In vitro vitamin K3 effect on conjunctival fibroblast migration and proliferation.

PURPOSE: To evaluate the dose effect of vitamin K3 on wound healing mechanisms. METHODS: Conjunctival fibroblasts were incubated for 24 hours. An artificial wound was made and the cells were incubated with fresh medium plus doses of vitamin K3 to be tested. Wound repair was monitored at 0, 18, 24, and 48 hours. Proliferation was measured in actively dividing cells by [(3)H]thymidine uptake. Six different groups were tested: group 1/no drugs added, group 2/ethanol 0.1%, group 3/vitamin K3 1 mg/L, group 4/vitamin K3 2 mg/L, group 5/vitamin K3 4 mg/L, and group 6/vitamin K3 6 mg/L. Each experiment was carried out in triplicate and 4 times. RESULTS: There were no differences among groups at the initial time. In vitro wound repair was slower in groups 4, 5, and 6. There were no differences between control and ethanol groups and between control and vitamin K3 1 mg/L groups. Fibroblast mitogenic activity was statistically decreased in all vitamin K groups; statistical differences were found among vitamin K3 1 mg/mL and higher doses too. In groups 5 and 6, cellular toxicity was presented. CONCLUSIONS: Vitamin K3 is able to inhibit fibroblast proliferation. Vitamin K3 2 mg/L or higher doses inhibit wound healing repair, exhibiting cellular toxicity at 4 and 6 mg/L.

Pentobarbital intoxication as a potential underlying cause for electrocution in a bearded vulture.

A bearded vulture (Gypaetus barbatus) found dead in northern Spain presented external lesions consistent with electrocution as the cause of death. During forensic examination, macroscopic lesions suggested potential comorbidity, so samples were collected for molecular and toxicological analyses. Gastric content and liver were analysed for toxic substances, and pentobarbital (a common pharmaceutical used for euthanasia in domestic animals) was detected at a concentration of 37.3 and 0.05 µg/g, respectively. Other toxicological, viral and endoparasite analyses (avian malaria, avian influenza and flaviviruses) were negative. Thus, although the cause of death was electrocution, pentobarbital intoxication likely impaired the equilibrium and reflexes of the individual, possibly causing the bird to contact energized wires that it would not have otherwise. These results underline the importance of comprehensive analysis of forensic cases of wildlife deaths and reveal barbiturate poisoning as an additional threat for the conservation of the bearded vulture in Europe.

Malaria quantitative POC testing using magnetic particles, a paper microfluidic device and a hand-held fluorescence reader.

A point-of-care (POC) device is reported for highly sensitive and selective detection of Plasmodium falciparum lactate dehydrogenase (Pf-LDH), a biomarker of malaria infection, based on a single-step magneto-immunoassay, a single-use microfluidic paper device and a customized hand-held fluorescence reader. The single-step magneto-immunoassay consists in a single 5-min incubation of immuno-modified magnetic particles (c-MAb-MPs), biotinylated detection antibody (bd-MAb), and an enzymatic signal amplifier (Poly-HRP). After on-chip MP concentration and washing, signal generation is achieved by adding a fluorescent enzymatic substrate (QuantaRed). Fluorescence signal is measured using a low-cost customized, portable, and sensible fluorescent detector. The POC affords quantitative Pf-LDH detection in <20 min, with a detection limit of 0.92 ng mL-1 (equivalent to 4.6 parasites µL-1). Furthermore, Pf-LDH quantitation in clinical samples correlates with that provided by the reference ELISA, is more sensitive than a commercial rapid diagnostic test (RDT) and entails little user intervention. These results show that fluorescent paper-based microfluidic devices can be exploited to simplify magneto-immunoassay handling, taking this type of test closer to the requirements of POC testing.

[Correlation between standard automated perimetry global indices and Heidelberg Retina Tomograph II parameters]. / Correlación entre los índices globales de la perimetría automatizada convencional y los parámetros topográficos papilares (Heidelberg Retina Tomograph II).

