The impact of cultural stress and gender norms on alcohol use severity among Latino immigrant men.

Alcohol misuse affects 15 million people in the United States. Compared to White men, Latino men have disproportionately higher rates of both alcohol misuse and negative alcohol-related consequences (e.g. drunk driving, liver disease, alcohol dependence, HIV/AIDS). This cross sectional study examined how cultural stressors [immigration stress and negative context of reception (NCR)] coupled with traditional Latino male gender norms (machismo and caballerismo) influences alcohol use severity (AUS) among adult Latino immigrant men. Data for the present study was collected between 2017 and 2018 from 279 Cuban, Central American, and South American adult Latino men who immigrated to the US approximately 10 years prior. Results from hierarchical multiple regression analysis revealed higher levels of perceived NCR (ß = 0.15, p = .01), and machismo (ß = 0.16, p = .02) were associated with greater AUS. Significant interaction effects were found between both cultural stressors and machismo [immigration stress x machismo (ß = 0.22, p < .001); NCR x machismo (ß = 0.22, p < .001)] whereby higher levels of machismo strengthened the association between cultural stress and AUS. Findings from the present study can inform culturally appropriate interventions aimed at mitigating alcohol use among Latino immigrant men.

COVID-19 Vaccine Acceptance Among Latino/a Immigrants: The Role of Collective Responsibility and Confidence.

Research on COVID-19 vaccine hesitancy has been sparse among Latino/a immigrants, a population at high risk for infection. This exploratory study examines rates of vaccine acceptance and its association with psychological antecedents of vaccination among Latino/a immigrants. A cross-sectional telephone survey on perceptions of COVID-19 was administered between October 2020 to February 2021 in South Florida to 200 adult Latino/a immigrants. Descriptive statistics, bivariate analysis, and logistic regression were employed to determine the influence of independent variables on vaccine acceptance. Most participants indicated a willingness to get vaccinated. Participants with higher confidence (aOR = 10.2, 95% CI: 4.8-21.8) and collective responsibility scores were (aOR = 3.1, 95%CI:1.3-6.9) more likely to report vaccine acceptance than those with lower scores. No other psychological antecedents or demographic variables were significantly associated with vaccine acceptance. Study results provide insights into motivating factors for vaccination that can inform culturally tailored education campaigns to increase vaccine acceptability in this population.

Dengue virus identification by transmission electron microscopy and molecular methods in fatal dengue hemorrhagic fever.

Dengue virus is the most significant virus transmitted by arthropods worldwide and may cause a potentially fatal systemic disease named dengue hemorrhagic fever. In this work, dengue virus serotype 4 was detected in the tissues of one fatal dengue hemorrhagic fever case using electron immunomicroscopy and molecular methods. This is the first report of dengue virus polypeptides findings by electron immunomicroscopy in human samples. In addition, not-previously-documented virus-like particles visualized in spleen, hepatic, brain, and pulmonary tissues from a dengue case are discussed.

Position statement relating ankle fractures in major outpatient surgery. / Documento de posicionamiento respecto a las fracturas de tobillo en cirugía mayor ambulatoria.

INTRODUCTION: The increase in the prevalence of osteoporosis associated with ageing, and sports and traffic accidents, are responsible for the increase in ankle fractures. This fact emphasises the need to protocolise their care in order to provide greater clinical benefit to patients, and better cost-benefit ratios to the health system. AIM AND METHOD: At present, there is no common framework for implementation of protocols and internal circuits of the Spanish centres for ankle fractures by means of major outpatient surgery (MOS), which is the final objective of this position paper. For this, the clinical and economic evidence of MOS, the local environment and the strategies for its implementation are reviewed, related to ankle fractures. CLINICAL AND ECONOMIC EVIDENCE: The results showed a better cost-benefit ratio in outpatients compared to traditional hospitalisation, with lower complications and readmission rates and therefore significant cost savings. BARRIERS AND STRATEGIES: General and specific barriers are reviewed, as well as strategies and circuits for proper implementation. RESULTS: The results show lower complication and readmission rates together with significant cost savings. It entails a better cost-benefit ratio in outpatient care compared to traditional hospitalisation. POSITION STATEMENT: The implementation of MOS contributes to improve the quality of care, and the satisfaction of both, patient and health care team, while optimising the utilisation of resources. Ankle fractures in patients selected for both the underlying pathology, anaesthetic risk, and the type of fracture can be operated satisfactorily under the MOS.

