Capsule Staining Research Articles

Isolation and identification of avirulent strains of Bacillus anthracis from environmental samples in Central Java, Indonesia

Background and Aim: Anthrax is a non-contagious infectious disease caused by Bacillus anthracis. The bacteria form spores that are resistant to extreme conditions and can contaminate the environment for decades. This study aimed to detect and characterize B. anthracis found in endemic areas of anthrax in Yogyakarta and Central Java province, Indonesia. Materials and Methods: Soil samples were collected from Gunungkidul regency, Yogyakarta province (n=315) and Boyolali regency, Central Java province (n=100). Additional soil samples (n=10) and straw samples (n=5) were obtained from Pati regency, Central Java province. The isolation and identification of B. anthracis were performed using conventional methods: Morphology of bacteria colony in solid media, Gram staining, capsule staining, spores staining, and motility test. Isolates were further identified using polymerase chain reaction (PCR) against Ba813, lef (pXO1), and capC (pXO2) gene. An avirulent vaccine strain of B. anthracis (strain 34F2) was used as a control. Results: Only four samples grew on blood agar with a ground-glass appearance, white-gray colony (Gunungkidul and avirulent strain) or yellowish (Boyolali and Pati). All were Gram-positive, presented chains, square-ended rods, spores, and were then identified as B. anthracis. Boyolali, Pati, and avirulent strain isolates had slightly different characteristics, including the growth of non-mucoid in the bicarbonate agar medium, and their uncapsulated form. The PCR showed two Gunungkidul isolates which amplified three genes, including Ba813, lef, and capC. Contrarily, the other isolates did not amplify the capC gene. Conclusion: Gunungkidul isolates were identified as virulent strains of B. anthracis while Boyolali and Pati isolates were proposed as avirulent strains. This is the first report of isolation and identification of avirulent strains of B. anthracis in Central Java, Indonesia.

Consequences of mechanical pupil dilation, a study based on the Swedish national cataract register.

To describe the outcomes and demographics of patients undergoing mechanical pupil dilation (MPD) during cataract surgery. All cataract procedures performed in Umeå University Hospital and Sunderbyn, Gällivare and Piteå hospitals reported to the Swedish National Cataract Register (NCR) during 2013-2019. Retrospective cohort study based on the Swedish NCR and electronic patient records. The number of control visits, pre- and postoperative visual acuities, surgical complications/intraoperative difficulties, ocular comorbidities and postoperative treatment regimens were retrieved for all cataract procedures with MPD. For each procedure, the consecutive procedure in the NCR from the same clinic without MPD was chosen to form a control group. A multinominal regression analysis with MPD as the dependent variable was performed to identify factors and outcomes independently associated with MPD. A total of 25 349 patients aged 18-97 years underwent cataract surgery in these hospitals during the study period. Of these, 653 (2.6%) had MPD. Factors such as pseudoexfoliations and capsule staining were over-represented among MPD eyes. As a group, eyes with MPD had more postoperative visits and more postoperative anti-inflammatory drops, and more frequently needed augmentation of the anti-inflammatory treatment at the first postoperative visit. MPD is independently associated with a more complicated intra- and postoperative course with more follow-up visits and requires more anti-inflammatory treatment postoperatively. This information could be added to the postoperative counselling, and more postoperative anti-inflammatory treatment could be considered in cases with MPD.

Characterization and Insilico Analysis of Cassia auriculata against Multi drug Resistant Klebsiella pneumoniae Isolated from Inanimate Origin

