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.

Perception of patients with retinal pathology on aspects of visual function and their management.

BACKGROUND AND OBJECTIVE: Age-related macular degeneration (AMD) and diabetic retinopathy (DR) are highly prevalent. OBJECTIVE: To explore perceptions of patients with AMD or DR about the impact of the disease and treatment on their daily living activities. MATERIALS AND METHODS: Semi-structured interviews with a questionnaire developed from validated patient reported outcomes questionnaires. The questionnaire consisted of 19 questions about the disease and 9 about the treatment. The questions (items) were answered on a scale from 1 to 9. In addition, the patient interviewed was invited to make free comments on each question. Nine patients with AMD and 9 with DR were interviewed by videoconference or telephone call. A quantitative analysis of the responses and a qualitative analysis of the comments were carried out. RESULTS: The most relevant item for patients with AMD or DR is "Recognize people when they are nearby", and "Read text in normal size font in a newspaper or book", followed, in patients with AMD, by "Do things what you would like" and, in patients with DR, "Feeling frustrated by the vision problems." Regarding the treatment, the most relevant aspects for both groups is that the treatment works and receiving appropriate information before and after the treatment. The qualitative comments were focused to the disease, the treatment, and to the role of doctors and the health system. CONCLUSION: Quantitative responses and free comments can be useful to improve the care of patients with AMD or DR by physicians and the health system.

[Corpus cavernosum fracture: the ultrasound in the emergency diagnosis]. / Fractura de cuerpo cavernoso: la ecografía en el diagnóstico de urgencia.

OBJECTIVE: The main objective of this work is to study usefulness of ultrasound (US) for the emergency diagnosis of the penile fracture. MATERIAL AND METHODS: We reviewed all the penile US studies registered in our Emergency Department between July 2007 and August 2009 with suspicion of a corpus cavernosum fracture. We compared US findings (subcutaneous haematoma, peri-albuginea haematoma, albuginea rupture and Bucks fascia rupture), and the clinical progress, with those of the surgery. We studied sensitivity, specificity, positive predictive value and negative predictive value of US in the diagnosis of rupture of the tunica albuginea. The epidemiological data of all the cases reviewed were collected. RESULTS: Twelve patients with a mean age of 37.8 years were reviewed. The most common cause of injury was sexual intercourse. A subcutaneous haematoma was found in nine patients, a peri-albuginea hematoma in eleven of the cases, and an albuginea rupture was seen in six of them. We found no Bucks fascia rupture. Seven patients underwent surgical treatment and in the remaining five patients, treatment was conservative. Clinical progress was good in all cases. US, as an emergency test to diagnose albuginea fracture gave 0.83 sensitivity, 1 specificity, 1 positive predictive value and 0.83 negative predictive value. CONCLUSION: We believe that US is a useful procedure in the diagnosis of acute penile fracture and that it could be proposed as the diagnostic method of choice to confirm the clinical suspicion of penile fracture. Identifying the exact site of a tear in the tunica albuginea facilitates the surgical procedure.

Retinal degeneration in two lines of transgenic S334ter rats.

Aim of this study was to examine synaptic connectivity changes in the retina and the location and rate of apoptosis in transgenic S334ter line-3 and line-5 rats with photoreceptor degeneration. Heterozygous S334ter-line-3 and line-5 at P11-13, P30, P60, P90 and several control non-dystrophic rats (Long Evans and Sprague-Dawley) at P60, were studied anatomically by immunohistochemistry for various cell and synaptic markers, and by PNA and TUNEL label.- S334ter line-3 exhibited the fastest rate of degeneration with an early loss of photoreceptors, with 1-2 layers remaining at P30, and only cones left at P60. Line-5 had 4-5 layers left at P30, and very few rods left at P60-90. In both lines, horizontal cell processes (including dendrites and axon) were diminished at P11-13, showing gaps in the outer plexiform layer (OPL) at P60, and at P90, almost no terminal tips could be seen. Bipolar cells showed a retraction of their dendrites forming clusters along the OPL. Synaptic terminals of A-II amacrine cells in the IPL lost most of their parvalbumin-immunoreactivity. The apoptosis rate was different in both lines. Line-3 rats showed many photoreceptors affected at P11, occupying the innermost part of the outer nuclear layer. Line-5 showed a lower number of apoptotic cells within the same location at P13. In summary, the S334ter line-3 rat has a faster progression of degeneration than line-5. The horizontal and bipolar terminals are already affected at P11-P13 in both models. Apoptosis is related to the mutated rhodopsin transgene; the first photoreceptor cells affected are those close to the OPL.