The percentage of CD39+ monocytes is higher in pregnant COVID-19+ patients than in nonpregnant COVID-19+ patients.

Current medical guidelines consider pregnant women with COVID-19 to be a high-risk group. Since physiological gestation downregulates the immunological response to maintain "maternal-fetal tolerance", SARS-CoV-2 infection may constitute a potentially threatening condition to both the mother and the fetus. To establish the immune profile in pregnant COVID-19+ patients, a cross-sectional study was conducted. Pregnant women with COVID-19 (P-COVID-19+; n = 15) were analyzed and compared with nonpregnant women with COVID-19 (NP-COVID-19+; n = 15) or those with physiological pregnancy (P-COVID-19-; n = 13). Serological cytokine and chemokine concentrations, leucocyte immunophenotypes, and mononuclear leucocyte responses to polyclonal stimuli were analyzed in all groups. Higher concentrations of serological TNF-α, IL-6, MIP1b and IL-4 were observed within the P-COVID-19+ group, while cytokines and chemokines secreted by peripheral leucocytes in response to LPS, IL-6 or PMA-ionomicin were similar among the groups. Immunophenotype analysis showed a lower percentage of HLA-DR+ monocytes in P-COVID-19+ than in P-COVID-19- and a higher percentage of CD39+ monocytes in P-COVID-19+ than in NP-COVID-19+. After whole blood polyclonal stimulation, similar percentages of T cells and TNF+ monocytes between groups were observed. Our results suggest that P-COVID-19+ elicits a strong inflammatory response similar to NP-COVID19+ but also displays an anti-inflammatory response that controls the ATP/adenosine balance and prevents hyperinflammatory damage in COVID-19.

Overview of autosomal dominant polycystic kidney disease in the south of Spain. / Panorámica de la poliquistosis renal autosómica dominante en una región del sur de España.

INTRODUCTION: Although autosomal dominant polycystic kidney disease is the most common hereditary kidney disease, available data tend to be limited to after initiation of renal replacement therapy. OBJECTIVE: To ascertain an overview of autosomal dominant polycystic kidney disease within the health area of Granada in southern Spain. MATERIAL AND METHODS: From January 2007 to December 2016, we collected clinical, family and demographic information about all patients with autosomal dominant polycystic kidney disease, irrespective of whether or not they were treated with RRT, in the Granada health area. The computer software SPSS 15.0 and GenoPro were used. RESULTS: 50.6% of the 1,107 diagnosed patients were men. 99.1% were Caucasian and 4-6 generations/family were studied. The geographical distribution was heterogeneous. There was no family history in 2.43%. The mean age of diagnosis was 34.0±17.80 years and the diagnosis was made after having offspring in 57.7% of cases. The main reason for diagnosis was family history (46.4%). The mean age of initiation of renal replacement therapy was 54.2±11.05 years. 96.3% of the deceased had some degree of renal failure at the time of death. The mean age of death was 60.9±14.10 years, the main cause of death being unknown in 33.5% of cases, followed by cardiovascular (27.8%). CONCLUSIONS: Cases and families were concentrated in certain geographical areas and a significant number of individuals were undiagnosed prior to cardiovascular death or diagnosed late after reproduction. Given that there is currently no curative treatment, the primary prevention strategy of preimplantation genetic diagnosis should play a leading role.

Long-term fluctuation of standard automatic perimetry, pulsar perimetry and frequency-doubling technology in early glaucoma diagnosis.

PURPOSE: Analyze the stability and accuracy of 3 perimetric techniques. METHODS: A total of 104 stable eyes (65 subjects) with ocular hypertension and early glaucoma [group G, mean defect = 1.08 dB, SD = 2.0, in standard TOP automatic perimetry (SAP)] were examined 5 times during 18 months using: (a) SAP; (b) Pulsar temporal modulation perimetry (T30W), and (c) frequency-doubling technology (FDT N30). Ninety eyes from 90 normal controls were compared with the first set of examinations of group G. RESULTS: The learning effect was minimal in the 3 techniques but higher in Pulsar (1.0 src, p < 0.05) than in SAP and FDT (0.4 dB). Long-term fluctuation (F) was significantly higher in FDT (3.1 dB, SD = 1.4, p < 0.0001) than in SAP (2.3 dB, SD = 1.1) and in Pulsar (1.9 src, SD = 0.7). Pulsar and FDT reduce F when increasing the number of examinations. F seems equivalent in SAP and FDT and lower in Pulsar, considering small-scale differences of the 3 perimeters. CONCLUSIONS: A slight learning effect would be expected on FDT and SAP in patients with previous experience with SAP. The stability and sensitivity of Pulsar is greater than on the other 2 systems. For early diagnosis of glaucoma it is essential to prove the reproducibility and coincidence of perimetric results.

