Biomarkers of treatment benefit with atezolizumab plus vemurafenib plus cobimetinib in BRAFV600 mutation-positive melanoma.

BACKGROUND: The phase III IMspire150 study (NCT02908672) demonstrated significantly improved progression-free survival (PFS) with atezolizumab, vemurafenib, and cobimetinib (atezolizumab group) versus placebo, vemurafenib, and cobimetinib (control group) in patients with BRAFV600-mutated advanced melanoma. We report exploratory biomarker analyses to optimize targeting of patients who are more likely to benefit from triplet combination therapy. PATIENTS AND METHODS: Five hundred fourteen patients were randomized to atezolizumab (n = 256) or control (n = 258). Outcomes were evaluated in subgroups defined by key biomarkers, including programmed death-ligand 1 (PD-L1) expression, lactate dehydrogenase (LDH) level, tumor mutational burden (TMB), and interferon-γ (IFN-γ) gene signature. Exploratory recursive partitioning analysis was then used to model associations between PFS and baseline covariates, including key biomarkers. RESULTS: PFS benefit for atezolizumab versus control was greater in patients with high TMB [≥10 mutations/Mb; hazard ratio (HR) 0.73; 95% confidence interval (CI) 0.52-1.02; P = 0.067] versus low TMB (<10 mutations/Mb; HR 0.92; 95% CI 0.65-1.30; P = 0.64) and similar between patients with strong IFN-γ (≥median; HR 0.76; 95% CI 0.54-1.06) versus weak IFN-γ (

Incidence of acute complications of herpes zoster among immunocompetent adults in England: a matched cohort study using routine health data.

BACKGROUND: Herpes zoster can cause rare but serious complications; the frequency of these complications has not been well described. OBJECTIVES: To quantify the risks of acute non-postherpetic neuralgia (PHN) zoster complications, to inform vaccination policy. METHODS: We conducted a cohort study among unvaccinated immunocompetent adults with incident zoster, and age-, sex- and practice-matched control adults without zoster, using routinely collected health data from the UK Clinical Practice Research Datalink (years 2001 to 2018). Crude attributable risks of complications were estimated as the difference between Kaplan-Meier-estimated 3-month cumulative incidences in patients with zoster vs. controls. We used Cox models to obtain hazard ratios for our primary outcomes in patients with and without zoster. Primary outcomes were ocular, neurological, cutaneous, visceral and zoster-specific complications. We also assessed whether antivirals during acute zoster protected against the complications. RESULTS: In total 178 964 incident cases of zoster and 1 799 380 controls were included. The absolute risks of zoster-specific complications within 3 months of zoster diagnosis were 0·37% [95% confidence interval (CI) 0·34-0·39] for Ramsay Hunt syndrome, 0·01% (95% CI 0·0-0·01) for disseminated zoster, 0·04% (95% CI 0·03-0·05) for zoster death and 0·97% (95% CI 0·92-1·00) for zoster hospitalization. For other complications, attributable risks were 0·48% (95% CI 0·44-0·51) for neurological complications, 1·33% (95% CI 1·28-1·39) for ocular complications, 0·29% (95% CI 0·26-0·32) for cutaneous complications and 0·78% (95% CI 0·73-0·84) for visceral complications. Attributable risks were higher among patients > 50 years old. Patients with zoster had raised risks of all primary outcomes relative to controls. Antiviral prescription was associated with reduced risk of neurological complications (hazard ratio 0·61, 95% CI 0·53-0·70). CONCLUSIONS: Non-PHN complications of zoster were relatively common, which may affect cost-effectiveness calculations for zoster vaccination. Clinicians should be aware that zoster can lead to various complications, besides PHN.

The association between partner bereavement and melanoma: cohort studies in the U.K. and Denmark.

