Pantetheinase activity of membrane-bound Vanin-1: lack of free cysteamine in tissues of Vanin-1 deficient mice.

Pantetheinase (EC 3.5.1.-) is an ubiquitous enzyme which in vitro has been shown to recycle pantothenic acid (vitamin B5) and to produce cysteamine, a potent anti-oxidant. We show that the Vanin-1 gene encodes pantetheinase widely expressed in mouse tissues: (1) a pantetheinase activity is specifically expressed by Vanin-1 transfectants and is immunodepleted by specific antibodies; (2) Vanin-1 is a GPI-anchored pantetheinase, and consequently an ectoenzyme; (3) Vanin-1 null mice are deficient in membrane-bound pantetheinase activity in kidney and liver; (4) in these organs, a major metabolic consequence is the absence of detectable free cysteamine; this demonstrates that membrane-bound pantetheinase is the main source of cysteamine in tissues under physiological conditions. Since the Vanin-1 molecule was previously shown to be involved in the control of thymus reconstitution following sublethal irradiation in vivo, this raises the possibility that Vanin/pantetheinase might be involved in the regulation of some immune functions maybe in the context of the response to oxidative stress.

Control of breathing and respiratory muscle strength in patients with multiple sclerosis.

In 11 patients with moderately severe multiple sclerosis, lasting 11.2 +/- 7.3 years, in stable condition, and in 10 age- and sex-matched control subjects, we investigated lung function, respiratory muscle strength, and ventilatory control system. Respiratory muscle strength was assessed by measuring maximal inspiratory and expiratory mouth pressures (Pimax and Pemax, respectively). Respiratory central drive was evaluated in terms of neuromuscular (P0.1) and ventilatory (Ve) output, breathing room air and during CO2 rebreathing. In the absence of any significant impairment of lung function, patients showed a reduction of Pimax and Pemax amounting to about 40 percent and 60 percent of the predicted value at functional residual capacity (FRC), respectively; a significant, inverse correlation was found between both Pimax and Pemax at FRC and the severity score of the disease. While at rest Ve was similar to that of control subjects, baseline P0.1 was significantly higher in patients (1.97 +/- 0.79 vs 0.97 +/- 0.20 cm H2O, p < 0.005). Compared with the control group, during CO2 rebreathing P0.1/PetCO2 slope, although less steep, was not dissimilar in patients (0.34 +/- 0.13 vs 0.46 +/- 0.19 cm H2O/mm Hg, NS); on the other hand, Ve/PetCO2 slope was much lower in the patient group (1.93 +/- 0.91 vs 3.27 +/- 1.11 L/min/mm Hg, p < 0.01) and was significantly related to the functional stage of disease and to Pimax and Pemax values at FRC. These results indicate that in patients with clinically stable, moderately severe multiple sclerosis, the respiratory muscle function is abnormal. Moreover, the inspiratory drive at rest is increased and the drive response to CO2 appears normal, while the ventilatory response to CO2 is significantly impaired. Respiratory muscle weakness (and/or lack of coordination) could explain, at least in part, the lower ventilatory response in these patients, whereas the mechanism of increased rate of the initial inspiratory force generation remains unclear.

Interferon-gamma (r-IFN-gamma) induced activation of alveolar macrophages (AM) from anergic patients with chronic obstructive pulmonary disease (COPD).

Forty-six anergic patients (37 males and 9 females, age range 55-79 yr) were selected from ninety-one patients suffering from COPD due to frequent exacerbations and impaired delayed cutaneous reactivity (43.9%). The phenotype of circulating lymphocytes, their proliferative response to a panel of polyclonal T-cell activators and the candidacidal activity (CA) of circulating PMNs (polymorphonuclear cells) were measured. In 13 patients presenting a defective CA of circulating PMNs, the in vitro response of alveolar macrophage CA to r-IFN-gamma was also determined. We found: 1) a significant reduction in the CL response to PHA in COPD patients vs controls; 2) a low PMN-CA in 23 (57%) COPD patients; 3) a non-significant difference in phenotype analysis in patients and controls; 4) lower CA of AMs in COPD patients than in controls; 5) restoration in vitro of CA by r-IFN-gamma in the group of anergic COPD patients presenting depressed CA. We conclude that a defective cell-mediated immunity could be the basis of the enhanced susceptibility to infectious exacerbations in many COPD patients and that, in vitro, it could be reversed by r-IFN-gamma treatment.