Topographical spatial summation in glaucoma.

PURPOSE: Stimulus luminance (L) and area (A) are related by the equation L x A(k)=constant. The authors evaluated the k value at 66 positions of the central visual field in patients with glaucoma, to modify L and A simultaneously in order to examine advanced glaucomas with a bigger dynamic range. METHODS: The luminance limitation of a computer screen with automatic photometric control was compensated for by increasing the stimulus area in the range between 0 and 17 dB, using the k topographic values previously calculated on normal subjects. Four initial series of 21, 12, 10, and 10 glaucomas were sequentially examined with the Octopus 311 in which the stimulus size cannot be freely changed during the examination, and with the experimental method (Pulsar-SAP) modifying stimulus sizes to equal the results. k Final estimation was verified in 60 new cases. RESULTS: k values increase progressively with defect deepness. Values higher than those of the normal population with equivalent topographic differences were obtained. Correlation between indices was as follows: MD: r=0.94 (p<0.0001); square root of the loss of variance (sLV): r=0.93 (p<0.0001). Frequency of local defects was similar in both procedures. Average topographic differences between thresholds were usually less than 1 dB. The average threshold difference favored Pulsar-SAP by 0.45 dB at those points where the average threshold of both examinations was less than 18 dB and 0.37 dB where such average was higher than or equal to 18 dB. CONCLUSIONS: k value is higher in patients with glaucoma than in normal subjects, although the topographic features are similar. It is feasible to design a scale combining stimulus luminance and sizes to use screens with relative low brightness as surfaces for visual field examination.

[Diagnostic capability of PULSAR, FDT y HRT-II in glaucoma suspects]. / Capacidad diagnóstica del HRT-II y de las perimetrías TOP, PULSAR y FDT en pacientes sospechosos de sufrir glaucoma.

PURPOSE: To determine the diagnostic capability of PULSAR-T30W, FDT-Threshold-N30 and HRT-II in glaucoma suspects. METHODS: Forty-seven eyes from 47 referred glaucoma suspects (GS) were examined twice with each technique. Cases with TOP-WW-MD>6dB were excluded. Results were compared with those of 70 eyes from 70 normal controls (C). RESULTS: Mean MD value using TOP-WW in the GS group (0.96dB. sd=1.7) was not significantly different from C (0.8dB. sd=1.77) (p>0.05). Disc area in GS group (2.12 mm(2). sd=0.34) was significantly greater than in C (1.97 mm2. sd=0.45) (p<0.01). For 95.7% specificity, PULSAR-sLV showed the highest sensitivity of 30.9% in individual examinations. The highest reproducible sensitivity in the two examinations was obtained using HRT-II maximum contour elevation (23.4%) and reference height (23.4%), and was 14.9% for various indices after correcting for the influence of disc area (cup area, cup/disc area ratio, maximum contour depression and mean RNFL thickness). Reproducible sensitivity of the perimetric indices was: PULSAR-MD=8.5%, PULSAR-sLV=17%, FDT-MD=6.4%, FDT-PSD=4.3%. The association of perimetric and HRT-II indices achieved high sensitivity but low diagnostic reproducibility. CONCLUSIONS: The most effective indices were maximum contour elevation, reference height and PULSAR-sLV, although the inclusion of the optic nerve head assessment in the selection of the GS sample may have favored the HRT-II results.

Ab initio determination of the crystal structure of cytochrome c6 and comparison with plastocyanin.

