Eight new Halophytophthora species from marine and brackish-water ecosystems in Portugal and an updated phylogeny for the genus.

During an oomycete survey in December 2015, 10 previously unknown Halophytophthora taxa were isolated from marine and brackish water of tidal ponds and channels in saltmarshes, lagoon ecosystems and river estuaries at seven sites along the Algarve coast in the South of Portugal. Phylogenetic analyses of LSU and ITS datasets, comprising all described Halophytophthora species, the 10 new Halophytophthora taxa and all relevant and distinctive sequences available from GenBank, provided an updated phylogeny of the genus Halophytophthora s.str. showing for the first time a structure of 10 clades designated as Clades 1-10. Nine of the 10 new Halophytophthora taxa resided in Clade 6 together with H. polymorphica and H. vesicula. Based on differences in morphology and temperature-growth relations and a multigene (LSU, ITS, Btub, hsp90, rpl10, tigA, cox1, nadh1, rps10) phylo-geny, eight new Halophytophthora taxa from Portugal are described here as H. brevisporangia, H. cele-ris, H. frigida, H. lateralis, H. lusitanica, H. macrosporangia, H. sinuata and H. thermoambigua. Three species, H. frigida, H. macrosporangia and H. sinuata, have a homothallic breeding system while the remaining five species are sterile. Pathogenicity and litter decomposition tests are underway to clarify their pathological and ecological role in the marine and brackish-water ecosystems. More oomycete surveys in yet undersurveyed regions of the world and population genetic or phylogenomic analyses of global populations are needed to clarify the origin of the new Halophytophthora species. Citation: Maia C, Horta Jung M, Carella G, et al. 2022. Eight new Halophytophthora species from marine and brackish-water ecosystems in Portugal and an updated phylogeny for the genus. Persoonia 48: 54 - 90. https://doi.org/10.3767/persoonia.2022.48.02..

The distribution of KIR-HLA functional blocks is different from north to south of Italy.

The killer cell immunoglobulin-like receptor (KIR)-human leukocyte antigen (HLA) interaction represents an example of genetic epistasis, where the concomitant presence of specific genes or alleles encoding receptor-ligand units is necessary for the activity of natural killer (NK) cells. Although KIR and HLA genes segregate independently, they co-evolved under environmental pressures to maintain particular KIR-HLA functional blocks for species survival. We investigated, in 270 Italian healthy individuals, the distribution of KIR and HLA polymorphisms in three climatic areas (from cold north to warm south), to verify their possible geographical stratification. We analyzed the presence of 13 KIR genes and genotyped KIR ligands belonging to HLA class I: HLA-C, HLA-B and HLA-A. We did not observe any genetic stratification for KIR genes and HLA-C ligands in Italy. By contrast, in a north-to-south direction, we found a decreasing trend for the HLA-A3 and HLA-A11 ligands (P = 0.012) and an increasing trend for the HLA-B ligands carrying the Bw4 epitope (P = 0.0003) and the Bw4 Ile80 epitope (P = 0.0005). The HLA-A and HLA-B KIR ligands were in negative linkage disequilibrium (correlation coefficient -0.1211), possibly as a consequence of their similar function in inhibiting NK cells. The distribution of the KIR-HLA functional blocks was different along Italy, as we observed a north-to-south ascending trend for KIR3DL1, when coupled with HLA-B Bw4 ligands (P = 0.0067) and with HLA-B Bw4 Ile80 (P = 0.0027), and a descending trend for KIR3DL2 when coupled with HLA-A3 and HLA-A11 ligands (P = 0.0044). Overall, people from South Italy preferentially use the KIR3DL1-HLA-B Bw4 functional unit, while those from the North Italy equally use both the KIR3DL2-HLA-A3/A11 and the KIR3DL1-HLA-B Bw4 functional units to fight infections. Thus, only KIR3DL receptors, which exert the unique role of microbial sensors through the specific D0 domain, and their cognate HLA-A and HLA-B ligands are selectively pressured in Italy according to geographical north-to-south distribution.

Description of the new allele HLA-DQB1*04:03:02.

