Microscopic and Biopharmaceutical Evaluation of Emulsion and Self-Emulsifying Oil with Cyclosporine.

Among the currently available commercial eye drops with cyclosporine A (Cs) there is a lack of long-acting dosage forms and products with a concentration of the drug substance higher than 0.1%, although Cs is widely used in ophthalmology. The aim of the research was to conduct the microscopic and biopharmaceutical evaluation of two formulations, an emulsion (EM) and a self-emulsifying oil (SEO), both with 0.5% of Cs, proposed for use in eye drops, and the comparison of both. SEO eye drops with Cs or any other drug substance are currently not available as marketed products, and the highest concentration of Cs in the ocular emulsion is only 0.1%. The microscopic evaluation of the emulsion and the SEO after emulsification with water was carried out using a high-resolution digital microscopy. The properties of both preparations were compared using the high dynamic range function or optical shadow effect mode. Images in the 3D composition mode were also recorded. The in vivo study of the Cs formulations was performed on male albino rabbits. The eye tolerance of the preparations was assessed using the ocular irritation test, which is a modified Draize test. Placebo carriers (without the drug substance) were also subjected to irritation testing. The concentration of Cs in the tissues (cornea and conjunctiva) and fluids (tear fluid and aqueous humor) of the rabbit eye was determined after multiple instillations of Cs-EM or Cs-SEO. The tested preparations were compared using the digital microscopy technique, which highlights the features of the formulations and eliminates the risk of unnoticeable properties that are difficult to observe in classical optical microscopy. Both tested Cs-loaded formulations are classified as practically non-irritating. There were also no significant differences when testing the placebo carriers. After a topical administration, Cs was widely distributed in all tissues (e.g., in cornea 1.3 ng/mg and 1.0 ng/mg) and fluids of the eye (e.g., in tear fluid 11.6 µg/mL and 4.3 µg/mL), after the administration of Cs-SEO and Cs-EM, respectively. The obtained results allow us to recognize both tested formulations, the emulsion and the self-emulsifying oil with 0.5% Cs content, as carriers safe for ophthalmic use and effective in delivering the drug substance to the structures of the eye.

Pilot Study of Eyesight Characteristics and Thickness of the Retinal Nerve Fiber and Ganglion Cell-inner Plexiform Layers in Adolescent Patients with Type one Diabetes Mellitus on an Insulin Pump.

AIM: Estimation of the ocular status in adolescents with diabetes mellitus type 1 (DM1) treated with continuous subcutaneous insulin infusion (CSII), assessment of the development of the diabetic retinopathy (DR) and nephropathy (DN) within 10 years. METHODS: 37 patients (74 eyes) aged 16-33 years, treated with CSII were enrolled to the study. Baseline, and a 10- year follow-up evaluation included: best corrected visual acuity (BCVA), tonometry, slit lamp exam and fluorescein angiography (FLA). Additionally, spectral-domain optical coherence tomography (SD-OCT) was done in the 7th year of observation to assess the thickness of the retinal nerve fiber (RNFL) and the ganglion cellinner plexiform layers (GCL-IPL) complex thickness. Glycated haemoglobin (HbA1) and albuminuria were also analysed. RESULTS: During the 10-year observation period DR (non-proliferative - NPDR, proliferative - PDR, diabetic macular edema - DME) was diagnosed in 3 (8%) patients. In the DR group: BCVA was significantly lower, intraocular pressure (IOP) levels and albuminuria were higher. There were no differences in HbA1 in both groups. The thinning of RNFL was observed in both groups. Macular RNFL, GCL-IPL complex thickness assessment showed a significantly higher number of borderline results in the group with DR. CONCLUSIONS: Diabetic patients treated with CSII are at a lower risk of developing vascular complications even with poor metabolic control. Increased albuminuria may be a predictive sign for early ocular complications, and requires intense observation. Diagnosis of RNFL and GCL-IPL decreased values is crucial prior to diabetic retinopathy development. SD-OCT is a non-invasive, easy-to-perform, relatively inexpensive procedure, and can be a useful tool to monitor neuropathy progression.