BACKGROUND: Electron transfer between cytochrome f and photosystem I (PSI) can be accomplished by the heme-containing protein cytochrome c6 or by the copper-containing protein plastocyanin. Higher plants use plastocyanin as the only electron donor to PSI, whereas most green algae and cyanobacteria can use either, with similar kinetics, depending on the copper concentration in the culture medium. RESULTS: We report here the determination of the structure of cytochrome c6 from the green alga Monoraphidium braunii. Synchrotron X-ray data with an effective resolution of 1.2 A and the presence of one iron and three sulfur atoms enabled, possibly for the first time, the determination of an unknown protein structure by ab initio methods. Anisotropic refinement was accompanied by a decrease in the 'free' R value of over 7% the anisotropic motion is concentrated at the termini and between residues 38 and 53. The heme geometry is in very good agreement with a new set of heme distances derived from the structures of small molecules. This is probably the most precise structure of a heme protein to date. CONCLUSIONS: On the basis of this cytochrome c6 structure, we have calculated potential electron transfer pathways and made comparisons with similar analyses for plastocyanin. Electron transfer between the copper redox center of plastocyanin to PSI and from cytochrome f is believed to involve two sites on the protein. In contrast, cytochrome c6 may well use just one electron transfer site, close to the heme unit, in its corresponding reactions with the same two redox partners.

[Patterns of susceptibility to antibiotics of Enterobacteriaceae causing intra-abdominal infection in Spain: SMART 2003 study outcomes]. / Patrones de sensibilidad a antimicrobianos de Enterobacteriaceae causantes de infecciones intraabdominales en España: resultados del estudio SMART 2003.

SMART (Study for Monitoring Antimicrobial Resistance Trends) is an ongoing global antimicrobial surveillance program focused on clinical isolates from intra-abdominal infections. The objective of this subanalysis was to assess antimicrobial susceptibility patterns among Entero-bacteriaceae recovered at 13 participating Spanish sites during 2003. Antimicrobial susceptibility testing was performed using broth microdilution techniques according to the CLSI (formerly NCCLS) guidelines for MIC testing. The presence of extended-spectrum beta-lactamases (ESBL) was confirmed in isolates with a MIC of ceftriaxone, ceftazidime, or cefepime>or=2 mg/l by comparing cefepime MICs with and with-out clavulanate. A total of 981 Enterobacteriaceae recovered from 840 patients were tested, of which 398 (41%) were community-acquired. Escherichia coli was the most common isolate (571 isolates; 58%), followed by Klebsiella spp. (153; 16% Enterobacter spp. (97; 10%), and Proteus spp. (63; 6%). A total of 191 isolates (19%) from 176 patients produced inducible beta-lactamases. The carbapenems and amikacin were the most consistently active agents against the Enterobacteriaceae (susceptibility>or=99%). Resistance rates for ceftazidime, cipro-floxacin, and levofloxacin exceeded 10%. ESBLs were detected phenotypically in 61 (6%) isolates, being the most common E. coli (61%), Klebsiella spp. (20%), and Enterobacter spp. (8%). Antimicrobial resistance among Enterobacteriaceae isolated from intra-abdominal infections is a problem in Spain. A significant proportion of inducible beta-lactamase and ESBL-producing Enterobacteriaceae causing intra-abdominal infection were acquired in the community. The carbapenems ertapenem, imipenem and meropenem and the aminoglycoside amikacin were highly active in vitro against Enterobacteriaceae isolated from intra-abdominal sites, including ESBL-producing organisms.

Diagnostic accuracy and reproducibility of tendency oriented perimetry in glaucoma.

PURPOSE: To evaluate the diagnostic capability of tendency oriented perimetry (TOP) in glaucoma. METHODS: A): The diagnostic accuracy of mean defect (MD), square-root of the loss variance (s LV), and number of pathologic points (NPP) was calculated in 295 normal and 414 glaucoma eyes (179 early, 112 moderate, and 123 advanced) examined with TOP. B): Threshold fluctuation (F) and its relationship with the loss variance (LV) was measured in 34 normal and 33 glaucoma eyes (mean MD=3 dB; SD=3.9) for TOP and for full-threshold perimetry (FT). C): Twenty-eight eyes with stable glaucoma (mean MD=9.5 dB; SD=7.2) were examined six times to quantify LV error. D): TOP and FT were tested with the simulation program PeriSim using different behavior models. RESULTS: A): The best diagnostic index in early glaucoma (MD<6dB) was sLV (specificity=90.2%, sensitivity=84.9). The three indices had similar precision in moderate and severe glaucoma. B): Threshold fluctuation and sLV were better correlated in TOP (r=0.72, p<0.01) than in FT (r=0.62, p<0.01). For normal subjects, in FT the incidence of F<2 dB was 8.82% and s LV<1.5 dB 5.88%. The same frequencies in TOP were 67.65% and 55.88%. C): Averaging six examinations reduced the sLV val ue by 22%. D): The threshold estimation error increased 1 dB in TOP in relation to FT for the same patient's behavior, but the error in TOP was lower than i n FT when the worst behavior was modeled. CONCLUSIONS: TOP is a good discriminator between glaucoma and normality. Perimetry results overestimate the real sLV value. TOP's high diagnostic ability is probably associated to the algorithm design and to less contaminating influences during the examination.