BACKGROUND: Psychological stress is commonly cited as a risk factor for melanoma, but clinical evidence is limited. OBJECTIVES: This study aimed to evaluate the association between partner bereavement and (i) first-time melanoma diagnosis and (ii) mortality in patients with melanoma. METHODS: We conducted two cohort studies using data from the U.K. Clinical Practice Research Datalink (1997-2017) and Danish nationwide registries (1997-2016). In study 1, we compared the risk of first melanoma diagnosis in bereaved vs. matched nonbereaved people using stratified Cox regression. In study 2 we estimated hazard ratios (HRs) for death from melanoma in bereaved compared with nonbereaved individuals with melanoma using Cox regression. We estimated HRs separately for the U.K. and for Denmark, and then pooled the data to perform a random-effects meta-analysis. RESULTS: In study 1, the pooled adjusted HR for the association between partner bereavement and melanoma diagnosis was 0·88 [95% confidence interval (CI) 0·84-0·92] across the entire follow-up period. In study 2, we observed increased melanoma-specific mortality in people experiencing partner bereavement across the entire follow-up period (HR 1·17, 95% CI 1·06-1·30), with the peak occurring during the first year of follow-up (HR 1·31, 95% CI 1·07-1·60). CONCLUSIONS: We found decreased risk of melanoma diagnosis, but increased mortality associated with partner bereavement. These findings may be partly explained by delayed detection resulting from the loss of a partner who could notice skin changes. Stress may play a role in melanoma progression. Our findings indicate the need for a low threshold for skin examination in individuals whose partners have died. What is already known about this topic? Psychological stress has been proposed as a risk factor for the development and progression of cancer, including melanoma, but evidence is conflicting. Clinical evidence is limited by small sample sizes, potential recall bias associated with self-report, and heterogeneous stress definitions. What does this study add? We found a decreased risk of melanoma diagnosis, but increased mortality associated with partner bereavement. While stress might play a role in the progression of melanoma, an alternative explanation is that bereaved people no longer have a close person to help notice skin changes, leading to delayed melanoma detection. Linked Comment: Talaganis et al. Br J Dermatol 2020; 183:607-608.

Partner bereavement and risk of psoriasis and atopic eczema: cohort studies in the U.K. and Denmark.

BACKGROUND: Stress is commonly cited as a risk factor for psoriasis and atopic eczema, but such evidence is limited. OBJECTIVES: To investigate the association between partner bereavement (an extreme life stressor) and psoriasis or atopic eczema. METHODS: We conducted cohort studies using data from the U.K. Clinical Practice Research Datalink (1997-2017) and Danish nationwide registries (1997-2016). The exposed cohort was partners who experienced partner bereavement. The comparison cohort was up to 10 nonbereaved partners, matched to each bereaved partner by age, sex, county of residence (Denmark) and general practice (U.K.). Outcomes were the first recorded diagnosis of psoriasis or atopic eczema. We estimated hazard ratios (HRs) and confidence intervals (CIs) using a stratified Cox proportional hazards model in both settings, which were then pooled in a meta-analysis. RESULTS: The pooled adjusted HR for the association between bereavement and psoriasis was 1·01 (95% CI 0·98-1·04) across the entire follow-up. Similar results were found in other shorter follow-up periods. Pooled adjusted HRs for the association between bereavement and atopic eczema were 0·97 (95% CI 0·84-1·12) across the entire follow-up, 1·09 (95% CI 0·86-1·38) within 0-30 days, 1·18 (95% CI 1·04-1·35) within 0-90 days, 1·14 (95% CI 1·06-1·22) within 0-365 days and 1·07 (95% CI 1·02-1·12) within 0-1095 days. CONCLUSIONS: We found a modest increase in the risk of atopic eczema within 3 years following bereavement, which peaked in the first 3 months. Acute stress may play a role in triggering onset of new atopic eczema or relapse of atopic eczema previously in remission. We observed no evidence for increased long-term risk of psoriasis and atopic eczema following bereavement.

Inequalities in zoster disease burden: a population-based cohort study to identify social determinants using linked data from the U.K. Clinical Practice Research Datalink.

BACKGROUND: Zoster vaccination was introduced in England in 2013, where tackling health inequalities is a statutory requirement. However, specific population groups with higher zoster burden remain largely unidentified. OBJECTIVES: To evaluate health inequalities in zoster disease burden prior to zoster vaccine introduction in England. METHODS: This population-based cohort study used anonymized U.K. primary care data linked to hospitalization and deprivation data. Individuals aged ≥ 65 years without prior zoster history (N = 862 470) were followed from 1 September 2003 to 31 August 2013. Poisson regression was used to obtain adjusted rate ratios (ARRs) for the association of sociodemographic factors (ethnicity, immigration status, individuals' area-level deprivation, care home residence, living arrangements) with first zoster episode. Possible mediation by comorbidities and immunosuppressive medications was also assessed. RESULTS: There were 37 014 first zoster episodes, with an incidence of 8·79 [95% confidence interval (CI) 8·70-8·88] per 1000 person-years at risk. In multivariable analyses, factors associated with higher zoster rates included care home residence (10% higher vs. those not in care homes), being a woman (16% higher vs. men), nonimmigrants (~30% higher than immigrants) and white ethnicity (for example, twice the rate compared with those of black ethnicity). Zoster incidence decreased slightly with increasing deprivation (ARR most vs. least deprived 0·96 (95% CI 0·92-0·99) and among those living alone (ARR 0·96, 95% CI 0·94-0·98). Mediating variables made little difference to the ARR of social factors but were themselves associated with increased zoster burden (ARR varied from 1·11 to 3·84). CONCLUSIONS: The burden of zoster was higher in specific sociodemographic groups. Further study is needed to ascertain whether these individuals are attending for zoster vaccination.

