"Inflammatory Pathways in Patients with Post-acute Sequelae of COVID-19 - The Role of the Clinical Immunologist".

As the SARS-CoV-2 pandemic progressed, some survivors noted prolonged symptoms following acute infection, termed post-acute sequelae of COVID-19 (PASC) or "long COVID." PASC is a significant clinical and public health concern that adversely impacts patients' quality of life, income, and health care expenses. PASC symptoms are highly heterogenous, the most common being fatigue and cognitive impairment, and they likely reflect a spectrum of clinical phenotypes. The proposed role of persistent inflammation is one of leading pathophysiological theories. This review article addresses these proposed mechanisms of persistent and aberrant inflammation, their clinical evaluation and theoretical approaches to management. A review of public databases was used to collect literature for the review. The literature supports a prominent role of persistent and aberrant inflammation as a major contributor to the symptoms of PASC. Proposed mechanisms for persistent inflammation include reactivation of latent viruses, viral persistence, loss of immunoregulatory pathways, autoimmune mechanisms, and/or mast cell dysregulation. Persistent inflammation may result in constitutional symptoms such as fatigue, brain fog, body aches, and/or organ specific dysfunction such as gastrointestinal dysregulation, myocardial inflammation and others. There are no approved or even proven therapies for PASC at this time, but some studies have identified therapeutic options that may either reduce the risk for progression to PASC or decrease symptom burden. Laboratory evaluation and therapeutic options are limited and require further investigation to establish their clinical value. A more refined definition of PASC is needed to address the wide variety of clinical presentations, pathophysiology, and therapeutic options.

Evaluation of the Amblyopia tracker app.

PURPOSE: The Amblyopia tracker app has been developed to be a tool for parents to monitor changes in vision at home during amblyopia treatment. The aims of this study were to evaluate the feasibility and repeatability of parents testing their children at home and to compare home test results to an assessment in clinic by an orthoptist. METHODS: Children (age < 18 years) with amblyopia (interocular acuity difference of ≥ 0.2logMAR) were recruited. Parents were asked to test their child with the app three times during a two week period followed by an online questionnaire about the usability. Participants also tested within 48 h of their appointment where the measurement was repeated by an orthoptist. RESULTS: Out of 277 potential participants contacted, 37 completed three home measurements, mean age 6.8 years (SD 2.94). Home tests comparisons were made between test two and three to ensure familiarity with the process. Paired t-tests showed no statistically significant difference for either eye or the interocular acuity difference (IAD). However, 29% had a difference in IAD of more than 0.1logMAR on repeated testing, with a maximum of 0.4logMAR difference in the IAD. Questionnaire responses from the parents who participated were predominantly positive with 97% of respondents saying they would use it if were available. Comparison of home and clinical measurements (n = 23, mean age 6.72 SD 2.60) showed no statistically significant differences for either eye or interocular acuity difference (paired t-test, p > 0.3 in all cases). CONCLUSION: Results show no statistically significant differences for the Amblyopia tracker app when used by parents at home on repeated testing, or between the home test by a parent and the test by a clinician. However, variability in the results does indicate that further improvements are required to ensure the results can be used as a reliable clinical tool.

The impact of foetal restrictions on mode of delivery in women with inherited bleeding disorders.

OBJECTIVES: Management of pregnancy in women with congenital bleeding disorders (CBD) is challenging and requires understanding of risks conferred to both the mother and the foetus. Some elements of labour management are considered to increase the risk of neonatal bleeding and are not recommended for neonates at risk of a significant bleeding disorder. The impact of these restrictions on obstetric outcomes in women with CBD is unknown. METHODS: We retrospectively reviewed obstetric outcomes in a large cohort of women with CBD attending a specialised obstetric/haematology antenatal clinic over a 6-year period. RESULTS: Ninety-four pregnancies in 76 women with a wide variety of CBDs were assessed. Foetal precautions were recommended in the majority of cases (88%). Twenty (21.2%) were delivered by elective Caesarean section (CS), predominantly for obstetric indications. Of the 63 women who laboured with foetal precautions in place, 6 (10%) had a CS that was performed because of these precautions. There was no neonatal bleeding but primary postpartum haemorrhage (PPH) occurred in 12.2% of women. CONCLUSIONS: These data show that foetal precautions in labour recommended for women with CBDs will influence mode of delivery in approximately 10% of cases. This is important information for counselling these women about labour and delivery.

