Melatonin attenuates thioacetamide-induced liver fibrosis in male rats through modulation of interleukin-6, interleukin-4, apoptosis and urokinase plasminogen activator receptor-associated protein/Endo180.

Liver fibrosis is a chronic progressive disease, its resolution still unclear, and the current study explored the role of melatonin in modulation of interleukin-6 (IL-6), interleukin-4 (IL-4), transforming growth factor beta1 (TGF-ß1) and urokinase plasminogen activator receptor-associated protein/Endo180 (uPARAP/Endo180) pathway in thioacetamide (TAA)-induced hepatotoxicity. Thirty two adult Sprague-Dawley rats were divided into four groups: vehicle control group, TAA-induced liver fibrosis group that was left untreated, melatonin administration before and along with TAA and melatonin along with TAA group. TTA-induced massive liver necrosis, fibrosis around portal tract and increases serum levels of liver enzymes and total bilirubin when compared with control vehicle group. While both melatonin pretreatment and treatment retained liver parenchyma and liver enzymes quite similar to control group and reduced TAA-induced liver injury. Notably, melatonin pretreatment and treatment increased collagen degradation in TAA liver injury by19, 31.7-fold respectively evidence by collagen percentage area. Melatonin also decreased the amount of thiobarbituric acid reactive compounds and retained the reduced glutathione and superoxide dismutase to basal level quite similar to control group. Additionally, melatonin significantly (P value ≤0.05) decreased the levels of TGF-ß1, epidermal growth factor (EGF), hydroxyproline, tissues IL-6, caspase-3, and receptor interacting serine/threonine kinase1 (RIPK1), fibrillin-1, and - smooth muscle actin in the liver tissues while significantly (P value ≤0.05) increasing the levels of IL-4 and uPARAP/Endo180. Due to its anti-inflammatory, anti-apoptotic, and antioxidant capabilities as well as its ability to decrease hepatic stellate cell activation and fibrogenesis, these data imply that melatonin has a powerful anti-fibrotic effect.

Impact of Adopting Routine Luminex-Based Pretransplant Assessment of HLA Antibodies on Clinical Practice and Outcomes in Kidney Transplantation.

BACKGROUND: Accumulating evidence suggests that detection of human leukocyte antigen (HLA) antibodies by solid phase Luminex assays predicts renal allograft outcomes. However, several controversies exist regarding the interpretation, reproducibility, impact and financial feasibility of global utilization of this assay in pretransplant assessment. METHODS: We studied short-term patient-centered outcomes, medical standards of care, and financial plausibility of using Luminex-based screening for HLA antibodies in renal allograft recipients compared to outcomes in nontested patients. RESULTS: We included 1808 patients assessed for transplantation from 2011 to 2018. Luminex-tested patients had lower rates of rejection in the first post-transplant week (OR 0.36, P < .001) and lower odds of antibody-mediated rejection in the first 6 months (OR 0.4, P = .004). Forty-four patients with preformed, donor-specific antibodies were transplanted, and everolimus was introduced into our protocols for low-risk patients based on risk stratification by Luminex results. The number of tests needed to be performed to prevent 1 episode of antibody-mediated rejection in the first 6 months was 28 (P = .004), which was financially plausible. CONCLUSIONS: Routine pre-transplant assessment of HLA antibodies using Luminex assays may allow for better patient-centered, short-term graft outcomes and objective tailoring of immunosuppression at a financially plausible, cost-effective rate.

Neurocognitive dysfunction in children with ß thalassemia major: psychometric, neurophysiologic and radiologic evaluation.