Late results of reconstructive surgery for renovascular hypertension.

In order to evaluate the late results of reconstructive surgery for renovascular hypertension, a review was made on a series of 120 consecutive patients who underwent operations over a 11 year period. There were 82 males (68.3%) and 38 females (31.7%) with a mean age of 48.4 years. Renal artery by-pass grafts were used in 90% (120/133), a thromboendarterectomy in 5.2% (7/133), and other surgical procedures were performed in 4.8% (6/133). Associated vascular procedures were performed in 38.3% (46/120) of patients. Operative mortality was 2.5% (3/120) overall; there was no mortality in the isolated renal artery reconstructions. There was a clinical success (after a mean follow-up of 48 months) in 80.4% of patients. The most important factors influencing clinical result after renal revascularization were: a generalized atherosclerosis (p less than 0.05), duration of hypertension (p less than 0.01) and the early post-operative response of the blood pressure (p less than 0.01). The overall five- and ten-year actuarial survival probabilities were 85 and 68%, respectively. The most common causes of death were myocardial infarction, stroke and cancer. Cox regression analysis for variables influencing survival indicated that persistence of severe hypertension was the major determinant of late survival (p less than 0.05). Hypertension in females is better tolerated, while younger patients appear to have better results and late survival after surgical treatment.

[Treatment of prostheto-digestive fistulas using in situ prosthetic bypass]. / Traitement des fistules prothéto-digestives par pontage prothétique in situ.

Secondary aorto-enteric fistula is one of the most serious complications of abdominal aortic reconstruction. Conventional management includes removal of all infected prosthetic graft, oversewing of aortic stump and restoration of lower limbs blood flow by extraanatomic bypass grafting, reporting high rates of mortality, limb loss, and even infection of the extraanatomic grafts. Dissatisfied by these results, frequently, due to aortic stump blowout or extraanatomic by-pass reinfection, some authors attempted a more conservative approach with au in situ replacement by a new synthetic graft. The aim of this paper was to verify the role of in situ graft replacement. From December 1989, 8 patients with secondary aorto-enteric fistula underwent in situ PTFE graft replacement. One patient (12.5%) died perioperatively for acute myocardial infarction. No limb loss occurred. One patient died after 44 months from pulmonary neoplasia without signs of graft infection. The others are doing well at 34 months follow-up. The authors suggest that, in selected patients, in situ prosthetic graft replacement provides better early and late results than extranatomic bypass.

[Renal artery stenosis and aorto-iliac restoration: is it necessary to do extensive evaluation?]. / Sténoses des artères rénales et restauration aorto-iliaque: faut-il faire un bilan extensif?

In patients with aortoiliac disease and a stenosis of one or two renal artery, renal hypertension is rarely the sole mechanism of the elevated blood pressure. The preservation of nephron mass being the aim of the renal revascularization, we firmly believe that only three tests are required for the operative decision: renal ultrasonography, nephroscintigraphy and global and selective aortography.

[Diagnostic methods and therapeutic options in vasculogenic impotence]. / Metodologia diagnostica e possibilità terapeutiche nell'impotenza vasculogenica.

Until a few years ago, the main cause of men's genital impotence was reputed to be psychogenic. On the other hand, the development of diagnostic techniques has been in aid in isolating the most frequent cause of impotence among the organic forms, and in particular those of vascular origin. The authors herein examine the diagnostic methods of vasculogenic impotence and evaluate the therapeutic options in relation to the various causes identified.

Cardiorespiratory comorbidity: a new challenge for physical and rehabilitation medicine specialist.