The Level of Cytokines in the Vitreous Body of Severe Proliferative Diabetic Retinopathy Patients Undergoing Posterior Vitrectomy.

OBJECTIVE: The objective of the study was to compare cytokine levels in the vitreous body of patients with proliferative diabetic retinopathy (PDR) undergoing posterior vitrectomy. PATIENTS AND METHODS: The study included 39 patients (39 eyes) undergoing pars plana vitrectomy (PPV). Patients were divided into three groups: patients with proliferative diabetic retinopathy (PDR) without aflibercept injection prior to the surgery, PDR patients administered aflibercept injection prior to the surgery, and patients without diabetes mellitus (control group). All patients underwent a comprehensive eye examination one day before and 3 weeks after the surgery, including measurements of: best-corrected visual acuity (BVCA) and intraocular pressure (IOP), slit-lamp examination and spectral domain optical coherence tomography (SOCT). Concentrations of cytokines: IL-6, IL-8, IL-12p70, TNF, IL-10, IL-1ß were measured in the vitreous body of patients with BD™ Cytometric Bead Array (CBA) Human Inflammatory Cytokines Kit. RESULTS: PDR patients who received pretreatment with aflibercept injection showed significantly lower concentrations of IL-12p70, TNF, IL-10 and IL-1ß in the vitreous body compared to the control group. Meanwhile, patients without prior aflibercept injection had a significantly higher concentration of IL-8. There was also a significant positive correlation between IOP before PPV and IL-8 concentration in both PDR patients' groups. CONCLUSION: Findings of our study suggest an important role of IL-8 in the development of severe PDR. Aflibercept administration on the day before elective vitrectomy facilitated the surgery.

The Influence of Silicone Oil on the Ganglion Cell Complex After Pars Plana Vitrectomy for Rhegmatogenous Retinal Detachment.

PURPOSE: Pars plana vitrectomy (PPV) and silicone oil endotamponade have been used as a treatment in rhegmatogenous retinal detachment (RRD). Improvement in the modality of spectral-domain optical coherence tomography (SD-OCT) allows for the assessment of ganglion cell layer-inner plexiform layer (GCL-IPL) in the macular region. Information about the GCL-IPL status may be a response to the question as to why the visual recovery after PPV with silicone oil tamponade is incomplete. The aim of the study was to evaluate the impact of silicone oil on GCL-IPL and compare it with other endotamponade types such as Sulfur hexafluoride gas (SF6), Perfluoropropane gas (C3F8) used during PPV performed due to RRD. PATIENTS AND METHODS: The study involved 57 eyes after PPV and 57 healthy, control eyes of patients with primary RRD who had undergone successful PPV. The patients were divided into three subgroups depending on the endotamponade type, and were tracked with complete ophthalmological examination during a period of 6 months. PPV with internal tamponade silicone oil, 24% SF6 or 14% C3F8 was performed. The medical records were reviewed and compared between the groups. RESULTS: SD-OCT analysis detected a significant reduction of average GCL-IPL thickness and reduction of GCLIPL parameter in almost all examined sectors in the group with silicone oil endotamponade during all follow-up visits (P<0.05). The study showed a significant vision deterioration in the silicone oil group in comparison with the SF6 group on all follow-up visits (P<0.05). CONCLUSION: This is the first in vivo SD-OCT study describing the toxic effect of silicone oil endotamponade on GCL-IPL complex after PPV for RRD. The study confirmed that the GCL-IPL complex value can be a predictive factor for assessing the final visual acuity. SD-OCT should be recommended as a clinical standard in the followup treatment of patients after PPV for RRD, especially with the use of silicone oil endotamponade. Plain Language Summary: The aim of the study was to examine the impact of silicone oil on the ganglion cell complex. The ganglion cell is a neuron type located in the retina and takes part in transmitting visual information from the retina to the brain. Silicone oil is a substance used during eye surgery called pars plana vitrectomy. This tamponade type acts to hold the retina in a proper position. However, during the retinal detachment, which is the separation of the retina from the layer underneath, a reduction of neurons is observed. We decided to use a new technology device called SD-OCT to determine the thickness of the ganglion cell complex. We compared the impact of silicone oil with other tamponade types. We found that silicone oil causes a reduction in the ganglion cell complex. Moreover, we observed vision deterioration in eyes treated with a silicone oil tamponade. The research describes the toxic effect of silicone oil on ganglion cells. Ganglion cell values may be used to determine visual improvement or deterioration after eye surgery with silicone oil tamponade. SD-OCT should be a clinical standard in monitoring patients with silicone oil tamponade.