The effect of induced fusional demand on static and dynamic stereoacuity thresholds: the digital Synoptophore.

BACKGROUND/AIMS: The ability to extract depth from disparity may be hindered under fusional stress, as alignment of the eyes may be more difficult to maintain consistently. Therefore we aim to determine the effect of fusional demand on stereoacuity in individuals with no known binocular vision impairments. METHODS: A novel static and dynamic binocular depth detection task, capable of assessing many discrete levels of stereoacuity, was presented on digital displays attached to each tube of the Synoptophore. Stereoacuity was measured with any latent deviation fully corrected and compared to that measured at the 'recovery' angle. This recovery angle is where single vision is restored after decompensation to diplopia, during vergence range assessment. RESULTS: Seventy-two subjects (50 Female, 22 Male; mean (SD) age 22 (6) years) were assessed. The amount of fusional demand was between 1 and 26 prism dioptres (PD), with a mean (SD) of 8(6)PD. Under zero fusion demand the mean (SD) static and dynamic depth detection thresholds were 322(53)" and 69(23)". Under fusional stress these were 224(40)" and 77(21)". There was no significant difference between thresholds in stressed and zero demand fusion (p = 0.08). Dynamic depth detection thresholds were significantly lower than static (P < 0.01). CONCLUSION: Fusional stress does not appear to impact on stereoacuity. The numerical value of the recovery point varied amongst individuals, but this represents a common point, where single vision is easily restored and binocularity well established. Due to individual differences in the ability to control a certain amount of fusional stress (e.g. vergences stress of 10PD, when recovery is 8PD, will perturb binocularity more than a person with a recovery of 20PD), previous reports may not accurately represent the effect of fusional stress. Whilst our findings are contrary to previous reports, we did not stress fusion beyond the recovery point and used a more accurate/repeatable method to measure stereoacuity.

MicroRNA-containing extracellular vesicles released from endothelial colony-forming cells modulate angiogenesis during ischaemic retinopathy.

Endothelial colony-forming cells (ECFCs) are a defined subtype of endothelial progenitors that modulate vascular repair and promote perfusion in ischaemic tissues. Their paracrine activity on resident vasculature is ill-defined, but mediated, at least in part, by the transfer of extracellular vesicles (EVs). To evaluate the potential of isolated EVs to provide an alternative to cell-based therapies, we first performed a physical and molecular characterization of those released by ECFCs. Their effects upon endothelial cells in vitro and angiogenesis in vivo in a model of proliferative retinopathy were assessed. The EVs expressed typical markers CD9 and CD63 and formed a heterogeneous population ranging in size from ~60 to 1500 nm by electron microscopy. ECFC EVs were taken up by endothelial cells and increased cell migration. This was reflected by microarray analyses which showed significant changes in expression of genes associated with angiogenesis. Sequencing of small RNAs in ECFCs and their EVs showed that multiple microRNAs are highly expressed and concentrated in EVs. The functional categories significantly enriched for the predicted target genes of these microRNAs included angiogenesis. Intravitreally delivered ECFC EVs were associated with the vasculature and significantly reduced the avascular area in a mouse oxygen-induced retinopathy model. Our findings confirm the potential of isolated EVs to influence endothelial cell function and act as a therapy to modulate angiogenesis. The functions associated with the specific microRNAs detected in ECFC EVs support a role for microRNA transfer in mediating the observed effects.

The rapid kinetics of optimal treatment with subcutaneous methotrexate in early inflammatory arthritis: an observational study.

