Conjunctivitis as a sign of persistent SARS-COV-2 infection? An observational study and report of late symptoms.

PURPOSE: To investigate if symptomatic conjunctivitis during the recovery phase of the disease could be associated to a persistent presence of SARS-CoV-2 in the upper respiratory tract. Secondary end points were to analyze the presence of SARS-CoV-2 in the conjunctiva of ocular symptomatic patients and to record the presence of ocular disturbances at this point of the disease. METHODS: An observational study including consecutive COVID19 patients treated at Humanitas Clinical and Research Hospital who were attending for nasopharyngeal swab to confirm the resolution of SARS-CoV-2 infection and end of isolation. We examined 129 consecutive patients from May to June 2020. The primary end point was to determine if symptomatic conjunctivitis at this point of the disease could be associated to a persistent presence of SARS-CoV-2 in the upper respiratory tract. Secondary end points were to analyze the presence of SARS-CoV-2 in the conjunctiva of ocular symptomatic patients and to record the presence of ocular disturbances at this point of the disease. RESULTS: One hundred twenty eight patients were included, 9.38% had conjunctivitis, none resulted positive to conjunctival PCR swab test, while two of them had positive nasopharyngeal result. Mean time elapsed since the first COVID-19 positive swab to the time of examination was 6 weeks ( ± 3). The only significant association was the presence of conjunctivitis with older age (65.3 ± 12.7 vs 56.7 + 13.5. p = 0.046). Nasopharyngeal swab resulted positive in 22 patients (17.19%). While 88 patients (68.2%) did not have any ocular complain during their COVID19 disease. The 40 patients (31.8%) reporting ocular disturbances complained about: redness (25.43%), tearing (19.53%), burning (18.35%), foreign body sensation (17.18%), itching (15.62%), and discharge (12.5%). CONCLUSION: This study showed that late conjunctivitis cannot be considered as a marker of persistent infection when patients are sent to confirm the resolution of SARS-CoV-2 infection.

Reducing rates of Clostridium difficile infection by switching to a stand-alone NAAT with clear sampling criteria.

Background: Clostridium difficile infection is an important cause of morbidity and mortality but the optimal method of diagnosis for both patient management and infection prevention remains controversial. Methods: Our hospital made a decision to switch from the use of toxin immunoassay to a stand-alone nucleic acid test. This change was accompanied by the provision of clear sampling guidance and rejection criteria and this study aimed to assess the impact of that change. We analysed sample numbers, numbers of positive results, and the proportion of cases assessed as healthcare acquired over a 6-year period during which the testing method was changed from a toxin A/B immunoassay to a stand-alone commercial nucleic acid test after the first two years. Results: Sample numbers and numbers of cases assessed as healthcare acquired fell following the introduction of the nucleic acid test and sampling guidance, while infection rates in other hospitals in the same region remained relatively stable. Conclusions: It is our opinion that the use of a highly sensitive assay together with clear sampling guidance offers the optimal approach to patient management and best use of isolation facilities.

Infections after T-replete haploidentical transplantation and high-dose cyclophosphamide as graft-versus-host disease prophylaxis.

BACKGROUND: Recently, a platform of T-cell replete haploidentical hematopoietic stem cell transplantation (haplo-HSCT) using post-transplant cyclophosphamide (Cy) has shown high reproducibility and acceptable safety profile. METHOD: This prospective cohort analysis allowed us to collect data on infections among 70 consecutive recipients of haplo-HSCT affected by various hematologic malignancies. RESULTS: After a median follow-up of 23 months, cumulative incidence of viral infections was 70% (95% confidence interval [CI] 59-81) at 100 days and 77% (95% CI 67-87) at 1 year; 35 of 65 patients at risk had CMV reactivation (54%) and the rate of polyomavirus-virus-associated cystitis was 19% (13/70). Cumulative incidence of bacterial and fungal infections at 1 year were 63% (95% CI 51-75) and 12% (95% CI 4-19), respectively. Of note, only 1 invasive fungal infection occurred beyond 1 year after transplant (day +739). CONCLUSION: In conclusion, despite a high rate of viral infections in the early period, present data suggest a satisfactory infectious profile after T-cell replete haplo-HSCT using post-transplant Cy. These results may help clinicians to improve both prophylactic and therapeutic antimicrobial strategies in this emerging haploidentical setting.

Italian validation of the Wagnild and Young Resilience Scale: a perspective to rheumatic diseases.