A new variant of HLA-DQB1*04:03 allele officially designated as HLA-DQB1*04:03:02 was detected in two unrelated Caucasoid individuals by polymerase chain reaction-sequence-specific primers and SBT. The new allele nucleotide sequence differs from HLA-DQB1*04:03:01 for a single silent point mutation in exon 2 at position 159, codon 21.

Bypass surgery for the treatment of upper limb chronic ischaemia.

OBJECTIVES: This study aims to evaluate the results and complications of surgical arterial revascularisation of the upper limb for treatment of chronic ischaemia using infrabrachial bypass. Results of limb salvage and follow-up with graft patency are analysed. DESIGN: This study is a retrospective analysis of 23 patients affected by chronic upper limb ischaemia and treated by surgical bypass. MATERIALS AND METHODS: We retrospectively analysed 23 patients with upper limb ischaemia treated between January 1998 and January 2008, by means of bypass graft revascularisation. After surgical revascularisation, eight patients (35%) with digital gangrene underwent minor amputations during the same surgical session, or within the following few days. Postoperatively, patients were followed up at regular intervals of 1, 3 and 6 months, and every 6 months thereafter, both clinically and with a duplex ultrasound scan. RESULTS: The mean 34 months' follow-up was 96% complete. Life table analysis revealed a primary patency of 82.6% and secondary patency of 91.3%. Limb salvage was 100%. During the follow-up period, four patients sustained graft occlusion and, of these, two underwent re-do revascularisation with success. CONCLUSIONS: We believe upper limb bypass surgery represents a valid treatment in this clinical setting, both for limb salvage and for relief of symptoms.

[The role of laparoscopic surgery in the treatment of occlusive and abdominal aortic aneurys. Review of the literature]. / Il ruolo della chirurgia laparoscopica nella patologia aneurismatica e occlusiva dell'aorta addominale. Revisione della letteratura.

UNLABELLED: The objective of this systematic review of the literature is to evaluate whether a laparoscopic operation can be performed on patients with occlusive or abdominal aortic aneurysm as a minimally invasive and durable alternative. For this purpose, the literature was reviewed and laparoscopic surgery results were compared with those of conventional and endovascular surgery. All series were included, even when containing also one case. Operative and clamping times, mortality and morbidity and hospital stay were evaluated. Thirty-five studies were identified about conventional (4), minilaparotomy (4), endovascular (4), total (12) and video-assisted (11) laparoscopic surgery. Operative and clamping times were shorter for video-assisted procedures than total-laparoscopic procedures. The mortality rate ranged from 3% to 4.5% for conventional surgery, from 0% to 3% for endovascular surgery, from 0% to 6% for total-laparoscopic surgery and from 0% to 4.2% for video-assisted laparoscopic surgery. A variable morbidity was described for all techniques, with a higher incidence in total-laparoscopic surgery. Mean hospital stay was similar for laparoscopic surgery procedures. The learning curve of a surgical team performing laparoscopic surgery improves the RESULTS: Laparoscopic abdominal aortic surgery is feasible and may offer good postoperative recovery with excellent mid-term patency. Shorter hospital stay and simple mid-term follow-up allow more comfort for the patient and probably monetary savings for the community. A steep learning curve is needed. For these reasons laparoscopic video-assisted technique can be considered a third means of treating severe occlusive and aortic aneurysm, but only new instruments for performing aortoprosthetic anastomoses can diffuse the total laparoscopic technique as a routine approach.

Endovascular repair of aortic lesions associated to video-assisted thoracoscopy surgery after chest trauma.

AIM: Trauma of the thoracic aorta for blunt trauma shows a very high incidence of mortality. Hospital mortality rate after aortic open surgery is between 15% and 30%. Endovascular management represents an alternative treatment Associated lesions are usually seen in those critical patients. Hemothorax may be present. The authors propose a combined treatment of endovascular repair for the aortic lesion and video-assisted thoracoscopy surgery (VATS) for the treatment of chest bleeding complications. METHODS: The authors report a series of three patients with post-traumatic aortic lesion and hemothorax. In two patients endovascular procedure was first performed, followed by VATS, few days later, for retained hemothorax. In the third patient the two procedures were performed at the same time because of the patient's critical conditions. RESULTS: There was technical success of stent-graft placement in all the treated cases. No postoperative mortality. No postoperative paraplegia. No VATS converted to thoracotomy. The postoperative follow-up time range between 10 and 19 months. CONCLUSION: Considering the relatively short procedural time and minimally invasive approach of both techniques, the concomitant use of them may represent an alternative to standard open surgery in cases of thoracic aorta lesions associated with hemothorax. Those procedures may be performed sequentially or together in emergency cases with intra-thoracic more active bleeding to exclude or to treat intra thoracic bleeding.