BACKGROUND: Methotrexate (MTX) is standard treatment for RA. Absorption is better in subcutaneous MTX (scMTX), which may impact speed of onset. In RA, earlier time to remission improves long-term results. Our objectives were to determine rapidity of response of subcutaneous methotrexate in early rheumatoid arthritis. METHODS: The change in several disease activity measures (including DAS28) from 0 to 6 weeks (early period) and 6 to 12 weeks (late period) was compared. The proportion achieving DAS28/CDAI/SDAI remission and/or low disease activity state was also compared. RESULTS: One hundred three patients were included from a single site between 2008 and 2014. All received MTX (98.0 % scMTX, 98 % 25 mg/week). There were no dropouts. There was a significantly greater early change in DAS28 (-1.9 vs. -0.2, p < 0.00); this effect was seen for several outcome measures. By 6 weeks, 59 % had achieved either DAS28 remission or low disease activity state, with 74 % achieving either state by 12 weeks. There were a larger proportion of patients achieving CDAI and DAS28 remission in the early versus late period (p < 0.0002 for both). There was significant improvement when using combination MTX and HCQ, however sample size was small (n = 9). The use of intra-articular steroids with MTX yielded the most disease measures that demonstrated early significant improvement. CONCLUSION: Subcutaneous MTX is rapid, as the change in many disease activity scores was significantly greater between 0-6 weeks compared to 6-12 weeks. Combination MTX + HCQ gave added value, although generalizability is limited by combination cohort sample size. Intra-articular steroid injections may contribute to the early effect.

Do Socioeconomic Inequalities Exist Within Ophthalmology and Orthoptics in the UK?: A Scoping Review.

Introduction: It is well documented that socioeconomic disadvantage adversely affects general health and ocular health worldwide. Within orthoptics, while clinicians recognise a relationship between socioeconomic situation and treatment outcome, no previous literature review was found to address this issue. Neither was a UK-specific literature review found to address the same issue for ophthalmology as a whole. Aim: This literature review evaluates evidence for an association between socioeconomic situation and ophthalmic/orthoptic conditions and their treatment outcomes, specifically within the UK. Methods: Keyword searches were conducted on Google Scholar and the University of Liverpool library catalogue. Results for the main analyses were limited to full papers, specific to the UK, written in English. Literature was only included from pre-2000 if more recent evidence was insufficient. Results: There is evidence of socioeconomic disadvantage being associated with the following: reduced visual acuity; reduced attendance at diabetic retinopathy screening appointments; and delayed presentation of glaucoma, cataracts, and diabetic retinopathy. However, evidence linking socioeconomic disadvantage to AMD is mixed. There is limited evidence of the increased prevalence of amblyopia and subsequent barriers to its treatment for socioeconomically underserved children. There is also evidence of a reduction in quality of life for socioeconomically underserved adults with strabismus. Conclusions: Health inequalities within ophthalmology and orthoptics are reported, but with confounding results for some conditions. Further research should explore the reasons behind the inequalities that are found and identify methods of reducing them.

Evaluating the Use of Contrast Sensitivity Tests By Orthoptists in the UK.

Introduction: The importance of the use of contrast sensitivity (CS) tests in orthoptic practice is well established. However, despite the clinical relevance the implementation within clinical care is known to be variable. There are no known studies that investigate the use of CS tests in Orthoptic clinics in the UK, therefore the aim of this study is to gather information from Orthoptists in the UK on their opinion of CS and use of CS testing in clinical practice, now and in the future. Methods: An online survey was distributed via JISC to the British and Irish Orthoptic Journal newsletter three times over a period of four weeks in June 2021 inviting practising orthoptists in the United Kingdom to complete. The questionnaire comprised of a series of questions regarding current use with free text responses for additional information. Results: There were 84 responses to the survey. The preferred test for adult and children testing is Pelli Robson with 50% reporting use of this test. 56% felt there is a need for a new CS test for young children, 12% said no and 32% were unsure. The highest percentage (57.1%) of participants were confident to some degree that their preferred test gave them useful clinical information. Conclusion: The result of the survey demonstrates the variability of CS testing currently in orthoptic practice in the UK. It also highlights the lack of currently available tests for children for CS testing, which may be addressed by the addition of the new Double Happy CS test.

A novel dual-fluorescence strategy for functionally validating microRNA targets in 3' untranslated regions: regulation of the inward rectifier potassium channel K(ir)2.1 by miR-212.