The 2.15 A crystal structure of a triple mutant plastocyanin from the cyanobacterium Synechocystis sp. PCC 6803.

The crystal structure of the triple mutant A42D/D47P/A63L plastocyanin from the cyanobacterium Synechocystis sp. PCC 6803 has been determined by Patterson search methods using the known structure of the poplar protein. Crystals of the triple mutant A42D/D47P/A63L, which are stable for days in its oxidized form, were grown from ammonium sulfate, with the cell constants a = b = 34.3 A and c = 111.8 A belonging to space group P3(2)21. The structure was refined using restrained crystallographic refinement to an R-factor of 16.7% for 4070 independent reflections between 8.0 and 2.15 A with intensities greater than 2 sigma (I), with root mean square deviations of 0.013 A and 1.63 degrees from ideal bond lengths and bond angles, respectively. The final model comprises 727 non-hydrogen protein atoms within 98 residues, 75 water molecules and a single copper ion. The overall tertiary fold of Synechocystis plastocyanin consists of a compact ellipsoidal beta-sandwich structure made up of two beta-sheets embracing a hydrophobic core. Each sheet contains parallel and antiparallel beta-strands. In addition to the beta-sheets, the structure contains an alpha-helix from Pro47 to Lys54 that follows beta-strand 4. The three-dimensional structure of Synechocystis plastocyanin is thus similar to those reported for the copper protein isolated from eukaryotic organisms and, in particular, from the cyanobacterium Anabaena variabilis, the only cyanobacterial plastocyanin structure available so far. The molecule holds an hydrophobic region surrounding His87, as do other plastocyanins, but the lack of negatively charged residues at the putative distant remote site surrounding Tyr83 could explain why the Synechocystis protein exhibits a collisional reaction mechanism for electron transfer to photosystem I (PSI), which involves no formation of the transient plastocyanin-PSI complex kinetically observed in green algae and higher plants.

Bone cadmium and lead in the ancient population from El Hierro, Canary Islands.

This study was performed to determine the levels of lead (Pb) and cadmium (Cd) in 63 bone samples of the prehispanic population of the island El Hierro, comparing them with the values obtained on 98 prehispanic samples from Tenerife, Fuerteventura, Gran Canaria, and La Palma, all of them in the Canary Islands, and with eight modern samples who served as controls. Prehispanic individuals from El Hierro showed the lowest bone Pb values of all the archipelago (0.72+/-1.01 mg/kg), significantly different (F=6.9, p<0.001) from the values obtained for the population of other islands such as Tenerife (4.87+/-5.36 mg/kg) or Fuerteventura (4.45+/-7.85 mg/kg) and also from those of the modern population (30.53+/-14.62 mg/kg). On the other hand, bone Cd, although slightly lower in the ancient population groups, was not significantly different when compared with the modern one. In addition, no differences were observed in bone Cd among the ancient population of the different islands. Bone lead but not cadmium kept an inverse significant relationship with the distance of the burial site both to south Spain (r=-0.31) and Atlantic Morocco (r=-0.28, p<0.001 in both cases).

[Spatial summation: its topography in the central visual field]. / Topografía de la sumación espacial en el campo visual central.

PURPOSE: Luminance (L) and area (A) of the stimulus are related by the equation LxAk=constant. We evaluated the k value in 66 positions of the central visual field. METHODS: Ten eyes of 10 healthy subjects were examined for conventional luminous thresholds at 66 positions of the central visual field with the TOP strategy using Goldmann sizes 4, 3.5, 3, 2.5 and 1.9 in the PULSAR perimeter. RESULTS: The k value increased in a lineal manner from the centre towards the periphery (slope=0.01 per degree, average k=0.616, r=0.98, p<0.01) but with unequal slopes at the different meridians. In the inferior hemi-fields it was higher (k=0.657) than in the superior hemi-fields (k=0.574). The k value at the supero-nasal hemifields is quite similar to that found at the paracentral region, and maximum at the temporal inferior region. CONCLUSIONS: Spatial summation in the central visual field has specific values for different positions.

