The Potential of Nanobody-Targeted Photodynamic Therapy to Trigger Immune Responses.

Nanobody-targeted photodynamic therapy (NB-PDT) has been recently developed as a more tumor-selective approach rather than conventional photodynamic therapy (PDT). NB-PDT uses nanobodies that bind to tumor cells with high affinity, to selectively deliver a photosensitizer, i.e., a chemical which becomes cytotoxic when excited with light of a particular wavelength. Conventional PDT has been reported to be able to induce immunogenic cell death, characterized by the exposure/release of damage-associated molecular patterns (DAMPs) from dying cells, which can lead to antitumor immunity. We explored this aspect in the context of NB-PDT, targeting the epidermal growth factor receptor (EGFR), using high and moderate EGFR-expressing cells. Here we report that, after NB-PDT, the cytoplasmic DAMP HSP70 was detected on the cell membrane of tumor cells and the nuclear DAMP HMGB1 was found in the cell cytoplasm. Furthermore, it was shown that NB-PDT induced the release of the DAMPs HSP70 and ATP, as well as the pro- inflammatory cytokines IL- 1ß and IL-6. Conditioned medium from high EGFR-expressing tumor cells treated with NB-PDT led to the maturation of human dendritic cells, as indicated by the upregulation of CD86 and MHC II on their cell surface, and the increased release of IL-12p40 and IL-1ß. Subsequently, these dendritic cells induced CD4+ T cell proliferation, accompanied by IFNγ release. Altogether, the initial steps reported here point towards the potential of NB-PDT to stimulate the immune system, thus giving this selective-local therapy a systemic reach.

Shedding of PECAM-1 during HIV infection: a potential role for soluble PECAM-1 in the pathogenesis of NeuroAIDS.

Human immunodeficiency virus (HIV) infection is characterized by viral entry into the central nervous system (CNS), which is mediated, in part, by the transmigration of HIV-infected monocytes into the brain. The elaboration of chemokines and other factors by these infected cells contributes to CNS inflammation and cognitive impairment in a significant number of HIV-infected individuals. Recently, we demonstrated that HIV-infected monocyte transmigration into the CNS is enhanced greatly by the chemokine CC chemokine ligand 2 (CCL2)/monocyte chemoattractant protein-1. Platelet endothelial cell adhesion molecule-1 (PECAM-1) plays an important role in leukocyte transmigration across the endothelium of the systemic vasculature by mediating homophilic interactions between endothelial cells (EC)-EC and EC-leukocytes, thus preserving vessel integrity. The role of PECAM-1 in HIV-infected leukocyte transmigration across the blood brain barrier (BBB) and NeuroAIDS has not been characterized. We demonstrate that in brain tissue from individuals with HIV encephalitis, there is an accumulation of cleaved, soluble forms of the extracellular region of PECAM-1 (sPECAM-1). In addition, HIV-infected individuals have elevated levels of sPECAM-1 in their sera. Our in vitro data demonstrate that HIV-infected leukocytes, when treated with CCL2, shed sPECAM-1, suggesting a mechanism of extracellular PECAM-1 cleavage and release dependent on HIV infection and CCL2. We hypothesize that sPECAM-1 production by HIV-infected leukocytes, resulting in the accumulation of sPECAM-1 within the CNS vasculature and the generation of truncated, intracellular forms of PECAM-1 within leukocytes, alters PECAM-1 interactions between EC-EC and EC-leukocytes, thus contributing to enhanced transmigration of HIV-infected leukocytes into the CNS and changes in BBB permeability during the pathogenesis of NeuroAIDS.

A prospective study of HIV disease progression in female and male drug users.

OBJECTIVE: To compare HIV disease progression and mortality in a cohort of female and male drug users. DESIGN: A prospective cohort study of 222 HIV-seropositive women and 302 HIV-seropositive men who attended a hospital-affiliated methadone maintenance program with on-site primary care. METHODS: Regression slopes of CD4+ cell decline were compared using the two sample t-test, and the distribution of AIDS-defining illnesses evaluated by Mantel-Haenszel chi2 test. Time to AIDS-defining clinical conditions and death were compared using the Kaplan-Meier log-rank test. Multivariate estimates of progression to clinical AIDS or death, for all participants, stratified by sex, were derived from Cox proportional hazards models. RESULTS: Ninety-five persons (43 women and 52 men) developed AIDS-defining conditions. Analyses of the rates of CD4+ cell decline, the distribution of first AIDS-defining illnesses, and the time to clinical AIDS did not differ by sex. In the multivariate model, sex was not associated with an AIDS outcome, whereas crack-cocaine use [hazards ratio (HR), 1.815; 95% confidence interval (CI), 1.151-2.863], CD4+ cell count (100 x 10(6)/l; HR, 0.589; 95% CI, 0.511-0.679), and two or more HIV-related symptoms (HR, 1.702; 95% CI, 1.125-2.576) were associated. Mortality rates (8.71 per 100 person-years in women and 9.85 per 100 person-years in men) were similar, using univariate or multivariate methods. CONCLUSIONS: There was little difference in clinical outcomes or mortality between HIV-seropositive female and male drug users with access to primary care. However, crack-cocaine use was independently associated with progression to clinical AIDS.