Gene targeting by microRNAs is important in health and disease. We developed a functional assay for identifying microRNA targets and applied it to the K(+) channel K(ir)2.1 [KCNJ2 (potassium inwardly-rectifying channel, subfamily J, member 2)] which is dysregulated in cardiac and vascular disorders. The 3'UTR (untranslated region) was inserted downstream of the mCherry red fluorescent protein coding sequence in a mammalian expression plasmid. MicroRNA sequences were inserted into the pSM30 expression vector which provides enhanced green fluorescent protein as an indicator of microRNA expression. HEK (human embryonic kidney)-293 cells were co-transfected with the mCherry-3'UTR plasmid and a pSM30-based plasmid with a microRNA insert. The principle of the assay is that functional targeting of the 3'UTR by the microRNA results in a decrease in the red/green fluorescence intensity ratio as determined by automated image analysis. The method was validated with miR-1, a known down-regulator of K(ir)2.1 expression, and was used to investigate the targeting of the K(ir)2.1 3'UTR by miR-212. The red/green ratio was lower in miR-212-expressing cells compared with the non-targeting controls, an effect that was attenuated by mutating the predicted target site. miR-212 also reduced inward rectifier current and K(ir)2.1 protein in HeLa cells. This novel assay has several advantages over traditional luciferase-based assays including larger sample size, amenability to time course studies and adaptability to high-throughput screening.

Test retest variability in stereoacuity measurements.

Background: A clinician's choice of stereotest is influenced by the robustness of the measurement, in terms of sensitivity, specificity and test-retest variability. In relation to the latter aspect, there are limited data on the test-retest variability of these new tests and how they compare to the more commonly used stereotests. Therefore, the aim of the study was to determine the test-retest variability of four different measures of stereoacuity (TNO, Frisby, Lang Stereopad and Asteroid (Accurate STEReotest On a mobIle Device)) and to compare the stereoacuity measurements between the tests in an adult population. Methods: Stereoacuity was measured twice using TNO, Frisby, Lang Stereopad and Asteroid. Inclusion criteria included adult participants (18 years and older), no known ophthalmic condition and VA (Visual Acuity) equal to or better than 0.3 logMAR (Logarithm of the Minimum Angle of Resolution) with interocular difference of less than 0.2 logMAR. Bland-Altman analysis was used to assess agreement within and between stereotests. Differences in stereo thresholds were compared using signed Wilcoxon tests. Results: Fifty-four adults (male: 23 and female: 31) with VA equal to or better than 0.3 logMAR in either eye and interocular difference less than 0.2 logMAR were assessed (mean age: 38 years, SD: 12.7, range: 18-72). The test-retest variability of all the clinical stereotests, with the exception of the Lang Stereopad (p = .03, Wilcoxon signed-rank test), was clinically insignificant as the mean bias was equal or less than 0.06 log seconds of arc (equivalent to 1.15 seconds of arc). While the Asteroid test had the smallest variation between repeated measures (mean bias: -0.01 log seconds of arc), the Frisby and Lang Stereopad tests had the narrowest and widest limits of agreement respectively. When comparing results between tests, the biggest mean bias was between Frisby and Lang Stereopad (-0.62 log seconds of arc), and 64.8% and 31.5% of differences were in the medium (21-100" of arc) and larger (>100" of arc) ranges respectively. Conclusion: The TNO and Frisby tests have good reliability but measure stereoacuity over a narrower range compared to the Asteroid which shows less variation on repeated testing but has a larger testing range. The data reported here show varying degrees of agreement in a cohort of visually normal participants, and further investigation is required to determine if there is further variability when stereoacuity is reduced.

Comparison of current paediatric contrast sensitivity assessments using simulated reduced contrast thresholds.