[Is H1N1 flu different from seasonal flu on initial plain chest films?]. / Gripe A ¿diferente de la gripe estacional?

OBJECTIVES: To determine whether there are differences in the findings on the initial plain chest films of patients with H1N1 influenza and those of patients with flu symptoms during the flu season. MATERIAL AND METHODS: All patients underwent plain-film chest radiography in the Emergency Department for flu symptoms; 95 patients had H1N1 influenza confirmed between July 2009 and December 2009 and 95 patients were attended for symptoms of seasonal flu in January 2009. We analyzed the views obtained, the distribution and location of the radiologic findings, and patients' age, sex, and previous disease. RESULTS: Patients with H1N1 influenza were younger than those with seasonal flu symptoms (mean 40.2 vs 50.9 years; p<0.001) and fewer had prior disease (48 vs. 63; p<0.001). Plain films were acquired with patients in the standing position in 75 patients in the H1N1 group and in 77 in the seasonal flu group; pathological findings were present in nearly 50% of the patients in each group. The most common findings in the H1N1 group were multifocal patchy consolidations (41.2%; p<0.001) and peribronchial-vascular opacities (16.3%), whereas in the seasonal flu group the most common finding was consolidation in a single lobe (43.9%). CONCLUSION: We found significant differences between the radiologic findings of patients with H1N1 influenza (severe) and those of patients with symptoms of flu during the flu season: the incidence of multifocal patchy consolidation was greater in H1N1 patients and H1N1 patients were younger.

Should we consider paranasal and chest computed tomography in severe asthma patients?

BACKGROUND: It is essential to recognize and treat findings that can simulate or worsen symptoms to improve asthma control and thereby to reduce costs. Guidelines highlight a paranasal (PS) and chest computed tomography (CT) scan as a tool for disease evaluation and, although they suggest its indication in patients whom presentation is atypical, there are not well-defined criteria. OBJECTIVES: To describe the most common findings in the PS and chest CT in severe asthma patients and to analyse the characteristics of asthmatics with the finding of nasal polyps or bronchiectasis. METHODS: We retrospectively reviewed the medical records of 161 adults with confirmed severe asthma who had undergone to PS and/or chest CT. Clinical data from their electronic health record and the findings from a PS and/or chest CT within the last five years were collected. RESULTS: In the PS CT, 70.5% of patients presented mucous thickening and 46.7% presented nasal polyps. Both findings were associated with male gender and level of blood eosinophils. In chest CT, 28% of individuals showed atelectasis, 16.5% air trapping, 17.7% affectation of the small airway, 11.6% pulmonary infiltrates and 10.4% emphysema. Bronchiectasis were identified in 60.4% of subjects, who were older and had poorer lung function. CONCLUSION: Paranasal and thoracic computed tomography are important tools in the treatment of severe asthma because they allow us to detect highly prevalent findings in this disease that can lead to poorer control of it.

[Retinal toxicity following chloroquine therapy]. / Toxicidad retiniana secundaria a tratamiento con cloroquina.

CASE REPORT: Two cases of bilateral ocular toxicity following chloroquine therapy in two women with systemic lupus erythematosus are presented. DISCUSSION: Retinal toxicity due to chloroquine is an uncommon iatrogenic disease. It is dose-dependent. New cases are still being described despite the application of ophthalmic screening techniques.

Partial preservation of rod and cone ERG function following subretinal injection of ARPE-19 cells in RCS rats.

We quantified rod- and cone-related electroretinogram (ERG) responses following subretinal injections of the human-derived retinal pigment epithelial (hRPE) cell line ARPE-19 at age P23 to prevent progressive photoreceptor loss in the Royal College of Surgeons (RCS) rat. Culture medium-injected eyes served as sham controls. At P60, in comparison with sham-injected eyes, all recordings from hRPE-injected eyes showed preserved scotopic a- and b-waves, oscillatory potentials, double-flash-derived rod b-waves and photopic cone b-waves, and flicker critical fusion frequencies and amplitudes. Although the actual preservation did not exceed 10% of a-wave and 20% of b-wave amplitude values in non-dystrophic RCS and deteriorated rapidly by P90, rod- and cone-related ERG parameters were still recordable up to P120 unlike the virtually unresponsive sham-injected eyes.

