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Congenital heart disease (CHD) is the most common birth defect and brings with it significant mortality and morbidity. The application of exome and genome sequencing has greatly improved the rate of genetic diagnosis for CHD but the cause in the majority of cases remains uncertain. It is clear that genetics, as well as environmental influences, play roles in the aetiology of CHD. Here we address both these aspects of causation with respect to the Notch signalling pathway. In our CHD cohort, variants in core Notch pathway genes account for 20% of those that cause disease, a rate that did not increase with the inclusion of genes of the broader Notch pathway and its regulators. This is reinforced by case-control burden analysis where variants in Notch pathway genes are enriched in CHD patients. This enrichment is due to variation in NOTCH1. Functional analysis of some novel missense NOTCH1 and DLL4 variants in cultured cells demonstrate reduced signalling activity, allowing variant reclassification. Although loss-of-function variants in DLL4 are known to cause Adams-Oliver syndrome, this is the first report of a hypomorphic DLL4 allele as a cause of isolated CHD. Finally, we demonstrate a gene-environment interaction in mouse embryos between Notch1 heterozygosity and low oxygen- or anti-arrhythmic drug-induced gestational hypoxia, resulting in an increased incidence of heart defects. This implies that exposure to environmental insults such as hypoxia could explain variable expressivity and penetrance of observed CHD in families carrying Notch pathway variants.

Purpose To report 2 cases with late postoperative Candida albicans interface keratitis and endophthalmitis after Descemet stripping automated endothelial keratoplasty (DSAEK) with corneal grafts originating from a single donor with a history of presumed pulmonary candidiasis. Methods Two patients underwent uncomplicated DSAEK by 2 corneal surgeons at different surgery centers but with tissue from the same donor and were referred to the Bascom Palmer Eye Institute with multifocal infiltrates at the graft-host cornea interface 6 to 8 weeks later, and anterior chamber cultures that were positive for the same genetic strain of C. albicans. Immediate explantation of DSAEK lenticules and daily intracameral and instrastromal voriconazole and amphotericin injections failed to control the infection. Thus, both patients underwent therapeutic penetrating keratoplasty with intraocular lens explantation, pars plana vitrectomy, and serial postoperative intraocular antifungal injection. Results Both patients are doing well at 2 years postoperatively with best-corrected vision of 20/20 and 20/30+ with rigid gas permeable lenses. One patient required repeat optical penetrating keratoplasty and glaucoma tube implantation 1 year after the original surgery. Literature review reveals that donor lenticule explantation and intraocular antifungals are often inadequate to control fungal interface keratitis, and a therapeutic graft is commonly needed. Conclusions Interface fungal keratitis and endophthalmitis due to infected donor corneal tissue is difficult to treat, and both recipients of grafts originating from the same donor are at risk of developing this challenging condition.

Birth defects occur in up to 3% of all live births and are the leading cause of infant death. Here we present five individuals from four unrelated families, individuals who share similar phenotypes with disease-causal bi-allelic variants in NADSYN1, encoding NAD synthetase 1, the final enzyme of the nicotinamide adenine dinucleotide (NAD) de novo synthesis pathway. Defects range from the isolated absence of both kidneys to multiple malformations of the vertebrae, heart, limbs, and kidney, and no affected individual survived for more than three months postnatally. NAD is an essential coenzyme for numerous cellular processes. Bi-allelic loss-of-function mutations in genes required for the de novo synthesis of NAD were previously identified in individuals with multiple congenital abnormalities affecting the heart, kidney, vertebrae, and limbs. Functional assessments of NADSYN1 missense variants, through a combination of yeast complementation and enzymatic assays, show impaired enzymatic activity and severely reduced NAD levels. Thus, NADSYN1 represents an additional gene required for NAD synthesis during embryogenesis, and NADSYN1 has bi-allelic missense variants that cause NAD deficiency-dependent malformations. Our findings expand the genotypic spectrum of congenital NAD deficiency disorders and further implicate mutation of additional genes involved in de novo NAD synthesis as potential causes of complex birth defects.