BACKGROUND: The cardiorespiratory comorbidity can reduce the participation in the rehabilitation project of patients with motor disorders. AIM: The first aim of the study was to assess the frequency of cardiopulmonary comorbidity in inpatient rehabilitation units in Italy. The second aim was to evaluate the influence of cardiorespiratory comorbidity on some process and outcome indicators. DESIGN: Data collection with a questionnaire sent to Physical and Rehabilitation Medicine specialists. SETTING: Inpatient Rehabilitation Units. POPULATION: Patients admitted to Rehabilitation Units with neurological and orthopedic disability. METHODS: A questionnaire was sent to 33 Rehabilitation Units in Italy. OUTCOME: Length of stay in hospital (LOS), percentage of transfer to acute wards, level of satisfaction of the Physicians related to the degree of functional recovery of patients with cardiopulmonary comorbidity. In a subgroup of patients with and without cardiopulmonary comorbidity indicators were also used to retrospectively assess functional recovery during the hospitalization. RESULTS: Analysis were made on 16 complete responses received. Data of 909 subjects were evaluated. The mean incidence of cardiovascular and respiratory diseases is high: 61.50%. The length of hospitalization (LOS) in patients with cardiac and respiratory comorbidity is significantly longer than in subjects without associated disease (46.55, SD 21.00 days vs. 37.26, SD 18.97; P<0.05). The number of transfers to acute wards is significantly higher in subjects with cardiorespiratory comorbidity (8.62% vs. 2.44%; P<0.05). Eleven out of 16 medical doctors (69%) said they were "quite satisfied" for the degree of functional recovery of patients with comorbidity. A group of patients with neurological disabilities associated with cardiorespiratory disease had significantly lower Functional Independence Measure (FIM) score at admission (55.36, SD 20.62, vs. 73.72, SD 22.15; P<0.05) than these without comorbidity. At discharge the subjects of the two groups assessed, independently from the presence of cardiorespiratory comorbidity, present no statistically significant difference of FIM scale values. CONCLUSION: The high frequency of cardiorespiratory comorbidity in patients with motor disorders negatively influenced the LOS and percentage of transfer to acute wards but not necessarily the functional results. It is, therefore, necessary to train the medical doctor who specializes in physical and rehabilitation medicine also in the clinical management of complex patients. CLINICAL REHABILITATION IMPACT: The results of the survey suggest that rehabilitation is useful even in patients with motor disorders and cardiorespiratory comordibity.

Is health-related-quality of life of stroke patients influenced by neurological impairments at one year after stroke?

AIM: The aim of this study was to evaluate which neurological impairments on discharge from rehabilitation services are associated with a worse outcome in terms of health related-quality of life (HRQoL) one year after stroke. METHODS: We enrolled 528 first-time stroke patients in a multicenter observational study involving 18 Italian inpatients rehabilitation centers. Neurological impairment-related factors, socio-demographic and general clinical variables and process indicators were considered independent variables at discharge. Outcome was represented by the following dependent variables of HRQoL: EuroQoL-5D questionnaire (5D-EQoL) and its derived index (Eq-Index) and a Visual Analogue Score (QoL-VAS). RESULTS: The strongest predictors of lower HRQoL were an incomplete limbs motor recovery. An incomplete lower limb motor recovery was associated with a lower HRQoL only in the "mobility" domains, whereas a incomplete upper limb motor recovery could predict a lower HRQoL in all domains except "mobility". Regarding cognitive impairments, the presence of neglect at discharge was associated with a lower HRQoL in "self-care", "pain-discomfort" and "anxiety-depression". Female gender, presence of urinary catheter seemed to exert a much lesser role in predicting HRQoL. CONCLUSION: The results of this study suggest that upper limb motor impairments may have a detrimental impact on the patient's perceived HRQoL one year after stroke.

Developing a minimum data set for stroke patients assessment: the " Protocollo di Minima per l'Ictus (PMIC) as a starting point towards an Italian stroke registry.

