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Purpose: We hypothesised that cleaning the internal limiting membrane (ILM) with a flexible nitinol loop following diabetic vitrectomy without peeling may reduce the common occurrence of postoperative epiretinal membrane (ERM) formation. Methods: Consecutive patients undergoing vitrectomy for proliferative diabetic retinopathy by one surgeon from 2015 to 2019 were studied and divided into 2 cohorts: the control group underwent standard surgery, and the ILM clean group underwent additional cleaning of the macular retina using a flexible nitinol loop after vitrectomy. Masked comparison of ERM on optical coherence tomography was performed at 3 months, and visual acuity (VA) was measured until 12 months postoperatively. Results: Baseline demographics, clinical features, and protein levels were similar between cohorts. The ILM clean group (n = 56) had fewer clinically significant ERM than the control group (n = 50; 4 vs. 20%; p = 0.01), and a significantly lower proportion of the ILM clean group required revision surgery (2 vs. 14%; p = 0.02). VA in the ILM clean group was significantly better than in the control group at 3 months (0.35 vs. 0.50 logMAR; p = 0.02) but not at 12 months (0.34 vs. 0.43 logMAR; p = 0.17). Conclusion: ILM cleaning with a flexible nitinol loop following diabetic vitrectomy resulted in significant reduction in ERM formation and reduced necessity for revision surgery. There was significant improvement in VA at 3 months but not over a longer follow-up.

BACKGROUND: Autistic masking is an emerging research area, and so far, research has suggested that masking has a negative effect on autistic people. Masking relates to general social practices (such as identity management) and is often driven by stigma avoidance. Many nonautistic people also experience stigma that might drive them to suppress aspects of their identity. In this study, we investigate similarities and differences in experiences of masking in autistic and nonautistic people. METHODS: We conducted an online survey about experiences and views of masking in autistic people (n = 144), neurodivergent people without an autism diagnosis (n = 49), and neurotypical people (n = 45) recruited via social media. We used thematic analysis to analyze responses to open-ended items about masking, using an inductive approach, at a semantic level, with a critical realist paradigm. RESULTS: Thematic analysis revealed that some aspects of masking are shared across autistic and nonautistic people, such as utilizing mimicry of others as a social strategy or feeling exhausted from masking. All groups reported that masking made them feel disconnected from their true sense of identity and had a negative effect on them. Other aspects of masking seemed more specific to autistic people, such as sensory suppression, and masking leading to suicidal ideation. CONCLUSIONS: Our findings suggest that many aspects of masking are experienced across different neurotypes and are likely related to outside perceptions of difference and stigma. It is likely that what we call “autistic masking” is similar to other forms of stigma management previously theorized. Some aspects of masking do seem more specific to the autistic neurotype (e.g., suppression of stimming) and should be explored further to provide support for autistic people recovering from the negative impact of masking. LAY SUMMARY: WHY WAS THIS STUDY DONE? Masking is the process of intentionally, or unintentionally, hiding aspects of yourself to avoid harm. Recent research has suggested that autistic people might mask a lot and that this can lead to long-term problems in mental health. At the moment, we do not know which parts of masking are specific to autistic people, and which parts might be experienced by others, especially other neurodivergent people who are not autistic, for example, people with attention-deficit hyperactivity disorder (ADHD). WHAT WAS THE PURPOSE OF THIS STUDY? The purpose of this study was to find out about masking in autistic and nonautistic people, to find out whether some parts of masking are specific to autistic people, or experienced by others. WHAT DID THE RESEARCHERS DO? We conducted an online survey about masking with three groups of people: autistic people, nonautistic people who have another neurodevelopmental or mental health diagnosis, such as dyslexia, ADHD, or depression, and nonautistic people who do not have any neurodevelopmental or mental health diagnoses. The survey included an open text box for people to write about their experiences of masking and their opinions about masking. We analyzed these responses using a method called “thematic analysis.” This method helps us look for ideas that people commonly talk about in their answers. WHAT WERE THE RESULTS OF THIS STUDY? We found out that autistic and nonautistic people think masking is very complicated and has a negative effect on them. Both autistic and nonautistic people said that masking made them exhausted and really unhappy and that it made them feel like people did not know the “real them.” Only autistic people mentioned that masking sometimes makes them feel suicidal and that masking includes things that other groups did not mention (such as trying to hide being upset by sensory things such as loud noise). WHAT DO THESE FINDINGS ADD TO WHAT WAS ALREADY KNOWN? Our findings suggest that some aspects of masking do not just affect autistic people (such as feeling like people do not know the real you), but other parts might be more unique to autistic people (such as hiding stims from other people). WHAT ARE POTENTIAL WEAKNESSES IN THIS STUDY? More women than men and nonbinary people took part, which means that we know more about women than other people. We also do not really know whether everyone in the nonautistic groups were really “not autistic.” They could be autistic people who do not have a diagnosis, which might affect our results. HOW WILL THESE FINDINGS HELP AUTISTIC PEOPLE NOW OR IN THE FUTURE? We hope our findings will help people to understand how masking affects autistic and nonautistic people. We also hope that people designing new studies will look at important things such as whether someone is autistic and nonbinary, or autistic and ADHD as these things might affect their experiences.