PURPOSE: To evaluate whether high-resolution optical coherence tomography (HR-OCT) can detect histologic tumor margins of ocular surface squamous neoplasia (OSSN). METHODS: Eight eyes of 8 patients with OSSN undergoing excision were studied prospectively. Immediately before surgery, the tumor was imaged using a commercially available HR-OCT (Bioptigen/Leica Microsystems, Buffalo Grove, IL) to identify the conjunctival margins of the neoplastic lesion. The tumor borders of the lesion determined by HR-OCT were mapped in relation to an anatomical reference point and transferred intraoperatively. The tumor was excised with 4 mm margins from the visible edge of the lesion with a “no-touch” technique. The specimens were sent for pathological analysis and the histologic tumor margin was compared to the HR-OCT predicted tumor border. RESULTS: Mean age of the 8 patients were 67±9.9 years. Seven (87.5%) were male, 7(87.5%) were white and ethnically, 3 (37.5%) were Hispanic. All eight tumors were bulbar and in the exposure zone. Seven tumors were limbal. Corneal extension was present in 5(62.5%). Mean tumor area was 17.5±11.1 mm(2). Clinically, two (25%) of the tumors were leukoplakic, 1(12.5%) papillomatous, and 3(37.5%) gelatinous. A conjunctival tumor margin identified with the HR-OCT coincided with the pathologically confirmed margin mark in all eyes. CONCLUSIONS: HR-OCT has the potential to predict histologic tumor margins in OSSN. Optical identification of tumor margins could potentially decrease the incidence of residual positive margins, and minimize healthy tissue removal. Advances in HR-OCT technology and integration into a microscope for ‘real-time” imaging are needed to further improve this technique.

Background Primary colonic epithelial defects leading to inflammatory responses are considered central to the development of ulcerative colitis (UC). However, a systematic analysis of various colonic subcompartments in the pathogenesis of UC before inflammation remains elusive. Here, we explored changes in colonic subcompartments and their associated niche signals in patient mucosal biopsies and in an animal model of colitis. Methods Analysis of mucosal biopsies obtained from uninvolved and involved regions of patients with UC and Crohn's disease was performed and compared with normal subjects. Temporal analysis of colonic subcompartments was performed in mice administered with 5% dextran sodium sulphate. Phenotypic enumeration of the crypt subcompartment was complemented with flow cytometric analysis. Members of Notch and Wnt signaling pathways were analyzed by molecular, biochemical, and colocalization studies. Results Phenotypic enumeration of colonocytes' subcompartments from patients revealed significant alterations of the lower crypt, enriched in stem cell and progenitors, independent of inflammation. These changes, unique to UC, were confirmed by immunohistochemistry and molecular analysis. In parallel, a defect in proliferation and Muc2 synthesis was observed. Animal data before inflammation recapitulated human studies. Mechanistic studies revealed that changes in signaling through Wnt primarily affected colonic stem cells, whereas Notch affected progenitor function. Conclusions Our results thus provide new insights into the development of inflammation and relapse in UC and suggest that the stem cell niche in the colon may influence pathogenesis of the disease.

Glaucoma may develop or worsen after ocular surface transplantation and often requires surgical management for adequate intraocular pressure control. Traditional glaucoma filtering procedures in patients with prior ocular surface transplant may be problematic for several reasons, which include mechanical disruption of the pre-existing graft, epithelial and stem cell toxicity induced by antifibrotic agents, and increased risk of future corneal transplantation failure. We describe the implantation of a glaucoma drainage implant via a limbal-based conjunctival incision with tube placement in the ciliary sulcus in three eyes of two patients with prior ocular surface transplantation. At a follow-up interval of 3-7 months, all three eyes have excellent postoperative control of intraocular pressure, stable vision, and healthy ocular surface grafts.

With the introduction of highly sensitive hepatitis B surface antigen immunoassay, transfusion associated HBV infection have reduced drastically but they still tend to occur due to blood donors with occult hepatitis B infection (OBI) and window period (WP) infection. Sera from, 24338 healthy voluntary blood donors were screened for HBsAg, HIV and HCV antibody using Vitros Enhanced Chemiluminescent Immunoassay. The median age of the donor population was 30 (range 18–54) with male preponderance (98%). All serologically negative samples were screened by nucleic acid testing (NAT) for viral DNA and RNA. NAT-positive samples were subjected to discriminatory NAT for HBV, HCV, and HIV and all samples positive for HBV DNA were tested for anti-HBc, anti-HBs, HBeAg. Viral load was determined using artus HBV RG PCR Kit. Of the 24,338 donors screened, 99.81% (24292/24338) were HBsAg negative of which NAT was positive for HBV DNA in 0.0205% (5/24292) donors. Four NAT positive donors had viral load of