AIM: The complex nature of stroke sequelae requires several assessment instruments to quantify correctly every residual symptom. As there was no general consensus on stroke evaluation among Italian Physiatrists, in 2004 the Italian Society of Physical Medicine and Rehabilitation and S. Lucia Foundation (a Scientific Institute for hospitalization and treatment) established a Project Group to propose a standardized assessment tool (''Protocollo di Minima per l'Ictus PMIC'') for acute, post-acute and community-living stroke patients. This tool aimed to be easy to use and comprehensive of all the elements necessary for accurately address the great range of different rehabilitation needs. The objective was to provide physiatrists with a standard assessment battery and to make prognostic factors available on large community samples. METHODS: From end 2004 to early 2006, the Project Group examined literature data on stroke assessment, prognostic factors and outcome and selected the specific data elements to be included in a data collection tool. RESULTS: A consensus was reached on a ''minimum'' core set of data. This protocol was peer submitted in early 2006, to test the burden of data collection, and to allow modifications and adjustments. Specific forms (file to download) for data collection and database to be shared (a dedicated ''Client'' software) are now freely offered by the Project Group for data collection. CONCLUSION: PMIC is an evaluation procedure manageable in every-day practice and in every setting, a quick screening instrument that, given its large diffusion, can be expanded from a National Database into a National Rehabilitation Stroke Registry.

Ovotransferrin expression and release by chicken cell lines infected with Marek's disease virus.

Mammals posses both serum transferrin and lactoferrin, whose functions are taken over in birds by ovotransferrin, displaying both iron transport and antibacterial activities. Ovotransferrin also exerts antiviral activity towards Marek's disease virus, an avian member of the herpes family of viruses. This virus infects lymphoid organs and induces the transcription of ovotransferrin in infected chicken embryo fibroblasts. However, it has not yet been established whether ovotransferrin gene transcription is linked to the release of the protein outside the cells or whether ovotransferrin expression and release also occurs in chicken lymphoblastoid cells in which the Marek's disease viral genome is integrated. Our results indicate that both serum and egg-white isoforms of ovotransferrin are expressed and released in the supernatants of chicken embryo fibroblast and lymphoblastoid cells in the absence of infection. Viral infection of chicken embryo fibroblasts caused a slight increase of ovotransferrin release, whereas viral reinfection of lymphoblastoid cells caused a remarkable ovotransferrin release in a virus concentration-dependent manner. These findings suggest that ovotransferrin release in vivo may play a crucial role in protecting the whole organism from viral infection spreading, and support the hypothesis that the antiviral activity of ovotransferrin is an important part of the innate immune response in birds, resembling the antiviral activity of lactoferrin in mammals.

Antiviral activity of ovotransferrin derived peptides.

Ovotransferrin and lactoferrin are iron-binding proteins with antiviral and antibacterial activities related to natural immunity, showing marked sequence and structural homologies. The antiviral activity of two hen ovotransferrin fragments DQKDEYELL (hOtrf(219-227)) and KDLLFK (hOtrf(269-301) and hOtrf(633-638)) towards Marek's disease virus infection of chicken embryo fibroblasts is reported here. These fragments have sequence homology with two bovine lactoferrin fragments with antiviral activity towards herpes simplex virus, suggesting that these fragments could have a role for the exploitation of the antiviral activity of the intact proteins towards herpes viruses. NMR analysis showed that these peptides, chemically synthetized, did not possess any favourite conformation in solution, indicating that both the aminoacid sequence and the conformation they display in the intact protein are essential for the antiviral activity.

Degradation of the epstein-barr virus latent membrane protein 1 (LMP1) by the ubiquitin-proteasome pathway. Targeting via ubiquitination of the N-terminal residue.