Laboratory markers and the risk of developing HIV-1 disease among injecting drug users.

OBJECTIVE: To characterize the progression to HIV-1 disease among injecting drug users (IDU) according to laboratory markers. DESIGN: Prospective study of cohort of HIV-1-seroprevalent IDU, with case-comparison component. METHODS: Different laboratory markers were examined as predictors of progression to HIV-1-associated diseases including AIDS in a cohort of 318 HIV-1-infected IDU. The cohort was enrolled from a methadone treatment program in the Bronx, New York, USA. The independent utility of non-CD4 cell markers was evaluated after adjustment for the association of low CD4 lymphocyte count with AIDS risk. Clinical events in the natural history of HIV-1 were related to changes in levels of two variables related to duration of infection, CD4 lymphocyte count and serum beta 2-microglobulin (beta 2M) concentration. RESULTS: On univariate analysis, AIDS incidence measured from baseline increased with declining CD4 lymphocyte number and percentage, increasing serum beta 2M level, low platelet count, low leukocyte count and p24 antigenemia. Among HIV-1-related outcomes prior to any AIDS diagnosis, the relative risk of pyogenic bacterial infections conferred by these markers was similar to the relative risk of AIDS. For all HIV-1 outcomes, the elevated risk encountered at CD4 lymphocyte number < or = 200 x 10(6)/l was entirely due to the high risk at < or = 150 x 10(6)/l. On multivariate analysis, control for CD4 lymphocyte count eliminated the association of any other marker with increased AIDS hazard. HIV-1-related outcomes tended to occur in this order: multiple constitutional symptoms, oral candidiasis, pyogenic bacterial infections and AIDS. CONCLUSIONS: In HIV-1-infected IDU, several laboratory markers may predict AIDS when analyzed individually. These are not, however, independently related to increased AIDS risk after adjustment for low CD4 lymphocyte count. A CD4 count < or = 150 x 10(6)/l is more strongly related to immediate risk of adverse outcome than a count of 200 x 10(6)/l. A progressive series of clinical events is associated with markers of duration of HIV-1 infection, prior to and including AIDS diagnosis.

Municipal solid waste management: identification and analysis of engineering indexes representing demand and costs generated in virtuous Italian communities.

The definition and utilisation of engineering indexes in the field of Municipal Solid Waste Management (MSWM) is an issue of interest for technicians and scientists, which is widely discussed in literature. Specifically, the availability of consolidated engineering indexes is useful when new waste collection services are designed, along with when their performance is evaluated after a warm-up period. However, most published works in the field of MSWM complete their study with an analysis of isolated case studies. Conversely, decision makers require tools for information collection and exchange in order to trace the trends of these engineering indexes in large experiments. In this paper, common engineering indexes are presented and their values analysed in virtuous Italian communities, with the aim of contributing to the creation of a useful database whose data could be used during experiments, by indicating examples of MSWM demand profiles and the costs required to manage them.

Body image in older men with or at-risk for HIV infection.

We performed a cross-sectional analysis of factors associated with negative body image among 550 older men with or at-risk for HIV infection, including demographics, depression, illicit drug use, and antiretroviral therapy adherence. Overall, 31 per cent of participants reported negative body image, which was independently associated with increased BMI, self-rated fair/poor health, depression, and erectile dysfunction, but not HIV status. Screening for and treating depression, sexual dysfunction, and obesity in older men should be considered.

Body image in middle-aged HIV-infected and uninfected women.

We assessed factors associated with negative body image among 225 HIV-infected and 207 uninfected women. Standardized interviews obtained sociodemographic data and elicited agreement with the statement: 'Overall I am satisfied with my body shape'. Height and weight were measured. Overall, 39% of the women were obese and 47% had a negative body image. Factors independently associated with negative body image were HIV-infection, BMI, and depression. Given the high prevalence of obesity and negative body image, interventions aimed at assisting women with weight loss are warranted.

[Magnetic resonance in postoperative evaluation of aortic dissection]. / La Risonanza Magnetica nella valutazione postoperatoria della dissecazione aortica.