Statin use and the risk of herpes zoster: a nested case-control study using primary care data from the U.K. Clinical Research Practice Datalink.

BACKGROUND: Statins are commonly prescribed worldwide and recent evidence suggests that they may increase the risk of herpes zoster (HZ). OBJECTIVES: To quantify the effect of statin exposure on the risk of HZ in the U.K. METHODS: A matched case-control study was conducted using data from U.K. primary care and hospital records. Patients > 18 years with an incident diagnosis of HZ were matched to up to four controls for age, sex and general practice. Patients were included in the statin exposure group if they had ever used a statin, and the daily dosage of the most recent statin prescription and the time since the most recent statin prescription were also recorded. The primary outcome was an incident diagnosis of HZ. Odds ratios (ORs) were estimated from conditional logistic regression and adjusted for potential confounders. RESULTS: A total of 144 959 incident cases of HZ were matched to 549 336 controls. Adjusted analysis suggested strong evidence for an increase in the risk of HZ related to statin exposure (OR 1·13, 95% confidence interval 1·11-1·15). There was also an increased risk when dosages were increased for patients who were currently or had recently been receiving statin treatment (Ptrend < 0·001), and we found an attenuation of the increased risk of HZ in previous statin users as the time since last statin exposure increased (Ptrend < 0·001). CONCLUSIONS: These findings are consistent with the hypothesis that statin therapy leads to an increase in the risk of HZ.

Variations in radiotherapy delivery in England - evidence from the national radiotherapy dataset.

AIMS, MATERIALS AND METHOD: Data in the national radiotherapy dataset for England for 2009-2011 is based upon downloads of activity from every linear accelerator in the country through its oncology management system linked to the local patient administration system to give a full overview of each patient episode. RESULTS: An analysis of this dataset shows that there is still a considerable variation in radiotherapy activity across the country, with a two-fold variation between the most and least active networks. Lower activity is seen in London and the southeast compared with the rest of the country, but when the data are split between the north and south of the country, no such variation is seen. Activity is higher in smaller centres and non-teaching centres. About half of all treatment is palliative and this proportion does not vary with geography, although there is considerable variation between individual centres in the proportion of radical radiotherapy given. There is a trend towards less use of radiotherapy, both radical and palliative, in the more deprived population groups, although no change in the relative use of palliative and radical treatment. CONCLUSION: It is important to emphasise that these data currently reflect activity patterns only and do not reflect quality of care or treatment outcomes, which will be achieved by linkage with cancer registry data in the future.

Relative torque profiles of elite male youth footballers: effects of age and pubertal development.

This study aimed to establish the effect of age and pubertal development on relative peak torque in youth footballers. One hundred and fifty-seven subjects were arranged into U12-U18 age groups; of these 133 also completed a self-reporting measure of pubertal development (PDS) and were grouped accordingly. Bilateral isokinetic testing included mean peak torque/body weight (PT/BW) and angle of peak torque (AoPT) for reps 2-4 of 5 in concentric quadriceps (CQ), concentric hamstrings (CH) and eccentric hamstrings (EH) at 60 degrees s(-1). Two minutes rest were provided between concentric and eccentric tests for both legs. A significant increase with age was noted for PT/BW in CQ, CH and EH (P<0.05), although POST HOC analyses revealed different patterns of significance dependent upon muscle and type of contraction. Specifically, the U18 s (CQ:2.20 nm/kg, CH:1.39 nm/kg, EH:2.16 nm/kg) were not significantly stronger than the U16 s and U15 s (CQ:2.42/2.29 nm/kg, CH:1.41/1.27 nm/kg, EH:2.22/2.15 nm/kg) even though they had entered full time training. The relationship between EH PT/BW and AoPT showed a weak but significant inverse correlation (P<0.001; R= -0.390 and -0.346 for dominant and non dominant legs). The findings of this study present unique football specific normative and age/PDS group data for PT/BW and AoPT, and the corresponding relationship between these variables.