PURPOSE To describe a method of surgical management for cases of localized epithelial downgrowth (EDG). METHODS Single-center retrospective chart review of patients with histologically confirmed localized EDG of stratified squamous epithelium. Each patient underwent a partial lamellar sclerokeratouvectomy encompassing the area of suspected EDG. The main outcome measure was recurrence of EDG. RESULTS At a mean of 51 months of postoperative follow-up (range: 31-86), no patient had evidence of recurrent EDG. One eye developed worsening of preexisting glaucoma. All 3 eyes developed localized corneal edema, but only 1 required corneal transplantation. CONCLUSIONS EDG is a potentially devastating complication of intraocular surgery or penetrating injury. In cases of localized involvement of the anterior chamber structures, definitive management of EDG can be achieved via resection of the involved scleral, corneal, and uveal tissue. A lamellar approach minimizes tissue loss and maintains a structurally sound, pressurized globe.

The primary aim of this study was to assess the usefulness of plasma hydrogen sulphide (H2S) level at admission as a predictor of severity of acute pancreatitis. The secondary aims were to examine whether the level of H2S after 48 h correlated with severity and whether level of H2S correlated with pulmonary, renal or infectious complications. Plasma hydrogen sulphide was measured within 24 h of admission and 48 h later, in patients with acute pancreatitis. Patients were classified as having mild or severe pancreatitis, and H2S levels in the two groups were compared. A total of 55 patients had H2S estimation carried out within 24 h of admission. H2S levels were similar in patients with mild (mean 31.8 ± 18.8, range 7.1 to 81.4 µmol/L) and severe pancreatitis (mean 28.2 ± 21.6, range 6.1 to 74.4 µmol/L; p = 0.339). There was no difference found between the groups after 48 h (mild n = 28, mean 26.8 ± 19.4 µmol/L, and severe n = 20, mean 34.6 ± 21.0 µmol/L; p = 0.127). There was also no difference in the levels between patients with or without lung injury, kidney injury or sepsis. Performing H2S estimation to predict severity in acute pancreatitis is not beneficial.

To determine the etiologic diagnosis of infectious corneal ulcers at Tilganga Institute of Ophthalmology, a tertiary teaching hospital in Kathmandu Nepal, from 2006-2009.This study involved a review of all microbiology records at Tilganga Institute of Ophthalmology from August 2006 through July 2009. Microbiologic records from the corneal scrapings of all patients suspected of having infectious corneal ulcers were included.Corneal scrapings were obtained from 468 patients. The average patient age was 52 years, and 55% of the affected cases were males. Microorganisms were grown from 185 of the corneal scrapings (40%). Pure bacterial cultures were obtained from 72 patients (39%), and pure fungal cultures were obtained from 113 patients (61%). Gram stain was 75% sensitive (95% confidence interval, 0.632-0.841) in identifying bacterial infection, whereas KOH prep was 80.5% sensitive (95% confidence interval, 0.718-0.871) in identifying fungal organisms. Of 72 bacterial isolates, 50 isolates (69%) were Streptococcus pneumoniae, the most common organism isolated in this study. Of 113 fungal isolates, 40 of isolates (35%) were identified as Aspergillus sp.Fungal organisms (61%) are the most common cause of infectious keratitis in this patient population. Of all organisms, S. pneumoniae was the most common organism identified. Smear microscopy is reliable in rapidly determining the etiology of the corneal infection and can be used to help guide initial therapy in this setting.