The practice of histopathological assessment of the uterus following hysterectomy for benign indications including pelvic organ prolapse (POP) surgery is common and often routine. While pathology is not anticipated, the finding of pathology requiring further action is always a concern, in particular CIN (cervical intraepithelial neoplasia) or cervical/uterine malignancy. We aimed to perform a systematic review to understand the prevalence of actionable uterine and cervical pathology in hysterectomy specimens performed for POP. A literature search was performed in January 2020 of MEDLINE, Embase and CINAHL using the Healthcare Databases Advanced Search platform. Included studies reported CIN and/or uterine/cervical malignancy in histological assessment of hysterectomy specimens performed purely for POP. Meta-analysis of prevalence was performed using the MetaXL ( www.epigear.com ) add-in for Microsoft Excel. Six hundred seventy-seven records were identified, out of which 34 studies were eligible. Overall prevalence (95% confidence interval [CI]) of endometrial cancer in 33 studies was 0.004 (0.003–0.006), I2 = 41%, number needed to treat (NNT) 1:250. Total actionable uterine pathology was 0.005 (0.003–0.006) in 33 studies, I2 = 35%, NNT = 1:200. Overall prevalence of cervical cancer in 19 papers was 0.001 (0.000–0.002), I2 = 18%, NNT = 1:1000. In 16 studies the overall prevalence of CIN was 0.013 (0.001–0.033), I2 = 95%, NNT = 1:77. Prevalence of total actionable pathology was 0.013 (0.006–0.0023), I2 = 86%, NNT = 1:77. The risk of actionable pathology is low, but not negligible. The variation between populations is wide. The prevalence of finding such pathology supports the routine practice of sending all hysterectomy specimens performed for POP for histological assessment.

Objectives\ud This is a protocol for a Cochrane Review (intervention). The objectives are as follows:\ud \ud To assess the effect of surgical interventions on postoperative visual and anatomical outcomes in people with a confirmed degenerative lamellar macular hole.