OBJECTIVES: Studying the characteristics of resilience may help to explain how, in the face of a chronic disease, people are able to cope in productive and effective ways. The Wagnild and Young Resilience Scale (RS) is an appropriate instrument to study resilience and has already been translated from the original English version into several languages. The aim of this study was to validate the Italian version of the RS, a 25-item scale ranging from 25 to 175 where higher scores indicate stronger resilience. METHODS: The Minimal Translation Criteria were followed to translate the scale which was then filled out by 1090 students to assess the reliability, stability, internal consistency and concurrent validity. RESULTS: Time stability was assessed in a sample of 117 students (M age=20.18 yr, SD 1.25) by test-retest correlation (r=0.78). RS reliability was evaluated in a second sample of 973 students (M age=16.95 yr, SD 1.50) with RS mean of 126.6 (SD 17.4). Concurrent validity was assessed by correlation with General Health Questionnaire (r=-0.51), Ego-Resilience Scale (r=0.63) and Beck Depression Inventory (r=-0.45). Internal consistency was evaluated by Cronbach alpha (α=0.84). Principal component analysis was performed on 24 out of the 25 items and resulted in six components. CONCLUSIONS: Our data indicated that the 24-item Italian version of the RS scale can be considered a useful instrument to measure resilience and can be used by healthcare staff to help patients cope effectively with stressful situations such as rheumatic and other chronic diseases.

Prolonged effect of two combined methods for Legionella disinfection in a hospital water system.

This study was conducted in our 650 bed general hospital, which is situated on the southern outskirts of Milan (Italy). After a first nosocomial case of pneumonia (caused by Legionella pneumophila serogroup 1), we first used a conventional method (heat shock) without success. To solve the problem we then tried a copper-silver ionization system combined with a chlorine dioxide device. During the four years after the installation of these two systems we recorded a significant (p < or = 0.05) reduction in Legionella colonization, and no new cases of Legionnaires' disease were observed. Our results suggest that the Cu-Ag ionization system, combined with a chlorine dioxide device, is a highly promising method for the control of Legionella pneumophila in a hospital water distribution system.

[From informed to shared: the developing process of consent]. / Il processo evolutivo del consenso: da informato a condiviso.

Based on the respect of the four well known ethical principles "autonomy, non-maleficence, beneficence and justice", of clinical cognitive psychology, social psychology and bioethics, and in the light of moral values, we wish to comment the clinical problems involving informed consent for diagnostic procedures, treatment and research in pediatrics. Studies of the issues in attitudes and shared consent in the clinical management involving children, their parents and the therapeutic team are still limited. Our suggestion is to manage the process of informed consent as a negotiated "shared consent" originated from the cognitive social representation theory, and taking into consideration the evolutive characteristics of the cognitive processes in children and adolescents, the ego defence mechanisms, the coping behaviour activated in the relationship among the pediatric patient, his/her family and the physicians. Many parents told us that the informed consent process is helpful though often confusing. Satisfaction was not related to ethnicity or education level. They found discussions more helpful than the consent documents. The more difficult process concerned their understanding of the concept of randomisation and the request of their consent to this procedure. The model we suggest has also the aim to give adequate and honest informations to children and adolescents through a continuous dialogue with the physician, until this become a routine part of their life in hospital care, to avoid confusion, to satisfy any request and curiosity, to be honest and helpful with any answer. We strongly believe that medical students, and in particular pediatricians, must be trained on "communication" and that they need to acquire, in addition to their medical capability, a good knowledge on this topic, including ethics and relational aspects. In our opinion, pediatricians must become expert also in the following topics: "Problem-related Learning", "The Family System Health Model", "The Theory of Social Representations", "Gadamerian Hermeneutics", and "The Communicative Skills".

The Impact-on-Family Scale: a test of invariance across culture.

OBJECTIVE: To test the cultural invariance of the Impact-on-Family Scale in order to make cross-cultural comparisons. METHODS: The Italian version of the scale was administered to mothers of children with chronic illnesses. Factorial invariance was examined to investigate whether the four factors found with the original United States (U.S.) scale could be replicated. RESULTS: The results clearly demonstrate the replicability of the first three factors, Financial Burden, Familial/Social Impact, and Personal Strain. In addition, internal consistency and the homogeneity of the items of the corresponding scales are satisfactory. However, the fourth factor, Mastery, could not be replicated, and the reliability of the corresponding scale is poor. Italian mothers scored significantly lower on the Financial Burden and on the Familial/Social Impact dimension, compared to the American sample. CONCLUSIONS: Three of the four factors of the Impact-on-Family Scale are useful for cross-cultural comparisons between U.S. and Italian samples.

Antioxidant enzymatic activities and lipid peroxidation in cultured human chondrocytes from vertebral plate cartilage.