Purpose To describe the morphological distribution of host tissue and microbial biofilms on the intraocular surfaces of Boston type I keratoprostheses (KPros) explanted because of corneal melt. Methods Retrospective study of scanning electron microscopy (SEM) images from 4 explanted Boston type I KPros composed of polymethylmethacrylate and titanium. SEM images of KPro-associated ocular surfaces were reviewed for the presence of inflammatory cells, microbes, and/or biofilm formation. One sterile type I KPro was also imaged to serve as a (device only) control. Results All 4 KPros were explanted because of culture-negative, clinically "sterile" donor corneal melt with impending KPro extrusion. In all cases, the rough, irregular surfaces of the device harbored more adherent corneal epithelium and stromacytes, inflammatory cells, and bacteria than the smooth, polished surface of the KPro optic. Two KPros showed not only evidence of prior bacterial colonization but marked biofilm formation. Conclusions SEM images of explanted KPros explanted for "sterile" corneal melt demonstrated evidence of biofilm formation despite negative donor corneal cultures and the absence of clinical suspicion for infection. These results suggest that "sterile" corneal melt may be due to inflammatory host responses to low microbial burdens as seen in biofilms and/or released antigens after antibiotic-induced lysis. There was increased adherence of host tissue cells and microbial biofilms on the nonpolished surfaces of the KPro. Polishing the intraocular polymethylmethacrylate and titanium KPro surfaces may decrease microbial adhesion and biofilm formation in human subjects with KPros, but what impact this will have on rates of postoperative endophthalmitis is unknown.

The present study highlights six cases of pneumococcusuria during the time period of May 2008 to May 2010. All the patients had a co-existing predisposing factor with the isolation of Streptococcus pneumoniae in urine. Five of the six patients having signs and symptoms of urinary tract infections (UTI) were treated and cured of the same. It becomes essential to consider pneumococcal UTI in the presence of clinical signs and symptoms associated with urinary tract abnormalities like hydronephrosis and renal stones. S. pneumoniae may be regarded as an emerging pathogen in UTI. Precise microbiological diagnosis must correlate with the clinical signs and symptoms for the administration of appropriate antibiotic therapy.

We describe a case of bacteraemia caused by Weissella confusa in a 48-year-old male who was operated on for adenocarcinoma of the gastro-oesophageal junction and maintained on total parenteral nutrition. Blood cultures were positive for a vancomycin-resistant streptococcus-like organism which was identified as W. confusa by 16S rRNA gene sequencing.

Infections caused by Nocardia farcinica are potentially lethal because of the organism's tendency to disseminate and resist antibiotics. Central nervous system involvement has been documented in 30% of infections caused N. farcinica.Case report and review of the literature.A case of primary brain abscess caused by N. farcinica, identified by 16SrRNA sequencing, is presented, and 39 cases reported previously in the literature are reviewed. Our patient underwent a neuronavigation-guided right frontal craniotomy and was treated with trimethoprim/sulfamethoxazole and amoxicillin-clavulanic acid for 12 mo. He showed marginal improvement in his prior left hemiparesis at the last review 14 months later.Cases of N. farcinica infections are being reported increasingly because of recent changes in taxonomy and diagnostic methodology. This change in epidemiology has implications for therapy because of the organism's pathogenicity and natural resistance to multiple antimicrobial agents, including third-generation cephalosporins. Any delay in starting appropriate antibiotic therapy can have adverse consequences.

Importance Retroprosthetic membrane (RPM) formation is the most common complication associated with the Boston type 1 keratoprosthesis and has been associated with corneal melt. Objective To identify the histological and immunohistochemical characteristics of RPMs associated with corneal melt. Design, Setting, and Participants Observational histopathological case series at a tertiary eye care referral center among patients who underwent Boston type 1 keratoprosthesis explantation because of donor corneal melt at the Illinois Eye and Ear Infirmary between January 1, 2011, and January 1, 2012. Exposures Seven RPM specimens from 7 eyes were stained with hematoxylin-eosin, cytokeratin 7, cytokeratin AE1/3, smooth muscle actin, vimentin, and CD34. Light microscopy was used to evaluate specimens for inflammation and epithelial ingrowth. XY-karyotyping using fluorescence in situ hybridization was performed on 4 specimens with known donor-recipient sex mismatch. Main Outcomes and Measures Histological and immunohistochemical characteristics of RPMs. Results Inflammatory cells were present in 4 of 7 RPMs. In 3 of 4 sex-mismatched specimens, tissue XY-karyotyping of the RPM interphase cells was consistent with the host sex karyotype. The fourth specimen showed a mixture of recipient-type and donor-type cells. Conclusions and Relevance Melt-associated RPMs show variable degrees of inflammation. Most membranes seem to originate from a proliferation of host cells, but donor tissue may contribute in some cases.