Purpose Determining which factors influence idiopathic macular hole (MH) size is important because it is a major prognostic indicator of treatment success. Foveal pit morphologic features are highly symmetrical within individuals and may influence idiopathic MH size. Using a series of patients with unilateral idiopathic MHs, we examined the foveal floor size of the fellow eye to evaluate its relationship with idiopathic MH size and postoperative outcomes. Design Retrospective observational study. Participants Two hundred forty-one participants with a unilateral idiopathic MH treated with surgery and a fellow eye with no ocular pathologic features. Methods Both eyes underwent spectral-domain (SD) OCT imaging at the time of surgery. Minimum linear diameter (MLD) and base diameter (BD) defined idiopathic MH size. Foveal floor width (FFW) and minimal foveal thickness defined foveal pit morphologic features of the fellow eye. Main Outcome Measures Baseline characteristics, SD OCT measurements, and preoperative variables were compared to determine their relationship with idiopathic MH size and postoperative visual acuity (VA) in logarithm of the minimum angle of resolution units. Results Foveal floor width was correlated with MLD (r = 0.36; P ≤ 0.001) and BD (r = 0.30; P ≤ 0.001), but not postoperative VA. Minimum linear diameter correlated with preoperative VA (r = 0.49; P ≤ 0.0001) and postoperative VA (r = 0.54; P ≤ 0.0001). A 2-stage regression model was developed to predict postoperative VA (r2 = 0.28): preoperative VA (β = 0.36; P = 0.002) explained 13% of variability and MLD (β = 0.29; P = 0.002), and idiopathic MH duration (β = 0.23; P = 0.004) explained a further 16%. Conclusions Foveal floor width of the fellow eye in patients with a unilateral idiopathic MH was correlated significantly with idiopathic MH size and may explain some of the variability in idiopathic MH size observed between individuals. However, FFW could not predict postoperative vision.

Background As of January 7, 2020, a total of 2558 hospitalized patients with nonfatal cases and 60 patients with fatal cases of e-cigarette, or vaping, product use-associated lung injury (EVALI) had been reported to the Centers for Disease Control and Prevention (CDC). Methods In a national study, we compared the characteristics of patients with fatal cases of EVALI with those of patients with nonfatal cases to improve the ability of clinicians to identify patients at increased risk for death from the condition. Health departments reported cases of EVALI to the CDC and included, when available, data from medical-record abstractions and patient interviews. Analyses included all the patients with fatal or nonfatal cases of EVALI that were reported to the CDC as of January 7, 2020. We also present three case reports of patients who died from EVALI to illustrate the clinical characteristics common among such patients. Results Most of the patients with fatal or nonfatal cases of EVALI were male (32 of 60 [53%] and 1666 of 2498 [67%], respectively). The proportion of patients with fatal or nonfatal cases was higher among those who were non-Hispanic white (39 of 49 [80%] and 1104 of 1818 [61%], respectively) than among those in other race or ethnic groups. The proportion of patients with fatal cases was higher among those 35 years of age or older (44 of 60 [73%]) than among those younger than 35 years, but the proportion with nonfatal cases was lower among those 35 years of age or older (551 of 2514 [22%]). Among the patients who had an available medical history, a higher proportion of those with fatal cases than those with nonfatal cases had a history of asthma (13 of 57 [23%] vs. 102 of 1297 [8%]), cardiac disease (26 of 55 [47%] vs. 115 of 1169 [10%]), or a mental health condition (32 of 49 [65%] vs. 575 of 1398 [41%]). A total of 26 of 50 patients (52%) with fatal cases had obesity. Half the patients with fatal cases (25 of 54 [46%]) were seen in an outpatient setting before hospitalization or death. Conclusions Chronic conditions, including cardiac and respiratory diseases and mental health conditions, were common among hospitalized patients with EVALI.

Objectives\ud To explore the impact of implementation of Community‐Acquired Pneumonia (CAP) quality care bundle.\ud \ud Setting\ud Eight acute hospitals in the North East of England and North Cumbria.\ud \ud Participants\ud ICD‐10 coded CAP aged >18 were identified. A total of 16 201 CAP patients were discharged 2016/2017 (15 707; 2015/2016 and 10 733; 2014/2015).\ud \ud Outcome measures\ud Secondary User Service (SUS) data were collected monthly from April 2014 to 2017. Data were pseudonymised and data flows governed by Data Sharing Agreements. CAP measures were based on British Thoracic Society guidance and agreed following clinician consultation. CAP admissions and individual organisational compliance with and impact of, CAP quality bundle measures was explored.\ud \ud Results\ud Average length of stay (LOS) was 10.4 days (median 6) 25% >13 days. Crude in‐hospital mortality rate was 17.6%, significantly lower (95% CI) than 19.1% in 2015/2016 and 19.3% in 2014/2015. Emergency readmissions within 28 days were 19.7% (19.2%; 2015/2016, 17.9%; 2014/2015). A total of 39.5% of patients received all appropriate care measures. Compliance has improved over time, although not for all hospitals. Most quality measures have higher mortality for those passing measures compared to those failing (P < .05 95% CI). Giving oxygen, had a significantly higher emergency readmission rate, 3.3% higher (95% CI 1.1% to 5.5%). Appropriate antibiotics and recording CURB‐65 scores reduced the emergency readmission rates (−2.7% (95% CI −4.5% to −0.8%) −2.6% (95% CI −3.8% to −1.4%), respectively, (P = ns)).\ud \ud Conclusion\ud CAP accounts for significant bed days, mortality and readmissions. Although mortality was lower, LOS and readmission rates were not, despite improvements in compliance after implementation of the care bundle. Care bundle use remained sub‐optimal.

