7-Keto hybrid steroidal esters of nitrogen mustard: cytogenetic and antineoplastic effects

Four newly synthesized antitumor steroidal compounds were compared, on a molar basis, regarding their ability to induce sister chromatid exchanges (SCEs) and cell division delays. The concept of designing and developing these compounds (1-4) is to enhance the anticancer activity of esteric steroidal derivatives of nitrogen mustard by introduction of a keto group at the 7-position of the D5 steroidal skeleton, and a double bond between positions 6 and 7 of the B ring of the steroidal nucleus. In our study, the cytogenetic and antileukemic effects of these newly synthesized compounds are reported. The four substances induced statistically significant enhancement of SCEs and of cell division delays, and in both schedules used, therapeutic effects. However, compounds 1 and 3 showed increased genotoxicity towards human lymphocytes (p<0.001) and antileukemic activity towards P388 leukemias (p<0.001), compared to compounds 2 and 4. It seems that the introduction of a keto group at the 7-position of the steroidal skeleton enhances the antitumor effect of these substances in comparison with our previous studies with the corresponding compounds characterized by the absence of the 7-keto group. Therefore, the in vivo antitumor effect of the four compounds appears to correlate well with the in vitro cytogenetic effect caused by these chemicals.     

Centrally administered galanin blocks morphine place preference in the mouse

Galanin is a neuropeptide with appetitive, antinociceptive and neuroendocrine functions. Galanin and galanin binding sites are present in brain areas that mediate reinforcement, such as nucleus accumbens and ventral tegmental area, as well as locus coeruleus, an area known to be involved in development of drug dependence and withdrawal. This localization, coupled with the observation that there is a strong interaction between morphine and galanin in spinal cord, made it of interest to study whether galanin might have effects on morphine reinforcement. Using the place preference paradigm we found that galanin (1 microg i.c.v.) alone does not possess reinforcing or aversive properties but attenuates the preference conditioned by peripheral administration of morphine (5 mg/kg s.c.). Quantitative receptor autoradiography showed that morphine treatment that could condition a place preference decreased galanin binding in the nucleus accumbens and increased galanin binding in the locus coeruleus. In contrast, acute naltrexone administration increased galanin binding in the nucleus accumbens, suggesting that levels of galanin binding are tonically regulated by opioid receptors in that area. Contrary to what is seen in the spinal cord, these results indicate that galanin and morphine have an antagonistic interaction in the brain that results in attenuation of morphine reinforcement by activation of the galaninergic system.     

Differential rates of replacement of human dermal dendritic cells and macrophages during hematopoietic stem cell transplantation

Animal models of hematopoietic stem cell transplantation have been used to analyze the turnover of bone marrow–derived cells and to demonstrate the critical role of recipient antigen-presenting cells (APC) in graft versus host disease (GVHD). In humans, the phenotype and lineage relationships of myeloid-derived tissue APC remain incompletely understood. It has also been proposed that the risk of acute GVHD, which extends over many months, is related to the protracted survival of certain recipient APC. Human dermis contains three principal subsets of CD45⁺HLA-DR⁺ cells: CD1a⁺CD14⁻ DC, CD1a⁻CD14⁺ DC, and CD1a⁻CD14⁺FXIIIa⁺ macrophages. In vitro, each subset has characteristic properties. After transplantation, both CD1a⁺ and CD14⁺ DC are rapidly depleted and replaced by donor cells, but recipient macrophages can be found in GVHD lesions and may persist for many months. Macrophages isolated from normal dermis secrete proinflammatory cytokines. Although they stimulate little proliferation of naive or memory CD4⁺ T cells, macrophages induce cytokine expression in memory CD4⁺ T cells and activation and proliferation of CD8⁺ T cells. These observations suggest that dermal macrophages and DC are from distinct lineages and that persistent recipient macrophages, although unlikely to initiate alloreactivity, may contribute to GVHD by sustaining the responses of previously activated T cells.     

SARS-CoV-2 Antigenemia as a Confounding Factor in Immunodiagnostic Assays: A Case Study

Humoral immunity has emerged as a vital immune component against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Nevertheless, a subset of recovered Coronavirus Disease-2019 (COVID-19) paucisymptomatic/asymptomatic individuals do not generate an antibody response, constituting a paradox. We assumed that immunodiagnostic assays may operate under a competitive format within the context of antigenemia, potentially explaining this phenomenon. We present a case where persistent antigenemia/viremia was documented for at least 73 days post-symptom onset using ‘in-house’ methodology, and as it progressively declined, seroconversion took place late, around day 55, supporting our hypothesis. Thus, prolonged SARS-CoV-2 antigenemia/viremia could mask humoral responses, rendering, in certain cases, the phenomenon of ‘non-responders’ a misnomer.     