Clinical effectiveness of a low-tech versus high-tech pressure-redistributing mattress.

OBJECTIVE: To compare the effectiveness of a high-specification foam mattress (control) with a high-tech (Duo2, Hill Rom) alternating/continuous low-pressure mattress (treatment) in the prevention of pressure ulceration. The study also evaluated if there is a difference in performance between the two working modalities (alternating and continuous low pressure) of the high-tech mattress in a comparable sample of patients. METHOD: Thirty-three patients were observed for two weeks in the control group. In the treatment group, 86 patients were randomised to receive alternating low pressure and 84 continuous low pressure. Incidence of pressure ulcers in both arms was recorded. Student's t-test was used to compare all Braden scores, and the chi-square test and Fisher's exact test to evaluate differences between groups. RESULTS: There was a high difference in the number of new pressure ulcers in the control group when compared with the treatment group. There was no difference in performance between the alternating and continuous low-pressure modes. However, the sample size is too small to prove or disprove a statistically significant difference between the two modalities. CONCLUSION: The high-tech mattress was markedly more effective than the high-specification foam mattress in preventing the onset of pressure ulcers. Initial data suggest that the use of alternating or continuous low pressure made little or no difference to the results.

Relation between heart rate and QT interval in exercise-induced myocardial ischemia.

The relation between heart rate and QT interval during dynamic upright exercise on a bicycle ergometer was investigated in control subjects (n = 18) and in patients with coronary artery disease (CAD), stable angina on effort, and angiographically documented significant coronary stenoses (n = 23). Both groups had a significant negative linear relation between heart rate and QT, with a higher correlation coefficient in control subjects (r = -0.78) than in patients with CAD (r = -0.64). This response may be a result of the nonhomogeneous response to ischemia in patients with CAD, particularly with regard to the different impact of exercise-induced ischemia. When the 2 regression lines were compared, a flatter slope was found in the CAD group (p less than 0.001) as a consequence of a faster decrease in the QT-increasing rate in control subjects. It is suggested that in control subjects exercise-induced increase in adrenergic tone causes a rapid and relevant decrease in QT-interval duration. In the CAD group, exercise-induced ischemia relatively prolonged the QT interval; this may have been the result of an impairment of myocardium in response to catecholamines release during exercise or the consequence of a direct effect of exercise-induced ischemia prolonging the duration of myocardial tension.

Relation between QT and QS2 intervals during exercise and recovery. Response in patient with coronary artery disease and age-matched control subjects.

We investigated the relationship between QT interval and QS2 (electromechanical systole) during exercise and recovery in patients with coronary artery disease (CAD) and exercise-induced myocardial ischemia (n = 12), and in age-matched controls (n = 20). Upright bicycle exercise was performed (50 watts/min + 20 watts/min every 2 min), recording electrocardiographic lead 2 (100 mm/sec) for QT and QS2 measurement at rest, at each step of uninterrupted exercise and every 60 sec during a 3-min recovery period. Resting data showed a QT less than QS2 finding in both groups; during exercise, QT and QS2 decreased. The values of QT and QS2, collected at each step of exercise and plotted against heart rate (HR) separately for both groups, showed a significant correlation coefficient. Comparing the regression lines of HR-QT and HR-QS2 separately for both groups, we found that both intervals decreased in parallel and the mean QT remained shorter than QS2 in both groups during exercise. The QT/QS2 ratio remained unchanged significantly during exercise and recovery in CAD. In control subjects, the ratio remained unchanged during exercise and the first min of recovery, while a significant change was detected in late (2,3 min) recovery from an adrenergic-induced effect. The mean exercise-induced response of QT-QS2 relationship includes a QT less than QS2 pattern in both groups. In CAD patients, an abnormal pattern was found in two patients during recovery by a relative prolongation of QT, suggesting the possibility of a risk factor for dangerous arrhythmias or sudden death.