In vitro models based on primary cultured human chondrocytes could be useful to study the ROS-mediated inflammatory processes that seem to involve chondrocytes in vivo. In this work, we studied the enzymatic antioxidative capability of human chondrocytes removed from vertebral plates during micro-discectomy and cultured 18 days, measuring total superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSHPx) activities. We also evaluated in the same cells the amount of malondialdehyde (MDA) in order to verify the effect of the variation of the cellular enzymatic antioxidative capability on the degree of membrane lipid peroxidation. Total SOD activity increased, even if not significantly, between the 12th and the 18th day. A significant variation of GSHPx (P<0.01) and of catalase (P<0.001) activity was observed between the 3rd and the 6th day with no further variation until the 18th day. A significant increase (P<0.001) of lipid peroxidation from the 3rd to the 18th day was also observed. These results seem to indicate that only fresh human cultured chondrocytes are suitable to study, through in vitro models, the in vivo behavior of the antioxidative status of these cells.

A longitudinal study of cognitive abilities and achievement status of children with myelomeningocele and their relationship with clinical types.

Cognitive abilities and achievement status of 178 children with myelomeningocele (MMC) were examined with neuropsychological techniques. These children as a whole scored below the population average. Children with MMC and shunted hydrocephalus performed worse than the arrested (unshunted) and non-hydrocephalic children. The greater deficit occurred on the Performance IQ. 80.7% of the subjects attended normal school while, 19.3% received remedial instruction. The results of our study are consistent with a large body of literature indicating that children with shunted hydrocephalus demonstrate poorer development of non-verbal relative to verbal cognitive abilities.

Deficiency of total collagen content and of deoxypyridinoline in intracranial aneurysm walls.

The tissue contents of total collagen and of 3-hydroxypyridinium cross-links, pyridinoline (PYD) and deoxypyridinoline (DPD), were measured in 15 samples of human aneurysms of Willis' Circle obtained at surgery and in 25 autopsy control samples of intracranial arteries of Willis' Circle obtained from 6 subjects who died of other causes than cerebral hemorrhage. PYD and DPD were detected fluorimetrically after HPLC separation. Total collagen content was significantly lower (P < 0.001) in aneurysm samples (mean +/- S.E.M. 2.50 +/- 0.33 nmol of alpha 1(I) collagen chain per mg of delipidated and dried material) than in controls (mean +/- S.E.M. 3.86 +/- 0.14). DPD, but not PYD, content appears to be lower in aneurysm walls. In the aneurysms, the tissue contents of PYD ranged from 212 to 587 pmol/nmol of alpha 1(I) collagen chain (mean +/- S.E.M. 430 +/- 31) while in control samples the values observed ranged from 292 to 642 (mean +/- S.E.M. 471 +/- 21). The tissue content of DPD was measurable only in 6 aneurysm samples (60%), ranging from 12 to 60 pmol/nmol of alpha 1(I) collagen chain (mean +/- S.E.M. 33 +/- 9), while in control samples, DPD content ranged from 30 to 123 (mean +/- S.E.M. 75 +/- 5).

Ovariectomy in the rat induces a rapid increase in the urinary excretion of hydroxylysine glycosides and non-reducible crosslink residues.

The ovariectomized rat is the most commonly used animal model of human postmenopausal osteoporosis, exhibiting a high rate of bone turnover with resorption exceeding formation. At present, bone turnover is quantified directly by dynamic histomorphometry. The aim of the present study was to determine whether the measurement of the urinary output of some specific bone collagen catabolites--pyridinolines and hydroxylysine glycosides--could be used to indirectly monitor the initial phase of bone turnover increase in ovariectomized 90-day-old rats. Ninety-day-old female rats were randomly divided into three groups (n = 6): ovariectomized, sham-operated and non-treated controls. Urine samples (24 h) were collected 6 days before surgery and twice weekly for the 4 weeks following ovariectomy. Urinary excretion of pyridinoline (PYD), deoxypyridinoline (DPD), glucosyl-galactosyl-hydroxylysine (GGHYL) and galactosyl-hydroxylysine (GHYL) were measured. As expected, ovariectomy was associated with a significant decrease in bone mineral density in both the proximal tibial and distal femoral metaphysis. Compared with both sham-operated and control animals, ovariectomized rats showed significant increases in PYD, GGHYL, and GHYL urinary output 8 days after surgery and in DPD output after 15 days. These changes were maintained throughout the study. The results confirm that measurement of the urinary excretion of pyridinolines and hydroxylysine glycosides represents a powerful tool for detecting the onset of bone turnover in ovariectomized 90-day-old rats.

Abnormalities of collagen cross-linkage in posterior communicating artery aneurysms: a preliminary study.