OBJECTIVE: To evaluate the impact of iron chelating drugs and serum ferritin on the neurocognitive functions of patients with ß thalassemia major (ß-TM), using psychometric, neurophysiologic and radiologic tests. METHODS: Eighty children with ß-TM were enrolled into the study and were compared to 40 healthy controls. All participants were evaluated by measuring serum ferritin, neurocognitive assessment by Benton Visual Retention Test, Wechsler Intelligence Scale for Children, Wisconsin Card Sort Test, P300 and magnetic resonance spectroscopy (MRS). RESULTS: WISC in our study showed that 40% of cases were borderline mental function as regards total IQ. Neurophysiologic tests were significantly impaired in patients compared to control group, with significant impairment in those receiving desferrioxamine (DFO). P300 amplitude was significantly lower in cases compared to controls (2.24 and 4.66 uv, respectively), recording the shortest amplitude in patients receiving DFO. Altered metabolic markers in the brain were detected by MRS in the form of reduced N-acetylaspartate to creatine ratio in 78.3% of our cases. There were significant correlations between psychometric tests and both neurophysiologic (P300) and radiologic (MRS) tests. CONCLUSION: ß-TM is associated with neurocognitive impairment that can be assessed by psychometric, neurophysiologic and radiologic tests. The role of hemosiderosis and iron chelation therapy on cognitive functioning still need more research. ABBREVIATIONS: ß-TM: beta thalassemia major; DFO: Dysferal; DFP: Deferiprone; DFX: Deferasirox; WISC: Wechsler Intelligence Scale for Children; VIQ: verbal IQ; PIQ: performance IQ; TIQ: total IQ; BVRT: Benton Visual Retention Test; WCST: Wisconsin Card Sort Test; MRS: Magnetic resonant spectroscopy; NAA/Cr ratio: N-acetylaspartate to creatine ratio.

Optical coherence tomography characteristics of different types of big bubbles seen in deep anterior lamellar keratoplasty by the big bubble technique.

PurposeTo define optical coherence tomography (OCT) characteristics of type-1, type-2, and mixed big bubbles (BB) seen in deep anterior lamellar keratoplasty.MethodsHuman sclero-corneal discs were obtained from UK (30) and Canada (16) eye banks. Air was injected into corneal stroma until a BB formed. UK samples were fixed in formalin before scanning with Fourier-domain (FD-OCT). One pair of each type of BB was scanned fresh. All BB obtained from Canada were scanned fresh with time-domain (TD-OCT). For each OCT machine used, type-1 BB from which Descemets membrane (DM) was partially peeled, were also scanned. The morphological characteristics of the scans were studied.ResultsFD-OCT of the posterior wall of type-1 (Dua's layer (DL) with DM) and type-2 BB (DM alone) both revealed a double-contour hyper-reflective curvilinear image with a hypo-reflective zone in between. The anterior line of type-2 BB was thinner than that seen with type-1 BB. In mixed BB, FD-OCT showed two separate curvilinear images. The anterior image was a single hyper-reflective line (DL), whereas the posterior image, representing the posterior wall of type-2 BB (DM) was made of two hyper-reflective lines with a dark space in between. TD-OCT images were similar with less defined component lines, but the entire extent of the BB could be visualised.ConclusionOn OCT examination the DM and DL present distinct features, which can help identify type-1, type-2, and mixed BB. These characteristics will help corneal surgeons interpret intraoperative OCT during lamellar corneal surgery.

Deep anterior lamellar keratoplasty--triple procedure: a useful clinical application of the pre-Descemet's layer (Dua's layer).

PURPOSE: To describe a technique which exploits the transparency and toughness of the pre-Descemets layer (Dua's layer) to safely perform deep anterior lamellar keratoplasty (DALK) and phacoemulsification at the same time. METHODS: Three DALK procedures combined with phacoemulsification were performed by the same surgeon using the big-bubble (BB) technique at the Research Institute of Ophthalmology, Cairo, Egypt. In two cases a type-1 BB, baring Dua's layer, was achieved and in one case a type-2 BB, baring the Descemet's membrane (DM), was achieved. The surgeries were video recorded and photo slit-lamp images were taken in the follow-up visits. RESULTS: DALK with phacoemulsification and lens implant was carried out in both patients where a type-1 BB was achieved. At a follow up of 18 months (first case) and 6 months (second case), the best corrected vision was 6/12 for each. In the case where a type-2 BB was achieved, the DM ruptured during injection of viscoelastic prior to capsulorhexis. The procedure was converted to a penetrating keratoplasty without phacoemulsification. CONCLUSION: When a type-1 BB is achieved simultaneous DALK and phacoemulsification can be safely accomplished. Dua's layer allows a clear view for performing phacoemulsification with the added benefit of its toughness, which can maintain a stable anterior chamber for cataract surgery. This should not be attempted when a type-2 BB is achieved.