Apolipoprotein L1 (ApoL1) is an intriguing protein and was shown to be involved in numerous diseases states such as chronic kidney disease (CKD), cardiovascular diseases(CVD), Human Immunodeficiency Virus associated Nephropathy (HIVAN), systemic lupus erythematosus (SLE) (associated collapsing glomerulopathy), and type 2 diabetes(especially for people of African descent). People of African ancestry were shown to have either one or two alleles which contribute to their susceptibility of diseases related to CKD. ApoL1 is characterized as a HDL binding protein. In this work we studied ApoL1 binding to HDL by using a gentle size fractionation technique, asymmetric flow field-flow fractionation (AF4), and liquid chromatography (LC) with quantitative tandem mass spectrometry (MS/MS) detection. We injected 50 uL of whole plasma onto the AF4 system, where smaller HDL particles eluted first followed by the larger HDL, and then by LDL, IDL and VLDL particles. The lipoproteins were separated into 40 fractions and quantitatively analyzed using on-line trypsin digestion coupled with LC-MS/MS analysis. We analyzed plasma samples both with and without purification by UC. When HDL particles were separated by UC, apoL1 was found in the higher density HDL fractions, as was also found by previous studies. When HDL separated using AF4 with or without UC, apoL1 eluted in the larger HDL size region (12-13 nm). With UC vs. without UC, the apoL1 peptide signal intensities were reduced by more than 50%, indicating significant loss of apoL1 from the surface of HDL particles during UC. Our data on apoL1 provides further evidence for the fact that size and density are not directly interconvertible physical characteristics of lipoproteins. Furthermore, our data shows that apoL1 is also an example of exchangeable apolipoproteins whose binding can be significantly diminished due to the effect of intense shear forces during UC separation. This work also demonstrates the advantages of utilizing AF4-IMER-UPLC-MS/MS methodology to separate lipoprotein particles and study of their apolipoprotein composition.

The number of caesarean sections at maternal request without medical indication is increasing. We aimed to explore the views of pregnant women, midwives and doctors using six hypothetical clinical scenarios and compare group views on: (a) perceived appropriateness of requests for caesarean section and (b) the reasons underlying these requests.A questionnaire was distributed to 166 pregnant women, 31 midwives and 52 doctors within maternity units at two hospitals in the North East region of England. Six hypothetical clinical scenarios for maternal requests were used: (1) uncomplicated first pregnancy, (2) one previous normal delivery, (3) one previous instrumental delivery, (4) one previous caesarean section, (5) one previous caesarean section with vaginal delivery since and (6) uncomplicated twin pregnancy. To highlight the differences in group responses, two main questions were asked for each scenario: 1. Should women be able to request a caesarean section? 2. What do you feel are the reasons for requesting a caesarean section? Data was analysed using Chi-squared or likelihood ratio as appropriate.In scenarios 1-3, professional groups were 'less likely' than pregnant women to always support a request (2.4% vs. 19.4%), (2.6% vs. 15.6%), (4.6% vs. 22%), (p0.001). No significant differences were shown between doctors and midwives except for scenario 6 (twins), where midwives more often felt maternal requests should be declined (26.1% vs. 1.9%) (p=0.001). Multiparous women (n=95) were more likely to agree 'sometimes' to maternal requests in scenarios 1, compared to nulliparous women (n=71) (21.1% vs. 4.2%) (p=0.04). 'Safety of the baby' was ranked highly with pregnant women in scenarios 1-3 (mean 24.4%, range [15.8-38%]) compared with healthcare professionals (7.6% [3.4-12.8%]). However in scenario 3, healthcare professionals attributed 'fear of injury to self' (29.6%) as the most likely reason compared to 14.6% of pregnant women.Healthcare professionals and pregnant women's views differ significantly. Multiparous patients' views differ from those who have not had children before. We should provide clearer information on risks and benefits which encompass areas that concern women most.

