementations. Femur weight and mineral contents (Ca, Mg) were significantly increased both by DAE supplementations and estrogen treatment compared to control. Serum estradiol levels were significantly elevated in DAE-fed groups than in the control group. In both DAE-fed and Est-treated groups, urinary hydroxyproline (OHPr) and deoxypyridinoline (DPD) excretion were significantly decreased in contrast to control group. Also serum interleukin-6 (IL-6) decreased both by DAE supplementations and Est treatment, suggesting that bone-sparing effect could be partly attributed to the modulation of osteoclastogenesis induced by IL-6. These results indicate that a DAE has beneficial effects on estrogen-dependent bone loss and can be considered as an alternative functional substance for estrogen replacement therapy (ERT) in treating postmenopausal osteoporosis.

High versus low body condition in mares: Interactions with responses to somatotropin, GnRH analog, and dexamethasone1

Mares that had previously been fed to attain body condition scores (BCS) of 7.5 to 8.5 (high) or 3.0 to 3.5 (low) were used to determine the interaction of BCS with the responses to 1) administration of equine somatotropin (eST) daily for 14 d beginning January 20 followed by administration of GnRH analog (GnRHa) daily for 21 d and 2) 4-d treatment with dexamethasone later in the spring when mares in low BCS had begun to ovulate. The majority of mares with high BCS continued to cycle throughout the winter, as evidenced by larger ovaries (P < 0.002), more corpora lutea (P < 0.05), greater progesterone concentrations during eST treatment (P < 0.04), and more (P < 0.05) large- and medium-sized follicles. Treatment with eST alone or in combination with GnRHa had no effect (P > 0.05) on ovarian activity or ovulation. Plasma leptin concentrations were greater (P < 0.002) in mares with high BCS; however, there was no effect (P > 0.10) of eST treatment. Plasma IGF-I concentrations were greater (P < 0.0001) in mares treated with eST compared with mares given vehicle, and mares with high BCS had greater IGF-I (P < 0.02) and LH concentrations (P < 0.02) than mares with low BCS. Plasma leptin concentrations in mares with high BCS were increased (P < 0.001) within 12 h of dexamethasone treatment; the leptin response (P < 0.001) in mares with low BCS was greatly reduced (P < 0.001) and transient. Glucose and insulin concentrations also increased (P < 0.0001) after dexamethasone treatment in both groups, and the magnitude of the response was greater (P < 0.0001) in mares with high BCS than in mares with low BCS. In summary,low BCS in mares was associated with a consistent seasonal anovulatory state that was affected little by eST and GnRHa administration. In contrast, all but one mare with high BCS continued to experience estrous cycles and(or) have abundant follicular activity on their ovaries. The IGF-I response to eST treatment was also reduced in mares with low BCS, as was the basal leptin concentration and leptin response to dexamethasone. Although low BCS and leptin concentrations were associated with inactive ovaries during winter and early spring, mares with low BCS eventually ovulated in April and May while leptin concentrations remained low.

Daily treatment of growing foals with equine somatotropin: pathologic and endocrinologic assessments at necropsy and residual effects in live animals.

