Structure/antileishmanial activity relationship study of naphthoquinones and dependency of the mode of action on the substitution patterns

A series of naphthoquinones was tested for activity against both extracellular promastigote and intracellular amastigote Leishmania major GFP in vitro. In parallel, the compounds were evaluated for cytotoxic effects against bone marrow-derived macrophages (BMM Φ) as a mammalian host cell control. Most of the compounds noticeably inhibited the growth of extracellular parasites (IC (50) 0.5 to 6?μM) and the intracellular survival of L. major GFP amastigotes (IC (50) 1 to 7?μM) when compared with the antileishmanial drug amphotericin B (IC (50) of 2.5 and 0.2?μM, respectively). In general, antiprotozoal activity and host cell cytotoxicity seemed to increase in parallel. Conspicuously, the cytotoxic effect was less pronounced on infected host cells when compared with that on noninfected cells. Concerning structure/activity relationships for the tested naphthoquinones, some interesting structural features emerged from this study. Introduction of a methyl or methoxyl group at C-2 of the parent 1,4-naphthoquinone slightly increased the antileishmanial activity against clinically relevant amastigotes, while the presence of a hydroxyl function in this position dramatically reduced the effectiveness. In contrast, hydroxylation at C-5 and dihydroxy substitution at C-5 and C-8 significantly enhanced the antiprotozoal activity. Similarly, the presence of a side chain hydroxyl group PERI to a carbonyl function as represented in the series of shikonin/alkannin derivatives increased the activity when compared with substituted analogs. Within the series of naphthoquinones tested, the dimeric mixture of vaforhizin and isovaforhizin showed the highest activity IN VITRO against the clinically relevant intracellular amastigote with an IC (50) of 1.1?μM. With IC (50) values mostly in the range of 1-3?μM, the shikonin/alkannin derivatives proved to be similarly considerably leishmanicidal. None of the compounds tested was capable to induce NO production known to play a crucial role in the host resistance against intracellular pathogens, excluding activation of microbicidal mechanisms in macrophages. The mode of action apparently depended on the substitution pattern, associated with the electrophilicity of the naphthoquinone or the efficiency of redox cycling. Conspicuously, members oxygenated in the quinone ring proved to be leishmanicidal when coincubated with glutathione, while the majority of the remaining compounds lost activity.     

Investigating the effect of antioxidant treatment on the protective effect of preconditioning in anesthetized rabbits

Reactive oxygen and nitrogen species are critical in preconditioning (PC). We sought to determine the effect of N-2-mercaptopropionyl glycine (MPG) on infarct size and on the oxidative status. Rabbits were exposed to 30-minute regional ischemia of the heart, which was followed by 3-hour reperfusion: (1) a control group without further intervention, (2) a PC1 group that was subjected to one cycle of PC, (3) a PC4 group that was subjected to 4 cycles of PC, (4) an MPG group that was treated with MPG for 60 minutes, starting 10 minutes before reperfusion, (5) MPG-PC1, and (6) the MPG-PC4 groups that were treated with the same dose of MPG and with 1 or 4 cycles of PC, respectively. Blood samples were drawn and collected for metabonomic analysis. In another series of experiments, 6 groups respective to the described ones were subjected to 30-minute regional ischemia of the heart and 20 minutes of reperfusion, after which pieces of heart tissue were quickly excised for malondialdehyde, nitrotyrosine, and glutathione content assessment. All PC and MPG groups developed smaller infarct size compared with control (16.5% ± 3.9%, 13.7% ± 3.1%, 18.6% ± 5.0%, 9.7% ± 2.0%, 15.0% ± 2.8% vs. 48.05% ± 7.2%; P < 0.05). MPG did not prevent lipid peroxidation and nitrotyrosine formation but enhanced the glutathione content. PC and MPG induced similar nuclear magnetic resonance changes. Long MPG infusion reduces the infarct size without abolishing the effect of PC, providing novel insights into the activity of MPG in PC.     