To improve awareness and recognition of chronic bacterial prostatitis (CBP) and chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) among non-specialists and patients. To provide guidance to healthcare professionals treating patients with CBP and CP/CPPS, in both non-specialist and specialist settings. To promote efficient referral of care between non-specialists and specialists and the involvement of the multidisciplinary team (MDT).The guideline population were men with CBP or CP/CPPS (persistent or recurrent symptoms and no other urogenital pathology for ≥3 of the previous 6 months). Consensus recommendations for the guidelines were based on a search to identify literature on the diagnosis and management of CBP and CP/CPPS (published between 1999 and February 2014). A Delphi panel process was used where high-quality, published evidence was lacking.CBP and CP/CPPS can present with a wide range of clinical manifestations. The four main symptom domains are urogenital pain, lower urinary tract symptoms (LUTS - voiding or storage symptoms), psychological issues and sexual dysfunction. Patients should be managed according to their individual symptom pattern. Options for first-line treatment include antibiotics, α-adrenergic antagonists (if voiding LUTS are present) and simple analgesics. Repeated use of antibiotics, such as quinolones, should be avoided if there is no obvious symptomatic benefit from infection control or cultures do not support an infectious cause. Early use of treatments targeting neuropathic pain and/or referral to specialist services should be considered for patients who do not respond to initial measures. An MDT approach (urologists, pain specialists, nurse specialists, specialist physiotherapists, general practitioners, cognitive behavioural therapists/psychologists, and sexual health specialists) is recommended. Patients should be fully informed about the possible underlying causes and treatment options, including an explanation of the chronic pain cycle.Chronic prostatitis can present with a wide variety of signs and symptoms. Identification of individual symptom patterns and a symptom-based treatment approach are recommended. Further research is required to evaluate management options for CBP and CP/CPPS.

The presence of diabetes mellitus in patients significantly increases the risk of cardiovascular disease, among many negative health outcomes. Several research groups have reported lipid species anomalies in subjects showing insulin resistance, including higher concentrations of sphingomyelin species relative to normal controls. Advances in lipidomic methodologies has allowed for the profiling of numerous lipid species in a single extraction and analytical run. We used the SCIEX Lipidyzer platform to determine the absolute concentration of 770 distinct lipid species from 52 subjects categorized into hyperglycemic (n=30) and normal subjects (n=22). Lipid species were determined from the following classes: cholesterol esters (CE), ceramides (CER), diacylglycerols (DAG), dihydroceramide (DCER), free fatty acids (FFA), hexosylceramides (HCER), lactosylceramides (LCER), lysophosphatidylcholines (LPC), lysophosphatidylethanolamines (LPE), phosphatidylcholines (PC), phosphatidylethanolamines (LPE), sphingomyelins (SM), and triacylglycerols (TAG). After normalizing by the sum of total apolipoprotein A and apolipoprotein B (to account for the possibility of reduced particle number due to medicinal intervention), lipids both by class and species were compared using one way analysis of variance. On a class level, relative to normal subjects, hyperglycemic patients showed increased levels of CER and FFA. Individual lipid species of these classes that were higher in hyperglycemic patients include CER(18:0), CER(20:0), CER(24:1), FFA(16:0), FFA(16:1), FFA(18:0), FFA(18:1), FFA(18:2), FFA(18:3), and FFA(20:3) Differences were also found in SM concentrations between hyperglycemic and normal patients resulting in a higher the SM/PC ratio, indicating changes in the lipid fluidity of the shell in lipoprotein particles of hyperglycemic patients. On a species level, SM(16:0), SM(18:1), and SM(24.1), were elevated in hyperglycemic subjects relative to normal subjects. This finding of elevated unsaturated SM species in hyperglycemic patients is at odds with other studies where it was found that the saturated SM moieties were elevated in insulin resistant subjects.