This experiment assessed the effects of 12 mo of daily treatment of young horses with recombinant equine somatotropin (eST) on 1) carcass and internal organ traits at necropsy and 2) residual effects in live horses for 60 d after cessation of treatment. Seven horses received eST daily at 20 microg/kg BW; seven others received vehicle (controls). Four horses from each group were killed at the end of treatment. There were few effects of eST treatment on hematologic assessments or histopathologic evaluations of internal organs. Treatment with eST increased the weights of the right adrenal gland (P = 0.090), left (P = 0.085) and right (P = 0.013) kidneys, liver (P = 0.012), tended to inrease the weights of pancreas (P = 0.082), spleen (P = 0.008), and heart (P = 0.102), and decreased (P = 0.032) somatotropin (ST) content in the adenohypophysis. Loin-eye area at the 10th rib was also greater (P = 0.01) in eST-treated horses than in controls. There was no difference (P > 0.15) between groups in left adrenal, brain, parathyroid glands, or thyroid gland weights or in 10th-rib fat thickness. In the remaining two control and three eST-treated horses (one control horse died), plasma IGF-I concentrations were higher (P = 0.001) in treated animals through d 6 after cessation of treatment and then dropped precipitously. Insulin concentrations in treated animals tended to be elevated (P = 0.08) only on d 0. There was a treatment x day interaction (P = 0.04) for plasma urea nitrogen levels, which increased in treated horses. A decrease (P < 0.05) in BW in the treated animals was observed by 21 d after treatment. There was no difference (P > 0.15) in insulin or glucose response to glucose tolerance tests given on d 0 through 60 after cessation of treatment. Overall ST response to secretagogue was reduced (P < 0.05) in eST-treated horses compared with controls. In summary, long-term treatment of growing horses with eST decreased endogenous ST response to secretagogue and increased plasma IGF-I concentrations and many internal organ weights but had little effect on hematologic or histopathologic characteristics at necropsy. The effects on IGF-I concentrations were lost within 6 d, and BW in treated horses decreased within 3 wk after cessation of treatment.

Bone density in the juvenile racehorse treated with exogenous somatotropin (eST)

Thirty juvenile horses were paired by age within sex, and one horse from each pair was randomly assigned to either eST treatment or to a control group. The horses were “broke” to ride and trained in a regimen typical for young racehorses. Radiographs were taken on days 0, 32, 50, 64, 82, 96 and 128 of the study. Increase in total radiographic bone aluminum equivalence (RBAE) was significantly greater in eST treated horses than in the control group (P<.003). Increase in medial RBAE was also significantly greater due to eST treatment (P<.0001). Trends were apparent for increases due to eST treatment in dorsal RBAE (P=.07) and lateral RBAE (P=.07). Bone density was selectively increased in the dorsal, lateral and medial vs. the palmar cortex in the eST treated horses.

Effects of exogenous equine somatotropin on mineral balance in two-year-old horses in race training

Summary This study examined the effects of exogenous equinesomatotropin (eST — Equigen) administration on mineral absorption and retention in two-year-old horses in race training. Sixteen Quarter Horse geldings were paired by age (ave.age=794 d), and one horse from each pair was assigned at random to either the eST treatment group or the control. The experiment was conducted over 112 days during which the horses were gentled to ride and trained on a dirt track in a regimen typical for race horses in training. At 28-day intervals, collections of total fecal and urinary output were made to determine effects on Ca, inorganic P, Mg, Cu and Zn mineral balance. Due to marginal and slightly deficient amounts (P .05) by treatment. The eST-treated horses increased (P

The effects of equine somatotropin (eST) on follicular development and circulating plasma hormone profiles in cyclic mares treated during different stages of the estrous cycle

The effects of exogenous equine somatotropin (eST) administration on ovarian activity and plasma hormone levels were evaluated on horse and pony mares. The objectives of this study were to determine the effects of eST on follicular development and circulating concentrations of leutinizing hormone (LH), estradiol, progesterone, and insulin-like growth factor I (IGF-I) in cyclic horse and pony mares. Sixteen mares received daily injections (im) of eST at a concentration of 25 μg/kg body weight on either Days 6 through 12 (Treatment A) or 13 through 19 (Treatment B) postovulation. In addition, contemporary mares were similarly given the carrier vehicle and served as controls (Treatments C and D). Blood samples were collected at 24-hr intervals and ultrasonographic evaluations were performed on the ovaries of each mare at 48-hr intervals beginning on the first day of treatment and ending either on the day of ovulation or 5 d postovulation. Circulating levels of insulin-like growth factor-I (IGF-I) were increased in treated mares by Day 3 post-treatment (P < 0.05). Also, mares in Treatment B exhibited a decrease in plasma estradiol concentrations (P < 0.05) when compared with control mares on Days 1 through 5 postovulation of the post-treated estrous cycle. In addition, circulating leutinizing hormone levels were different for mares in Treatment A compared with controls on Days −8 through −1 pre-ovulation (P < 0.05). All follicles present on the ovaries of each mare were measured and placed into one of five categories based on their diameter. Neither the mean number of follicles per size category ⩾8 mm in diameter nor the mean follicular diameter within each size category differed among treatment and control mares. However, eST treatment significantly increased the number of follicles ⩽7 mm on the ovaries of mares treated early in the estrous cycle when compared with control mares on Days 3 and 7 post-treatment and at the onset of standing estrus.