The value of MRI was investigated in the demonstration of residual alterations and postoperative complications of aortic dissection. October 1988 to December 1992, fifty-nine patients were examined with MRI. The series consisted of 53 patients affected with type A and 6 with type B aortic dissection, all of them surgically treated. The following parameters were studied: 1) aortic dilatation above and 2) below the prosthesis, 3) redissection, 4) persistent intimal flap, 5) origin of abdominal vessels from the false lumen and 6) study of supra-aortic vessels. Twelve of 59 patients were considered normal since all parameters were negative. The aorta was dilated in 19 patients distal to the graft and in 4 proximal to it. Redissection was observed in 33 of 59 patients. Residual intimal flap was clearly demonstrated in 33 of 59 patients; the state of the false lumen was clearly depicted in 12 patients with SE images alone and in 18 of the extant 21 with phase imaging. In 11 of 59 patients abdominal vessels originated from the false lumen. In 53 of 59 patients supra-aortic vessels were clearly demonstrated and appeared to be involved in 10 patients. In 6 cases MRI failed to yield enough information. In our experience MRI is the method of choice for monitoring the aorta after surgical dissection to detect changes and complications and therefore choose the most appropriate treatment.

Emergency physician workload: a time study.

STUDY HYPOTHESIS: Physician service time varies with patient service category, length of stay, and intensity of service. DESIGN: Prospective time study of emergency physician services. Physicians recorded the beginning and ending times of each service episode offered to a patient (whether at the bedside or occurring elsewhere in the department). Each episode was defined as an "interaction," with the total service time offered to a patient being the sum of all interactions for that patient. Length of stay was the time interval from when the patient registered in the emergency department to when the patient was released. Intensity of service was calculated as service time divided by length of stay. SETTING: University-affiliated community teaching hospital. TYPE OF PARTICIPANTS: One thousand three hundred forty-seven ED patients were entered into the study for nonselected (514), walk-in (637), observation (52), laceration repair (102), or critical care (42) services. Six of 12 physicians in the group staffing the ED participated in the study. Patient data were entered onto study cards when the service was offered. Patients were entered into the study consecutively except when the physician became too busy to see one patient at a time and accurately enter time data; such interruptions occurred for 18% of the patients. RESULTS: Physician service time for nonselected service patients (24.2 minutes per patient; 95% CI, 23.1-25.3) was consistent with ACEP's findings for nonselected services offered by emergency physicians (22 minutes per patient). Physician service time did not vary significantly from the standard for laceration repair patients (25.0 minutes per patient; 95% CI, 22.6-27.4) but did vary significantly from the standard for walk-in (9.8 minutes per patient; 95% CI, 9.3-10.3; P < .05), observation (55.6 minutes per patient; 95% CI, 50.7-60.5; P < .05), and critical care patients (31.9 minutes per patient; 95% CI, 26.2-37.6; P < .05). Walk-in and laceration repair patients had a single physician-patient interaction (1.3 per patient and 1.1 per patient, respectively), consistent with a discrete service offered during episodic care. Observation and critical care patients had multiple physician-patient interactions (6.3 per patient and 2.6 per patient, respectively) over an extended period, which is consistent with additional services being offered during their period of observation/holding. CONCLUSION: Case mix of patient services affects emergency physician workload and should be considered in planning departmental staffing needs.

[Intravenous leiomyomatosis with uterine onset and extending to the heart. Value of the radiological study]. / Leiomiomatosi intravenosa a partenza uterina con estensione cardiaca. Valore dello studio radiologico.

Two cases are reported of intravenous leiomyomatosis with extension of neoplastic thrombus to the right atrium; an histological documentation of each patient has been successfully obtained during surgical procedure. International literature is reviewed and a useful diagnostic protocol for the correct evaluation of these patients is suggested, in order to choose the best surgical approach. The important role of CT with intravenous injection of contrast medium is pointed out. The advantages resulting from the use of echotomography and from angiographic study of the inferior vena cava are stressed.

Temporal trends in the progression of human immunodeficiency virus disease in a cohort of drug users.

We evaluated changes over time in rates of progression to AIDS, mortality, and distribution of AIDS-defining illnesses in 524 human immunodeficiency virus (HIV)-seropositive injection drug users enrolled between 1986 and 1995 in a prospective study of HIV infection in the Bronx, NY. At enrollment, participants attended a hospital-affiliated methadone maintenance program with on-site primary care. Using the 1993 clinical definition of AIDS, we found that the hazard ratio (HR) of progression to AIDS declined for enrollees over time in comparison with the referent group of persons enrolled in 1986-1987. For program enrollees in 1988-1989, the HR was 1.0 [95% confidence interval (CI) = 0.6-1.6]; for enrollees in 1990-1991, the HR was 0.3 (95% CI = 0.1-0.9); for enrollees in 1992-1993, the HR was 0.5 (95% CI = 0.3-0.9); and for enrollees in 1994-1995, the HR was 0.2 (95% CI = 0.1-0.7), after controlling on initial CD4+ cell counts and age. Nevertheless, the greater AIDS-free time of later study entrants was not associated with reduced mortality. The study provides evidence that drug users with access to primary care likely benefited from improved management of HIV disease in prolonging AIDS-free time but, through 1996, did not experience greater survival.