• Benign prostatic hyperplasia (BPH) is a common cause of bothersome lower urinary tract symptoms. In the past, the aim of drug treatment was to relieve symptoms until surgery became necessary, predominantly using an α-blocker or a 5α-reductase inhibitor (5ARI) as monotherapy. • Together with improving knowledge about the pathogenesis of BPH, there is now strong evidence from large randomized trials that risk stratification and appropriate treatment with combined α-blocker/5ARI therapy can significantly reduce the risk of disease progression and avoid long-term complications such as acute urinary retention and surgery. • BPH will increasingly be managed in primary care in the future and, if new management strategies based on this evidence are to be implemented cost effectively, there is a need to introduce shared care between the primary and secondary care sectors to optimise use of resources and expertise.

Aims: Analysis of pre-operative spectral domain optical coherence tomography (SD-OCT) characteristics of full-thickness macular holes (FTMH) and effect on optimum management. Methods: We retrospectively reviewed SD-OCT characteristics of a consecutive cohort of patients waitlisted for FTMH surgery and categorized them by current evidence-based treatments. Results: Out of the 106 holes analysed, 36 were small, 40 medium and 30 large. Initially, 33 holes had vitreomacular adhesion (VMA). 41 holes were analysed for change in characteristics with a median duration of 8 weeks between the scans. The number of small or medium holes decreased from 20 to 6 and that of large holes doubled. The number of holes with VMA halved. Smaller hole size (p = 0.014) and being phakic (p = 0.048) were associated with a larger increase in size. The strongest predictor of hole progression into a different surgical management category was the presence of VMA. Conclusion: FTMH characteristics can change significantly pre-operatively and affect optimal treatment choice.

Botulinum neurotoxins (BoNTs) are a family of seven toxin serotypes that are the most toxic substances known to humans. Intoxication with BoNT causes flaccid paralysis and can lead to death if untreated with serotype-specific antibodies. Supportive care, including ventilation, may be necessary. Rapid and sensitive detection of BoNT is necessary for timely clinical confirmation of clinical botulism. Previously, our laboratory developed a fast and sensitive mass spectrometry (MS) method termed the Endopep–MS assay. The BoNT serotypes are rapidly detected and differentiated by extracting the toxin with serotype-specific antibodies and detecting the unique and serotype-specific cleavage products of peptide substrates that mimic the sequence of the BoNT native targets. To further improve the sensitivity of the Endopep–MS assay, we report here the optimization of the substrate peptide for the detection of BoNT/A. Modifications on the terminal groups of the original peptide substrate with acetylation and amidation significantly improved the detection of BoNT/A cleavage products. The replacement of some internal amino acid residues with single or multiple substitutions led to further improvement. An optimized peptide increased assay sensitivity 5-fold with toxin spiked into buffer solution or different biological matrices.

Botulinum neurotoxin (BoNT) is one of the most toxic substances known. BoNT is classified into seven distinct serotypes labeled A through G. Among individual serotypes, researchers have identified subtypes based on amino acid variability within a serotype and toxin variants with minor amino acid sequence differences within a subtype. BoNT subtype identification is valuable for tracing and tracking bacterial pathogens. A proteomics approach is useful for BoNT subtyping since botulism is caused by botulinum neurotoxin and does not require the presence of the bacteria or its DNA. Enzymatic digestion and peptide identification using tandem mass spectrometry determines toxin protein sequences. But with the conventional one-step digestion method, producing sufficient numbers of detectable peptides to cover the entire protein sequence is difficult and incomplete sequence coverage results in uncertainty in distinguishing BoNT subtypes and toxin variants because of high sequence similarity. We report here a method of multiple enzymes and sequential in-gel digestion (MESID) to characterize the BoNT protein sequence. Complementary peptide detection from toxin digestions has yielded near-complete sequence coverage for all seven BoNT serotypes. Application of the method to a BoNT-contaminated carrot juice sample resulted in the identification of 98.4% protein sequence which led to a confident determination of the toxin subtype.