Role of computed tomography and magnetic resonance imaging in aortobronchial fistula diagnosis following aortic coarctation reparative surgery. Report of two cases.

Aortobronchial fistula (ABF) is an uncommon complication of aortic coarctation repair which may occur years after successful coarctation correction. It is invariably fatal if not diagnosed and treated. ABF diagnosis poses a challenge for clinicians and radiologists because of the difficulty in detecting the fistula and the risks associated with some of the diagnostic procedures. Two cases of ABF occurring 1 and 20 years after reparative surgery of aortic coarctation are reported. The advantages and disadvantages of different imaging procedures for the evaluation of patients with suspected ABF are reviewed and the role of computed tomography angiography and magnetic resonance imaging is underlined.

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.

Enhanced cone dysfunction in rats homozygous for the P23H rhodopsin mutation.

The heterozygous P23H transgenic rat is a model of autosomal dominant retinitis pigmentosa, in which a mutation in the rhodopsin gene leads to a rapid loss of rods and a more protracted loss of cones. It has been suggested that rods play an essential role in preserving cones. We tested this hypothesis by examining whether higher levels of dysfunctional rhodopsin in rats homozygous for the P23H mutation would result in exacerbated cone dysfunction when compared with heterozygous P23H rats. Electroretinogram (ERG) responses were recorded from P21 to P250 in Sprague-Dawley (SD) and homozygous P23H rats. Both scotopic and photopic intensity response ERGs were severely depressed already at P21 when compared with age-matched SD rats. Furthermore, flicker amplitudes and critical fusion frequencies were also lower in P23H compared with SD rats at P21. Scotopic and photopic intensity responses as well as flicker amplitude and critical fusion frequencies declined rapidly up to P60, reaching a steady state that was maintained up to P200. We conclude that in rats homozygous for P23H rhodopsin mutations, the severe loss of rod function already seen by P21 is accompanied by substantial cone functional loss at that age. While rod-related responses are more severely affected than cone-related responses at all ages, their actual rate of decline with age is surprisingly similar. Both undergo a biphasic temporal pattern of decline: very rapid (P21-P60) followed by very slow (P60-P200) deterioration in response parameters, implying a tight link between rod and cone functional deterioration.

Measuring the cone contribution to the ERG b-wave to assess function and predict anatomical rescue in RCS rats.

Subretinal injections of human retinal pigment epithelial (RPE) cells early in the course of retinal degeneration in Royal College of Surgeons (RCS) rats can rescue photoreceptors. Fourteen injected animals were studied using a double flash electroretinogram (ERG): 10 were examined longitudinally and four terminally with immunohistochemistry. The proportion of cone contribution to the ERG b-wave rather than the absolute size of isolated cone response proved to be a reliable indicator of function over time and a predictor of the proportion of cones identified anatomically in the area of optimal photoreceptor rescue.

Regressive and reactive changes in the connectivity patterns of rod and cone pathways of P23H transgenic rat retina.

We have used the P23H line 1 homozygous albino rat to study how progressive photoreceptor degeneration affects rod and cone relay pathways. We examined P23H retinas at different stages of degeneration by confocal microscopy of immunostained sections and electroretinogram (ERG) recordings. By 21 days of age in the P23H rat retina, there is already substantial loss of rods and reduction in rod bipolar dendrites along with reduction of metabotropic glutamate receptor 6 (mGluR6) and rod-associated bassoon staining. The cone pathway is relatively unaffected. By 150 days, when rods are absent from much of the retina, some rod bipolars remain and dendrites of rod and cone bipolar cells form synaptic complexes associated with cones and horizontal cell processes. These complexes include foci of mGluR6 and bassoon staining; they develop further by 270 days of age. Over the course of degeneration, beginning at 21 days, bipolar axon terminals atrophy and the inner retina undergoes further changes including a reduced and disorganized AII amacrine cell population and thinning of the inner plexiform layer. Electroretinogram (ERG) results at 23 days show reductions in a-wave amplitude, in rod and cone-associated b-waves (using a double flash paradigm) and in the amplitude of oscillatory potentials (OPs). By 38 days, rod scotopic a-wave responses and OPs are lost. B-wave amplitudes decline until 150 days, at which point they are purely cone-driven and remain stable up to 250 days. The results show that during the course of photoreceptor loss in the P23H rat, there are progressive degenerative changes, particularly in the rod relay pathway, and these are reflected in the changing ERG response patterns. Later reactive changes involving condensation of cone terminals and neurotransmitter receptors associated with rod and cone bipolar dendrites and with horizontal cell processes suggest that at this stage, there are likely to be complex changes in the relay of sensory information through the retina.