Chromoblastomycosis is a chronic subcutaneous mycosis commonly caused by Fonsecaea, Phialophora, and Cladophialophora spp. Out of these, Fonsecaea pedrosoi is the most common etiological agent, implicated in 70%–90% of the cases reported worldwide. The histopathological diagnosis of chromoblastomycosis is based on visualization of medlar or sclerotic bodies in the tissue. These sclerotic bodies divide by planar division. Rarely, budding is seen in these sclerotic bodies. As this entity can be confused with phaeohyphomycosis, it is important to be aware of such a presentation also. We report two cases of chromoblastomycosis that showed budding sclerotic bodies.

Systemic endemic mycoses, such as blastomycosis, are rare in Asia and have been reported as health risks among travelers who visit or reside in an endemic area. Adrenal involvement is rarely seen in blastomycosis and has never been reported from Asia. We report the first case of blastomycosis with bilateral involvement of the adrenals in a diabetic patient residing in the state of Arunachal Pradesh, India.

Intracranial infections, especially subdural empyema, due to salmonella are rare. Subdural empyema caused by Salmonella paratyphi A has been documented only once earlier in the literature. Hence, we report a case of subdural empyema and osteomyelitis of cranial vault due to S. paratyphi A. A 42- year-old male presented with headache and purulent discharge from right parietal burr hole wound site. Patient gave a history of head injury two years ago. He underwent burr hole evacuation of chronic subdural haematoma, excision of outer membrane and right parietal craniectomy. The cultures grew S. paratyphi A. Recovery was uneventful following surgical intervention and antibiotic therapy.

Diving into lakes and ponds is a common activity of rural children. We present two cases of penetrating orbital injuries from plant matter sustained in this manner. Such injuries pose a particular challenge because wooden foreign bodies are often missed during orbital exploration, and current imaging modalities cannot reliably identify retained organic material. When a patient presents with orbital penetration after a high-risk mechanism of injury, such as freshwater diving, the clinician must maintain a very high index of suspicion for retained wooden foreign body.

The authors report a rare case of renal cell carcinoma (RCC) metastasis to the paranasal sinuses. The authors review RCC and its potential for sinonasal metastasis and discuss the variable presentation and need for clinical suspicion for early diagnosis and treatment. A 74-year-old man presented with numbness of the left side of the face, reduced visual acuity and ptosis 12 years after nephrectomy for RCC. Imaging studies showed a lesion in the left pterygopalatine fossa and the histological features supported the diagnosis of metastatic RCC. RCC metastasis to the paranasal sinuses is very rare and can present with various symptoms depending on the affected organ. These symptoms occasionally are the initial manifestation of renal RCC and it is very important to recognise them so that the patient receives the appropriate therapy to improve survival.

Intraocular gnathostomiasis is a rare parasitic infection caused by the third-stage larvae of spiruroid nematode Gnanthostoma spp. seen mostly in tropical and subtropical regions. It is a food-borne zoonosis caused by ingestion of raw or undercooked freshwater fish, amphibians, reptiles, birds, and mammals, all of which are known to harbor advanced third-stage larvae of Gnanthostoma spp. To date, 74 cases of intraocular gnathostomiasis have been reported from 12 different countries. Only four countries have reported more than 10 cases each, and India shares the rare distinction of being one of them, with 14 cases. Surprisingly, not a single case of cutaneous gnanthostomiasis has ever been reported from India. We present one such case of intraocular gnathostomiasis in a 41-year-old male who presented with an actively motile worm attached to the iris, and we review the pertinent literature of all such cases reported from India.

Coryneform bacteria belonging to the genus Brevibacterium have emerged as opportunistic pathogens. Of the nine known species of Brevibacterium isolated from human clinical samples, Brevibacterium casei is the most frequently reported species from clinical specimens. We report the first case of B. casei brain abscess in an immunocompetent patient successfully treated by surgery and antimicrobial therapy.

Pneumonia due to non-typhoidal Salmonella is a rarely reported entity. A fatal case of Salmonella pneumonia is reported here where Salmonella Typhimurium was isolated from the endotracheal aspirate and blood culture.