Botulinum neurotoxins (BoNTs) cause the disease botulism, which can be lethal if untreated. There are seven known serotypes of BoNT, A–G, defined by their response to antisera. Many serotypes are distinguished into differing subtypes based on amino acid sequence, and many subtypes are further differentiated into toxin variants. Previous work in our laboratory described the use of a proteomics approach to distinguish subtype BoNT/A1 from BoNT/A2 where BoNT identities were confirmed after searching data against a database containing protein sequences of all known BoNT/A subtypes. We now describe here a similar approach to differentiate subtypes BoNT/B1, /B2, /B3, /B4, and /B5. Additionally, to identify new subtypes or hitherto unpublished amino acid substitutions, we created an amino acid substitution database covering every possible amino acid change. We used this database to differentiate multiple toxin variants within subtypes of BoNT/B1 and B2. More importantly, with our amino acid substitution database, we were able to identify a novel BoNT/B subtype, designated here as BoNT/B7. These techniques allow for subtype and strain level identification of both known and unknown BoNT/B rapidly with no DNA required. Figure Identification of an existing or new BoNT/B can be accomplished through MS/MS analysis of digestion fragments of the protein.

Bordetella pertussis (Bp) is the etiologic agent of pertussis or whooping cough, a highly contagious respiratory disease occurring primarily in infants and young children. Although vaccine preventable, pertussis cases have increased over the years leading researchers to re-evaluate vaccine control strategies. Since bacterial outer membrane proteins, comprising the surfaceome, often play roles in pathogenesis and antibody-mediated immunity, three recent Bp circulating isolates were examined using proteomics to identify any potential changes in surface protein expression. Fractions enriched for outer membrane proteins were digested with trypsin and the peptides analyzed by nano liquid chromatography-electrospray ionization-mass spectrometry (nLC-ESI-MS), followed by database analysis to elucidate the surfaceomes of our three Bp isolates. Furthermore, a less labor intensive non-gel based antibody affinity capture technology in conjunction with MS was employed to assess each Bp strains' immunogenic outer membrane proteins. This novel technique is generally applicable allowing for the identification of immunogenic surface expressed proteins on pertussis and other pathogenic bacteria.

Purpose.: To describe the intraoperative staining pattern of the internal limiting membrane (ILM)-specific dye Brilliant Blue G (BBG) in a cohort of patients with idiopathic macular holes; to analyze the associations of the staining pattern with pre- and postoperative variables and to correlate the staining pattern with transmission electron microscopy (TEM) of the excised ILM.\ud \ud Methods.: Fifty-five consecutive patients were studied. The staining pattern was divided into three subtypes based on the intraoperative appearance. The presence of a narrow rim of nonstaining around the macular hole (MH) edge was noted and measured. In the final 21 patients, the excised ILM was examined with TEM.\ud \ud Results.: The pattern of staining observed was categorized as uniform in 33 patients (60%), patchy nonstaining in 17 (31%), and no visible staining in 5 (9%). The staining pattern correlated with the MH stage. In the patients with uniform or patchy staining, a nonstaining rim was observed in 26 (52%) of the 50. The presence of a rim was associated with a greater hole diameter and lower postoperative visual acuity. The stain pattern correlated significantly with the amount of cellular tissue on the vitreous side of the ILM on TEM, with a greater proportion of multicellular layer membranes and new collagen in the incomplete staining groups.\ud \ud Conclusions.: A variety of nonstaining patterns around macular holes can be observed using BBG, and these patterns correlate to the amount of cellular tissue on the vitreous side of the ILM seen histologically. These patterns could be used to guide the ILM peeling requirement or extent in future studies.