The collagen of tissues submitted to mechanical load (i.e. arterial wall) is characterised by the presence of intermolecular covalent cross-links (hydroxylysyl pyridinoline or pyridinoline: PYD; and lysyl-pyridinoline or deoxypyridinoline: DPD) which stabilise the molecular structure. In this preliminary study we look for quantitative or qualitative alterations of collagen cross-linkage in intracranial aneurysms of posterior communicating artery (PCoA) which may be considered a site of elective weakening in the intracranial arterial circulation, being one of the most frequent locations of intracranial aneurysms and of so called "infundibular widening'. We analysed the collagen cross-linkage in 6 autopsy samples of intracranial arterial segments of the Circle of Willis which were removed from patients whose cause of death was other than cerebral hemorrhage, and in 6 samples of intracranial PCoA aneurysms obtained at surgery. The analysis of cross-links showed that there was no significant difference in collagen and cross-link content between autopic and aneurysmatic samples except for PCoA. In autoptic nonaneurysmatic samples of PCoA a lower content of PYD than in internal carotid artery (ICA) and a lower content of DPD than in all other arterial segments has been demonstrated; moreover a lower content of cross-links (DPD + PYD/ Collagen) was evident in nonaneurysmatic PCoA samples when compared to other segments (ICA and Anterior Communicating Artery). On the other hand, the mean content of DPD was significantly lower in PCoA aneurysms than in nonaneurysmatic samples of the artery and moreover, a significantly low content of cross-links (DPD + PYD/Collagen) is overemphasised in PCoA aneurysms, suggesting that the peculiar lower content of DPD in PCoA arteries may be considered the expression of minor resistance of the arterial wall at this site, and may be related to the higher incidence of aneurysms or infundibular widening of this arterial segment.

Urinary output of hydroxylysine glycosides and pyridinium cross-links in detecting rat bone collagen turnover rate.

Glucosylgalactosylhydroxylysine (GGHYL), galactosylhydroxylysine (GHYL), pyridinoline (PYD) and deoxypyridinoline (DPD) were measured in the urine (6 h serial specimens over 96 and 24 h urine specimens for 4 days) collected from four adult Sprague Dawley rats and in the femoral and tibia] bone as well as in the dorsal skin of the same rats. No significant daily variations were found in the urine excretion of GGHYL, GHYL, PYD and DPD but significant diurnal variations. The GGHYL/GHYL ratio in rat urine (0.46 +/- 0.1) reflected neither the bone collagen ratio (1.9 to 2.4) nor the skin collagen ratio (1.22 +/- 1.07), a finding that may reflect GGHYL conversion into GHYL. The content of both pyridinolines was very low in the skin and high in the bone collagen and the urinary PYD/DPD ratio (1.46 +/- 0.15) reflected essentially the bone collagen ratio (0.8-3.0). These results suggest the usefulness of measuring GGHYL, GHYL, PYD and DPD in 24 h urine specimen and, based on the inter-animal variations, the necessity to consider each animal as its own control when bone turnover needs to be monitored.

High-performance liquid chromatographic method for measuring hydroxylysine glycosides and their ratio in urine as a possible marker of human bone collagen breakdown.

Glucosyl-galactosyl-hydroxylysine (GGHYL) and galactosyl-hydroxylysine (GHYL) are constituents of collagen protein. The ratio of the two hydroxylysine glycosides varies with the collagen type and, moreover, for a given collagen type, it also varies according to the connective tissue. For example, in type I collagen (the most abundant in the body), the GGHYL/GHYL ratio tends to be greater in soft connective tissues and lower in bone. The hydroxylysine glycosides are not recycled during collagen turnover and are excreted in the urine. Therefore, the urinary GGHYL/GHYL ratio, which reflects the proportion of the two metabolites in the various collagens, may indicate the type of connective tissue affected by pathological turnover, and may thus be a promising marker of bone metabolism. In this paper a method is described for the measurement of urinary hydroxylysine glycosides by reversed-phase liquid chromatography after purification of the sample by solid-phase extraction. The method presented is analytically reliable and suitable for routine use in a clinical laboratory.

Isotopic and nonisotopic assays for measuring somatotropin compared: re-evaluation of cutoff value in provocative tests.

Measurement of human growth hormone (hGH; somatotropin) concentrations in serum after provocative tests is crucial for diagnosing deficiencies in production of this hormone. Serum hGH can be measured by various immunoassays, isotopic and nonisotopic, with monoclonal or polyclonal antibodies: a cutoff value of 10 micrograms/L after provocative testing is usually used to distinguish normal from hGH-deficient children. Previous studies demonstrated discrepancies in hGH measurement by different radioisotopic immunoassays. Here we evaluated the responses of six different commercial assays, radioisotopic and nonisotopic, with monoclonal or polyclonal antibodies in a series of 16 provocative tests (stimulation with clonidine) in short children. A wide range of discrepant values was obtained with the different kits. A cutoff of 10 micrograms/L produced discordance of diagnosis among assays for two children, whereas complete agreement was reached for a cutoff value of 7 micrograms/L. Parallelism tests performed with hGH international standard, pure recombinant hGH, and a serum with high hGH content suggest that heterogeneity of the antibodies used by the manufacturers, even among monoclonal antibodies, is the main source of discordant results. Cutoff values and reference values must be established separately for each method proposed for routine use.