PURPOSE: There are limited tests of contrast sensitivity (CS) for use in children. The Hiding Heidi (HH) is suitable for all cognitive abilities, but has a ceiling effect. The Double Happy (DH) test has comparable thresholds to the Pelli Robson (PR), however the ability to detect changes in contrast has not been established. This study aims to compare contrast thresholds and agreement between HH and the DH, comparing to the PR chart in normal conditions and under reduced visual and lighting conditions. METHODS: Tests were repeated under different conditions to reduce the contrast. Room illumination was 20,900{plus minus}2% lux in bright conditions and 2,000{plus minus}2% lux in dim conditions, both conditions were repeated with the addition of simulation spectacles to reduce the clarity of vision. Participants' CS was measured uniocularly using the PR, HH and DH tests. RESULTS: 50 participants, age 18-62 years (mean{plus minus}standard deviation: 24.5{plus minus}7.98), were assessed. On HH 94% (n = 47) reached the maximum score, with the DH it was 18% (n = 9). The difference in reduction between conditions was smaller with HH in comparison to PR and DH, but significantly different from baseline conditions. Under dim conditions the reduction in PR and DH was -0.21 logCS units, but only -0.04 logCS for HH. CONCLUSION: The DH test has better agreement with PR than HH and is better at detecting CS changes, highlighting the advantages of use in clinical practice.

Selective extracellular vesicle-mediated export of an overlapping set of microRNAs from multiple cell types.

BACKGROUND: MicroRNAs (miRNAs) are a class of small RNA molecules that regulate expression of specific mRNA targets. They can be released from cells, often encapsulated within extracellular vesicles (EVs), and therefore have the potential to mediate intercellular communication. It has been suggested that certain miRNAs may be selectively exported, although the mechanism has yet to be identified. Manipulation of the miRNA content of EVs will be important for future therapeutic applications. We therefore wished to assess which endogenous miRNAs are enriched in EVs and how effectively an overexpressed miRNA would be exported. RESULTS: Small RNA libraries from HEK293T cells and vesicles before or after transfection with a vector for miR-146a overexpression were analysed by deep sequencing. A subset of miRNAs was found to be enriched in EVs; pathway analysis of their predicted target genes suggests a potential role in regulation of endocytosis. RT-qPCR in additional cell types and analysis of publicly available data revealed that many of these miRNAs tend to be widely preferentially exported. Whilst overexpressed miR-146a was highly enriched both in transfected cells and their EVs, the cellular:EV ratios of endogenous miRNAs were not grossly altered. MiR-451 was consistently the most highly exported miRNA in many different cell types. Intriguingly, Argonaute2 (Ago2) is required for miR-451 maturation and knock out of Ago2 has been shown to decrease expression of other preferentially exported miRNAs (eg miR-150 and miR-142-3p). CONCLUSION: The global expression data provided by deep sequencing confirms that specific miRNAs are enriched in EVs released by HEK293T cells. Observation of similar patterns in a range of cell types suggests that a common mechanism for selective miRNA export may exist.

Deep sequencing reveals predominant expression of miR-21 amongst the small non-coding RNAs in retinal microvascular endothelial cells.

The retinal vascular endothelium is essential for angiogenesis and is involved in maintaining barrier selectivity and vascular tone. The aim of this study was to identify and quantify microRNAs and other small regulatory non-coding RNAs (ncRNAs) which may regulate these crucial functions. Primary bovine retinal microvascular endothelial cells (RMECs) provide a well-characterized in vitro system for studying angiogenesis. RNA extracted from RMECs was used to prepare a small RNA library for deep sequencing (Illumina Genome Analyzer). A total of 6.8 million reads were mapped to 250 known microRNAs in miRBase (release 16). In many cases, the most frequent isomiR differed from the sequence reported in miRBase. In addition, five novel microRNAs, 13 novel bovine orthologs of known human microRNAs and multiple new members of the miR-2284/2285 family were detected. Several ∼30 nucleotide sno-miRNAs were identified, with the most highly expressed being derived from snoRNA U78. Highly expressed microRNAs previously associated with endothelial cells included miR-126 and miR-378, but the most highly expressed was miR-21, comprising more than one-third of all mapped reads. Inhibition of miR-21 with an LNA inhibitor significantly reduced proliferation, migration, and tube-forming capacity of RMECs. The independence from prior sequence knowledge provided by deep sequencing facilitates analysis of novel microRNAs and other small RNAs. This approach also enables quantitative evaluation of microRNA expression, which has highlighted the predominance of a small number of microRNAs in RMECs. Knockdown of miR-21 suggests a role for this microRNA in regulation of angiogenesis in the retinal microvasculature.