Immune reconstitution after bone marrow transplantation for combined immunodeficiencies: down-modulation of Bcl-2 and high expression of CD95/Fas account for increased susceptibility to spontaneous and activation-induced lymphocyte cell death.

We have studied the regeneration of T cell subsets and function after BMT in 21 children affected by combined immunodeficiency after BMT. In the first months, the striking predominance of CD4+ cells displayed the primed CD45R0+ phenotype and a high number of activated (HLA-DR+) T cells were observed. Regeneration of naive CD4+CD45RA+ cells correlated with the recovery of proliferative responses to mitogens (r = 0.64, P<0.001). Peripheral blood lymphocytes circulating after BMT undergo an increased process of in vitro cell death, resulting from two mechanisms: spontaneous apoptosis (SA), a consequence of defective production of IL-2 and down-regulation of Bcl-2 (P = 0.02 vs. healthy controls), and high susceptibility to activation-induced cell death (AICD) after restimulation with mitogens. In accordance with the role of CD95/Fas in this latter process, we have observed a high level of CD95 expression (P<0.001 vs. healthy controls), correlated with AICD (P<0.001) but not with SA, and decreasing with time after BMT (P<0.001). Both SA and AICD levels correlated with the presence of activated T cells and decreased with the progressive recovery of T cell proliferative response. Therefore, the lymphocyte hyperactivated status might explain their susceptibility to apoptosis and contribute to the genesis of immunodeficiency that follows BMT.

Cytotoxic T-lymphocyte responses in melanoma through in vitro stimulation with the Melan-A peptide analogue A27L: a qualitative analysis.

Modifications in tumour antigen-derived epitopes that stabilize the major histocompatibility complex (MHC)-peptide complex result in enhanced stimulatory capacity and improved immunogenicity of the altered peptide. These epitope analogues are attractive candidates for the development of peptide-based vaccine trials. Any modification, however, in tumour antigens may induce T-cell responses that could either fail to react against the naturally occurring peptides or represent only a subset of the total antigen-specific repertoire. In the present study, we performed a critical analysis of the ability of cytotoxic T-lymphocyte (CTL) clones, derived from two melanoma patients through stimulation with the A27L peptide analogue, to cross-react with the naturally processed Melan-A/MART-1 (Melan-A) peptides in terms of T-cell receptor (TCR) affinity, functional avidity and fine antigen specificity. We found that all the A27L-specific clones analysed possessed a very low avidity for the natural Melan-A peptides, and that their binding affinity for human leukocyte antigen (HLA) tetramers complexed with both the modified and the natural Melan-A peptides did not strictly correlate with their functional avidity. We also observed that these clones were able to cross-recognize both natural Melan-A peptides in one patient, but only one peptide in the second patient. We discuss the capability of the A27L peptide analogue to stimulate all the available Melan-A-specific repertoire.

CD4+ and CD8+ subsets: naive and memory cells in the peripheral blood of patients with systemic sclerosis.

Systemic Sclerosis (SSc; scleroderma) is associated with several immunological abnormalities, including altered proportion between lymphocyte subsets. Peripheral blood lymphocyte subsets from 25 patients with SSc were studied by two-colour flow cytometry using monoclonal antibodies against CD45RA and CD29 markers, which allow a dissection of CD4+ and CD8+ populations into 'naive' and 'memory' subsets. A decrease of the percentage of CD8+ (p < 0.05) and of CD8+CD29+ (p < 0.001) cells was observed compared to that in 20 age and sex-matched controls. These abnormalities were not significantly associated with the extension of cutaneous disease or other clinical features of SSc nor with treatment, pattern of autoantibodies or HLA phenotype.

Systolic and diastolic time intervals during spontaneous angina.