Long-term management of homozygous protein C deficiency: replacement therapy with subcutaneous purified protein C concentrate.

We present the case of a full-term newborn in whom purpura fulminans developed shortly after birth. A diagnosis of homozygous protein C deficiency was established based upon undetectable plasma protein C activity and antigenemia in the newborn infant, and was later confirmed by protein C gene analysis. Specific replacement therapy with intravenous protein C concentrate was started 9 days after birth. This rapidly led to the complete regression of cutaneous lesions and consumption coagulopathy. After stabilization, oral anticoagulation was initiated in association with prophylactic treatment with intravenous protein C concentrate. However, oral anticoagulation was finally abandoned as the patient presented several thrombotic and hemorrhagic episodes clearly related to difficulties with anticoagulation. Due to the hazards related to prolonged venous access, we are currently using subcutaneous infusion of protein C concentrate for the long-term management of this condition, with satisfactory results.

Penicillium brevicompactum as the cause of a necrotic lung ball in an allogeneic bone marrow transplant recipient.

The non-Candida, non-Aspergillus fungal infections are being reported with increasing frequency in BMT patients. One of these agents is Penicillium which has rarely been implicated as a pathogen in these patients. Only a few cases of isolated fungemias have been reported to date. We present the first documented case of invasive lung infection due to Penicillium brevicompactum in an allogeneic BMT recipient. As this case shows, the diagnosis of non-Candida, non-Aspergillus fungal infections may be incorrect if only histologic findings are available, mainly because misdiagnosis with other more common fungus can occur. A positive culture is required in order to make an accurate diagnosis.

Long-term liver dysfunction after allogeneic bone marrow transplantation: clinical features and course in 61 patients.

This retrospective study has aimed at determining the prevalence, aetiology and clinical evolution of chronic liver disease (CLD) after allogeneic bone marrow transplantation (BMT). A total of 106 patients who had been transplanted in a single institution and who had survived for at least 2 years after BMT were studied. The prevalence of CLD was 57.5% (61/106). In 47.3% of cases more than one aetiopathogenic agent coexisted. The causes of CLD were iron overload (52.4%), chronic hepatitis C (47.5%), chronic graft-versus-host disease (C-GVHD) (37.7%), hepatitis B (6.5%), non-alcoholic steatohepatitis (NASH) (4.9%), autoimmune hepatitis (AIH) (4.9%) and unknown two (3.3%). Twenty-three patients with iron overload underwent venesections which were well tolerated. An improvement in liver function tests (LFTs) was observed in 21 (91%) patients. All six patients with siderosis as the only cause of CLD normalized LFT as well as three patients with HCV infection. Clinical evolution was satisfactory for patients with GVHD, AIH, NASH and hepatitis B. At the last visit 23 patients continued with abnormal LFTs, and 19 of them were infected by the HCV. A sustained biochemical and virologic response was achieved in only one case out of six patients with CHC who received interferon. We have found that CLD is a common complication in long-term BMT survivors. The aetiology is often multifactorial, iron overload, CHC and C-GVHD being the main causes. The CLD followed a rather 'benign' and slow course in our patients as none of them developed symptoms or signs of liver failure and we did not observe an increase in morbidity or mortality in these patients, but a longer follow-up is necessary in HCV infected patients based on the natural history of this infection in other populations.

Clinical results of a stringent policy on prophylactic platelet transfusion: non-randomized comparative analysis in 190 bone marrow transplant patients from a single institution.

