Characteristics affecting survival after locally advanced colorectal cancer in Quebec.

BACKGROUND: We estimated the relations of sociodemographic, organizational, disease, and treatment variables with the risk of death from colorectal cancer (crc) in a Quebec population-based sample of patients with locally advanced crc (lacrc) who underwent tumour resection with curative intent. METHODS: Information from medical records and administrative databases was obtained for a random sample of 633 patients surgically treated for stages ii-iii rectal and stage iii colon cancer and declared to the Quebec cancer registry in 1998 and 2003. We measured personal, disease, and clinical management characteristics, relative survival, and through multivariate modelling, relative excess rate (rer) of death. RESULTS: The relative 5- and 10-year survivals in this cohort were 67.7% [95% confidence interval (ci): 65.8% to 69.6%] and 61.2% (95% ci: 58.3% to 64.0%) respectively. Stage T4, stage N2, and emergency rather than elective surgery affected 18%, 24% and 10% of patients respectively. Those disease progression characteristics each independently increased the rer of death by factors of 2 to almost 5. Grade, vascular invasion, and tumour location were also significantly associated with the rer for death. Receiving guideline-adherent treatment was associated with a 60% reduction in the rer for death (0.41; 95% ci: 0.28 to 0.61), an effect that was consistent across age groups. Clear margins (proximal-distal, radial) and clinical trial enrolment were each associated with a nonsignificant 50% reduction in the rer. Of patients less than 70 years of age and 70 years of age and older, 81.3% and 42.0% respectively received guideline-adherent treatment. CONCLUSIONS: This study is the first Quebec population-based examination of patients with lacrc and their management, outcomes, and outcome determinants. The results can help in planning crc control strategies at a population level.

Soft Tissue Sarcomas in Octogenarian Patients: Are Treatment Options and Oncological Outcomes Different? A SEER Retrospective Study.

AIMS: As the US population continues to age, oncological strategies and outcomes for soft tissue sarcomas (STSs) should continue to be examined for varying age groups. The aim of this study was analyse and compare treatment strategies and oncological outcomes for octogenarian patients with STSs. MATERIALS AND METHODS: Data from the Surveillance, Epidemiology and End Results (SEER) national database were used. Varying treatment modalities were studied when utilised for specific tumour staging with respect to the eighth edition of the American Joint Committee on Cancer. RESULTS: In total, 24 666 patients were included for analysis, where 3341 (14%) were 80 years old or older. The octogenarian group was diagnosed with more advanced disease (stages II-IV), relative to their younger counterparts (85% versus 75%, P < 0.001). However, a smaller proportion of the older patients underwent surgical resection (74% versus 86%, P < 0.001). Likewise, the octogenarians received less chemotherapy (4% versus 21%, P < 0.001) and radiotherapy (29% versus 42%, P = 0.010). Surgical resection and chemotherapy significantly improved overall survival for those older patients with stage II STS, whereas surgical resection and radiotherapy improved mortality in this cohort with both stage III and IV STS. Overall survival at 1 and 5 years of follow-up was lower within the octogenarian group compared with the younger group (1 year: 68% versus 88%, P < 0.001 and 5 years: 7% versus 58%, P < 0.001). CONCLUSIONS: Octogenarian patients, in most cases, are diagnosed with stage III or metastatic disease. Surgical resection of the primary tumour was beneficial in both age cohorts, with radiotherapy correlating to better overall survival when used in those patients with higher stage STS. Chemotherapy was associated with better mortality in the younger cohort with respect to tumour stage. The octogenarian overall survival at 1 and 5 years was lower than for younger patients.

Protective effect of pyridoxal-5-phosphate (MC-1) on perioperative myocardial infarction is independent of aortic cross clamp time: results from the MEND-CABG trial.