Infection with carbapenem-resistant Enterobacteriaceae (CRE) or carbapenemase-producing Enterobacteriaceae is emerging as an important challenge in health-care.50 swabs were collected from inanimate origins located in Chennai and the organisms isolated were identified by performing various preliminary tests like gram staining, capsule staining, motility, catalase, oxidase and biochemicals. antibiotic sensitivity testing was done for all the isolates using Ampicillin (AMP), Tetracycline (TE), Nitrofurantoin (NIT), Colistin (CL), Piperacillin (PIT), Noroxin (NX), Cefepime (CPM), Cefeparazone (CPZ), Ceftazidime (CFZ), Ciprofloxacin (CIP), Cotrimazole (COT), Ceftriaxone (CFX), Tigecycline (TGC), Tobramycin (TOB), Gentamicin (GEN), Amikacin (AK) and Imipenem (IMP) drugs and highest resistant against all the gram negative drugs was shown by Klebsiella which was further confirmed by MIC. The isolate was analysed for species by 16S rRNA sequencing which was reported as Klebsiella pneumoniae. The main objective of the present study is to identify the genes responsible for showing MDR and given for PCR reported as NDM, OXA-48 and K2A are the genes. Molecular docking was done by retrieving the gene structures of OXA-48 NDM and K2A of K.pneumoniae from protein data bank with proteins present in the leaf part of Cassia auriculata in patchdock and the binding interactions were interpreted and visualized using pyMOL to give a breakpoint by designing a drug for MDR bacterial strains.

Biocompatible pH-Degradable Functional Capsules Based on Melamine Cyanurate Self-Assembly.

Development of adaptive self-regulating materials and chemical–biological systems—self-healing, self-regulating, etc.—is an advanced modern trend. The very sensitive pH-controlled functionality of supramolecular assemblies is a very useful tool for chemical and biochemical implementations. However, the assembly process can be tuned by various factors that can be used for both better functionality control and further functionalization such as active species, e.g., drugs and dyes, and encapsulation. Here, the effect of a dye, sodium fluorescein (uranine) (FL), on the formation of a self-assembled melamine cyanurate (M–CA) structure is investigated and calculated with density functional theory (DFT) and molecular dynamics. Interestingly, the dye greatly affects the self-assembly process at early stages from the formation of dimers, trimers, and tetramer to nucleation control. The supramolecular structure disassembly and subsequent release of trapped dye occurred under both high- and low-pH conditions. This system can be used for time-prolonged bacterial staining and development of supramolecular capsules for the system chemistry approach.

First report on genetic characterization, cell-surface properties and pathogenicity of Lactococcus garvieae, emerging pathogen isolated from cage-cultured cobia (Rachycentron canadum).

The diseased cage-cultured cobia (Rachycentron canadum) displayed clinical signs, haemorrhagic eyes, dorsal darkness and gross pathological lesions, enlargement of spleen and liver. Haemorrhages were found in brain, heart and liver with cumulative mortality rates ranging from 20% to 50%. Extensive congestion in the heart, liver, spleen, kidney and brain was observed histopathologically. Epicarditis and meningitis were also revealed in diseased cobia. All isolates recovered from the organs (liver, spleen, head kidney, posterior kidney, brain and muscle) of cobia were found to be gram-positive, non-motile, ovoid cocci, short-chain-forming (diplococci) and α-haemolytic. The API 32 strep system together with the polymerase chain reaction assay for species-specific primers (pLG1 and pLG2) and the internal transcribed spacer (ITS) region (G1 and L1 primers) confirmed all four selected isolates as Lactococcus garvieae. Partial 16S rDNA nucleotide sequence (~1,100bp) of one representative L.garvieae isolate AOD109191 (GenBank accession number, MW328528.1) shared 99.9% identities with the 16S rDNA nucleotide sequence of L.garvieae (GenBank accession numbers: MT604790.1). Transmission electron microscopy (TEM) evaluation of one representative L.garvieae isolate (AOD109191) and the results of multiplex PCR did not reveal the presence of the capsular gene cluster (CGC), thus categorizing the isolate as the KG+ phenotype. Capsule staining and TEM observations confirmed the presence of a hyaluronic acid-like capsule, a possible virulence factor in KG+ phenotype L.garvieae isolates. The pathogenic potential of the representative isolate (AOD109191) was assessed through intraperitoneal injection challenges in cobia. The gross lesions and histopathological changes found in experimentally infected cobia were similar to those seen in naturally infected fish. This is the first report that confirms L.garvieae-induced 'warm water lactococcsis' can cause outbreaks of diseases in cage-cultured cobia.