Physiologic and skeletal response to exogenous equine somatotropin (eST) in two-year-old Quarter Horses in race training

This study was conducted to determine the physiologicand skeletal response to exogenous equine somatotropin (eST) in two-year-old horses in race training. Sixteen Quarter Horse geldings were paired by age (ave. age=26 months), and one horse from each pair was assigned at random to either the eST treatment group or the control. The experiment was conducted over 112 days during which the horses were broke to ride and trained on a dirt track in a regimen typical for racehorses in training. Radiographs were taken of the left third metacarpal on day 0,28, 42, 56, 70, 84 and 112. Jugular blood samples were collected every 14 days and analyzed for calcium (Ca), phosphorous (P), osteocalcin, 25-dihydroxyvitamin D (Vit D), parathyroid hormone (PTH), insulin-like growth factor-I (IGF-I) and growth hormone (GH). Daily injections of eST were well tolerated and resulted in no observed side effects. There was a trend (P= .14) for increased plasma GH concentrations in the treated horses as the trial progressed. Plasma IGF-I concentrations were increased (P<.01) in the treated horses by Day 14 and remained higher than the control horses for the duration of the study. As seen in other studies, there was a pattern of decreasing and subsequent increasing of both medial (P<.02) and lateral (P<.01) cortical bone density during the training period, but treatment differences were not significant at any day. Serum osteocalcin was affected by day (P <.01) and was higher (P<.04) in the eST-treated horses on Day 42. Serum Vitamin D, calcium, and phosphorus were affected by day (P<.01) but not by treatment (P>.05). In this study, administration of eST was effective in elevating plasma IGF-I, which is stimulatory to osteoblasts, and serum osteocalcin, which is a product of osteoblastic activity.

Feed intake, body weight, body condition score, musculation, and immunocompetence in aged mares given equine somatotropin.

Sixteen 20- to 26-yr-old mares were given 0, 6.25, or 12.5 mg/d equine somatotropin (eST) to determine whether aged mares respond to ST with changes in feed intake, body weight, body condition score (based mostly on fat cover), or immunocompetence. Neither dry matter intake, body weight, nor body condition scores were altered during the 6 wk of eST injection. However, based on photographs taken to evaluate musculation before and after treatment (scores 0 to 4), mares given eST developed greater (P < .07) muscle definition (1.8 +/- .6 and 2.5 +/- .6 for 6.25 and 12.5 mg eST/d, respectively) than control mares (.7 +/- .4). Total circulating leukocytes increased (P < .05) in both of the eST-treated groups during the 6-wk injection period, caused by an increase (P < .05) in granulocytes. Lymphocyte numbers were not altered. Granulocyte oxidative burst activity was not altered by eST treatment. Although lymphocyte proliferative responses to phytohemagglutinin, pokeweed mitogen, or lipopolysaccharide were not altered during the treatment period, lymphocyte proliferation in response to phytohemagglutinin and pokeweed mitogen increased twofold in eST-treated horses at 2 wk after eST treatment. In overview, the increased musculation and the increase in granulocyte numbers in mares given eST suggest that eST supplementation may improve the health and well-being of aged mares.