The threshold for prophylactic platelet transfusion remains controversial. Usually the decision is based on arbitrary numerical criteria. The classical 20 x 10(9)/l trigger could be safely reduced with considerable benefits. Few studies have evaluated the clinical impact of stringent policies. We have performed a retrospective analysis comparing major haemorrhages during hospitalization in 190 patients undergoing BMT in two different periods. In 87 patients transplanted from 1990 to 1991, the 20 x 10(9)/l trigger was used for prophylactic platelet transfusion. In 103 other patients transplanted from 1993 to 1994, we adopted a stringent prophylactic policy: < 10 x 10(9)/l for stable patients and < 20 x 10(9)/l when higher platelet consumption factors were present. In the stringent group, 12 patients presented 13 major haemorrhages and four died from haemorrhage. In the classical group 12 patients presented 14 major haemorrhages and three died from haemorrhage. Platelet consumption factors were present in 12 of 13 haemorrhages in the stringent group and in 12 of 14 in the classical group. By contrast, stable patients presented less haemorrhages (2/14 and 1/13, respectively). A statistically significant reduction in the use of platelet units was observed when comparing both groups: the median of platelet units administered in the first 100 days of transplant was 73 (3-943) and 54 (0-647) in the classical and in the stringent group, respectively (P < 0.01); and the median of platelet units received per day was 0.8 (0.03-30) and 0.5 (0-6.94) (P < 0.01). Our results emphasize the safety of a stringent prophylactic platelet transfusion policy after BMT, reducing the overall use of platelet transfusions. Further studies are necessary to confirm these results and to define optimal transfusion strategies.

Short-wavelength automated perimetry and retinal nerve fiber layer evaluation in suspected cases of glaucoma.

OBJECTIVES: To determine if short-wavelength automated perimetry (SWAP) provides evidence that indicates early functional losses in ocular hypertensive subjects and to establish a direct comparison with early structural abnormalities in the retinal nerve fiber layer (RNFL). METHODS: A total of 160 eyes belonging to 83 patients with ocular hypertension (intraocular pressure >21 mm Hg and normal results on standard automated perimetry evaluation), on which a SWAP and RNFL study were performed, were examined. One hundred twenty-eight age-matched subjects without ocular hypertension were evaluated to establish the 95% and 99% confidence intervals at each of the 76 exploration points of the SWAP test. RESULTS: The RNFL study results were normal in 83 cases (51.8%) and pathologic in 77 cases (48.1%). The SWAP results were pathologic in 57 cases (35.6%). Significant differences (P<.001) were observed when comparing the distribution of normal and pathologic SWAP results among the types of defects in the RNFL (focal wedge, diffuse atrophy, and mixed atrophy). CONCLUSIONS: Short-wavelength automated perimetry is a useful test for the early detection of visual field losses. It is more sensitive than standard automated perimetry and provides a high association with RNFL assessment, which has proved capable of detecting signs of glaucomatous damage several years before the onset of the typical visual field defects.

Contribution of rod and cone pathways to the dark-adapted electroretinogram (ERG) b-wave following retinal degeneration in RCS rats.

Although the RCS rat is widely used as a model of progressive photoreceptor loss, it is unclear how the relative rod and cone functions change with age. Rod and cone b-waves were isolated using a double flash ERG paradigm. In contrast to cones, rods never reached normal functional maturity levels, and the ERG b-wave changed from being predominantly rod-driven to being purely cone-driven by age 74 days, at which point, b-waves were progressively replaced by negative STR-like (scotopic threshold response) waves that persisted up to age 180 days. A double flash commonly abolished this wave and unveiled a b-wave.

Correlation of functional and structural measurements in eyes suspected of having glaucoma.

PURPOSE: This study was conducted to determine the correlation between structural changes in the retinal nerve fiber layer (RNFL) and functional loss detected on short-wavelength automated perimetry (SWAP) in a population of patients with suspected glaucoma. METHODS: With a selection criteria of intraocular pressure (IOP) more than 21 mmHg and normal results of conventional automated perimetry, 49 eyes of 49 patients with ocular hypertension were enrolled in the study. The SWAP was performed with a modified Humphrey field analyzer, and visual field indexes (mean deviation [MD], corrected pattern standard deviation [CPSD]) were calculated. Semiquantitative RNFL scores were given separately to diffuse and localized defects of the RNFL. RESULTS: The MD increased significantly with higher diffuse and total RNFL scores, with good correlation coefficients. A weak correlation was found between CPSD and diffuse, total, and localized RNFL scores. CONCLUSION: Diffuse RNFL loss are associated with abnormalities in visual field indexes (MD), whereas focal structural damage showed no correlation with visual field loss.