AIM: Aortic cross-clamp time remains a significant marker of mortality and morbidity after coronary artery bypass graft (CABG) surgery. Pyridoxal-5-phosphate (MC-1), blocking purinergic receptors and intracellular influx of calcium, was shown to decrease the incidence of perioperative myocardial infarction in the prospective, randomized, double-blinded MC-1 to Eliminate Necrosis and Damage in CABG (MEND-CABG) clinical trial. METHODS: We studied the relationship between treatment with MC-1 and aortic cross-clamping relative to the incidence of cardiovascular (CV) death and myocardial infarction (MI) in the trial that enrolled 901 high-risk patients undergoing CABG with cardiopulmonary bypass. Patients were randomized to receive either placebo, MC-1 250 mg/day or MC-1 750 mg/day starting 3-10 h before CABG and continued for 30 days after surgery. Serial creatine kinase-myocardial band (CK-MB) determinations, ECGs and clinical evaluations were performed. RESULTS: Cross-clamping time increased the event rate of death and MI with an odds ratio (95% confidence interval) of 1.67 (1.17-2.37, P=0.0044). Treatment with MC-1 decreased the rate of events (P=0.0073) with odds ratios of 0.52 (0.31-0.88 for MC-1 250 mg/day versus placebo) and 0.48 (0.29-0.82 for MC-1 750 mg/day versus placebo). There was no interaction between cross-clamp time and treatment (P=0.61) on the occurrence of the combined endpoint. CONCLUSION: MC-1 decreased the incidence of CV death and MI (CK-MB >or=100 ng/mL) during the first 90 days after CABG in the MEND-CABG trial. Although longer aortic clamping time increased the risk of cardiovascular events, the protective effect of MC-1 was independent of ischemic time during CABG.

Maternal factors implicated in fetal bradycardia after combined spinal epidural for labour pain.

BACKGROUND AND OBJECTIVE: Combined spinal epidural analgesia is effective for fast relief of severe labour pain but has been associated with worrisome decreases in fetal heart rate. Since the reasons for this phenomenon remain elusive, some anaesthesiologists may abstain from using this technique. We postulated that factors unrelated to the neuraxial technique could play a role in the decrease in fetal heart rate. To our knowledge, no prospective study has previously looked into this possibility. METHODS: We collected prospective data on 223 consecutive patients who received combined spinal epidural analgesia (123) or epidural analgesia (100). Maternal blood pressure, analogue pain scores, exogenous infusion of oxytocin, cervical dilatation, maternal age, parity and ethnicity were collected and correlated with the occurrence of decreases in fetal heart rate post combined spinal epidural. RESULTS: Univariate analysis showed a correlation between the incidence of fetal bradycardia and higher maternal pain scores, older maternal age, and combined spinal epidural analgesia. Multivariate analysis revealed that only pain scores and maternal age were independent predictors of fetal bradycardia post neuraxial blockade. CONCLUSIONS: Maternal pain scores and older maternal age are factors unrelated to the neuraxial technique that are independent predictors of fetal bradycardia after neuraxial analgesia for labour.

Enhancer and promoter elements directing activation and glucocorticoid repression of the alpha 1-fetoprotein gene in hepatocytes.

Mutations were introduced in 7 kilobases of 5'-flanking rat alpha 1-fetoprotein (AFP) genomic DNA, linked to the chloramphenicol acetyltransferase gene. AFP promoter activity and its repression by a glucocorticoid hormone were assessed by stable and transient expression assays. Stable transfection assays were more sensitive and accurate than transient expression assays in a Morris 7777 rat hepatoma recipient (Hepa7.6), selected for its strong AFP repression by dexamethasone. The segment of DNA encompassing a hepatocyte-constitutive chromatin DNase I-hypersensitive site at -3.7 kilobases and a liver developmental stage-specific site at -2.5 kilobases contains interacting enhancer elements sufficient for high AFP promoter activity in Hepa7.6 or HepG2 cells. Deletions and point mutations define an upstream promoter domain of AFP gene activation, operating with at least three distinct promoter-activating elements, PEI at -65 base pairs, PEII at -120 base pairs, and DE at -160 base pairs. PEI and PEII share homologies with albumin promoter sequences, PEII is a near-consensus nuclear factor I recognition sequence, and DE overlaps a glucocorticoid receptor recognition sequence. An element conferring glucocorticoid repression of AFP gene activity is located in the upstream AFP promoter domain. Receptor-binding assays indicate that this element is the glucocorticoid receptor recognition sequence which overlaps with promoter-activating element DE.

Functional analysis of developmentally regulated chromatin-hypersensitive domains carrying the alpha 1-fetoprotein gene promoter and the albumin/alpha 1-fetoprotein intergenic enhancer.