Intracranial haemorrhage is linked to late onset vitamin K deficiency in infants aged 2-24 weeks.

AIM: Late vitamin K deficiency bleeding (VKDB) can be serious and manifest as early onset intracranial haemorrhage (ICH). This study aimed to determine the frequency of ICH in relation to vitamin K deficiency and the outcome in infants aged two to 24 weeks. METHOD: A hospital-based study was conducted in two main tertiary hospitals in Cairo, Egypt, from May 2011 to May 2012 with 40 patients with ICH and 50 age-matched controls without ICH. RESULTS: Forty patients with ICH were recruited, 19 were excluded for clinical reasons and the remaining 21 had a significantly low vitamin K level. Exclusive breast feeding (81% of patients), diarrhoea lasting more than 1 week (38.1%) and antibiotic consumption within a week before the development of ICH (57.1%) were more common in the patients than in the control group (p value>0.05, <0.01 and <0.01, respectively). CONCLUSION: A high frequency of ICH due to late VKDB was reported in Egyptian infants aged two to 24 weeks, with poorer outcomes than international studies. A national survey is required to evaluate the timing and protective value of a second booster vitamin K dose to reduce ICH, especially in high-risk patients in this age group.

Sequential selective same-day suture removal in the management of post-keratoplasty astigmatism.

AIMS: In a previous study, we proposed that corneal topography performed 30-40 min after the initial suture removal can identify the next set of sutures requiring removal, for the treatment of post-keratoplasty astigmatism. The aim of this study was to evaluate the effect of removing subsequent sets of sutures at the same sitting. METHODS: 10/0 nylon interrupted sutures were placed, to secure the graft-host junction, at the time of keratoplasty. Topography was performed using Pentacam (Oculus) before suture removal. The sutures to be removed in the steep semi-meridians were identified and removed at the slit-lamp biomicroscope. Topography was repeated 30-40 min post suture removal, the new steep semi-meridians determined, and the next set of sutures to be removed were identified and removed accordingly. Topography was repeated 4-6 weeks later and the magnitude of topographic astigmatism was recorded. A paired-samples t-test was used to evaluate the impact of selective suture removal on reducing the magnitude of topographic and refractive astigmatism. RESULTS: Twenty eyes of 20 patients underwent sequential selective same-day suture removal (SSSS) after corneal transplantation. This study showed that the topographic astigmatism decreased by about 46.7% (3.68 D) and the refractive astigmatism decreased by about 37.7% (2.61 D) following SSSS. Vector calculations also show a significant reduction of both topographic and refractive astigmatism (P<0.001). CONCLUSION: SSSS may help patients to achieve satisfactory vision more quickly and reduce the number of follow-up visits required post keratoplasty.

Intra-lesional 5 fluorouracil for the management of recurrent pterygium.

AIM: Recurrence is the most common complication arising from pterygium surgery. The aim of this study was to investigate the effectiveness of 5 fluorouracil (5FU) in halting the recurrence of pterygium after surgical excision. METHODS: A retrospective review of patients treated for pterygium recurrence was carried out. Patients with recurrent (secondary) pterygium were treated with multiple weekly intra-lesional injections of 0.1-0.2 ml (2.5-5 mg) 5FU post-operatively depending on the size of the recurrence. The treatment was started within 1 month from the date of recurrence. The time from surgery to start of recurrence, previous treatment modalities, and number of recurrences were documented. The number of injections required to induce arrest of progression and/or regression of vascularity and fleshiness of the pterygium and any complications related to 5FU treatment were examined. RESULTS: Fifteen eyes from 14 patients with recurrent pterygium treated with intra-lesional 5FU injections were analysed. Three of the 15 eyes had undergone a secondary excision and 12 had undergone a primary excision. In all, 93.3% of patients showed regression of the fibrovascular tissue (thickness and vascularity) and arrest of progression following a dose of 0.1-0.2 ml (2.5-5 mg) 5FU. Twelve eyes required three injections or fewer, whereas one patient required eight injections. This beneficial effect was maintained over an average follow-up period of 17 months. No complications from 5FU were observed. CONCLUSION: The use of weekly intra-lesional 5FU injections for the treatment of recurrent pterygium is safe and effective in limiting the progression and inducing the regression of recurrent pterygium. The number of injections can be tailored according to clinical need.