Ocular irritation and cyclosporine A distribution in the eye tissues after administration of Solid Lipid Microparticles in the rabbit model.

The aim of this study was to investigate the in vivo effect of Solid Lipid Microparticles (SLM), proposed for topical ocular administration of cyclosporine, on the rabbit eye. SLM carrier is an aqueous dispersion of lipid microparticles (20% w/w) with a size up to 15 µm. Cyclosporine was dissolved in the formulation in the concentration of 0.5 or 2.0% (w/w). Ocular tolerance of microsphere dispersion was assessed in rabbit model by the Draize eye test (SLM was compared with emulsion and oily solution), and cyclosporine distribution in ocular tissues was evaluated after multiple application of tested formulations (SLM dispersions, emulsions and oily solution) for 7 days. Good tolerance of cyclosporine-SLM formulation was demonstrated in the rabbit model. Concentration of cyclosporine in the precorneal tissues, such as cornea and conjunctiva, was much higher than the therapeutic value (8.4 ng/mg and 3.2 ng/mg, respectively). After SLM administration, the cyclosporine concentrations determined in the anterior ocular tissues, were also significantly higher compared to those obtained after the application of other tested carriers (emulsions and oily solution). The obtained results prove that the recognized SLM dispersions are safe formulations for ophthalmic use. It can be concluded that lipid microparticles are highly promising for an efficient ophthalmic drug delivery, when compared to other conventional dosage forms.

A 10-year monitoring of the eyesight in patients after kidney transplantation.

A pilot study of a 10-year analysis of the eyesight characteristics in patients after renal transplantation with a view to a later wider study of the same population.The study encompassed 50 eyes in 25 patients who underwent renal transplantation in the years 2007 and 2008. All patients underwent: visual acuity measurement, tonometry, slit lamp examination, and spectroscopic optical coherence tomography.Changes in the eyes observed during the 10-year observation period included mostly: cataract (48%), hypertensive angiopathy (28%), diabetic macular edema (16%), and glaucoma (16%). Ten years after the renal transplant visual acuity declined in 15 patients (60%). In 67% of those with eyesight deterioration the cause was cataract, while in patients with no changes in the eyesight (n = 10) cataract was diagnosed only in one. Patients with cataracts had been more often treated with cyclosporine, and that difference was statistically significant (73% vs 21%; P < .05). Comparing patients with hypertensive angiopathy with controls has shown that in the first group creatinine levels were statistically significantly higher (1.6 vs 1.16 mg/dL; P < .05). Patients with angiopathy had been also longer on renal replacement therapy before transplant (57 vs 26 months, P > .05), and this group included also statistically more persons after retransplantation (43% vs 5%, P < .05).Most frequent ophthalmological diagnoses in patients after a kidney transplant include cataract, diabetic retinopathy, and hypertensive angiopathy. Visual acuity deterioration was seen in 60% of patients and was mainly the effect of cataract progress. The effect of cyclosporine on cataract progress was significant. The diagnosis of hypertensive angiopathy corresponded with poorer function of the transplanted kidney.

[Pigmented paravenous retinochoroidal atrophy--a case report]. / Barwnikowy okolozylny zanik siatkówkowo-naczyniówkowy--opis przypadku.