The latent membrane protein 1 (LMP1) of the Epstein-Barr virus is a constitutively active receptor essential for B lymphocyte transformation by the Epstein-Barr virus. It is a short-lived protein, but the proteolytic pathway involved in its degradation is not known. The ubiquitin pathway is a major system for specific protein degradation in eukaryotes. Most plasma membrane substrates of the pathway are internalized upon ubiquitination and delivered for degradation in the lysosome/vacuole. Here we show that LMP1 is a substrate of the ubiquitin pathway and is ubiquitinated both in vitro and in vivo. However, in contrast to other plasma membrane substrates of the ubiquitin system, it is degraded mostly by the proteasome and not by lysosomes. Degradation is independent of the single Lys residue of the protein; a lysine-less mutant LMP1 is degraded in a ubiquitin- and proteasome-dependent manner similar to the wild type protein. Degradation of both wild type and lysine-less protein is sensitive to fusion of a Myc tag to the N terminus of LMP1. In addition, deletion of as few as 12 N-terminal amino acid residues stabilizes the protein. These findings suggest that the first event in LMP1 degradation is attachment of ubiquitin to the N-terminal residue of the protein. We present evidence suggesting that phosphorylation is also required for degradation of LMP1.

General anaesthesia versus cervical block and perioperative complications in carotid artery surgery.

PURPOSE: To compare the influence of anaesthetic technique on perioperative complications in patients undergoing carotid endarterectomy. MATERIAL AND METHODS: In a retrospective study of 1020 consecutive patients who underwent carotid artery surgery over 10 years, perioperative neurologic and cardiologic complications and the use of an internal carotid artery shunt were compared in 337 patients (33%) treated under general anaesthesia and 683 (67%) under cervical block. The two groups had similar characteristics. The most frequent surgical indication was symptomatic carotid artery disease (91.5%). The remaining patients had asymptomatic severe internal carotid lesions (> 70%). RESULTS: The overall perioperative stroke rate was 1.9%, the death-stroke rate 0.7% and the cardiac complication rate 0.8%. The perioperative stroke rate was higher in the general anaesthesia group than in the cervical block group (3.2% vs 1.3%, p = 0.01). Cardiac complication rates were similar in the two groups. A carotid artery shunt was used in 75 patients (22%) receiving general anaesthesia and in 92 patients (13%) receiving cervical block (p = 0.0004). The causes of stroke in the cervical block group were intraoperative embolism (4 cases, 26%), perioperative thromboembolism (7 cases, 58%) and clamping ischaemia (1 case, 16%). Mechanisms causing stroke in the general anaesthesia group remained unidentified or uncertain. CONCLUSIONS: Cervical block anaesthesia yields better perioperative results than general anaesthesia probably because it allows more reliable cerebral monitoring, reducing or even eliminating perioperative strokes related to clamping ischaemia. It facilitates detection of the mechanism underlying intraoperative stroke allowing surgical techniques and intraoperative management to be modified accordingly. Cervical block anaesthesia significantly reduces the need for internal carotid artery shunting.

Energy cost of exercise in multiple sclerosis patients with low degree of disability.

In 10 patients (five females) suffering from multiple sclerosis with mild degree of disability, (EDSS ranging from 0 to 2) and in 10 age and sex matched control subjects we investigated lung function, respiratory muscles strength and cardiorespiratory response to incremental exercise in order to assess the metabolic cost of exercise. In the absence of any impairment of lung volumes and flows and in- and expiratory maximal mouth pressures, at peak of exercise oxygen consumption (VO2max = 1886 +/- 145 ml/min) and workload (Wmax = 137 +/- 9.8 watts) were slightly diminished in patients, as compared with controls (VO2max = 2246 +/- 196 ml/min and Wmax = 164 +/- 14.7 watts). These findings were associated with an increased heart rate (HR) and reduced oxygen pulse (VO2/HR) at the same workloads. During the whole exercise, however, the slope of the linear relationship between VO2 and work exhibited by the patients, amounting to 9.9 +/- 0.6 ml/min/watt, was similar to that of the controls (10.9 +/- 0.42 ml/min/watt). Incidentally, both at rest and during exercise, the patients showed a significantly greater minute ventilation (VE) due to a faster respiratory rate, associated with an augmented dead space (P < 0.05). We conclude that an increase of metabolic cost of exercise does not occur in multiple sclerosis patients with mild disability, suggesting a lack or a low degree of spasticity and/or ataxia elicited by the effort. Thus, their exertional capacity appears to be limited mainly by a poor training. The tachypnea observed in these patients at rest and during exercise was unexpected and the reason for adopting such a pattern of breathing is unclear.