During liver development, the tandem alpha 1-fetoprotein (AFP)/albumin locus is triggered at the AFP end and then asymmetrically enhanced; this is followed by autonomous repression of the AFP-encoding gene. To understand this regulation better, we characterized the two early developmental stage-specific DNase I-hypersensitive (DH) sites so far identified in rat liver AFP/albumin chromatin: an intergenic DH-enhancer site and the AFP DH-promoter site. Mutation-transfection analyses circumscribed the DH-enhancer domain to a 200-bp DNA segment stringently conserved among species. Targeted mutations, DNA-protein-binding assays, and coexpression experiments pinpointed C/EBP as the major activatory component of the intergenic enhancer. Structure-function relationships at the AFP DH-promoter site defined a discrete glucocorticoid-regulated domain activated cooperatively by HNF1 and a highly specific AFP transcription factor, FTF, which binds to a steroid receptor recognition motif. The HNF1/FTF/DNA complex is deactivated by glucocorticoid receptors or by the ubiquitous factor NF1, which eliminates HNF1 by competition at an overlapping, high-affinity binding site. We propose that the HNF1-NF1 site might serve as a developmental switch to direct autonomous AFP gene repression in late liver development. We also conclude that the intergenic enhancer is driven by C/EBP alpha primarily to fulfill albumin gene activation functions at early developmental stages. Factor FTF seems to be the key regulator of AFP gene-specific functions in carcinoembryonic states.

Difference in the action of ethyl and methyl methane sulfonates on DNA template activity for RNA synthesis in vitro.

RNA produced in vitro from alkylated T7 DNA has been characterized by polyacrylamide gel electrophoresis. Methylation of T7 DNA by methyl methane sulfonate reduces RNA chain length. In contrast, ethylation of T7 DNA by ethyl methane sulfonate, while reducing RNA synthesis to the same extent, does not alter chain length.

A new culture and quantitation system for megakaryocyte growth using cord blood CD34+ cells and the GPIIb/IIIa marker.

A new culture and quantitation system has been established for growth of megakaryocyte-lineage cells from human progenitor cells. CD34+ progenitor cells were enriched from umbilical cord blood using an avidin-biotin immunoadsorption process. These cells were preincubated in bulk liquid culture for 3 to 4 days in the presence of the growth factors interleukin-3 (IL-3) and IL-6. The cells were then washed and seeded at 5000 cells/well in 96-well plates that contained a variety of test samples. The plates were incubated for 7 days, and the cells were then washed, transferred to ELISA plates, and fixed. Megakaryocyte growth was determined by an ELISA for the platelet glycoprotein (GP) IIb/IIIa, an abundant membrane protein found on cells committed to the megakaryocyte lineage. The growth factor IL-3 was found to produce a very strong signal in this assay. The addition of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-6, stem cell factor (SCF), or leukemia inhibitory factor (LIF) to low levels of IL-3 also stimulated megakaryocyte growth, as measured by IIb/IIIa expression. Plasma from patients with aplastic anemia was also stimulatory in this assay, and showed marked synergy with IL-3. This progenitor cell culture system, due to its judicious use of progenitor cells and an automated, 96-well quantitation method, allows for screening large numbers of test samples and multiple combinations and concentrations of growth factors.

Gold suppression of human neutrophil function in vitro.

The effect of myochrysine and Auranofin on leukocyte function were measured using quantitative leukocyte iodination. Both suppressed iodination at concentrations achieved in patients. Under conditions of leukocyte submaximum stimulation, enhanced gold suppression was observed. The active portion of Myochrysine appeared to be protein bound while the active portion of Auranofin appeared to be free. Preincubation experiments indicated suppression of the myeloperoxidase-halide system. Inhibition of this probable mediator of inflammation may be one of the modes of action of gold.

Should we patch corneal erosions?

OBJECTIVE: To study the effect of patching on the speed of reepithelialization, slit-lamp signs of epithelial wound healing, and patient discomfort following a corneal abrasion. METHODS: Forty-eight eyes of 46 patients with corneal erosion sparing Bowman membrane were randomized into 2 groups: with or without patching. Slit-lamp examination and photographs of the fluorescein-stained cornea were performed on a daily basis until reepithelialization was complete. Photographs were analyzed using computer-assisted planimetry. RESULTS: No statistically significant difference was found between patched (n = 25) and nonpatched (n = 22) eyes for the mean size of the initial erosion (patched eyes, 23.7 mm2; nonpatched eyes, 18.9 mm2; P = .42), linear speed of reepithelialization (reduction over time of the radius of the largest circle included in the erosion: patched eyes, 0.0375 mm/h; nonpatched eyes, 0.0353 mm/h; P = .78), and surface speed of reepithelialization (reduction over time of the erosion area: patched eyes, 0.6510 mm2/h; nonpatched eyes, 0.5657 mm2/h; P = .60). The power to detect a 12-hour delay of epithelial closure was 95%. There were no significant differences between the 2 groups for pain, analgesia, insomnia, aspect of the epithelial border, intensity and duration of stromal edema, Descemet folds, anterior uveitis, and filaments. CONCLUSIONS: Patching a corneal erosion does not significantly accelerate reepithelialization and does not alter the epithelial wound healing pattern. It does not reduce the incidence and severity of inflammation nor relieve pain when compared with treatment without patching.