Pigmented paravenous retinochoroidal atrophy is a rare dystrophy, which is usually asymptomatic and limited to the paravenous area. Due to the slow progression of eye fundus changes, typically without the macular involvement, pigmented paravenous retinochoroidal atrophy is not thought to be a vision-threatening ocular disease. We report a case of a 64-year-old man with sudden unilateral reduction of vision and binocular diplopia, diagnosed with bilateral pigmented paravenous retinochoroidal atrophy. The diagnosis was based on a characteristic fundus appearance, additionally accompanied by the bilateral mild macular changes, minor optic disc pallor and moderate retinal artery narrowing. The presented case posed a diagnostic challenge due to the sudden onset and atypical manifestation.

Five-month observation of persistent diabetic macular edema after intravitreal injection of Ozurdex implant.

AIMS: This retrospective analysis was aimed at evaluating the effectiveness of treatment of persistent diabetic macular edema with intravitreal injections of 0.7 mg dexamethasone implant Ozurdex. The study comprised three male patients (6 eyes). RESULTS: The average thickness of the retina at baseline was 632 µm, the medial BCVA was 0.8 logMAR, and corrected intraocular pressure was 13.7 mmHg. The maximum decrease in mean retinal thickness was observed at four weeks following the treatment and was 365 µm (-267 µm) and visual acuity improved by an average of two lines and was 0.6 logMAR. The largest increase in mean retinal thickness to average of 528 µm (+164 µm) occurred at 16 weeks and the average BCVA was 0.614 lines BCVA logMAR. In one eye, there was a steroid cataract development after the third dose of dexamethasone implant of 0.7 mg. CONCLUSIONS: The intravitreal dexamethasone implant treatment of patients with persistent diabetic macular edema in whom laser photocoagulation proved to be ineffective and as a result they required a monthly injection of anti-VEGF factors (Ranibizumab, Bevacizumab) may be a good alternative to extending the interval of injections. However, reinjections involve a high risk of developing poststeroid cataracts, which is not without significance in middle-aged patients.

Biomarkers in diabetic retinopathy and the therapeutic implications.

The main problem both in type 1 (T1DM) and type 2 (T2DM) diabetes is the development of chronic vascular complications encompassing micro- as well as macrocirculation. Chronic complications lower the quality of life, lead to disability, and are the cause of premature death in DM patients. One of the chronic vascular complications is a diabetic retinopathy (DR) which leads to a complete loss of sight in DM patients. Recent trials show that the primary cause of diabetic retinopathy is retinal neovascularization caused by disequilibrium between pro- and antiangiogenic factors. Gaining knowledge of the mechanisms of action of factors influencing retinal neovascularization as well as the search for new, effective treatment methods, especially in advanced stages of DR, puts special importance on research concentrating on the implementation of biological drugs in DR therapy. At present, it is antivascular endothelial growth factor and antitumor necrosis factor that gain particular significance.

Relationship between serum transforming growth factor ß 1 concentrations and the duration of type 1 diabetes mellitus in children and adolescents.

The aim of this study was to evaluate the relationship between serum transforming growth factor ß 1 (TGF- ß 1) concentrations and the duration of type 1 diabetes mellitus (T1DM) in children and adolescents. One hundred and sixteen patients with T1DM and 19 healthy controls were examined. Serum TGF- ß 1 concentrations were measured using the cytometric bead array (CBA). A positive association between the time of diabetes duration and higher serum TGF- ß 1 concentrations was observed. Similarly, the prevalence of microvascular complications, such as retinopathy and nephropathy, increased with the duration of diabetes. Logistic regression analysis showed that serum TGF- ß 1 concentrations and the duration of the disease are independent risk factors of microangiopathy development. Higher serum TGF- ß 1 concentrations were associated with a significant risk of microangiopathy development after 10 years of T1DM duration. In the successive years of the disease, the effect was even stronger. The results of our study indicate that serum TGF- ß 1 concentrations are one of the factors that may have an impact on the progression of vascular complications in children and adolescents with T1DM.

Serum concentrations of transforming growth factor-Beta 1 in predicting the occurrence of diabetic retinopathy in juvenile patients with type 1 diabetes mellitus.