In 5 patients who suffered spontaneous angina during cardiac catheterization, aortic pressure and electrocardiographic lead (V5) were recorded at rest, at the onset of anginal pain, 5 and 10 min after 0.6 mg sublingual nitroglycerin (NTG). Heart rate, systemic arterial pressure, systolic, and diastolic time intervals were measured. Heart rate and systemic arterial pressure rose significantly immediately after the onset of angina and declined progressively within 10 min from NTG administration. Preejection period did not change during angina, while left ventricular ejection time and electromechanical systole lengthened. As a consequence, diastolic time, expressed as percent of cardiac cycle, shortened sharply. All parameters considered went back to basal values within 10 min from NTG administration, and were preceded by relief or reduction of anginal pain. We concluded that a fall in diastolic time, secondary to a prolongation of electromechanical systole occurring during angina, may further increase the degree of ischemia resulting in a vicious cycle than can be interrupted by NTG administration.

Exercise-induced changes of left ventricular ejection time in noninvasive evaluation of chest pain.

Significance and clinical usefulness of exercise-induced changes of noninvasive left ventricular ejection time, recorded by mechanocardiography, in detecting coronary artery disease is still controversial. We investigated the changes of the left ventricular ejection time (LVET), corrected for heart rate as ETI (LVET/square root RR), after a standard 4-min exercise by bicycle ergometer (50 W/min for 2 min, increased by 20 W after 2 min) in 56 male volunteers. They were invasively studied for typical or atypical chest pain. Thirty-four had coronary artery disease (CAD) and the others served as controls. Immediately after exercise LVET shortened according to the rise in heart rate in both groups. ETI increased similarly. After 2 minutes from exercise ETI increased only in the CAD group, according to a longer LVET, in spite of the rise in heart rate. In contrast, ETI was unchanged in controls, according to a shortened LVET. Totally, 27/34 CAD patients and 9/22 controls had a greater ETI than at rest. Employing this delta ETI as a "marker" of CAD we found a predictive accuracy of 74%. We suggest postexercise ETI could be a simple and inexpensive support to electrocardiogram in basic evaluation of subjects with chest pain.

Exercise time: a possible source of misleading results during long-term pharmacological studies by multiple stress testings in coronary artery disease.

The assessment of chronic pharmacological treatment of stable angina requires serial exercise stress testings. It is well known that exercise tolerance can be improved by the training effect of performing repeated testings. Our study investigated the values of heart rate, systolic blood pressure, rate-pressure product, and duration of exercise at 0.1 mV ST depression during exercise and the same parameters plus the maximal ST-segment depression at peak exercise, collected from three different tests. The first and second were performed at one-week intervals before, and the third (75 days after the first), was performed after a double-blind study with a drug versus placebo. We found a significant increase of exercise duration at 0.1 mV ST depression and at peak exercise, while 6 of 12 patients increased exercise duration from the second to the third test. Individual variability of exercise duration showed increasing values, ranging from 0 to 71% (first vs. third test). In contrast, the ratio of heart rate and systolic blood pressure did not differ between the tests. Our data criticized the use of mean values of exercise time for pharmacological studies; moreover, individual variability could affect results independently of drug or placebo administration. These findings should be taken into account in order to exclude misleading results.

Introduction to apoptosis in ophthalmology.

Apoptosis represents a mode of cellular death genetically programmed to maintain homeostasis of tissues. In specific pathologic circumstances, the death program may be activated by various environmental factors such as exposure to toxic substances or bacteria or deprivation of nutrients. From this point of view, apoptosis is considered the final event in several pathologies. In ophthalmology, experimental evidence has confirmed that apoptosis is a type of cellular death involved in various pathologic processes including glaucoma, retinitis pigmentosa, ischemic retinopathy, corneal reparative processes, cataract, and retinoblastoma. The aim of this article is to review the most recent results published in this field and to describe some of the molecular mechanisms responsible for the activation of the apoptotic program in some important ocular disorders. The understanding of such mechanisms could outline new therapeutic strategies for the prevention of cellular death in ophthalmology.

Prevention of corneal keratocyte apoptosis after argon fluoride excimer laser irradiation with the free radical scavenger ubiquinone Q10.