Efficacy of Natural Blue Dye from Butterfly Pea Flower for Anterior Capsule Staining during Cataract Surgery: An Ex Vivo Study

Efficacy of Natural Blue Dye from Butterfly Pea Flower for Anterior Capsule Staining during Cataract Surgery: An Ex Vivo Study

Staining of subfossil chironomid head capsules: a method for improving the extraction process from lake sediments and peat

Subfossil chironomids are regarded as a useful biological proxy for palaeoenvironmental research, but picking chironomid head capsules from aquatic sediments is extremely time-consuming. This is often the case for finding and isolating small head capsules from the deflocculated sediment. In this study, two stains (aniline blue and cotton blue) were used to dye chironomid head capsules in lake and peat sediments to improve the traditional pretreatment method and boost the picking efficiency. The results suggested that: (1) there were sharp contrasts between chironomids and residues after staining, which could shorten picking time for both lake and peat samples; (2) different parts of head capsules showed distinctive colour in stained samples and thus facilitated identification and taxonomy of subfossil chironomids; (3) the dyeing effects of two stains were comparable and no significant effect of staining on chironomid composition has been observed compared with unstained samples. This study demonstrates that aniline blue and cotton blue stain can promote picking efficiency of subfossil chironomids, and can be widely applied to pretreatment of sedimentary chironomids in palaeoenvironmental studies on lakes and peatlands.

Occurrence and Distribution of Cryptococcus Species in Environmental Sources from Lower Assam Belt of India

Cryptococcus is a non-motile, gram positive, non-fermenting Basidiomycetous encapsulated yeast like fungus that causes respiratory, neurological and other systemic diseases in both humans and animals. Present study delineates the possible distribution of Cryptococcus species in pigeon droppings, excreta of other avian species, eucalyptus tree and contaminated soil specimens collected from different geographical co-ordinates of six geographical regions of the lower Brahmaputra Valley of Assam, India. The fungi were isolated through conventional methods of Sabouraud Dextrose Agar (SDA) and Bird Seed Agar (BSA) media and identified through negative staining of capsule as well as performing classical bio-chemical tests. Identity of the isolates was further confirmed through sequence analysis of ITS-1 and ITS-4 region of the 18S rDNA. Two pathogenic species of Cryptococcus were isolated from 67 (15.40%) of the 435 specimens. Of these positive isolates 41 (9.43%) belonged to Naganishia albida (Cryptococcus albidus) while 26 (5.98%) represented Papiliotrema laurentii (Cryptococcus laurentii). Both the species were recovered from 58 (18.35%) dry and 9 (7.56%) moist specimens. The percentage of prevalence of Naganishia albida in dry and moist specimens were 35 (11.07%) and 6 (5.04%) respectively. Contrary to this, the percentage of prevalence of Papiliotrema laurentii in dry and moist were 23 (7.28%) and 3 (2.52%) respectively. The findings indicate that Cryptococcus species have established a better ecological sustenance in dry specimens than moist. The findings of the investigation demonstrated that the prevalence of Cryptococcus albidus in attics, dovecotes / houses of pigeon fanciers, contaminated soil, eucalyptus tree and droppings of other birds were 11(12.36%) out of 89, 23(14.11%) of 163,2(3.23%) of 62,4(7.84%) of 51 and only 1(1.43%) out of 7 specimens respectively. The recovery of Papiliotrema laurentii in the above specimens were 3(3.37%), 20(12.27%), 1(1.61%), 1(1.96%) and 1(1.42%) respectively. The findings revealed that the prevalence of Naganishia albida is more than that of Papiliotrema laurentii in natural substrates. The notorious pathogenic fungi, Cryptococcus neoformans could not be isolated, indicative of the fact that the region selected for the study is not environmentally favorable for growth and sustenance of the species. Findings of the study clearly demonstrate the ecological and epidemiological significance of the non-neoformans species of the genus cryptococcus that needs further comprehensive studies to access the prevalence of the genus from public health point of view.