We studied the aerobic bacterial isolates from bile and their susceptibilty pattern in patients with biliary tract disease. Samples of bile collected during endoscopic retrograde cholangio pancreatography (ERCP) and surgery, were inoculated on standard media. Bacterial identification and susceptibility were done by standard techniques. A total of 209 samples were cultured, out of which 128 samples showed growth. Total number of isolates obtained was 221. Poly-microbial infection was detected in 67 patients. Predominant aerobic bacterial isolates obtained were Escherichia coli 30% (67), Klebsiella species 23.98% (53), Enterococcus species 12.21% (27). Multi-drug resistance was noted in 57%. Higher resistance rate was noted among Gram negative bacilli for ampicillin (92.4%), cephalexin (82.46%), ciprofloxacin (68.42%), piperacillin (64.33%). Sensitivity to meropenem was 90.64% and amikacin was 76.61%. Gram positive bacteria showed high resistance to gentamicin (39.53%). Sensitivity to ampicillin was 86.05% and penicillin was 81.4%. Vancomycin and teicoplanin showed 100% sensitivity. From our study we conclude that E. coli, Klebsiella species and Enterococcus species are common pathogens infecting biliary tract. Poly-microbial infection and multi-drug resistance warrants culture and sensitivity to guide antimicrobial therapy. We recommend combination of amikacin and ampicillin for empirical therapy at our institution.

Purpose To evaluate whether retro-backplate retroprosthetic membrane is correlated with risk of melt in patients with a type I Boston Keratoprosthesis (KPro). Design Retrospective, observational case series. Methods Study of 50 eyes of 47 patients with type I Boston KPro and postoperative anterior segment optical coherence tomography (AS OCT) imaging performed at the University of Illinois at Chicago. Main outcome measures were presence of retro-backplate membrane and development of melt requiring explantation. For eyes with melt, membrane thickness was measured using the AS OCT images obtained at the last visit before melt occurred. For eyes without melt, the last available AS OCT images were used for measurement. Results AS OCT evidence of a retro-backplate membrane was observed in 100% of eyes that melted and in 34.1% of eyes that did not ( P = .0034; risk ratio, 2.9; 95% confidence interval, 1.9 to 4.4). Retro-backplate membrane thickness in the melt group was 278 μm versus 193 μm in the nonmelt group ( P = .025). Conclusions The retro-backplate portion of a retroprosthetic membrane is to be differentiated from the retro-optic portion seen at the slit lamp. The retro-backplate membrane as shown by AS OCT imaging is correlated with an increased risk of sterile keratolysis, possibly because of impedance of nutritional support from the aqueous humor. Further studies are needed to better standardize the AS OCT measurements of retro-backplate membranes as well as to identify early interventions to prevent progression of thin membranes once identified on AS OCT.

Purpose To evaluate the efficacy of quantitative real-time polymerase chain reaction (qPCR) in the diagnosis of postoperative bacterial endophthalmitis among patients who underwent cataract surgery at a tertiary care center. Design Prospective experimental study. Methods This was a single-center study of 64 eyes of 64 patients presenting with clinical signs and symptoms of endophthalmitis within 1 year of cataract surgery. Patients with glaucoma filtering or cornea surgery in the past year, postoperative trauma, fungal endophthalmitis, or preoperative inflammatory conditions were excluded. Vitreous samples were obtained during vitreous tap or vitrectomy and sent for both culture and qPCR with sequencing. Vitreous samples obtained from 50 patients undergoing vitrectomy for noninflammatory indications served as controls. The main outcome measures were the sensitivity of qPCR compared to culture and concordance of results of pathogen identification with sequencing vs phenotypic speciation. Results qPCR detected 16s bacterial DNA in 37 patients (66%), compared to 19 (34%) with traditional culture. Only 1 patient had a positive result by culture ( Nocardia species) but negative result by qPCR. For the 18 samples positive by both qPCR and culture, there was a 100% concordance in pathogen identification between sequencing and phenotypic speciation. Conclusion In cases of suspected bacterial endophthalmitis, qPCR offers an improved diagnostic yield and may be a useful adjunct to traditional culture. Further large-scale clinical studies are needed to elucidate the full clinical utility of qPCR.