In the present study, we have decided to evaluate if serum transforming growth factor-beta 1 (TGF- ß 1) concentrations may have diagnostic value in predicting the occurrence of diabetic retinopathy (DR) in juvenile patients with type 1 diabetes mellitus (T1DM). The study included 81 children and adolescents with T1DM and 19 control subjects. All study participants had biochemical parameters examined, underwent an eye examination, and 24-hour blood pressure monitoring. Moreover, serum concentrations of TGF- ß 1 were measured. The group of patients with T1DM and nonproliferative diabetic retinopathy (NPDR) had statistically significant higher serum levels of TGF- ß 1 (P = 0.001) as compared to T1DM patients without retinopathy as well as the healthy control subject. The threshold serum TGF- ß 1 concentrations which had a discriminative ability to predict the presence of DR were calculated using the receiver operating characteristic (ROC) curves analysis and amounted to 443 pg/ml. The area under the ROC curve (AUCROC) was 0.80, and its population value was in the range of 0.66 to 0.94. The sensitivity and specificity were calculated to be 72% and 88%, respectively. Our results suggest that TGF- ß 1 serum concentrations may be an additional parameter in predicting the occurrence of DR in juvenile patients with T1DM.

Evaluation of corneal endothelium in children and adolescents with type 1 diabetes mellitus.

PURPOSE: To evaluate the systemic and local factors that contribute to the damage of endothelial cells in diabetic patients and to compare the endothelial structure of the cornea in diabetic and nondiabetic patients. MATERIALS AND METHODS: The endothelial cell density (ECD) and central corneal thickness (CCT) were investigated in 123 eyes of type 1 diabetic patients and in 124 eyes of nondiabetic patients. The mean diabetic patients age was 15.34 ± 3.06 years versus 14.58 ± 2.01 years in the control group. The mean duration of diabetes was 8.02 ± 3.9 years. The corneal endothelium was imaged by the Topcon SP-2000P. RESULTS: The mean ECD in diabetic eyes was 2435.55 ± 443.43 cells/mm(2) and was significantly lower than in control group (2970.75 ± 270.1 cells/mm(2)). The mean CCT was 0.55 ± 0.03 mm in diabetic group versus 0.53 ± 0.033 mm in control group. ECD and CCT significantly correlated only with duration of diabetes. There was no correlation between ECD and CCT and patient age, sex, HbA1C level, and plasma creatinine level. CONCLUSIONS: ECD is decreased and CCT is increased in children and adolescents with diabetes mellitus. Duration of diabetes is the factor that affects ECD and CCT.

Threshold serum concentrations of tumour necrosis factor alpha (TNFα) as a potential marker of the presence of microangiopathy in children and adolescents with type 1 diabetes mellitus (T1DM).

The aim of this study was to estimate the threshold serum concentrations of advanced glycation end products (AGEs) and their soluble receptors (sRAGE) as well as tumour necrosis factor alpha (TNFα), vascular endothelial growth factor(165) (VEGF(165)) and interleukin-12 (IL-12) in predicting the occurrence of microangiopathy in children and adolescents with type 1 diabetes mellitus (T1DM). We studied 88 children and adolescents (age range: 6-20 yrs) with T1DM and 32 control subjects (age range: 7-20 yrs). All study participants had their daily urinary albumin excretion, HbA1c and serum creatinine levels measured, and underwent an eye examination and 24-h blood pressure monitoring. Moreover, serum concentrations of AGEs, sRAGE, TNFα, VEGF(165) and IL-12 were measured. In order to calculate the threshold values of the studied parameters, the Receiver Operating Characteristic (ROC) curve analysis was used. The results of our study have shown that among all the studied parameters a discriminative ability was found for TNFα, VEGF(165), duration of the disease, serum AGEs concentrations and daily urinary albumin excretion. However, the highest value of the area under the ROC curve (AUC(ROC)) in predicting the occurrence of diabetic microangiopathy was found for serum TNFα concentrations with its threshold value of 1.7 pg/ml [AUC(ROC) = 0.88 (95% CI: 0.79-0.97)]. The sensitivity and specificity for this variable was at the level of 85.7% and 94.3%, respectively. In conclusion, according to our results serum TNFα concentrations over 1.7 pg/ml may point to the presence of diabetic microangiopathy in children and adolescents T1DM.