Detection of carbapenem resistant Klebsiella species isolated from clinical samples

The dissemination of carbapenem resistance among the family of Enterobacteriaceae is one of the most potent intimidation to an individual’s health nowadays. In this research, the foremost focus is on carbapenemase-producing Klebsiella species. Carbapenems are a genre of beta-lactam ring containing antibiotics which includes meropenem imipenem, ertapenem, biapenem, doripenem & panipenem. Carbapenems are highly effective antibiotics as they arrest the cell wall of bacteria so if the resistance against carbapenem emerges than it will subsequently lead to narrow therapeutic options. The foremost purpose of this research is the determination of carbapenem resistance among Klebsiella species. Fifty (n=50) gram-negative samples were collected from the Centralized Diagnostic Laboratory of Karachi, Pakistan. All the samples were sub-cultured on EMB (eosin methylene blue) agar to differentiate them from other Enterobacteriaceaes. MacConkey agar was used to check lactose fermentation. Gram staining and capsule staining for morphological identification, biochemical tests (Indole, Methyl red, Voges Proskauer, Citrate, (IMViC), Triple Sugar Iron (TSI)) were carried out to differentiate among Klebsiella species. Carbapenem resistance against Imipenem and Meropenem was identified by the disc diffusion method based on the Clinical Laboratory Standard Institute (CLSI) guidelines. Out of which 26 isolates were identified as Klebsiella spp. In these 26 isolates, 10 isolates were identified as Klebsiella pneumoniae. Further results showed that K. pneumoniae isolates were more resistant to Meropenem as compared to the Imipenem. All ten isolates of K. pneumoniae were found resistant to Meropenem while only two isolates of K. pneumoniae were resistant to Imipenem. This study was done to determine the carbapenem resistance among Klebsiella so that data can be provided which will help in rapid and appropriate treatment of patients so that in the future we can use alternative drugs or cocktail of drugs to treat infections caused by Klebsiella. Keywords: Beta-lactamases; Carbapenems; Emergence; Klebsiella pneumoniae; Resistance http://dx.doi.org/10.19045/bspab.2020.90175

Insights into myxozoan composition and physiology revealed by histochemical properties of myxospores.

Myxozoa (phylum Cnidaria) are a diverse group of metazoan parasites that predominately infect fish. Little is known regarding the composition and physiology of their myxospore life stage. The objective of this work was to investigate the composition of myxospores and extrasporogonic stages of nine myxozoan species infecting various teleost fish using histochemical staining techniques. Thirty histochemical stains were applied to formalin-fixed, paraffin-embedded tissues processed routinely for light microscopic evaluation. The polar capsules were the most consistent stain target across the taxa examined. Polar capsule staining with Alizarin red, von Kossa and methyl green-pyronin suggests the presence of intracapsular calcium and phosphate, which may contribute to polar filament discharge or pathogenesis of host invasion. The shell valves and suture lines of most myxozoans were stained with Luna and phosphotungstic acid haematoxylin stains, consistent with the presence of chitin and microfibrils, respectively. Vacuoles were consistently highlighted by diastase-susceptible periodic acid-Schiff and Grocott's methenamine silver staining, indicating glycogen. Other histochemical stains exhibited inconsistent staining across the taxa, suggesting differences in myxospore composition potentially reflective of physiologic variations and tissue tropisms. This work provides some information on conserved features and taxa-associated composition of myxospores and lends insight into myxozoan physiology and host-parasite interactions.

Commentary: Capsule staining and its aftermath.

Commentary: Capsule staining and its aftermath.

Fortuitous corneal stromal staining during wound exploration and repair with cataract extraction

The use of trypan blue in staining of the anterior capsule during cataract surgery plays a very important role in per operative visualization of the anterior capsule for anterior capsulorhexis and phacoemulsification. It is used in staining the anterior capsule during cataract surgeries, internal limiting membrane during vitreo-retinal surgeries or can be used as part of Siedel’s test to detect a leak in cornea. This dye is non-toxic and is incorporated in intra ocular surgeries for many years. We report a case of Traumatic corneal perforation with traumatic cataract in which the corneal stroma was accidently stained by trypan blue. The corneal staining in our case was result of improper port construction as it was difficult on account of hypotony, the anterior chamber being shallow and continuous leaking of air from the corneal wound. In our opinion the complication can be prevented by proper visualization and use of sharp instruments in construction of the port.

Phacoemulsification in eyes with corneal opacities after deep anterior lamellar keratoplasty.