Breast density, scintimammographic 99mTc(V)DMSA uptake, and calcitonin gene related peptide (CGRP) expression in mixed invasive ductal associated with extensive in situ ductal carcinoma (IDC + DCIS) and pure invasive ductal carcinoma (IDC): correlation with estrogen receptor (ER) status, proliferation index Ki-67, and histological grade

Background

We evaluated the variation of calcitonin gene related peptide (CGRP) expression in patients with mixed invasive with extensive in situ ductal carcinomas (IDC + DCIS) and pure IDC, in relation to mammographic breast density (%BD), proliferation-seeking radiotracer ^99mTc(V) dimercaptosuccinic acid (DMSA) uptake (scintimammographic-SMM), proliferation index Ki-67, and estrogen receptor (ER) status. We also assessed CGRP expression with histological grade.

Methods

We studied retrospectively 24 women with suspicious findings on mammography who were evaluated preoperatively with ^99mTc(V)DMSA scintimammography. Histology revealed 12 IDC (grade II in 8, grade III in 4 patients; mean size ± SD, 2.6 ± 1.3 cm; mean age ± SD, 66.5 ± 13.1 years) and 12 IDC + DCIS (grade II in 6, grade III in 6 patients; DCIS component mean size ± SD, 5.3 ± 1.8 cm; IDC component mean size ± SD, 2.5 ± 1.1 cm; mean age ± SD, 58.5 ± 15.1 years). Immunohistochemistry for CGRP, Ki-67, and ER status was performed in all 24 surgical specimens. BD and SMM were calculated by computer-assisted methods and were statistically correlated with CGRP expression. BD, SMM, Ki-67, and ER were statistically compared between IDC and IDC + DCIS, whereas CGRP, Ki-67, and ER were compared between patients with BD >25 and <25%. CGRP was also compared (t test) between grade II and grade III in both groups.

Results

Overall positive correlation was found between BD and CGRP (r = 0.577, P < 0.001). Positive correlation was established between SMM and CGRP only in IDC + DCIS (r _{SMM(IDC+DCIS)–CGRP} = 0.634, P < 0.05). CGRP and Ki-67 were significantly higher in patients with BD >25% compared with <25% BD patients (P = 0.00008 and P = 0.014, respectively). BD and SMM were significantly higher in CGRP(+) than in CGRP(?) patients as well as in IDC + DCIS compared with IDC. Ki-67 was significantly higher, whereas ER was significantly lower, in IDC + DCIS than in IDC. In all patients, CGRP was significantly higher in grade II as compared with grade III (P = 0.005). In the mixed group (IDC + DCIS), grade II cancers had also significantly higher CGRP values as compared with grade III ones (P = 0.004). In pure IDC, no statistical difference was found between grade II and III (P = 0.4).

Conclusions

ΒD, SMM, CGRP, and Ki-67 were significantly increased, whereas ER was significantly decreased, in IDC + DCIS as compared with IDC, indicating that IDC + DCIS is an entity that is more aggressive, ER independent, and possibly associated with a pathway linked to stromal involvement and CGRP activity.     

Organochlorine Pesticide Residues in Human Breast Milk from Southwest Greece: Associations with Weekly Food Consumption Patterns of Mothers

Breast milk samples were collected from 112 mothers who resided in southwest Greece during the years 1995-1997. The following chemicals, which were present in some of the milk samples, were quantified by gas chromatography: residues of lindane (i.e., α-hexa-chlorocyclohexane [BHC], (β-BHC, γ-BHC), δ-BHC, 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (p,p-DDE), 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane (p,p'-DDD, and 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (p/p'-DDT). With the assistance of a qualified dietitian, the mothers also completed a 7-d food-frequency questionnaire and provided additional personal information (e.g., educational level, profession, previous or present residence, use of pesticides, treatment of dermal scabies). Concentrations of γ-BHC (i.e., lindane) were present in 57.1 % of the samples (mean concentration = 0.58 μg/l [whole milk], range = nondetectable to 10.86 μg/l). Concentrations of α-BHC, β-BHC, δ-BHC, p,p'-DDD, and p,p'-DDT were detected in 16%, 39.3%, 11.6%, 41%, and 55.3% of the samples, respectively. Moreover, p,p'-DDE, which was detected in all samples, ranged from 0.33 to 278 μg/l. Typically, the amounts of γ-BHC and Σ-DDTs in breast milk were below the established acceptable daily intake value. Nonetheless, 3 of the 112 samples contained concentrations of Σ-DDT derivatives that exceeded the acceptable daily intake value established by the World Health Organisation in 1987. Concentrations of insecticides in breast milk were affected by dietary habits. Some women who consumed 7 (or more) portions of fresh vegetables per week had γ-BHC concentrations in whole milk that exceeded 0.15 μg/l (odds ratio = 1.23 [95% confidence interval = 1.05, 1.44]; p = .006). The concentrations of DDT derivatives were associated with the portions of fish, chicken, fruits, milk, and potatoes consumed each week.     