Using a Computerised Staircase and Incremental Optotype Sizes to Improve Visual Acuity Assessment Accuracy.

Background: Given the impact of visual acuity results on diagnosis and management, it is essential that the test is accurate, determined by factors such as test-retest variability. Standardisation improves accuracy, which can be performed via a computerised staircase methodology. Standard clinical tests with scoring of 0.02 per optotype implies an incremental score per optotype despite optotype size remaining constant on each line. The aim of this study is to establish if near continuous incremental optotype display and scoring improves test-retest variability compared to current testing methods. Methods: A computerised three up, one down adaptive staircase was used to display Kay Picture optotypes on an LCD monitor. Three methods of visual acuity assessment were undertaken: ETDRS, Kay Pictures and computerised Kay Pictures. Tests were performed twice under standard clinical conditions. Results: One hundred nineteen adults were tested. Test-retest variability for computerised Kay pictures was 0.01 logMAR (±0.04, p = 0.001). Good levels of agreement were observed for computerised Kay pictures in terms of test-retest variability, where the test had the smallest mean bias (0.01 logMAR compared to 0.03 and 0.08 logMAR for Kay Pictures and ETDRS respectively) and narrowest limits of agreement. Participants performed better in computerised Kay pictures than Kay Pictures by 0.03 logMAR, and better in ETDRS than computerised Kay pictures by 0.1 logMAR. Conclusion: Computerised Kay pictures exhibited a low test-retest variability, demonstrating it is reliable and repeatable. This repeatability measure is lower than the test-retest variability of the ETDRS and Kay Pictures tests.

Corrigendum: Using a Computerised Staircase and Incremental Optotype Sizes to Improve Visual Acuity Assessment Accuracy.

[This corrects the article DOI: 10.22599/bioj.271.].

Changes to Visual Parameters Following Virtual Reality Gameplay.

Introduction: Virtual reality (VR) gameplay is popular with a range of games and educational resources available. However, it puts high demands on the visual system. Current evidence shows conflicting impacts on visual parameters. Therefore, this study explores the changes to vision following VR gameplay. Methods: The study was conducted at the School of Health Sciences, University of Liverpool. All participants had binocular vision with good visual acuity and no manifest strabismus. Participants were assessed before and after playing 15 minutes of the VR game Beat Saber, which incorporated convergence and divergence movements. Clinical assessments including near point of convergence (NPC) and near point of accommodation (NPA) using the RAF rule; accommodative convergence to accommodation (AC/A) ratio; motor fusion using the prism fusion range (at 33cm), accommodation facility using +2.00/-2.00DS flipper lenses, and stereoacuity using the Frisby stereo test were assessed before and after playing. Results: Seventy-eight participants (19-25 years old) were included in the study, with 16 males and 41 females respectively. The breakpoint of convergence reduced by 0.5 cm (p = 0.001). The binocular accommodative facility improved by 2 cycles per minute (cpm); p = 0.004. The mean, near horizontal prism fusion range (PFR) base break and recovery points both worsened by of 5.0 dioptres (p = 0.003), whereas the mean near horizontal PFR base in recovery point improved by of 4.0 dioptres (p = 0.003). Discussion: The study validated previous findings as VR gameplay over-exercised and fatigued convergence muscles, but to a small degree. The VR experience improved the participants' ability to change focus quickly and improve accommodation, as well as the divergence function of the eye. However, as the participants were retested directly after the VR gameplay, the findings were limited to short term effects on vision.

Developmental Regulation and Functional Prediction of microRNAs in an Expanded Fasciola hepatica miRNome.