To evaluate the maneuverability and efficacy of phacoemulsification and intraocular lens (IOL) implantation in eyes with corneal opacities after deep anterior lamellar keratoplasty (DALK), twelve eyes of 12 patients with mild to moderate corneal opacities after DALK and coexisting cataracts were analyzed retrospectively. Phacoemulsification and IOL implantation assisted with anterior capsule staining, as well as non-invasive optical fiber illumination, were performed on all eyes. No intraoperative or postoperative complications were noted. Mean corrected distance visual acuity (logMAR) improved from 1.24±0.17 to 0.73±0.22. Post-phaco intraocular pressure was maintained between 13 to 20 mm Hg in all cases throughout the follow-up period. Mean endothelial cell density decreased from 2258.42±205.94 to 1906.25±174.23 cells/mm2. Phacoemulsification and IOL implantation are safe and valid in eyes with mild to moderate corneal opacities after DALK and coexisting cataracts when assisted with anterior capsule staining and non-invasive optical fiber illumination.

Core needle biopsy in the management of thyroid nodules with an indeterminate fine-needle aspiration report.

Ultrasonography (US)-guided fine-needle aspiration biopsy (FNAB) cytology is widely used but is limited due to its pathologically indeterminate results in diagnosing thyroid nodules. Recently, US-guided core-needle biopsy (CNB) was introduced as an effective and safe technique for diagnosing indeterminate thyroid nodules. Using CNB, information about architectural histologic structure such as nodule capsule or more immunochemical staining can be obtained which lead to a more accurate diagnosis. Up to 98% of indeterminate thyroid lesions can be classified as malignant or benign when CNB is used for follow-up analysis. Other evidences revealed the effectiveness of CNB in reducing inconclusive results and improving the diagnostic performance of thyroid nodules initially diagnosed as AUS/FLUS by FNAB. In this review, we investigate how to deal with indeterminate thyroid nodules diagnosed by FNAB and determine how CNB has a role in diagnosing these indeterminate thyroid nodules.

Differential Gene Expression in Capsules Derived From Smooth and Textured Silicone Implants

PURPOSE: Capsular contracture is the most common complication of implant-based breast surgery in the United States, leading to abnormal breast contour, increased firmness, and pain to touch. It is well established that smooth implants have higher rates of capsular contracture than textured implants. Assessing the effect of implant texture on contracture pathogenesis may help identify therapeutic targets for this disease process. The purpose of this study is to elucidate the pathogenesis of capsular contracture by examining RNA expression in a rat model of capsular formation around textured and smooth silicone implants, with pathologic correlation. METHODS: A small animal model of capsular contracture was developed using Fischer rats. Ten animals underwent miniature smooth or textured silicone implant insertion in the submammary gland position. Six weeks postoperatively, implant capsules were harvested for histologic and molecular analysis. RNA sequencing was performed to identify target genes expressed in extracted capsules. Selected gene expression levels were confirmed with quantitative reverse transcriptase polymerase chain reaction and immunohistochemistry (IHC). Additionally, implant capsules collected from patients with and without capsular contracture were analyzed and correlated with results from our animal model. RESULTS: RNA sequencing data were subjected to the Probability of Positive Log Ratio (PPLR) algorithm. Transcripts were identified for further characterization using cutoff values of PPLR ≥0.975 (2-fold increase) or a PPLR ≤0.025 (2-fold decrease). We identified 18,555 transcripts that met PPLR inclusion criteria. Quantitative reverse transcriptase polymerase chain reaction was performed for matrix metalloproteinase-3 (MMP-3), troponin T3 (TNNT-3), and neuregulin-1 (NRG-1). Expression of MMP-3 and TNNT-3 was upregulated in textured implant capsules compared to smooth implant capsules with a mean relative fold change of 8.79 (P = 0.0059) and 4.81 (P = 0.0056), respectively. Expression of NRG-1 was downregulated in textured implant capsules with a mean relative fold change of 0.40 (P < 0.0001) compared to smooth implant capsules. IHC staining of capsules extracted from our animal model was consistent with differential expression patterns, with smooth implant capsules expressing less MMP-3 and TNNT-3, and more NRG-1 than textured implant capsules. Similarly, IHC staining of human specimens revealed that contracted capsules had lower expression of MMP-3 and TNNT-3 and greater expression of NRG-1 compared to healthy capsules. CONCLUSION: We demonstrated that capsules around smooth and textured implants have different histologic appearances and different patterns of gene expression. Importantly, pathologic correlation reveals that contracted capsules have expression patterns for MMP-3, TNNT-3, and NRG-1 that are consistent with capsules derived from smooth implants, which are known to have higher contracture rates. These results may help elucidate the mechanism of capsular contracture and identify potential genes for the development of future therapeutic targets for this condition.