Predictors of quality of life outcomes in chronic rhinosinusitis after sinus surgery

The predictive value of olfaction for quality of life (QoL) recovery after endoscopic sinus surgery (ESS) in chronic rhinosinusitis (CRS) is still underestimated. The aim of this study was to explore the proportion of patients suffering from CRS who experience clinically significant QoL improvement after ESS and identify pre-operative clinical phenotypes that best predict surgical outcomes for QoL, focusing mainly on the role of patients’ olfaction. One hundred eleven patients following ESS for CRS and 48 healthy subjects were studied. Olfactory function was expressed by the combined “Threshold Discrimination Identification” score using “Sniffin’ sticks” test pre-treatment and 12 months after treatment. All subjects completed validated, widely used QoL questionnaires, specific for olfaction (Questionnaire of Olfactory Deficits: QOD), for assessing psychology (Beck Depression Inventory: BDI) and for general health (Short Form-36: SF-36). Statistically significant improvement of olfactory function by 41.8 % and of all QoL questionnaires scores (all p < 0.001) was observed on the 12-month follow-up examination. Clinically significant improvement for QoL was measured in a proportion of 56.8 % of patients on QOD, 64.9 % on SF-36 and 49.5 % on BDI scales results. Although olfactory dysfunction, nasal polyps, female gender, high socio-economic status and non-smoking habits were significantly associated with better QoL results, multivariate logistic regression analysis revealed that only olfactory dysfunction and nasal polyps were independent predictors significantly associated with higher likelihood of clinically significant improvement in all QoL questionnaire results. Olfactory dysfunction and nasal polyps were independent pre-operative predictors for surgical outcomes with regard to QoL results.     

Comparison of Two Radiotherapeutic Hypofractionated Schedules in the Application of Tumor Bed Boost

AimEvaluation of related radiation toxicity and efficacy in terms of local control of 2 radiotherapeutic hypofractionated schedules in the application of tumor bed boost by using 2 different planning techniques.MethodEighty-one patients with stage I-II disease were retrospectively selected with either concomitant (group A) or sequential (group B) boost for the tumor bed. In group A, 27 patients were treated with a total dose of 46 Gy to the whole breast and 54 Gy to the tumor bed in 20 concomitant fractions. In group B, 54 patients were treated with a total dose of 42.4 Gy in 16 fractions to the whole breast and 53 Gy to the tumor bed by 4 sequential fractions. The boost was administered with multiple photon-beam fields. The median follow-up time was 24 months.ResultsThe statistical analysis for the 2 groups of the study showed that skin toxicity was significantly worse in group A (P < .05, Kruskal-Wallis H test). For groups A and B at the completion of radiation therapy, grade 1 skin toxicity was observed in 18/27 patients (66.6%) and 13/54 patients (24.1%), respectively, whereas grade 2/3 was observed in 9/27 patients (33.3%) vs. 5/54 patients (9.3%), respectively (P < .001). One year after irradiation, in group A and in group B, the skin toxicity was of grade 1 in 6/27 patients (22.2%) vs. 2/54 patients (3.7%), respectively (P = .008). Within 2 years, the breast returned to its original form in all patients. No patient showed local disease recurrence.ConclusionsThe accelerated hypofractionated schedules in the application of the tumor bed boost by using the 2 different planning techniques appears to be effective and well tolerated.     