Anterior stromal punctures for bullous keratopathy.

OBJECTIVE: To evaluate the therapeutic effects of anterior stromal punctures (ASP) in patients with bullous keratopathy (BK). PATIENTS AND METHODS: Twenty-seven patients awaiting penetrating keratoplasty with a diagnosis of BK were examined. They were seen before treatment with ASP and 1, 4, and 12 weeks after treatment. The examination included slit-lamp examination, photography of the cornea, ultrasonic pachymetry, central esthesiometry, and pneumotonometry. Subjective evaluations of pain, discomfort, and photophobia were also done using a visual scale model. Photographs were analyzed by computer-assisted planimetry and used to measure the corneal surface covered by bullae and microcysts. Pretreatment and posttreatment values (mean +/- SEM) were compared using the Student paired t test. RESULTS: At 3 months, a significant reduction in pain was noted. A decrease in the mean corneal surface covered by bullae (BKPreASP = 2733 +/- 553 microns2; BK3mo = 1006 +/- 356 microns2, P = .004) was observed. A decrease in the esthesiometry (E) measurement (EPreASP = 3.5 +/- 0.4 cm; E3mo = 1.3 +/- 0.3 cm, P < .001), an increase in corneal thickness ([CT] CTPreASP = 869 +/- 24 microns; CT3mo = 902 +/- 21 microns, P < .001), and a decrease in the number of quadrants through which iris (I) details could be seen (IPreASP = 1.7 +/- 0.3; I3mo = 1.2 +/- 0.3, P = .015) were also noted. These findings corroborate the clinical observation of increased subepithelial fibrosis following ASP. CONCLUSIONS: Anterior stromal punctures reduce bullae formation and alleviate pain in patients with BK, and they constitute a valuable alternative to penetrating keratoplasty should surgery be delayed or contraindicated.

Oncodevelopmental and hormonal regulation of alpha 1-fetoprotein gene expression.

The main features of the oncodevelopmental biology of alpha 1-fetoprotein (AFP) are reviewed. Progress made in the molecular biology of AFP gene regulation is discussed and we present our recent data on the mechanisms of AFP suppression by glucocorticoid hormones. The relationship between AFP gene transcription and cell replication is examined, and it is suggested that the degree of methylation of the AFP gene (or of co-methylated regulatory DNA sequences) conditions its response to hormones.

Corneal transplant tolerance of cryopreservation.

PURPOSE: To study the outcome of corneal transplants performed with cryopreserved tissue. METHODS: Maisonneuve-Rosemont Hospital medical records of all corneal transplantations performed with cryopreserved tissue by one surgeon (M.L.F.) between March 1978 and April 1991 were reviewed. The Kaufman--Capella cryopreservation technique was used. Corneas were cryopreserved for periods of 3 days to 16.8 years (mean, 4.6 years) before transplantation. RESULTS: We report a mean follow-up of 54 months (range, 2.8--151.3 months). Survival analysis showed the probability of a clear graft to be 76% at 1 year and 73.2% at 2 years. At the time of the last visit, visual acuity was 20/40 or better in 61 eyes (49.2%). The mean postoperative pachometry was 0.58 mm (range, 0.50--0.75 mm). Specular microscopy performed in 57 eyes showed a mean endothelial cell count of 938 cells/mm(2) 55.1 months (range, 2.9--151.3 months) after surgery. For comparison purposes, the outcome of a subgroup of cryopreserved (n = 33) and noncryopreserved (n = 26) corneas transplanted by the same surgeon between April 1986 and April 1990 was studied. CONCLUSION: Despite an increase in the primary failure rate and higher initial endothelial cell loss, cryopreserved transplants are viable. Although we do not recommend cryopreservation of corneas for elective surgery, we consider that cryopreserved corneas can be very useful in emergency situations when tissue availability is a problem.

Hydroxyl radical production in the cortex and striatum in a rat model of focal cerebral ischemia.