Cataract Surgery in Very Old Patients

In recent years, there has been an increase in the number of very old patients requiring a cataract operation. However, there is little information on the intraoperative complications and safety in these patients. For various reasons, operations on patients from age 85 may be demanding: dense nuclear cataract, narrow pupils, low count of endothelial cells, loose zonula, and other ocular or systemic diseases. The aim of this retrospective study was to compare very old to younger cataract patients with respect to intraoperative complications and the maturity of the cataract. This was a retrospective study on 4065 cataract patients treated in our department with IOL implantation with the corneal small incision technique between January 2015 and January 2018. The patients were split by age (from 85 and under 85). Of the 4065 cataract operations, 10.6% (431/4065) were performed on patients aged at least 85, 1.7% (69/4065) on patients aged at least 90 and 0.1% (4/4065) on patients of at least 95 years. There were statistically significant differences between the younger and older patients with respect to pupil dilatation (4.6 vs. 6.0%), use of the capsule tension ring (0.4 vs. 0%) and in capsule staining (5.5 vs. 7.0%). There were no intraoperative complications (e.g. anterior capsule laceration) in patients aged at least 85 and in 0.71% of patients aged under 85; loss of vitreous fluid was recorded in none of the patients in the older group and in 0.41% of patients in the younger group. Our study confirms that advanced age alone is not a contraindication for a cataract operation and is not associated with a greater rate of intraoperative complications. Cataract operations on very old patients are generally successful, but should be undertaken early - particularly on multimorbid patients or those with dementia - in order to support the psychological status. The age of the patient is irrelevant.

Micro-staining microbes: An alternative to traditional staining of microbiological specimens using microliter volumes of reagents

Microbial staining techniques are widely employed in clinical and academic laboratories for classifying and identifying microorganisms derived from clinical, food and environmental samples. Staining allows for the rapid visualization and determination of many morphological characteristics of microorganisms, used for their identification and classification. Over the past century, staining techniques such as the Gram stain, the Capsule stain, the Acid-fast stain and the Endospore stain, have seen few advances, and manual staining remains the gold standard. Typical instructions for these staining procedures recommend ‘flooding’ glass slides with milliliter volumes of dye, resulting in large volumes of hazardous waste. Here we present micro-staining, a simple alternative to flooding that utilizes microliter volumes of dye. Micro-staining minimizes the volume of waste generated, leads to significant cost savings for the laboratory, requires limited training, and produces results with equivalent quality to traditional stains.

Phenotypic Detection and Biofilm Formation among Pseudomonas aeruginosa Isolated from Different Sites of Infection

