The HIV-positive patient in intensive care--psychological profile.

AIM: This paper aims to outline the profile of HIV-positive patients in intensive care, in terms of psycho-emotional and vital parameters. MATERIAL AND METHODS: We evaluated the HIV-positive patients that required intensive care (IC), from January 2011 to December 2013, in the HIV/AIDS Regional of the "Sf. Parascheva" Infectious Diseases Clinical Hospital Iasi. RESULTS: From January 2011 to December 2013, the HIV/AIDS Regional Centre in Iasi recorded 2649 hospitalizations, of which 0.67% (18 cases) required intensive medical care. Of these 10 were males and 8 females, aged between 24 and 65 years with a median of 24 years. There were 29 deaths (1.09% of all hospitalizations), 11 of which in intensive therapy (38% of all deaths)--7 men and 4 women. CD4 counts in persons requiring IC care were between 1 and 112/mm3, and most naive patients who died were late-presenters. The main diseases diagnosed were pulmonary tuberculosis and pneumocystosis, the main cause of death being multiple organ failure. The duration of hospitalization ranged between 4.5 and 30 days. Treatment success rate was correlated with the CD4 and biological status: liver and renal failure, respiratory failure, meningeal coma, hypoproteinemia, diselectrolitemia. From a psychological perspective, patients that arrived in the intensive care showed a history of non-compliance and non-adherence, a personality structure often marked by a lack of respect for them, indifference or ignorance regarding the factors that generate well-being. CONCLUSIONS: HIV-positive patients in the position of requiring intensive care showed a marked immunological collapse due to abandonment of therapy or late detection.

Clinical features and evolution of organ dysfunctions in sepsis.

UNLABELLED: In sepsis, the systemic inflammatory response is adapted to the etiologic agent and the increase in the level of mediators is associated with organ dysfunction. Currently, a rapid assessment of patient ability to develop an adequate immune response is not possible, the response mechanisms being similar in the context of different etiological agents. AIM: To find statistical arguments for the evolution of laboratory parameters in sepsis patients. MATERIAL AND METHODS: This retrospective study included 90 patients diagnosed with sepsis. The clinical, etiological, and laboratory data, and Carmeli and APACHE II prognostic scores were analyzed. The data were processed using SPSS version 16.0. RESULTS: The causative agents was identified in 16 cases; organ involvement and systemic response varied, and no statistical correlations were found between the inflammatory syndrome parameters and Carmeli or APACHE II prognostic scores or identification of the causative agent. CONCLUSIONS: Statistical correlations were found between maximum blood glucose levels and the presence of organ dysfunction in the studied sepsis patients. No correlations were found between sepsis severity and the presence of anemia or thrombocytopenia, or between fever syndrome and inflammatory syndrome.

Infective endocarditis in non-HIV immunosuppressed patients.

UNLABELLED: Infective endocarditis is a septic illness with sudden or insidious onset that causes heart lesions or aggravates the preexisting ones; during its course it can affect other organs, mortality being high. AIM: To identify non-HIV immunosuppressed patients with infective endocarditis, namely those with associated diabetes, hepatitis, cancer, or old age with multiple comorbidities, and to assess the nosocomial risk by Carmeli score. MATERIAL AND METHODS: This is a retrospective study using epidemiological, clinical, laboratory and treatment data from 56 patients admitted to and diagnosed with infective endocarditis at the lasi Infectious Diseases Hospital in the interval January 2008 - May 2012. To these patients we looked for non-HIV immunosuppressed character, having in their pathology association diabetes, chronic hepatitis, malignancy, or other comorbidities. RESULTS: From the infective endocarditis patients, 92.8% were found non-HIV immunosuppressed, 25% of them presenting renal failure, and 21.1% having diabetes. The most common Carmeli score obtained was score 2 in 57.1% of the cases. In these patients, the most frequent etiologic agent of infective endocarditis was Enterococcus faecalis (16%), followed by Staphylococcus aureus (10.7%). CONCLUSIONS: It was noted that in addition to endocardial damage due to the multiple etiologic agents involved, non-HIV immunosuppression in these patients plays an important role. The association of Carmeli score in patients diagnosed with infective endocarditis is significant, especially in case of adequate therapy initiation for a right treatment of this pathology.

Antibiotic therapy in severe sepsis in HIV-positive patients.