Efficacy and safety of deferasirox at low and high iron burdens: results from the EPIC magnetic resonance imaging substudy.

The effect of deferasirox dosing tailored for iron burden and iron loading based on liver iron concentration (LIC) was assessed over 1 year in less versus more heavily iron-overloaded patients in a substudy of the Evaluation of Patients' Iron Chelation with Exjade®. Deferasirox starting dose was 10-30 mg/kg/day, depending on blood transfusion frequency, with recommended dose adjustments every 3 months. Therapeutic goals were LIC maintenance or reduction in patients with baseline LIC <7 or ≥7 mg Fe/g dry weight (dw), respectively. Changes in LIC (R2-magnetic resonance imaging) and serum ferritin after 1 year were assessed. Adverse events (AEs) and laboratory parameters were monitored throughout. Of 374 patients, 71 and 303 had baseline LIC <7 and ≥7 mg Fe/g dw, respectively; mean deferasirox doses were 20.7 and 27.1 mg/kg/day (overall average time to dose increase, 24 weeks). At 1 year, mean LIC and median serum ferritin levels were maintained in the low-iron cohort (-0.02 ± 2.4 mg Fe/g dw, -57 ng/mL; P = not significant) and significantly decreased in the high-iron cohort (-6.1 ± 9.1 mg Fe/g dw, -830 ng/mL; P < 0.0001). Drug-related gastrointestinal AEs, mostly mild to moderate, were more frequently reported in the <7 versus ≥7 mg Fe/g dw cohort (39.4 versus 20.8 %; P = 0.001) and were not confounded by diagnosis, dosing, ethnicity, or hepatitis B and/or C history. Reported serum creatinine increases did not increase in low- versus high-iron cohort patients. Deferasirox doses of 20 mg/kg/day maintained LIC <7 mg Fe/g dw and doses of 30 mg/kg/day were required for net iron reduction in the high-iron cohort, with clinically manageable safety profiles. The higher incidence of gastrointestinal AEs at lower iron burdens requires further investigation.

Adrenal function in thalassemia major adolescents.

BACKGROUND: Several studies reported a significant prevalence of adrenal insufficiency, ranging from 18-45%, in patients with thalassemia. Evidence for dissociation of cortisol and adrenal androgen secretion in patients with beta-thalassemia was previously reported. AIM: We measured adrenal androgen response along with cortisol to the standard (250 mg) dose ACTH test. METHODS: Forty five beta-thalassemia major (TM) patients were enrolled. Their ages ranged between 12 and 20 years (14.9 ± 2.2 years). All patients underwent the 250 mg cosyntropin test in the morning before blood transfusion. Blood samples for total cortisol, dehdroepiandrosterone (DHEA) and androstendione (A) measurements were collected before and 60 min after IV injection of 250 mg cosyntropin. Adrenal insufficiency was observed in 7 of 45 (15.5%) patients. Adrenal androgen levels decreased significantly with advancing Tanner stage. No difference was noted between patients with and without adrenal insufficiency regarding anthropometric and laboratory parameters. CONCLUSION: Adrenal insufficiency is not a rare complication in thalassemia. Adrenal androgen production declines with advancing puberty in thalassemic adolescents and might explain the poor development of pubic and axillary hair observed in this condition.

Persistent factor VIII inhibitors and orthopaedic complications in children with severe haemophilia A.