[Idiopathic, bilateral panuveitis--efficacy of general steroidotherapy and intravitreal dexamethason injection. Case report]. / Idiopatyczne, obustronne zapalenie blony naczyniowej--efektywnosc steroidoterapii ogólnej oraz iniekcji doszklistkowej dexamethazonu. Opis przypadku.

This paper presents the comparison of different schemes of two kinds of general steroidotherapy in one case of idiopathic, bilateral panuveitis and effectiveness of intravitreal dexamethason to one eye. The effectiveness of the treatment was mainly concerned on best corrected visual acuity, the illustration of macular retinal area in OCT (optic coherence tomography), where the reduction of cystic macular oedema was detected and the pictures of fluorescein angiography. The observation and the treatment has lasted for 3 years. At the beginning, the patient received general steroidotherapy (medrol), which lasted seventy four days. Improvement of clinical symptoms lasted only four months. The similar therapy the patient obtained once more. The symptoms of illness returned, after the same period of time. Then the intravitreal dexamethason (Posurdex) was injected. The better visual acuity and improvement of clinical symptoms was observed for three months. The patient couldn't obtain posurdex once more. The physician decided to apply the different scheme of general steroidotherapy. The observation after finished treatment, lasted over one year. Clinical symptoms of the illness retracted. The fluorescein angiography of the eyes were correct and the structure of the macular retinal area presented in optical coherence tomography were regular. The observation of the patient is continued.

Expansion of CD14+CD16+ monocytes producing TNF-α in complication-free diabetes type 1 juvenile onset patients.

We concentrated on the complication-free phase of juvenile onset type 1 diabetes mellitus (T1DM) searching for associations between concentration of inflammatory factors TNF-α, CRP and VEGF and two monocyte subsets the CD14(++)CD16(-) and CD14(+)CD16(+). We analysed a randomly selected group of 150 patients without complications (disease duration 2.74 ± 2.51 years) at the start of the project and 5 years later. They were compared with 24 patients with retinopathy (6.53 ± 3.39 years of disease) and 30 healthy volunteers. Our results indicate that in the complication-free period the concentration of TNF-α significantly increased and continued to increase after retinopathy was established. After 5 years the percentage and absolute number of CD14(+)CD16(+) monocytes doubled in complication-free patients. Our study indicates that the size of CD14(+)CD16(+) monocyte subset may be used alternatively to CRP values as an indicator of inflammation grade. Our results imply the necessity of trials using anti-TNF-α therapy in the complication-free phase of the disease.

Decreased angiogenin concentration in vitreous and serum in proliferative diabetic retinopathy.

Diabetic retinopathy is the most common cause of vision loss in young adults in developed countries. The disease therapy with anti-vascular endothelial growth factor (VEGF) agents gives some positive results, but is associated with retinal ischemia and vasoconstriction. Therefore, determination of factors involved in the physiological and pathological angiogenesis in the diabetic eye is of great importance for understanding of the pathogenesis of diabetic retinopathy and its effective treatment. Previously, we found that diabetic patients were characterized by increased serum concentration of VEGF, but decreased levels of other proangiogenic factor-angiogenin. The involvement of VEGF in pathogenesis of diabetic retinopathy is well established, but there is lack of data regarding angiogenin in retinopathy. Therefore, in the present study we measured angiogenin concentration in vitreous and serum samples of the patients with type 1 diabetes to determine its role in diabetic retinopathy. In addition, in each time, we compared the level of angiogenin with level of VEGF as a known factor involved in the pathogenesis of the disease. Angiogenin was found to be significantly more abundant in serum than in vitreous in both diabetic groups. In addition, patients with retinopathy had twofold lower vitreous angiogenin levels than diabetic individuals without complications. On the contrary, vitreous concentration of VEGF was dramatically increased only in participants with retinopathy. Patients without diabetic complications had significantly lower VEGF levels in vitreous than in serum and were characterized by high local and systemic concentration of angiogenin. These data suggest a local imbalance between two proangiogenic factors-VEGF and angiogenin in retinopathy. Low vitreous concentration of angiogenin in diabetic patients suggests that this factor is not responsible for pathological neovascularization in diabetic eye. Further studies will elucidate if angiogenin can be used to improve the insufficient angiogenesis in diabetes and prevent retinal ischemia after retinopathy treatment with anti-VEGF agents.