The liver fluke, Fasciola hepatica, is a global burden on the wellbeing and productivity of farmed ruminants, and a zoonotic threat to human health. Despite the clear need for accelerated discovery of new drug and vaccine treatments for this pathogen, we still have a relatively limited understanding of liver fluke biology and host interactions. Noncoding RNAs, including micro (mi)RNAs, are key to transcriptional regulation in all eukaryotes, such that an understanding of miRNA biology can shed light on organismal function at a systems level. Four previous publications have reported up to 89 mature miRNA sequences from F. hepatica, but our data show that this does not represent a full account of this species miRNome. We have expanded on previous studies by sequencing, for the first time, miRNAs from multiple life stages (adult, newly excysted juvenile (NEJ), metacercariae and adult-derived extracellular vesicles (EVs)). These experiments detected an additional 61 high-confidence miRNAs, most of which have not been described in any other species, expanding the F. hepatica miRNome to 150 mature sequences. We used quantitative (q)PCR assays to provide the first developmental profile of miRNA expression across metacercariae, NEJ, adult and adult-derived Evs. The majority of miRNAs were expressed most highly in metacercariae, with at least six distinct expression clusters apparent across life stages. Intracellular miRNAs were functionally analyzed to identify target mRNAs with inversely correlated expression in F. hepatica tissue transcriptomes, highlighting regulatory interactions with key virulence transcripts including cathepsin proteases, and neuromuscular genes that control parasite growth, development and motility. We also linked 28 adult-derived EV miRNAs with downregulation of 397 host genes in F. hepatica-infected transcriptomes from ruminant lymph node, peripheral blood mononuclear cell (PBMC) and liver tissue transcriptomes. These included genes involved in signal transduction, immune and metabolic pathways, adding to the evidence for miRNA-based immunosuppression during fasciolosis. These data expand our understanding of the F. hepatica miRNome, provide the first data on developmental miRNA regulation in this species, and provide a set of testable hypotheses for functional genomics interrogations of liver fluke miRNA biology.

Gestational age-specific hematological patterns in preterm infants following necrotizing enterocolitis.

OBJECTIVE: To investigate gestational age (GA) specific hematological and transfusion response patterns in preterm infants following necrotizing enterocolitis (NEC). DESIGN: A retrospective study comparing hematological/transfusion information in three GA groups' infants: Group A ≤ 28 weeks. Group B 28-32 weeks, Group C > 32 weeks following necrotizing enterocolitis. RESULTS: Group A infants responded with significantly higher WBC count, thrombocytopenia, higher absolute neutrophil, and higher absolute monocyte and lower absolute lymphocyte counts following NEC onset, received more blood transfusions before NEC onset (59.8 versus 30.0%; p = .007), and had higher odds of surgical NEC (OR 3.39 [95% CI 1.19-10.38]; p = .02) than group C. One unit increase in absolute lymphocyte count on the day, and 24 h following NEC was significantly associated with lower surgical NEC odds than groups C. CONCLUSION: The infant's in-group A had significantly different hematological response patterns following NEC than infants with higher gestational age (groups B and C).

Orthoptic assessment of vision in patients with complex disability: the importance of understanding the patient's communication methods.

Care for children with acquired brain injury requires a multi-disciplinary team (MDT) to maximize rehabilitation. These children frequently present to the orthoptic clinic with complex difficulties impacting visual functions and eye movements. We report on one case which led to a reevaluation of our current assessment methods, clinical care pathways and the contents of clinic correspondence. We present a case report of a 14-year-old girl who suffered a cardiac arrest due to underlying Wolff-Parkinson-White syndrome; MRI revealed hypoxic brain injury with extensive white matter and basal ganglia volume loss. On presentation, it was evident there was a significant impact on visual function. Conventional optotype acuity testing was not possible. Her inability to maintain fixation made observational responses unreliable. Observations revealed no steady fixation, exotropia and roving eye movements. Although she was reported to be unable to speak, progress with communication was reported and she was able to give repeatable, reliable responses to indicate "yes" or "no." These cues were used and a response of 1.70 cycles per degree was obtained under binocular conditions. This communication method was also utilized to assess visual fields by confrontation. However, the clinic letter written by the ophthalmologist based on the orthoptic assessment simply provided the acuity score with no interpretation of the information or how it was obtained. This case highlighted the importance of understanding a child's communication methods so that assessment can be appropriately adapted. The referral letter was lacking in detail regarding the patient's abilities but equally the ophthalmology letter provided limited detail impacting on the MDT practitioner's ability to understand and apply the information. This case has highlighted the importance of good communication: professionals need to acknowledge and work with an individual's communication methods so that assessment can be completed. Professionals also need to improve communication of relevant findings to others involved in the patient's care.