Isokinetic thigh muscle ratios in youth football: effect of age and dominance.

This study aimed to analyse the effect of age on muscle peak torque (PT), and Hamstring (H): Quadricep (Q) ratio in elite youth footballers. To date, no study has considered age-group playing level and pubertal development in this population. One hundred and fifty-seven elite youth footballers in the age groups U12 to U18 volunteered to participate in this study, 133 of these were further grouped for pubertal development. Prior to testing subjects completed separate familiarisation, a three minute cycle ergometer warm up (resistance 50-60W), and two sub-maximal repetitions. Concentric and eccentric isokinetic PT measures for reps 2-4 of H and Q muscle action were taken at 60 degrees s (-1). From this, conventional and functional H: Q ratio was calculated along with dominant: non dominant ratio for the concentric Q and H, and eccentric H conditions. Significant main effects were observed for the age/pubertal development group and PT in all muscles and conditions (P<0.05). Of particular interest was a significant main effect for age and Functional H: Q (P<0.05), which suggested a move away from equality at U18. Our study provides normative data for coaches, trainers and clinicians working with youth footballers and may also have connotations for injury prevention and performance.

Social deprivation and outcomes in colorectal cancer.

BACKGROUND: The aim of this study was to examine the influence of social deprivation on postoperative mortality and length of stay in patients having surgery for colorectal cancer. METHODS: Data were extracted from the Association of Coloproctology of Great Britain and Ireland database of patients presenting between April 2001 and March 2002. The effect of social deprivation, measured by the Townsend score, on 30-day postoperative mortality and length of stay was evaluated by two-level hierarchical regression analysis. RESULTS: A total of 7290 (86.8 percent) patients underwent surgery. Operative mortality was 6.7 percent and median length of stay 11 days. Deprivation indices were significantly higher in patients with Dukes' 'D' cancers, undergoing emergency surgery and with higher American Society of Anesthesiologists (ASA) grades (P<0.005). Worsening deprivation was associated with higher operative mortality and longer stay (P=0.014). For each unit increase in deprivation, there was 2.9 (95 percent confidence interval 0.5 to 5.2) percent increase in 30-day mortality. On multifactorial analysis, social deprivation was an independent predictor of length of stay, but its effect on operative mortality was explained by differences in ASA grade, operative urgency and Dukes' classification. CONCLUSION: Social deprivation was an independent risk factor of postoperative length of stay and associated with higher postoperative mortality. These results have important implications for risk modelling of postoperative outcomes.

A national audit of radiotherapy in head and neck cancer.

AIMS: To undertake a national audit of radiotherapy practice in head and neck cancer to estimate compliance with published guidelines and national standards. METHODS: A two-part electronic data entry form was distributed to all U.K. radiotherapy centres in September 2000. The first part examined the centres' policies for managing interruptions, the second collected summaries of the management of 50 consecutive patients treated in each centre for head and neck cancer. The outcome measures were: frequency and causes of interruptions to therapy: policy and compliance with policy for managing interruptions; prolongation; and time between first visit to clinic and start of treatment. RESULTS: Fifty-five out of 56 centres returned data on a total of 2553 patients. Overall, 1467 (55%) patients had one or more treatment interruptions. Of patients whose treatment was interrupted, 56% still completed on time due to compensatory steps, but in 32% no attempted compensation was undertaken. Seven centres had no policy for dealing with treatment interruptions. Centres whose policies included treatment on bank (public) holidays achieved higher compliance and fewer prolonged cases than those whose policies did not. Average time from first visit to head and neck oncology clinic to starting radiotherapy was 40 days; six centres had an average wait of less than 28 days. CONCLUSIONS: This audit demonstrates wide variations in the quality of care between centres, failure to comply with guidelines for compensation for gaps and failure to meet national targets (for waiting times) that have serious implications both for patient outcomes and for the success of the National Cancer Plan.

Students' perceptions of learning outcomes from group-based, problem-based teaching and learning activities.