Increased spontaneous production of VEGF by CD4+ T cells in type 1 diabetes.

In the present study we report that CD4(+) T cells from patients with type 1 diabetes produce significantly higher amounts of VEGF than respective cells from the healthy individuals. Among CD4(+) T cells memory subsets were the main source of VEGF. In addition, memory CD4(+) T subsets were the most numerous in patients with diabetic retinopathy (DR). DR was also characterized by significant increase of VEGF concentration in serum and culture supernatants. Hence, these data indicate that there is a sustained spontaneous production of VEGF by CD4(+) T cells in type 1 diabetes, which is additionally exacerbated in DR. In our opinion alterations in the proportions of CD4(+) T cell subsets and their VEGF production may be a useful tool for early assessment of the risk of DR onset and progression.

Glycemic control influences serum angiogenin concentrations in patients with type 2 diabetes.

OBJECTIVE: Because diabetes is the most frequent factor responsible for microvascular and macrovascular disease, we investigated angiogenin serum levels within the diabetic patient group. RESEARCH DESIGN AND METHODS: We investigated 49 patients who met the criteria to be in the diabetic group. Forty nondiabetic patients were included in the control group. We set A1C <7% as well-controlled diabetes. Serum angiogenin level was measured using the enzyme-linked immunosorbent assay method. RESULTS: Serum angiogenin levels of poorly controlled patients with type 2 diabetes were significantly lower than those of group with well-controlled diabetes (361.23 +/- 126.03 ng/ml vs. 446.37 +/- 134.10 ng/ml; P = 0.001). Moreover, they were characterized by a significantly longer duration of the disease (P = 0.006), higher BMI (P = 0.0003), and higher systolic blood pressure (P = 0.01). Levels of total cholesterol, triglycerides, LDL, and HDL were not significantly different in both groups. CONCLUSIONS: Patients with poorly controlled type 2 diabetes (A1C >7%) have lower angiogenin levels than patients with well-controlled diabetes.

Membrane potential of CD4+ T cells is a subset specific feature that depends on the direct cell-to-cell contacts with monocytes.

CD4(+) T cells can be divided into three subsets: naive (Tn), central memory (Tcm), and effector memory (Tem) lymphocytes. These subpopulations differ in phenotype, migratory capacity, pattern of secreted cytokines, and activation threshold. T-cell activation is associated with changes in membrane potential, which provide an electrical driving force for calcium entry into the cytosol. These phenomena were shown to precede lymphocyte proliferation, cytokine synthesis, migration, and apoptosis. Hence the aim of the study was the analysis of these early activation events in the subsets of CD4(+) T cells. We measured the membrane potential and intracellular calcium concentration ([Ca(2+)](i)) in CD4(+) Tn, Tcm, and Tem cells as well as the dependency of these parameters in CD4(+) T cells on their cell-to-cell contacts with other leukocyte subsets. The data indicate that membrane potential of CD4(+) T cells is a subset specific feature maintained by direct contact with monocytes. In addition, monocytes were found to control Ca(2+) influx in CD4(+) T cells.

The potential use of the eye temperature decrease in determining the time of death in the early postmortem period: studies in pigs.

The experiment was performed in pigs during which the postmortem body temperature was continuously measured. The study was focused on a period of time starting 0.5 hours after the animals' termination and consisted in computer recording of the cooling process of the eyeball (vitreous humor), orbit soft tissues, muscles, and the rectum. The experiment revealed the absence of a plateau phase of temperature decrease in eyeball and orbit soft tissues contrary to muscles and rectum, what demonstrates that temperature measurements in the eye could become an alternative method of determining the time of death during the initial phase of postmortem cooling. During this early postmortem period determination of the time of death based on measurements of the rectum or muscles temperatures due to existing plateau is not possible.