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Huntington's chorea is a rare neurodegenerative disorder caused by the dominant inheritance of the mutated huntingtin gene, housing poly-CAG or polyglutamine repeats. It is characterized by a wide variety of symptoms ranging from chorea and hypokinetic movements to behavioural and cognitive decline, followed by dementia and inevitable death. Over the past 120 years, all available therapeutics have been for the symptomatic management of Huntington's disease (HD) and require supportive physiotherapy and counselling to maximize the efficacy of the treatment. Several animal models have been employed to help elucidate and decrypt the pathophysiology of the disease, and also screen potential therapeutic candidates. In the last few decades, a deeper understanding of the cellular and molecular dynamics associated with HD has helped shed light on the mechanisms involved in disease progression. Genetic intervention for early detection, spreading awareness about HD and its symptoms, and training professionals in the nuances of the disease condition can significantly improve the lifestyle of patients. This article aims at summarizing the complex pathogenesis of HD at the cellular level using various disease models and available therapeutics. [ABSTRACT FROM AUTHOR]

Frontal fibrosing alopecia (FFA) and lichen planopilaris (LPP) are both lymphocytic cicatricial alopecias but with different clinical presentation. Patients seen at the University of Sao Paulo with biopsy-proven LPP ( I n i = 10) and FFA ( I n i = 28), both gender, with similar age and disease duration underwent biopsies from affected (anterior) and unaffected (posterior) scalp. AhR immunohistochemical staining of FFA affected scalp (a), FFA unaffected scalp (b), LPP affected scalp (c), LPP unaffected scalp (d), anterior scalp/control (e), posterior scalp/control (f). [Extracted from the article]

Mucosal lesions of paracoccidioidomycosis (PCM) are frequently described and clinically important. Macrophages are classified as M1 or M2. M1 are proinflammatory and M2 are related to chronicity. Dectin-1 recognizes β-glucan and plays an important role against fungal cells. The objective was to verify the presence of M1, M2, and dectin-1 and a possible correlation with Th1/Th2 cytokines in mucosal PCM lesions. In sum, 33 biopsies of oral PCM were submitted to histological and immunohistochemistry analysis, and positive cells were quantified. Eleven biopsies were characterized by compact granulomas (G1), 12 with loose granulomas (G2), and 10 with both kind of granulomas (G3). pSTAT-1 was equally increased in the three groups. G1 was characterized by an increased number of CD163+ macrophages. G2 presented similar number of arginase 1, iNOS, and CD163 expressing cells. G3 presented an increased number of cells expressing arginase 1 and CD163 over iNOS. G1 and G3 presented high number of cells expressing interferon (IFN)-γ; interleukin (IL) 5 was increased in G2 and G3; the expression of IL10 was similar among the three groups, and the expression of tumor necrosis factor (TNF)-α was higher in G3. G1 correlates to Th1 cytokines and pSTAT-1 and G2 correlates to Th2 cytokines. G3 presents both kinds of cytokines. We could not associate the expression of arginase-1, CD163, iNOS, and dectin-1 with the pattern of cytokines or kind of granuloma. [ABSTRACT FROM AUTHOR]

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Background: Malignant otitis externa is an inflammatory condition of the external ear which has the propensity to spread to the skull base. It can be a difficult entity to treat as clinical presentation varies and response to treatment differs between patients. We reviewed cases of malignant otitis externa in our setup to document the epidemiology and outcome of management. Methods: This is a retrospective case review observational study from January 2013–December 2017. Fourteen patients diagnosed with malignant otitis externa in our tertiary referral centre were included in the study. Based on hospital protocol, empiric treatment was started. After discharge, the patients follow up visits to the hospital were also documented. Results: Otalgia was the most common symptom. Edema and congestion of the external auditory canal were observed in most cases. Diabetes was present in all patients. Three cases had associated facial palsy, and one patient had involvement of 7th, 9th, 10th, 11th and 12th cranial nerve. Two patients with facial palsy recovered. Pseudomonas aeuroginosa was the most common organism isolated (50%). Conclusions: In our series, malignant otitis externa invariably presented with severe otalgia. Lower cranial palsies were also seen. Methods to evaluate complete eradication of disease should be centered on clinical symptoms and signs, but the measurement of erythrocyte sedimentation rate or radiological imaging may be used as a useful adjunct when there is uncertainty. Keywords: Malignant otitis media, Granulation tissue, Pseudomonas aeuroginosa, Diabetes