[Nomogram for ocular hypertension progression risk based on the ocular hypertension treatment study]. / Nomograma de riesgo de progreso de hipertensión ocular basado en el ocular hypertension treatment study.

INTRODUCTION: A practical nomogram has been designed in order to present the results obtained from the Ocular Hypertension Study (Gordon et al. Arch Ophthalmol 2002; 120: 714-720), where the relation between intraocular pressure (IOP) and corneal thickness becomes apparent, involving the risk of evolution from ocular hypertension into glaucoma within a 6 year period. MATERIAL AND METHODS: We used a multiple logarithmic regression for the nine parameters shown in figure 1 of the above mentioned paper. RESULTS: A correlation coefficient of 0.91 (p<0.001) permits to establish the following equation: Probability of evolution (%) = 13539.5 x (1.1385IOP) x (0.9818(CORNEAL THICKNESS)). This implies that a variation of 10 microns on corneal thickness leads to an IOP's modification of 1.5 mmHg in the same sense. From these data, we designed the nomogram included in this paper. CONCLUSIONS: IOP and pachymetry together allow an estimation of the risk of evolution from ocular hypertension into glaucoma in a graphical practical way. From this indirect estimation, the influence of corneal thickness on IOP's measure seems to be much higher than previously estimated.

Post-immigration Changes in Social Capital and Substance Use Among Recent Latino Immigrants in South Florida: Differences by Documentation Status.

Changing social capital among recent Latino immigrants (RLIs) influences substance use post-immigration. This was a longitudinal study of 476 South/Central American RLIs examining social capital and substance use changes pre to post-immigration. Self-reported measures of social capital and substance use were compared between surveys administered within 1 year of immigration and 2 years post-immigration. Post-immigration, social capital, hazardous drinking and illicit drug use decreased. Women were less likely to engage in hazardous drinking [adjusted odds ratio (AOR) .32, p < .001], and less likely to use illicit drugs (AOR .67, p = .01). Documented individuals with higher levels of 'business' social capital had increased odds of illicit drug use (AOR 2.20, p < .05). Undocumented individuals with higher levels of 'friend and others' social capital had decreased risk for hazardous drinking and illicit drug use (AOR .55, p < .01; AOR .56, p < .05). Documentation status moderated the relationship between social capital and substance use. RLIs can be targeted for primary prevention of substance abuse.

Synechocystis 6803 plastocyanin isolated from both the cyanobacterium and E. coli transformed cells are identical.

Native plastocyanin from Synechocystis 6803 has been isolated and purified to electrophoretic homogeneity. The corresponding gene (petE) has been cloned and expressed in E. coli, thus leading to a protein completely identical to plastocyanin purified from the cyanobacterial cells. The petE gene product is correctly processed in E. coli as deduced from the N-terminal amino acid sequences. These results, along with the identical physicochemical and kinetic properties of the two protein preparations, confirm that expression of petE in E. coli is an adequate tool to address the study of Synechocystis plastocyanin by site-directed mutagenesis.

Cloning and correct expression in E. coli of the petJ gene encoding cytochrome c6 from Synechocystis 6803.

Cytochrome c6 from the cyanobacterium Synechocystis 6803 has been isolated and purified to electrophoretic homogeneity. The gene coding for such a heme protein (petJ) has been cloned and properly expressed in E. coli. This procedure yields a protein preparation completely identical to that obtained from the cyanobacterial cells. The N-terminal amino acid sequences of cytochrome c6 synthesized in both organisms are the same, thus allowing us to conclude that the petJ gene product is correctly processed in E. coli. To the best of our knowledge, this is the first time that any cytochrome c6 is produced in the enterobacterium. The identical physicochemical and kinetic properties of the proteins isolated from both sources confirm that expression of the petJ gene in E. coli is an adequate tool to address the study of Synechocystis cytochrome c6 by site-directed mutagenesis in a parallel way to that carried out with plastocyanin from the same organism.