PURPOSE: To assess in vitro the potential of the free radical scavenger ubiquinone Q10 in preventing keratocyte apoptosis after argon fluoride (ArF) excimer laser irradiation. METHODS: Cultured rabbit keratocytes were irradiated at very low single-pulse laser fluences. The cumulative effects generated by three total fluence doses between 12 and 45 mJ/cm2, representative of single-pulse subablative doses during photorefractive keratectomy (PRK) in humans, were evaluated. We employed the following parameters to compare pretreated (10 microM ubiquinone Q10) and untreated samples: 1) number and morphology of living cells by Trypan blue test and ultramicroscopy, respectively; 2) level of free-radical formation assessed by malonaldehyde quantitation; 3) cellular energy level evaluated by ATP assay. RESULTS: Excimer laser irradiation kills cultured keratocytes by inducing apoptosis. The effect increases with the cumulative fluence dose. In the samples pretreated with ubiquinone Q10 there were significantly fewer cumulative apoptotic events than in the untreated ones. Quantitative analysis of malonaldehyde cellular levels suggested this protective action of ubiquinone Q10 was connected with its ability to scavenge laser-generated free radicals. ATP assay also confirmed that it raised cellular energy levels. CONCLUSIONS: The treatment of corneal keratocytes with relatively low concentrations of ubiquinone Q10 can prevent apoptosis after ArF excimer laser irradiation. If these findings are confirmed on human keratocytes this treatment could be usefully exploited in the PRK surgical procedure. That might lead to a reduction in the occurrence of haze and curvature regression triggered by programmed cell death.

Abnormal recovery systolic blood pressure response for detecting coronary artery disease in men and women investigated by upright bicycle exercise.

We investigated the clinical significance of recovery systolic blood pressure (SBP) ratio, obtained dividing the recovery SBP at 1st (R1/A) or 3rd min (R3/A) by the peak exercise SBP (before stopping), during upright bicycle exercise in 530 subjects (ranging from 17 to 73 years). Our results may be summarized as follows: 1) we found a higher value of R1/A in control subjects with exercise induced ST depression; 2) the normal range in women was higher than in men; 3) the use of recovery SBP ratios gives a lower sensitivity and a higher specificity than ST segment analysis in detection of CAD; 4) this pattern may be useful particularly in patients with previous myocardial infarction and not detectable ST segment analysis during exercise.

[Polyarteritis nodosa and pulmonary neoplasms. Report of a clinical case]. / Poliarterite nodosa e neoplasia polmonare. Descrizione di un caso clinico.

A case of rare association between PAN and cancer is presented. The tumour was a bronchogenic carcinoma (large cell type) diagnosed about 14 years after the beginning of PAN, that in this case had a very long course increasingly affecting the principal arteries of the legs.

[Arrhythmias in the elderly]. / Le aritmie nell'anziano.

A survey was made on a population of 6059 subjects aged more than 60 years with the aim to assess 1. the prevalence of heart arrhythmias and 2. the relationships between arrhythmias and some other ecg alterations. Arrhythmias resulted present in 29.0% of the whole population with a significantly higher prevalence among males (30.7% vs 28.1%, P less than 0.05) and among subjects over 75 years of age (33.2% vs 23.9%, P less than 0.001). Supraventricular extrasystoles (SE, 11.55%), atrial fibrillation (AF, 10.44%) and ventricular extrasystoles (VE, 8.91%) were the most frequent arrhythmias, followed by sinus bradycardia (SB, 2.04%), sinus arrhythmia (SA, 1.35%), atrial flutter (AFL, 1.09%) and junctional rhythms (JR, 0.20%). AF and AFL resulted significantly more frequent among females, whilst SE, VE and SB were more frequent among males. All the above arrhythmias, with the exception of AFL and JR resulted significantly more frequent among subjects over 75. A significantly higher prevalence of ecg signs of left ventricular hypertrophy, ischemia, previous myocardial infarction (MI) and of the so-called "minor" T-wave changes (MTC) was found among the subjects with arrhythmia as compared with those free from rhythm disturbances. Ecg signs of MI and MTC were significantly more frequent among males and MTC were more frequent among females and among subjects over 75. It is concluded that in an old person the presence of an arrhythmia should lead to a careful evaluation of the general and cardiological clinical situation in order to avoid 1. to prescribe an unnecessary and potentially dangerous antiarrhythmic treatment, and 2. to misdiagnose an underlying clinical condition liable to a decisive improvement under adequate treatment.