BACKGROUND: Increases in hydroxyl radical production have been used as evidence of oxidative stress in cerebral ischemia/ reperfusion. Ischemia can also induce increased dopamine release from the striatum that may contribute to hydroxyl radical formation. We have compared hydroxyl radical production in the cortex and striatum as an index of oxidative stress in a rat model of focal cerebral ischemia with cortical infarction. METHODS: Using a three vessel occlusion model of focal cerebral ischemia combined with bilateral microdialysis, hydroxylation of 4-hydroxybenzoate (4HB) was continuously monitored in both hemispheres in either the lateral striatum or frontoparietal cortex. The ischemia protocol consisted of one hour equilibration, 30 min of three vessel occlusion, then release of the contralateral common carotid artery (CCA) for 2.5 h. RESULTS: Induction of ischemia resulted in a 30-fold increase in dopamine release in the lateral striatum. Compared to the nonischemic striatum, the ratio of the hydroxylation product 3,4-dihydroxybenzoate (34DHB) to 4HB (trapping agent) in the ipsilateral striatum increased significantly 30 min after ischemia induction. In contrast, during the 30 min of three vessel occlusion there was no increase in the ratio in the cortex. Following the release of the contralateral CCA, the ratio from the ischemic cortex increased significantly compared to sham-operated animals. However, under all circumstances, the 34DHB/4HB ratio was greater in the striatum than in the cortex. CONCLUSION: The increase in the 34DHB/4HB ratio in the lateral striatum coincides with the increased dopamine release suggesting a role for dopamine oxidation in the increased production of hydroxyl radicals. The significant increase in the ratio from the ischemic cortex compared to that from the sham-operated animals is consistent with increased oxidative stress induced by ischemia. However, the lower 34DHB/4HB ratio in the cortex which does not receive dopaminergic innervation compared to the striatum suggests a different mechanism for hydroxyl radical production. Such an alternate mechanism may represent a more toxic oxidative insult that contributes to infarction.

[Support psychotherapy or support in psychotherapy : a necessary reflection on the work in sector psychiatry.]. / Psychothérapie de soutien ou soutien en psychothérapie : une réflexion nécessaire sur le travail en psychiatrie de secteur*

This article deals with the notion of supportive psychotherapy. This overall approach is often discredited even if, in the mental health network, it is widely used. Therefore, it appears important to analyze this approach and to define the idea of support in psychotherapy; this article reviews secondly when it is opportune to offer such an approach, what are the techniques related to this approach and finally, what are the results we can hope for? This article wants to initiate a reflexion while integrating some readings on the matter and using a clinical experience evolved from a practice in sector psychiatry.

Determinants Of Oral corticosteroid Responsiveness in Wheezing Asthmatic Youth (DOORWAY): protocol for a prospective multicentre cohort study of children with acute moderate-to-severe asthma exacerbations.

INTRODUCTION: Oral corticosteroids are the cornerstone of acute asthma management in the emergency department. Recent evidence has raised doubts about the efficacy of this treatment in preschool-aged children with viral-induced wheezing and in smoking adults. The aims of the study were to: (1) document the magnitude of response to oral corticosteroids in children presenting to the emergency department with moderate or severe asthma; (2) quantify potential determinants of response to corticosteroids and (3) explore the role of gene polymorphisms associated with the responsiveness to corticosteroids. METHODS AND ANALYSIS: The design is a prospective cohort study of 1008 children aged 1-17 years meeting a strict definition of asthma and presenting with a clinical score of ≥4 on the validated Pediatric Respiratory Assessment Measure. All children will receive standardised severity-specific treatment with prednisone/prednisolone and cointerventions (salbutamol with/without ipratropium bromide). Determinants, namely viral aetiology, environmental tobacco smoke and single nucleotide polymorphism, will be objectively documented. The primary efficacy endpoint is the failure of emergency department (ED) management within 72 h of the ED visit. Secondary endpoints include other measures of asthma severity and time to recovery within 7 days of the index visit. The study has 80% power for detecting a risk difference of 7.5% associated with each determinant from a baseline risk of 21%, at an α of 0.05. ETHICS AND DISSEMINATION: Ethical approval has been obtained from all participating institutions. An impaired response to systemic steroids in certain subgroups will challenge the current standard of practice and call for the immediate search for better approaches. A potential host-environment interaction will broaden our understanding of corticosteroid responsiveness in children. Documentation of similar effectiveness of corticosteroids across determinants will provide the needed reassurance regarding current treatment recommendations. RESULTS: Results will be disseminated at international conferences and manuscripts targeted at emergency physicians, paediatricians, geneticists and respirologists. TRIAL REGISTRATION NUMBER: This study is registered at Clinicaltrials.gov (NCT02013076).