The effects of equine somatotropin (met-eGH) on follicular development in cycling mares

The effects of exogenous equine somatotropin (eST) administration on ovarian activity and plasma hormone levels were evaluated on horse and pony mares. The objectives of this study were to determine the effects of eST on follicular development and circulating concentrations of leutinizing hormone (LH), estradiol, progesterone, and insulin-like growth factor I (IGF-I) in cyclic horse and pony mares. Sixteen mares received daily injections (im) of eST at a concentration of 25 μg/kg body weight on either Days 6 through 12 (Treatment A) or 13 through 19 (Treatment B) postovulation. In addition, contemporary mares were similarly given the carrier vehicle and served as controls (Treatments C and D). Blood samples were collected at 24-hr intervals and ultrasonographic evaluations were performed on the ovaries of each mare at 48-hr intervals beginning on the first day of treatment and ending either on the day of ovulation or 5 d postovulation. Circulating levels of insulin-like growth factor-I (IGF-I) were increased in treated mares by Day 3 post-treatment (P < 0.05). Also, mares in Treatment B exhibited a decrease in plasma estradiol concentrations (P < 0.05) when compared with control mares on Days 1 through 5 postovulation of the post-treated estrous cycle. In addition, circulating leutinizing hormone levels were different for mares in Treatment A compared with controls on Days −8 through −1 pre-ovulation (P < 0.05). All follicles present on the ovaries of each mare were measured and placed into one of five categories based on their diameter. Neither the mean number of follicles per size category ⩾8 mm in diameter nor the mean follicular diameter within each size category differed among treatment and control mares. However, eST treatment significantly increased the number of follicles ⩽7 mm on the ovaries of mares treated early in the estrous cycle when compared with control mares on Days 3 and 7 post-treatment and at the onset of standing estrus.

Evaluation of combined coramine-electroshock therapy in the treatment of schizophrenia.

1. Thirteen schizophrenic female patients from among consecutive new admissions were given 5 c. c. coramine in conjunction with their regular series of EST, and a roughly comparable group of 13 patients received saline injections with their EST. 2. These patients were given the Wechsler-Bellevue Intelligence Scale and the Bender-Gestalt test just prior to their shock series and were retested 5 weeks following their last treatment. 3. Results indicate that EST used in conjunction with coramine significantly decreased the time spent in EST treatment. Patients receiving coramine returned to a higher level of intellectual functioning and more adequately perceived reality than did their controls as measured by psychological tests. 4. It is hypothesized that coramine-EST brings about an increased blood flow and thalamic stimulation occurs with most of the results being similar in quality, but greater in quantity, to those with standard EST. The one major qualitative difference noted in contrast with standard EST treatment was the lowering of inadequate defenses. The use of coramine-EST makes "intensivetreatment EST" for excited patients unnecessary, thereby avoiding the undesirable post treatment confusion and clinical organic symptoms. 5. Coramine-EST patients experience less fear and appear more receptive to psychotherapy. Acutely ill new admissions benefit most. 6. This is a pilot study and the results should be considered tentative in view of the size of our sample. A 2-year follow-up of these patients would be necessary to draw any definite conclusions as to the permanency of the effects of coramine-EST. The marked tendency in favor of the coramine-EST group does, however, warrant further investigation of coramine as a variation in EST.

A REVISION OF THE PSYCHIATRIC RATING SCALE

In a previous paper(I) we described a psychiatric rating scale and demonstrated its use as a method for quantitative recording of psychiatric clinical findings and the changes that occur in them during the course of the illness. It was pointed out that a recording method of this type was particularly essential in any scientific investigation, the purpose of which is to search for possible correlation between clinical symptoms and physiological or biochemical data. It was also found that when patients were rated with this scale by several psychiatrists there was a satisfactory correlation in scores so that independent determination by individual workers could be regarded as reliable. In a subsequent paper(2) the application of this scale in an investigation of a number of patients suffering from agitated depression was reported. It was found that it was possible to use this scale in correlating changes in the clinical picture produced by electric shock treatment with the concomitant neurophysiological and biochemical findings. During the last 15 months, since the conclusion of the work reported in the abovementioned papers, we have continued our studies on this rating scale, our main purpose being to render it a more adequate tool for evaluation of changes in the clinical picture and extend its applicability in research investigations. These studies have resulted in the correction of certain faults in the original scale, and we now present the revised scale and our experiences in the use of it. First however we would like to point out some of the more important changes that were made and the reasons for introducing them. In the first place it was important to establish a more adequate base line against which the deviations in the clinical picture of our patients could be compared. In our briginal