Severe sepsis has become one of the most frequent causes of hospitalization in intensive care units for patients diagnosed with HIV infection. The difficulty in setting a sepsis diagnosis in HIV-positive patients led to the systematic exclusion of these patients from studies on sepsis, which limited the understanding of its impact on the evolution of the disease. Our study aims to evaluate the etiology of sepsis in immunocompromised HIV-positive patients and the evolution after antibiotic therapy. 30 patients diagnosed with HIV infection and sepsis, admitted to our clinic between January 2008 and April 2012, were followed. Severity of illness, time since diagnosis, CD4 count, antiretroviral treatment, incidence of severe sepsis, and organ dysfunctions were registered. Patients were between 1 and 61 years of age, most of them were classified into stages B2, B3 and C3, requiring hospitalization for a period ranging from 14 to 28 days, with an average of 16.7 days and a median of 18 days, while 8 required monitoring in the intensive care unit. In about 40% of cases, the starting point was an infection of the lower respiratory tract, but also of the upper urinary tract and skin infections. Evolution and mortality in sepsis associated with HIV/AIDS infection depend on the presence of organ failure and are less influenced by the level of immunodepression, complex antibiotic therapy being the cornerstone in controlling patients.

Ligands playing musical chairs with G-quadruplex DNA: a rapid and simple displacement assay for identifying selective G-quadruplex binders.

We report here the details of G4-FID (G-quadruplex fluorescent intercalator displacement), a simple method aiming at evaluating quadruplex-DNA binding affinity and quadruplex- over duplex-DNA selectivity of putative ligands. This assay is based on the loss of fluorescence upon displacement of thiazole orange from quadruplex- and duplex-DNA matrices. The original protocol was tested using various quadruplex- and duplex-DNA targets, and with a wide panel of G-quadruplex ligands belonging to different families (i.e. from quinacridines to metallo-organic ligands) likely to display various binding modes. The reliability of the assay is further supported by comparisons with FRET-melting and ESI-MS assays.

Influence of the matrix on analyte fragmentation in atmospheric pressure MALDI.

In this paper, we report the measurement of the degree of analyte fragmentation in AP-MALDI as a function of the matrix and of the laser fluence. The analytes include p-OCH3-benzylpyridinium, three peptides containing the sequence EEPP (which cleave very efficiently at the E-P site), and three deoxynucleosides (dA, dG, and dC), which lose the neutral sugar to give the protonated base. We found that the matrix hardness/softness was consistent when comparing the analytes, with a consensus ranking from hardest to softest: CHCA >> DHB > SA approximately THAP > ATT > HPA. However, the exact ranking can be fluence-dependent, for example between ATT and HPA. Our goal here was to provide the scientific community with a detailed dataset that can be used to compare with theoretical predictions. We tried to correlate the consensus ranking with different matrix properties: sublimation or decomposition temperature (determined using thermogravimetry), analyte initial velocity, and matrix proton affinity. The best correlation was found with the matrix proton affinity.

Gas phase thermal denaturation of an oligonucleotide duplex and its complexes with minor groove binders.

Electrospray ionization with in-source collisionally induced dissociation has been used to probe the gas phase stability of an oligonucleotide duplex and its complexes with some minor groove binding drugs. On the basis of the arguments developed in detail by Drahos et al. (J. Mass Spectrom. 1999; 34:1373), this type of experiment can also be described as 'thermal denaturation in the gas phase'. We found that the gas phase denaturation curves were very similar to the solution phase denaturation curves determined by the traditional UV spectrophotometric method and, by analogy with the melting temperature T(m) which characterizes the stability in solution, we define a melting voltage V(m) to characterize the stability in the gas phase. A comparison of the T(m) and V(m) relative values suggests that the structure of the complexes is conserved during the electrospray process which transfers the ions from the solution to the gas phase.

Interaction between antitumor drugs and a double-stranded oligonucleotide studied by electrospray ionization mass spectrometry.

Electrospray ionization mass spectrometry was used to investigate the complex formation between a double-stranded oligonucleotide and various antitumor drugs belonging to two categories: intercalators (ethidium bromide, amsacrine and ascididemin) and minor groove binders (Hoechst 33258, netropsin, distamycin A, berenil and DAPI). The goal of this study was to determine whether the relative intensities in the mass spectra reflect the relative abundances of the species in the solution phase. The full-scan mass spectra suggest non-specific binding for the intercalators and specific binding for the minor groove binders. The preferential stoichiometries adopted by each minor groove binder were determined by studying the influence of the drug concentration on the spectra. We obtained 2:1 > 1:1 for distamycin, 1:1 > 2:1 for Hoechst 33258 and DAPI and only the 1 : 1 complex for netropsin and berenil. These features reflect their known behavior in solution. The compared tandem mass spectra of the 1 : 1 complexes with Hoechst 33258 and netropsin, when correlated with published crystallographic data, suggest the possibility of inferring some structural information. The relative binding affinities of the drug for the considered duplex were deduced with two by two competition experiments, assuming that the relative intensities reflect the composition of the solution phase. The obtained affinity scale is netropsin > distamycin A > DAPI > Hoechst 33258 > berenil. These examples show some of the potential uses of mass spectrometry as a useful tool for the characterization of specific drug binding to DNA, and possibly a rapid drug screening method requiring small amounts of materials.