Clinical manifestations and predictors of disease progression in drug users with human immunodeficiency virus infection.

BACKGROUND AND METHODS: To examine the clinical course of human immunodeficiency virus (HIV) infection in injection-drug users, we conducted a prospective study of a cohort of patients in a methadone-treatment program in New York City from July 1985 through December 1990. The patients underwent standardized evaluations at base line and semiannually thereafter and received on-site primary medical care. Rates of progression to the acquired immunodeficiency syndrome (AIDS) and major outcomes before the development of AIDS were examined by univariate analyses; the risk of AIDS was also assessed by product-limit survival analysis and proportional-hazards regression. RESULTS: Of 318 HIV-seropositive patients who did not yet have AIDS (171 men and 147 women), 90 were black, 179 were Hispanic, and 49 were white; the median age was 33 years. Over a median of 3.0 years of follow-up, 55 (17 percent) received a diagnosis of AIDS (incidence per 100 person-years, 5.8). Major outcomes before the development of AIDS included oral candidiasis (incidence per 100 person-years, 11.2), pyogenic bacterial infections including pneumonia and sepsis (8.0), pulmonary tuberculosis (1.2), multiple constitutional symptoms (13.6), and herpes zoster (1.3). There were 41 deaths from AIDS, and 13 seropositive patients without AIDS (4.1 percent) died of bacterial infections, as compared with only 1 of 411 seronegative patients studied (P < 0.001). The incidence of AIDS was 62 percent lower among those who took zidovudine than among those who did not (P = 0.02). In the multivariate analysis, progression to AIDS was best predicted by low numbers and percentages of CD4+ lymphocytes, nonuse of zidovudine, and the presence of oral candidiasis, bacterial infections, or tuberculosis. There was no consistent relation between progression to disease and the continued use of injection drugs. CONCLUSIONS: HIV-infected injection-drug users have progression to AIDS at rates comparable to those of other HIV-infected groups, but they have substantial pre-AIDS morbidity and mortality, particularly from bacterial infections, which also appear to predict disease progression.

Effects of HIV infection on the serologic manifestations and response to treatment of syphilis in intravenous drug users.

OBJECTIVE: To describe the effects of human immunodeficiency virus (HIV) infection on the serologic manifestations and response to treatment of syphilis in intravenous drug users. DESIGN: Cohort study of intravenous drug users. SETTING: Medical clinic in a hospital-based methadone maintenance treatment program in New York City. PATIENTS: Fifty patients with syphilis, of whom 31 were HIV seropositive and 19 HIV seronegative. MEASUREMENTS: Serologic tests for syphilis and clinical manifestations. RESULTS: Stage of syphilis at presentation was not associated with HIV serologic status. No unusual or fulminant manifestations of early syphilis or neurosyphilis were noted among HIV-seropositive cases. Maximum nontreponemal titers were higher among HIV-seropositive (median, 1:128) than among HIV-seronegative (median, 1:32) patients with syphilis (P = 0.05); this difference was present only among patients with first-episode syphilis. All 26 evaluable, HIV-seropositive patients treated for syphilis responded appropriately, including 13 patients given standard or less-than-standard doses of penicillin. Seven of 43 patients (16%) showed reversion to negative treponemal antibody assay results after treatment for syphilis; this finding was not associated with HIV infection, CD4 count, or stage of syphilis. Low nontreponemal titer was weakly associated with treponemal test reversion. CONCLUSIONS: Infection with HIV did not alter the stage at presentation, clinical course, serologic manifestations, or response to treatment of syphilis in this cohort of intravenous drug users.

Antiretroviral therapy adherence and viral suppression in HIV-infected drug users: comparison of self-report and electronic monitoring.

To compare electronically monitored (MEMS) with self-reported adherence in drug users, including the impact of adherence on HIV load, we conducted a 6-month observational study of 67 antiretroviral-experienced current and former drug users. Adherence (percentage of doses taken as prescribed) was calculated for both the day and the week preceding each of 6 research visits. Mean self-reported 1-day adherence was 79% (median, 86%), and mean self-reported 1-week adherence was 78% (median, 85%). Mean MEMS 1-day adherence was 57% (median, 52%), and mean MEMS 1-week adherence was 53% (median, 49%). One-day and 1-week estimates were highly correlated (r>.8 for both measures). Both self-reported and MEMS adherence were correlated with concurrent HIV load (r=.43-.60), but the likelihood of achieving virologic suppression was greater if MEMS adherence was high than if self-reported adherence was high. We conclude that self-reported adherence is higher than MEMS adherence, but a strong relationship exists between both measures and virus load. However, electronic monitoring is more sensitive than self-report for the detection of nonadherence and should be used in adherence intervention studies.