OBJECTIVE: A study aimed at exploring the variation in perceptions of learning outcomes reported by undergraduate nursing students enrolled in a problem-based learning subject in a pre-registration Bachelor of Nursing course (BN). METHOD: Students were asked to respond to four open-ended questions which focussed on their learning outcomes in the different teaching/learning modalities of the subject. Data were analysed in two phases using a modified phenomenographic analysis. In the first phase a set of categories of description were developed from the student responses to questions related to the learning modalities. In the second phase the individual responses were classified in terms of the categories. Finally, correlations between the learning modalities were identified. In this paper the approach to analysis, the process of category identification and the correlations between the learning modalities will be described and the implications for further research and teaching will be discussed. RESULTS: The findings indicated that there were two distinct groups of student responses. Inward focussed students who described outcomes in terms of their own learning and students whose focus was outward i.e. describing learning in terms of patient care and how learning relates to that care. Another important result shows the relationship between the learning modalities and outcomes. From the students' perspective, the most sophisticated outcomes of the lectures and laboratories were ideas and skills to be used and applied in clinical settings. Whereas, the group-based activities in which clinical problems were presented to the students in the form of Situation Improvement Packages (SIPS) focussed their attention on the clinical setting which constituted a preparation for the realities of clinical practice. CONCLUSION: The findings from this study indicate that students perceive their learning in the group based teaching/learning modality (SIPS) as effective in focussing them on the reality of their role in the clinical practice environment while lectures and laboratories provided the skills and knowledge required for this setting.

Cell wall-anchored CshA polypeptide (259 kilodaltons) in Streptococcus gordonii forms surface fibrils that confer hydrophobic and adhesive properties.

It has been shown previously that inactivation of the cshA gene, encoding a major cell surface polypeptide (259 kDa) in the oral bacterium Streptococcus gordonii, generates mutants that are markedly reduced in hydrophobicity, deficient in binding to oral Actinomyces species and to human fibronectin, and unable to colonize the oral cavities of mice. We now show further that surface fibrils 60.7 +/- 14.5 nm long, which are present on wild-type S. gordonii DL1 (Challis) cells, bind CshA-specific antibodies and are absent from the cell surfaces of cshA mutants. To more precisely determine the structural and functional properties of CshA, already inferred from insertional-mutagenesis experiments, we have cloned the entire cshA gene into the replicative plasmid pAM401 and expressed full-length CshA polypeptide on the cell surface of heterologous Enterococcus faecalis JH2-2. Enterococci expressing CshA exhibited a 30-fold increase in cell surface hydrophobicity over E. faecalis JH2-2 carrying the pAM401 vector alone and 2.4-fold-increased adhesion to human fibronectin. CshA expression in E. faecalis also promoted cell-cell aggregation and increased the ability of enterococci to bind Actinomyces naeslundii cells. Electron micrographs of negatively stained E. faecalis cells expressing CshA showed peritrichous surface fibrils 70.3 +/- 9.1 nm long that were absent from control E. faecalis JH2-2(pAM401) cells. The fibrils bound CshA-specific antibodies, as detected by immunoelectron microscopy, and the antibodies inhibited the adhesion of E. faecalis cells to fibronectin. The results demonstrate that the CshA polypeptide is the structural and functional component of S. gordonii adhesive fibrils, and they provide a molecular basis for past correlations of surface fibril production, cell surface hydrophobicity, and adhesion in species of oral "sanguis-like" streptococci.

Dietary supplements used in the treatment of depression, anxiety, and sleep disorders.

Dietary supplement use has increased during the past decade. Epidemiologic studies suggest that patients turn to dietary supplements because of a reluctance to take prescription medications or a lack of satisfaction with the results. They often perceive dietary supplements to be a safer or more natural alternative. Patients with mental health conditions, including depression, anxiety, and sleep disorders, are among those who use dietary supplements. St. John's Wort is used to treat depression. Clinical studies comparing dietary supplements with low-dose antidepressants (maprotiline, amitriptyline, or imipramine at 75 mg/day) or high-dose antidepressants (imipramine at 150 mg/day) find no significant difference between treatments. Kava kava is used to treat anxiety. Clinical trials demonstrate it to be superior to placebo, and roughly equivalent to oxazepam 15 mg/day or bromazepam 9 mg/day. Agents discussed for use in sleep disorders include melatonin, valerian, 5-hydroxytryptamine, catnip, chamomile, gotu kola, hops, L-tryptophan, lavender, passionflower, skullcap, and valerian. Familiarity with the evidence for use and the possible resulting risks can help health professionals to guide patient decisions regarding use of dietary supplements.

Spinal cord monitoring in scoliosis surgery. Experience with 1168 cases.