Persistence of inhibitors against factor VIII (FVIII) may be a risk factor that increases physical disability in haemophilia A (HA) patients. This study aimed to evaluate prevalence of FVIII inhibitors in previously treated children with severe HA and the impact of persistent inhibitors on knee joint status and lumbar bone mineral density (BMD). Fifty children with severe HA, FVIII <1%; aged 5-16 years were enrolled in this study; they received plasma-derived FVIII on-demand treatment for 50-250 exposure days (EDs). Inhibitors were checked at basal visit and were followed up for 1 year, using Bethesda assay. Cross-sectional clinical scoring and radiological evaluation of the knee joint (by Arnold-Hilgartner staging and Pettersson score), along with lumbar BMD by Dual Energy X-ray Absorptiometry (DEXA) were performed. Patients with persistent inhibitors for 1 to 5 years, median 2.5 years, were 10 (20%). Six had high titre and none of them had completely normal knees, seven had advanced knee arthropathy and six had low lumbar BMD in comparison to 2 and 8 of the 40 patients without inhibitors respectively (P < 0.05). Persistence of inhibitors for more than 2 years without immuno-prophylaxis was a risk factor for joint damage. Low lumbar BMD was found in 88.9% of patients with stages four and five knee arthropathy and in 66.7% of patients with positive hepatitis C. Severe HA children in this Egyptian study had a relatively low prevalence of persistent FVIII inhibitors, which, if not treated, may increase the risk of knee arthropathy and lumbar osteopenia.

Evaluation of the oral health situation of a group of Egyptian haemophilic children and their re-evaluation following an oral hygiene and diet education programme.

Haemophilic children in Egypt have received minimal dental intervention and their dental needs required assessment. The purpose of this study was to assess the oral health needs of a sample (n = 60) of Egyptian haemophilic children (6-12 years), so as to develop, implement and evaluate an oral hygiene education programme over an 8-month period on the experimental group (n = 30) vs. the control group (n = 30). The oral hygiene index simplified (OHI-S) index was used for baseline data and at the end of the study, while DMFS and defs were used to collect caries experience baseline data on each subject. The results showed that the DMFT and deft were significantly higher than those of the non-haemophilic population in Egypt and also higher than those of haemophilic children in developed countries and that the decayed component represented most of the index values. At phase I, the mean value of the OHI-S of experimental and the control groups was 2.67 +/- 0.45 and 2.53 +/- 0.53, respectively, but the difference was not significant (P > 0.05), both values were in the 'fair' category (1.3-3.0). At phase II, the end of the 8 months follow-up period and after the application of a strict oral care programme in the experimental group, there was a significant decrease from 2.67 to 1.20 (P < 0.001), a shift of values occurred from the 'fair' category to the 'good' category (0.1-1.2) while there was no significant difference in the control group. It can be concluded that professional plaque control, education and access to oral hygiene aids is paramount to improve oral health of these children.

Prospective evaluation of high-cost management of severe chronic ITP in children and adolescents<16 years.

Chronic ITP rarely presents with severe bleeding episodes (SBE). Number and duration of SBE were evaluated in relation to the cost of management. Out of 157 chronic ITP patients attending our institution from 1994 to 2003, 37 patients, <16 years with persistent thrombocytopenia (>6 months), suffering from SBE or platelet count<10x10(9)/L were prospectively randomized to receive either intravenous immunoglobulins (IVIG), anti-D immunoglobulin (anti-D) or high-dose methyl prednisolone (HDMP). Sixty-one patient-years were followed, during which 351 SBE were documented. The high-cost management (IVIG and anti-D) showed insignificantly better platelet recovery, less frequent SBE with shorter duration per patient, higher rate of CR, and less splenectomy in contrast to the steroid groups. The effectiveness of high-cost management compared with methyl prednisolone could not be documented in this study.

Childhood ITP: 12 months follow-up data from the prospective registry I of the Intercontinental Childhood ITP Study Group (ICIS).