Increased apoptosis in the alveolar microenvironment of the healthy human lung

BACKGROUND: Apoptosis represents a physiological clearance mechanism in human tissues. The role of apoptosis has not been examined in normal lung cell populations, such as alveolar macrophages and polymorphonuclear cells. What is the percentage, as well as the role, of apoptosis in the alveolar microenvironment of the healthy human lung? PATIENTS AND METHODS: Bronchoalveolar lavage was obtained from 21 volunteers without lung disease. The specimens were analyzed using: Annexin V binding, DNA laddering, light microscopy and immunohistochemistry for bcl-2 expression. RESULTS: Apoptosis of the total bronchoalveolar lavage cell population was 51.2%. Both alveolar macrophages and polymorphonuclear cells had a high apoptotic rate (62.1% and 48.3%, respectively) as determined by Annexin V binding. These findings were further confirmed using morphological criteria for apoptosis and gel electrophoresis for DNA fragmentation. In the majority of the individuals examined, (8 out of 21), the bcl-2 gene was expressed in the lymphocyte population mainly. CONCLUSIONS: The percentage of apoptosis in lung cells of healthy humans is high. Apoptosis plays a key role in normal lung cell death. It appears to be the mechanism that opposes cell proliferation by eliminating, aged or damaged cells thus facilitating the process of lung remodeling.     

The role of the kidney in blood volume regulation: the kidney as a regulator of the hematocrit

The kidney plays a pivotal role in the regulation of blood volume by controlling the plasma volume and red blood cell (RBC) mass. Further, it is proposed that the kidney coordinates the relative volumes of these 2 blood components and in so doing regulates the hematocrit. This novel function as proposed is a functional concept whereby the kidney does not simply produce erythropoietin, but that the kidney regulates the hematocrit is termed the critmeter function. The kidney is unique in that it can indirectly report on blood volume as a tissue oxygen signal. It is proposed that the kidneys detect small changes in tissue oxygen tension for erythropoietin production at the critmeter, a functional unit of marginal oxygen tension within the kidneys. As the production of erythropoietin is modulated by angiotensin II, the renin-angiotensin system entrains the production of erythropoietin as part of the effector signals of the feedback loop of blood volume regulation. Collectively, the consideration of these points generates a paradigm shift in our understanding of blood volume regulation in that the role of the kidney may be expanded from simply "producing" erythropoietin to regulating the hematocrit. Further, this concept broadens the scope of the traditionally identified effector mechanisms of plasma volume regulation to include the modulation of erythropoietin production and hence RBC mass. The inclusion of both plasma volume and RBC mass as factors targeted by the effector signals recapitulates that whole blood volume is sensed and reported in the afferent signals. In summary, distinct sensing and effector mechanisms for regulating the volume of the two components of whole blood (plasma and red cell mass) are recognized. The coupling of the regulation of these 2 components of blood volume is highlighted.     

Quality of life and social-economic characteristics of greek male patients on long-term oxygen therapy

BACKGROUND: Chronic obstructive pulmonary disease (COPD) profoundly impacts patients' functional status, especially in the advanced stages, when long-term oxygen therapy (LTOT) is implemented. OBJECTIVE: To determine the health-related quality of life (HRQOL) in patients with COPD and using LTOT, and assess the relationship of socioeconomic characteristics and pulmonary function test results to HRQOL scores. METHODS: We studied a group of 85 patients with COPD and hypoxemia who were on LTOT, and a control group of 48 patients with stable COPD but without hypoxemia. All subjects were asked to rate their dyspnea on the Modified Medical Research Council dyspnea scale, and to take the Medical Outcomes Study Short Form (SF-36), the General Health Questionnaire (30 questions), and a questionnaire (which we developed for this study) to measure their independence in activities of daily living (ADL). We also conducted pulmonary function tests and arterial blood gas analyses, and recorded socioeconomic characteristics. RESULTS: The subjects' socioeconomic status was moderate to low. HRQOL was impaired in patients on LTOT, especially in the physical function domain, and most of the examined dimensions correlated with the severity of dyspnea and psychological status. There was a significant association between ADL score and SF-36 score in the vitality and physical domains, but there was no significant association between HRQOL score and spirometry or blood gas values. CONCLUSIONS: HRQOL in patients with COPD and on LTOT is low and is influenced by dyspnea, mental status, and incapacity, rather than by physiological variables. We recommend a multidimensional therapeutic approach that targets symptom-control and ADL support to improve the patient's overall HRQOL.     

R7BP modulates opiate analgesia and tolerance but not withdrawal

The adaptor protein R7 family binding protein (R7BP) modulates G protein coupled receptor (GPCR) signaling and desensitization by controlling the function of regulator of G protein signaling (RGS) proteins. R7BP is expressed throughout the brain and appears to modulate the membrane localization and stability of three proteins that belong to R7 RGS family: RGS6, RGS7, and RGS9-2. RGS9-2 is a potent negative modulator of opiate and psychostimulant addiction and promotes the development of analgesic tolerance to morphine, whereas the role of RGS6 and RGS7 in addiction remains unknown. Recent studies revealed that functional deletion of R7BP reduces R7 protein activity by preventing their anchoring to the cell membrane and enhances GPCR responsiveness in the basal ganglia. Here, we take advantage of R7BP knockout mice in order to examine the way interventions in R7 proteins function throughout the brain affect opiate actions. Our results suggest that R7BP is a negative modulator of the analgesic and locomotor activating actions of morphine. We also report that R7BP contributes to the development of morphine tolerance. Finally, our data suggest that although prevention of R7BP actions enhances the analgesic responses to morphine, it does not affect the severity of somatic withdrawal signs. Our data suggest that interventions in R7BP actions enhance the analgesic effect of morphine and prevent tolerance, without affecting withdrawal, pointing to R7BP complexes as potential new targets for analgesic drugs.