Walking with the advanced reciprocating gait orthosis (ARGO) in thoracic paraplegic patients: energy expenditure and cardiorespiratory performance.

The aim of this study was to quantify the energy cost and the cardiorespiratory response while walking at free speed with an Advanced Reciprocating Gait Orthosis (ARGO). The study was conducted on a group of six male paraplegic patients, age range 16-31 years (median = 22.50), with complete traumatic injuries ranging from T3 to T12 and a median time lapse from the trauma of 21 months. The data was recorded 6-8 weeks after the patients were fitted and trained to use the orthosis. Maintaining a standing position produced a significant increase of both the heart rate (HR) and the respiratory rate (RR), whilst the increase of energy consumption was not significant. The data for median speed, energy consumption and energy cost observed during free walking was similar to that of the Reciprocating Gait Orthosis (RGO) in thoracic level paraplegic individuals. This study indicates that in the utilisation of the ARGO the workload is not excessive when it is limited to maintaining a standing position. In contrast, walking results in early anaerobic conditions and unsustainable fatigue after short distances. The great energy cost recorded is considered to be a main reason for the frequent abandonment or the low utilisation of the orthosis at follow-up.

Extraperitoneal approach for repair of inflammatory abdominal aortic aneurysm.

Inflammatory abdominal aortic aneurysms are characterized by dense perianeurysmal fibrosis involving the adjacent organs. Attempts to isolate the aneurysm can lead to operative injuries of these structures, thus increasing the rates of complications and mortality. In the last 12 years 45 patients with inflammatory abdominal aortic aneurysms underwent aneurysm resection at the Department of Vascular Surgery of the University of Rome. The aneurysm was resected through a standard, midline transperitoneal approach in 39 patients, through a thoracophrenolaparotomy in two patients, and through a left-flank extraperitoneal approach in the last four patients. The extraperitoneal approach simplified aneurysm dissection and aortic clamping with no cases of postoperative morbidity or death. In addition, we reviewed the CT scan findings of 12 patients surgically treated for inflammatory abdominal aortic aneurysm. The amount of fibrosis in the anterior wall of the aneurysm was greater than in the left posterolateral aspect (p = 0.008). We conclude that the left-flank extraperitoneal approach is the most anatomically advantageous route for repair of inflammatory abdominal aortic aneurysm.

Bacterial and clinical criteria relating to the outcome of patients undergoing in situ replacement of infected abdominal aortic grafts.

OBJECTIVES: In a retrospective non-randomised study we assessed the outcome after in situ replacement of infected knitted Dacron abdominal aortic grafts in patients without septicaemia or retroperitoneal abscesses. We also assessed whether the specific bacterial infection influenced outcome. MATERIALS AND METHODS: Over the 5 years studied, 18 patients (9 with perigraft infection and 9 with aortoenteric erosion) underwent in situ replacement of aortofemoral grafts. All patients were haemodynamically stable, none required emergency treatment. Preoperative assessment included CT, MRI, leukocyte-labelled scintigraphy, and bacterial cultures whenever possible. Infected grafts were totally excised and replaced in situ with standard PTFE prostheses. Bacterial diagnosis included intraoperative Gram-staining and postoperative graft cultures. None of the patients had retroperitoneal collections or proximal anastomotic dehiscence. All patients had 6 week intravenous antibiotic therapy. RESULTS: One patient died of myocardial infarction, and another of haemorrhagic shock from proximal anastomotic dehiscence, accounting for a graft-related mortality of 6%. Dehiscence resulted from a polymicrobial infection. Mean 37 month surveillance showed no amputations and no graft-related infections. CONCLUSIONS: In clinically and bacteriologically selected patients, total in situ replacement of infected abdominal aortic grafts offers an excellent outcome.