BACKGROUND: Acute and chronic idiopathic thrombocytopenic purpura (ITP) is traditionally based on the duration of thrombocytopenia at the cut-off point of 6 months after diagnosis. Registry I evaluated the diagnosis, definition, management, and follow-up of childhood ITP. This report focuses on children with thrombocytopenia persisting more than 6 months. PROCEDURE: Data were collected by questionnaires to the physicians caring for children with ITP, at diagnosis, 6, and 12 months later. Data were compared regarding initial features and follow-up with emphasis on children with persistent thrombocytopenia, and those with ITP who recovered their platelet counts between 7 and 12 months from diagnosis. RESULTS: At 12 months from diagnosis, 79 of 308 (25.6%) evaluable children recovered from ITP and 229 had ongoing ITP. Children with recovered ITP were younger than children with ongoing ITP (P = 0.043) and exhibited a lower frequency of bleeding symptoms during the first 6 months after diagnosis (P = 0.018). Frequency of hospitalization, bone marrow aspiration, and drug treatment differed regionally. CONCLUSIONS: The high rate of recovery from ITP between 7 to 12 months demonstrates, that the cut-off point of 6 months for the definition of chronic ITP does not adequately differentiate chronic from acute ITP. The majority of children with ITP have variable time to recovery with gradual improvement of platelet counts and disappearance of bleeding signs. ITP is a heterogeneous disorder with a diverse natural history and diverse pattern of treatment response.

Frequency of inhibitor development in severe haemophilia A children treated with cryoprecipitate and low-dose immune tolerance induction.

The frequency of factor VIII inhibitor development was evaluated in a hundred severe haemophilia A patients < 18 years of age (mean 10.4 +/- 5.1 years); 25 were previously untreated patients (PUPs), with a mean age of 11.2 +/- 2.9 months. All were followed up for 3 years from December 1996. Immune tolerance (IT) was induced with low-dose factor VIII (FVIII); 25-50 IU kg(-1) every other day for the 10 haemophiliacs who developed persistent inhibitors. The incidence of inhibitors for PUPs was 3/25 (12%; 95% confidence interval [CI], 0. 7-24.7%) and were detected after 4, 15 and 20 exposure days (mean 13 +/- 8.2 days; 95% CI, 3.7-22.2%). Children with maximum inhibitor levels of > 40 Bethesda units (BU) per mL (n=4) received IT therapy as 25 U kg(-1) FVIII in the form of cryoprecipitate every other day for 1-4 months (mean 2.4 +/- 1.6 months; 95% CI, 0.8-3.9%), which was successful in all of them. FVIII (50 U kg(-1)) was given every other day for six patients with maximum inhibitor level > 40 BU mL(-1) for 3-9 months (mean 5.4 +/- 3.2 months; 95% CI, 2.9 -7.9%) with success in 4/6 (66.6%; 95% CI, 28.8-104.3%). Patients who showed a good IT response had an inhibitor level < or = 30 BU mL(-1), were < or = 9 years of age at inhibitor development with few exposure days to FVIII and had an early immune tolerance. In conclusion, inhibitor development in severe haemophilia A children exclusively treated with cryoprecipitate is low. Early low-dose IT induction for high responders may be achieved successfully if inhibitor level is < or = 50 BU mL(-1).

Oxidant damage to erythrocyte membrane in glucose-6-phosphate dehydrogenase deficiency: correlation with in vivo reduced glutathione concentration and membrane protein oxidation.

Chronic nonspherocytic hemolytic anemia has been observed in a recently described glucose-6-phosphate dehydrogenase (G6PD) variant, G6PDWayne. The mechanical properties of these erythrocytes and other G6PD variants were examined. The deformability of G6PD-deficient erythrocytes was normal, as determined by osmotic scan ektacytometry, and was not significantly affected by hemolytic crisis. In the common varieties of G6PD deficiency, the mechanical stability of the red blood cell (RBC) membrane was greater than normal, but G6PDWayne membranes were abnormally susceptible to shear-induced fragmentation. There was no evidence for a concurrent genetic defect in spectrin, because self-association constants and tryptic digests were normal. The fragility of G6PDWayne membranes appeared to be a consequence of oxidative damage to membrane thiol groups associated with a low glutathione (GSH) level in these RBCs. Associations among GSH level, thiol oxidation, and membrane instability were also found when a larger group of G6PD-deficient RBCs were examined. In normal erythrocytes, 1-chloro-2,4-dinitrobenzene was used to reduce GSH levels by 50%. Membrane thiol oxidation and membrane fragility both increased when these cells were kept at 4 degrees C for 3 to 5 days. Our findings suggest that chronic depletion of GSH leads to the destabilization of membrane skeleton through oxidation of membrane protein thiols.