Efficacy of brimonidine 0.2% in controlling acute postoperative intraocular pressure elevation after phacoemulsification

PURPOSE: To determine the efficacy of brimonidine tartrate 0.2% drops given 2 times a day in reducing intraocular pressure (IOP) spikes during the first 24 hours after phacoemulsification cataract surgery. SETTING: Department of Ophthalmology, General Hospital of Patras Agios Andreas, Patras, Greece. METHODS: In this prospective double-blind placebo-controlled study, 1 eye of 40 consecutive normotensive cataract patients having small-incision cataract surgery was randomized into 1 of 2 treatment arms. Twenty patients received a placebo (artificial tears) and 20 patients received brimonidine tartrate 0.2% drops 2 times a day the day before and the day of surgery. Diurnal IOP variation was the primary efficacy variable; IOP was measured at baseline, before surgery, and 4, 6, 12, and 24 hours postoperatively. RESULTS: The placebo group had higher IOPs at every time point after surgery. Peak elevation of IOP occurred 6 hours after surgery. The mean IOP in the placebo group (27.71 mm Hg +/- 3.75 [SD]) was statistically significantly higher than in the brimonidine group (21.45 +/- 1.32 mm Hg) (P<.001). A major IOP rise (>/=20 mm Hg above baseline IOP) occurred in 1 patient (5%) in the placebo group who required emergency hypotensive therapy. Twenty-four hours after surgery, 11 eyes (55%) in the brimonidine group and 4 eyes (20%) in the placebo group had an IOP lower than baseline. CONCLUSION: Prophylactic treatment with brimonidine tartrate 0.2% 2 times a day for 2 days was effective in reducing IOP peaks throughout the first 24 hours after phacoemulsification surgery.     

Predictive value of amniotic fluid cystatin C levels for the early identification of fetuses with obstructive uropathies

Objective To compare the diagnostic accuracy of cystatin C with that of creatinine in discriminating renal function in fetuses without ultrasononographic evidence of renal malformations from those with obstructive uropathies.
Design Prospective, observational cohort study.
Setting Prenatal morphologic and functional evaluation of fetal obstructive uropathies throughout pregnancy.
Population A total of 96 healthy pregnant women at different stages of pregnancy, without any pregnancy-related maternal disease. Eighty-one pregnant women without clinical and ultrasonographic evidence of any fetal anomaly, confirmed at birth, were defined as controls; 15 pregnant women with various fetal obstructive uropathies, evidenced by repeated ultrasound examinations and confirmed at birth, were defined as cases.
Methods Creatinine was measured by a kinetic Jaffe picric acid method and cystatin C by a nephelometric immunoassay. Variables were analysed by applying conventional statistical tests; the non-parametric receiver operating curves (ROC) analysis was used to evaluate the diagnostic efficiencies of the biochemical markers.
Main outcome measures Incidence of confirmed, diagnosed, neonatal obstructive uropathy by measuring baseline levels of cystatin C and creatinine in amniotic fluid.
Results Baseline levels of cystatin C in amniotic fluid were significantly higher (   P = 0.0015  ) among cases than in controls with comparable gestational age; no significant difference was found for creatinine levels (   P = n.s.  ). The maximum diagnostic accuracy of serum cystatin C in discriminating controls from fetal uropathies was 96%, while that of creatinine was 62%.
Conclusion Cystatin C may be considered a sensitive biochemical marker for the early identification of fetuses with obstructive uropathies.     

An Overview of Corneal Collagen Cross-Linking (CXL)

Corneal collagen cross-linking (CXL) was first described over a decade ago and is now considered to be one of the most important surgical innovations of modern ophthalmology. Prior to its introduction, no interventions were available to arrest, or slow down ectatic disease progression, with corneal transplantation required in the majority of cases. Unlike earlier treatments of corneal ectasias that attempted to only improve the consequences of the disease, CXL aims to address the corneal biomechanical weakening itself. The long-term safety and efficacy of CXL have been established in several studies that have documented significant improvements in all outcome measures (visual acuity, spherical equivalent, astigmatism, and keratometric findings). The emerging combination of CXL with other interventions (termed ‘CXL plus’) optimizes the visual and topographic outcomes. This, along with the expansion of the techniques’ indications for other clinical conditions, such as microbial keratitis, highlights the continuous improvement of the initial technique and confirms its wide acceptance. Overall, CXL has already demonstrated much promise and has several clinical indications, representing a clear example of recent advances in ocular therapy.