Since 1981, during operations for spinal deformity, we have routinely used electrophysiological monitoring of the spinal cord by the epidural measurement of somatosensory evoked potentials (SEPs) in response to stimulation of the posterior tibial nerve. We present the results in 1168 consecutive cases. Decreases in SEP amplitude of more than 50% occurred in 119 patients, of whom 32 had clinically detectable neurological changes postoperatively. In 35 cases the SEP amplitude was rapidly restored, either spontaneously or by repositioning of the recording electrode; they had no postoperative neurological changes. One patient had delayed onset of postoperative symptoms referrable to nerve root lesions without evidence of spinal cord involvement, but there were no false negative cases of intra-operative spinal cord damage. In 52 patients persistent, significant, SEP changes were noted without clinically detectable neurological sequelae. None of the many cases which showed falls in SEP amplitude of less than 50% experienced neurological problems. Neuromuscular scoliosis, the use of sublaminar wires, the magnitude of SEP decrement, and a limited or absent intra-operative recovery of SEP amplitude were identified as factors which increased the risk of postoperative neurological deficit.

Effect of whole-body pH changes on thoracic duct lymph flow in anesthetized sheep.

Physiologic alterations of pH in vitro produce alterations of the oncotic pressure of both plasma and interstitial proteins, the effect being more marked in the latter. Therefore we postulated that by changing an animal's pH we could produce alterations in fluid distribution and affect the whole-body lymph flow rate. To test this hypothesis, we infused seven acutely nephrectomized anesthetized sheep with 2% body weight/volume isosmotic lactated Ringer's solution and bicarbonate (200 mEq) after a 30-minute control period. The fluid was infused over a 10-minute period with 20-minute recovery intervals. To another group of seven ewes, we infused the same volume of lactated Ringer's solution and hydrochloric acid (40 mEq). Throughout the experiment, we continuously measured arterial and venous pressures and lymph flow rate. Every 10 minutes we obtained samples for arterial pH, for hematocrit, and for plasma and lymph protein and osmolality. The infusion of bicarbonate was associated with a comparatively lower peak lymph flow rate (383% over baseline compared with 757% for acid infusion). Also the lymph flow rate after acid infusion started to increase approximately 5 minutes after the beginning of the infusion, as compared with approximately 10 minutes after the base infusion. Acid infusion did not increase arterial pressure, in contrast to the other infusions. The changes in pH obtained with the infusions were insignificant for the Ringer's infusion, +0.17 for the base, and -0.16 for the acid. The plasma/lymph protein concentration ratios at the end of the infusion were no different for acid or base: 112.4% +/- 4.4% and 101.9% +/- 5% (difference from control, mean +/- SEM) respectively (p greater than 0.1). In conclusion, minimal alterations of pH produced alterations in the cardiovascular and lymph flow rate responses to fluid challenge in anesthetized animals.

Inhibition of the catalytic activity of bovine pineal gland hydroxyindole-O-methyltransferase (EC 2.1.1.4) by estrone, estradiol, progesterone, and testosterone but not pteridines.

The effect of pteridines and sex hormones on hydroxyindole-O-methyltransferase (HIOMT) catalysis was examined. Equivalent quantities of partially purified bovine HIOMT was incubated in the presence and absence of the various test substances and the results fitted to the Michaelis-Menten equation. The pteridines (triampterene and xanthopterin) had no noticeable effect on HIOMT catalysis, while all the sex hormones examined were found to be competitive inhibitors of the enzyme with respect to the hydroxy-substrate. Testosterone concentrations causing noticeable inhibition were above physiological while estrone, estradiol, and progesterone all caused inhibition within the physiological range; estrone was the most potent and progesterone the least potent inhibitor. The possible significance of these findings are discussed.

Probable mechanism of catalysis of pineal gland hydroxyindole-O-methyltransferase (HIOMT) from rainbow trout (Salmo gairdneri).

Activity of trout pineal HIOMT was found to increase with increase in incubation temperature from 5 to 40 degrees C although the activation energy remained constant over this range. From examination of the effects of the products of HIOMT catalysis on the enzyme it was apparent that the catalytic mechanism was ordered Bi-Bi with S-adenosylmethionine as the obligatory first substrate. Trout HIOMT was found to methylate all the common pineal hydroxyindoles with hydroxytryptophol having the greatest affinity for the enzyme. The pH optimum for trout HIOMT was found to be about pH 9.0 although routine use of a pH of 7.9 is recommended to limit potentially deliterious effects caused by degradation of S-adenosylmethionine at elevated pHs.