Drosophila heat shock system as a general model to investigate transcriptional regulation.

Whereas the regulation of a gene is uniquely tailored to respond to specific biological needs, general transcriptional mechanisms are used by diversely regulated genes within and across species. The primary mode of regulation is achieved by modulating specific steps in the transcription cycle of RNA polymerase II (Pol II). Pol II "pausing" has recently been identified as a prevalent rate-limiting and regulated step in the transcription cycle. Many sequence-specific transcription factors (TFs) modulate the duration of the pause by directly or indirectly recruiting positive transcription elongation factor b (P-TEFb) kinase, which promotes escape of Pol II from the pause into productive elongation. These specialized TFs find their target-binding sites by discriminating between DNA sequence elements based on the chromatin context in which these elements reside and can result in productive changes in gene expression or nonfunctional "promiscuous" binding. The binding of a TF can precipitate drastic changes in chromatin architecture that can be both dependent and independent of active Pol II transcription. Here, we highlight heat-shock-mediated gene transcription as a model system in which to study common mechanistic features of gene regulation.

Left ventricular end-diastolic pressure is a predictor of mortality in cardiac surgery independently of left ventricular ejection fraction.

BACKGROUND: Several risk factors have been shown to increase mortality in cardiac surgery. However, the importance of left ventricular end-diastolic pressure (LVEDP) as an independent risk factor before cardiac surgery is unclear. Method. This observational study investigated 3024 consecutive adult patients who underwent cardiac surgical procedures at the Montreal Heart Institute from 1996 to 2000. The primary outcome was in-hospital mortality with 99 deaths (3.3%) among these patients. RESULTS: Of the 35 variables subjected to univariate analysis, 23 demonstrated a significant association with mortality. Stepwise multivariate logistic regression identified LVEDP as an independent predictor of mortality after cardiac surgery. The area under the receiver operating characteristic curve of the model predicting mortality was 0.85. CONCLUSIONS: Elevated LVEDP is an independent predictor of mortality in cardiac surgery. This variable is independent of left ventricular ejection fraction.

Purification and spectroscopic characterization of a recombinant chloroplastic hemoglobin from the green unicellular alga Chlamydomonas eugametos.

Hemoglobins (Hb), which have the important task of delivering molecular oxygen by facilitating its reversible binding to the heme, are now thought to have evolved in all groups of organisms including prokaryotes, fungi, plants and animals. Our recent finding of a light-inducible chloroplastic Hb in the green unicellular alga Chlamydomonas eugametos has further extend this idea, while raising questions about the function that an Hb could play in a high oxygen environment such as in the chloroplast. In order to understand the role played by this new Hb, we have undertaken its biochemical characterization. To facilitate the characterization of Chlamydomonas Hb, which represents less than 0.01% of the soluble protein in the green alga, the protein has been expressed in Escherichia coli and purified to apparent homogeneity. The purified recombinant protein possesses a non-covalently bound iron-protoporphyrin IX heme. The oxy form of the recombinant Hb. purified directly from bacterial cells, is very stable, with a measured half-life of 7 days at pH 8 and has an ultraviolet/visible spectrum similar to those of the related cytoplasmic Hbs of the ciliated protozoa Paramecium and Tetrahymena and of the cyanobacterium Nostoc commune. In contrast to what has been reported for oxymyoglobins and oxyhemoglobins, the dioxygen molecule bound to the L1637 Hb can be reduced by the electron-transfer mediator phenazine methosulfate in the presence of NADPH, indicating that the heme pocket of Chlamydomonas Hb may be more accessible to small molecules. With regard to this we found that when the small reducing agent sodium dithionite is used to reduce the met form, it must be removed anaerobically from the Hb prior to oxygenation of the protein to stably produce the oxy form. Otherwise, the oxy form is obtained readily from the met form under an oxygenic atmosphere when ferredoxin and ferredoxin NADP+ reductase are used to enzymically reduce the Hb. Finally, the spectra of the deoxy and met forms were unusual, the heme being partly low-spin at physiological pH. These results confirm the existence of a reversible oxygen-binding protein in the chloroplast of C. eugametos. The unusual spectral and biochemical properties of the protein may reflect a specialized function for this Hb.