PURPOSE: To correlate the optic nerve head topographic parameters measured by the Heidelberg Retina Tomograph II (HRT) with the perimetric indices of standard automated perimetry (SAP). METHODS: This study included 101 normal subjects, 247 ocular hypertensive eyes (increased intraocular pressure with normal SAP) and 102 glaucomatous subjects (IOP above 21 mm Hg and abnormal standard automated perimetry). Only one eye was randomly chosen from each subject for the study. The visual field was evaluated by means of Humphrey Field Analyzer (24-2 full threshold strategy). The HRT II (Heidelberg Engineering) was used to acquire and measure the optic disc topographic parameters. Pearson correlations between topographic data and perimetric indices were performed for the total sample and each group of patients. The distribution of values obtained in the samples was normal. RESULTS: A significant correlation was found between several optic disc parameters and the global indices of SAP. Rim area, rim volume, cup/disc area ratio, rim/disc area ratio, cup shape measurement, RNFL cross-sectional area, and discriminant functions FSM and RB, showed the strongest correlation with the visual field indices in the total and glaucoma groups (RIM AREA: total group: r=0.32; p=4.14x10(-11)/glaucoma group: r=0.28; p=0.004. RIM VOLUME: total group: r=0.26; p=1.55x10(-7)/glaucoma group: r=0.26; p=0.006). The ocular hypertensive group showed few significant correlations. CONCLUSIONS: The correlations found between standard automated perimetry and HRT defined topographic parameters allow a better understanding of glaucomatous damage and make decision-making easier.

[Pseudophakic cystoid macular edema. Assessment with optical coherence tomography]. / Edema macular quístico pseudofáquico. Detección mediante "optical coherence tomography".

PURPOSE: To determine the incidence of cystoid macular edema (CME) by means of clinical evaluation and subclinical assessment by means of ocular coherence tomography (OCT), and to compare the incidence between diabetic and non-diabetic groups of patients. METHODS: Prospective study of 260 consecutive cataract surgeries operated from September 2004 to March 2005. The procedures were performed by means of phacoemulsification plus intraocular acrylic lens implantation. Group A: 208 eyes of non-diabetic patients; Group B: 42 eyes of patients with diabetes and Group C: 10 eyes of diabetic patients with macular edema that received an intravitreal injection of triamcinolone at the end of surgery. Postoperative follow-up visits were performed 6 days (basal visit), 5 weeks and 12 weeks after surgery. Each visit included posterior pole biomicroscopy and OCT. RESULTS: Central macular thickness measured by OCT was significantly increased in group B compared with group A (241.6 versus 204.6 microm; p<0.001). No clinical evidence of CME was found in group A, although 4 eyes (1.92%) showed macular thickness equal to or greater than 43.74 microm (2 standard deviations of the basal value for group A). In group B, clinical evidence of CME was found in 6 eyes, with decreased visual acuities (14.2%). The differences between these groups were statistically significant (p<0.001). The 10 eyes that received an intravitreal injection of triamcinolone after the surgical procedure showed a mean decrease in central retinal thickness of 77 microm after 12 weeks postoperative. CONCLUSIONS: This study has shown a low incidence of clinical CME. OCT showed increased macular thickness in both groups of patients in a small percentage of cases, and significantly increased macular thickness in diabetic patients.

Retinal Sensitivity in Patients with Type I Diabetes without Retinopathy or with Minor Retinal Changes.

Objective: To evaluate the ability of short-wavelength automated perimetry (SWAP) for the detection of visual impairment in patients with type I diabetes without retinopathy or with minor retinal vascular changes. Design: Comparative cross-sectional study. Participants: 30 eyes of 30 healthy subjects and 73 eyes of 73 patients with type I diabetes mellitus were studied. Methods: Ophthalmic examination of diabetic patients showed no retinopathy or minimal changes (less than 5 microaneurisms in each eye) with no previous laser treatment. All patients were examined by means of the SWAP 24-2 strategy. Mean Deviation (MD) and Pattern Standard Deviation (PSD) were compared between both groups. Results: There were differences in the clusters of altered points between both groups (p=0.004). SWAP MD was lower in the diabetic group than in the controls (-2.89 dB vs. -0.20 dB, p<0.001). SWAP PSD also differed between both groups (2.50 dB in control group, 3.12 dB in the diabetic group, p=0.003). In the diabetic group, mean period from the onset of diabetes was 12.6±6.7 years and minimal vascular changes were observed in the retina of 18 eyes (24.7%), while 55 had no lesions (75.3%). No differences in SWAP changes were found between patients without and with minimal diabetic retinopathy. Conclusions: Retinal sensitivity assessed by SWAP is depressed in patients with type I diabetes regardless of the presence of retinal vascular changes.