In The present study, included the collection of (100) samples from different clinical sites. Clinical samples were collected from patients who were visit and admitted All-Hilla teaching hospital at the period from November (2017) to February (2018). Cultural, biochemical and VITEK2 system were used for identification, and depending on the VITEK2 system (VITEK-2 GN Kit), revealed that twenty one (21) Pseudomonas aeruginosa isolates were recovered, The percentage conformational identification of P. aeruginosa was performed using VITEK2 system of (21) P. aeruginosa was (99%). Nine(42.8%) samples were isolated from burns, 5(23.8%) samples from wound, 3(14.2%) from urine, 2(9.5%) from ear swab, and 1(4.7%) sample was isolated from both blood and sputum. The phenotypic detection of some virulence factors for all isolates were detected. Detection of capsule was done by using capsule staining technique was carried out for P. aeruginosa isolates; it was found that all P. aeruginosa isolates (100%) have a capsule surrounding the bacterial cell. Hemolysin production by P. aeruginosa was studied; it was found that 12(57.1%) isolates were able to produce extracellular hemolysin on blood agar. P. aeruginosa isolates were also investigated for their ability to produce siderophores. The results showed that 9(42.8%) isolates of P. aeruginosa were able to produce siderophores. Protease production by P. aeruginosa isolates was studied; it was found that all these isolates (100%) have this enzyme as appear as a zone around the colony when being grown on (M9) media after adding of (3ml) of (5%) Trichloroacetic acid and incubation for (24 hrs.). Ability of P. aeruginosa to produce lipase has been investigated; it found that all these isolates (100%) were able to produce lipase after incubation for (48 hrs.) on egg yolk agar. Also, bacterial biofilms cause chronic diseases that are difficult to control and in the present study, differentiation of bacteria as biofilm producers and non-biofilm producers was done by using (ELISA) TCP method, a total of (21) isolates were tested for their ability to produce biofilm. From these isolates, (19) isolates were form strong biofilm, (2) isolates were form moderate biofilm.

Safety of Trypan Blue Capsule Staining to Corneal Endothelium in Patients with Diabetic Retinopathy.

Purpose To study the potential corneal endothelial cell toxicity of trypan blue (TB) when used for phacoemulsification to stain the anterior capsule in patients with diabetic retinopathy. Methods This was a single-center, prospective, randomized, individual cohort study. One eye in each patient with diabetic retinopathy underwent phacoemulsification without trypan blue capsule staining (control eye), while the other eye underwent phacoemulsification with trypan blue capsule staining (study eye). Both eyes underwent intraocular lens implantation. Preoperative and four-week postoperative quantitative and qualitative morphometric endothelial cell analyses of the cornea were performed using noncontact specular microscopy. Results There were no significant differences in endothelial cell density (mean ± SD for the study group: 2506.74 ± 413.99 cells/mm2; mean ± SD for the control eyes: 2466.34 ± 369.12 cells/mm2; P=0.316) or endothelial cell density (CD) loss% (mean CD loss% was 7.23 ± 13.31 for the study eyes and 9.94 ± 9.36 for the control eyes; P=0.157) four weeks after the operation. Additionally, no significant differences were seen in the percentage of hexagonal cells, coefficient of variation, or corneal thickness between the two groups preoperatively and 4 weeks postoperatively. Conclusions Direct administration of trypan blue into the anterior chamber for staining of the anterior capsule during cataract surgery did not result in any significant corneal endothelial changes on specular microscopy in patients with severe nonproliferative diabetic retinopathy or high-risk proliferative diabetic retinopathy at 4 weeks postoperatively. This trial is registered with NCT03755752.

Intraoperative Complications of Cataract Surgery Using Intracameral Illumination in the Elderly over 75 Years.

Purpose To evaluate intraoperative complications and utilization of adjunctive devices between microscope and intracameral illuminations during cataract surgery in the elderly over 75 years. Design A retrospective, consecutive, interventional case series Participants. Two hundred eighty-six eyes of 184 patients older than 75 years who underwent cataract surgery using microscope and intracameral illuminations. Methods A chart review was performed on an advanced cataract surgery group of 141 consecutive cases in which the intracameral illumination was used and on a standard cataract surgery group of 145 consecutive cases in which the intracameral illumination was not used. Main Outcome Measures Intraoperative complications (posterior capsule rupture, radial tear of the anterior capsule, dropped nucleus, or sulcus-implanted/sclera-fixated IOL) and utilization of adjunctive devices (pupil expansion device or anterior capsule staining). Results The frequency of use of the pupil expansion device was lower in the advanced cataract surgery group than that in the standard cataract surgery group (0.7% vs 6.9%; p=0.007). Furthermore, the rates of a posterior capsule rupture and at least one intraoperative complication were lower in the advanced cataract surgery group than those in the standard cataract surgery group (0.7% vs 4.8%; p=0.067) (0.7% vs 7.6%; p=0.004). Conclusions In the current cohort of patients over 75 years, the rate of intraoperative complications was lower when using the intracameral illumination than that when using the conventional method. Cataract surgery using intracameral illumination would be good option for elderly people.