An experiment was conducted with broilers from 22 to 33 days of age to evaluate the efficiency of six microbial phytases supplemented in diets (1500 FTU/kg) that were formulated with three different calcium:available phosphorus (Ca:Pavail) ratios (4.5:1.0, 6.0:1.0 and 7.5:1.0). A positive control diet without phytase was formulated with a Ca:Pavail ratio of 7.5:3.4 to meet the nutritional requirements of the broilers. The P and ash contents of the tibia, magnesium in the plasma, performance, balance and retention of phytate phosphorus (Pphyt), intake of total P and nitrogen (N), nitrogen-corrected apparent metabolizable energy and apparent digestibility of dry matter of the diets were not influenced (p > 0.05) by the type of phytase or the dietary Ca:Pavail ratio. However, there was an interaction (p < 0.05) between the phytase type and the Ca:Pavail ratio for the retention coefficients of total P, Ca and N. Phytase B resulted in the highest Ca deposition in the tibia (p < 0.01). Phytases D, E and F reduced the Ca concentrations in the tibia (p < 0.01) and plasma (p < 0.05). Phytase D increased the P level in the plasma and decreased the total P excretion (p < 0.01). Phytases E and F increased Ca excretion, while phytase A reduced it (p < 0.01). Regardless of the phytase type, increasing the dietary Ca:Pavail ratio reduced (p < 0.05) the plasma P concentration and the excretion of total P and N and, conversely, increased (p < 0.05) the plasma concentration, intake and excretion of Ca. For the rearing period evaluated, it is possible to reduce the Pavail of the diet to 1.0 g/kg when Ca is maintained at 7.5 g/kg, and the diet is supplemented with 1500 FTU of phytase A, C, D or E/kg. This diet allows the maintenance of performance and adequate bone mineralization, and it improves the Ca, total P and Pphyt utilization in addition to reducing the excretion of N and P into the environment. [ABSTRACT FROM AUTHOR]

PurposeTo evaluate the long-term visual outcome of type 2 diabetic patients receiving early vitrectomy and endolaser for severe vitreous haemorrhage (VH).Materials and methodsRetrospective case note review of 88 eyes (69 type 2 diabetics and 19 type 1 diabetics) of 80 patients who underwent vitrectomy and endolaser within 6 months of VH. Post-operative and most recent VA, in addition to long-term retinopathy grading, were analysed. A subset of patients fulfilling the criteria for the Diabetic Retinopathy Vitrectomy Study was compared with this study.ResultsMean pre-operative visual acuity (VA) in the type 2 group was 0.64 logMAR, with 1 eye showing perception light (PL), 10 eyes detecting hand movements (HMs), and 7 eyes counting fingers (CFs). At the 2-week post-operative visit, the mean VA had improved to 0.46 logMAR, with two eyes showing PL, two eyes detecting HM, and one eye CF (P=0.0002); at the last review, mean VA score was 0.36 logMAR, with three eyes showing PL and four eyes detecting HM (P=0.0008). Mean pre-operative VA in the type 1 group was 0.47 logMAR, with one eye showing PL, one eye detecting HM, and two eyes CF. At the 2-week post-operative visit, the mean VA had improved to 0.37 logMAR, with one eye showing PL (P=0.002), and at the latest review, the mean VA was 0.20 logMAR (P=0.027).ConclusionOur study shows that type 2 DM patients can observe improvement in VA and stabilisation of their proliferative retinopathy after early vitrectomy and endolaser for vitreous haemorrahage, which is maintained after long-term follow-up. [ABSTRACT FROM AUTHOR]

Background: Annual influenza vaccine uptake is recommended by the Portuguese General-Directorate of Health to certain priority groups. In 2020, amidst the COVID-19 pandemic, some exceptional measures were adopted to operationalize the Portuguese influenza vaccination campaign, like the extension and phasing of the vaccination period. No major changes were introduced in the vaccination of healthcare workers. In Baixo Tâmega health cluster (Portugal), it was perceived that these workers’ vaccine uptake had markedly increased in 2020-2021. This study aims to compare their influenza vaccination coverage rates in 2020-2021 to the last 10 seasons. Methods: A cross-sectional study of the vaccine uptake among the health workers of Baixo Tâmega health cluster was developed. A descriptive analysis was carried out, calculating annual influenza vaccination coverage rates from 2010-2011 to 2020-2021, in doctors, nurses and other health workers. Inferential analysis was performed through chi-squared tests in IBM SPSS Statistics 27.0.1.0., considering significant p-values<0.05. Results: The difference between health workers 2020-2021’s influenza coverage rate and the last season’s average rates was 22.81%. Before 2020-2021, average vaccination coverage rates were 51.60% (standard deviation 7.34%, 44.26-58.94%), 60.44% (standard deviation 4.12%, 56.32-64.56%) and 51.20% (standard deviation 7.66%, 43.54-58.86%), in doctors, nurses and other health workers, respectively. In 2020-2021, vaccination rates significantly increased to 79.34%, 79.52% and 74.24% in doctors, nurses and others, respectively (p < 0.001). Conclusions: Influenza vaccination uptake increased in 2020-2021 among health workers. Further studies should be developed to evaluate this tendency on a larger scale and to better understand its associated factors, which could have been unstructured awareness-raising campaigns, perception of protection against COVID-19 or avoidance of COVID-19- like symptoms and coinfection. Key messages: Influenza vaccination coverage rates increased in 2020-2021 among the healthcare workers of a Portuguese health cluster. The COVID-19 pandemic may have contributed to the increase in the influenza vaccination coverage rates. [ABSTRACT FROM AUTHOR]

This work aimed at evaluating the effects of 4 digestible Met+Cys levels on the diet of commercial layers and their influence on the productive performance, quality, and amino acid profile of eggs and economic viability of the activity. A total of 576 white Lohmann LSL-Lite layers was distributed into 6 replicates of 24 birds for each diet. The experimental design was completely randomized, with 4 treatments defined by levels evaluated in the feed (0.465, 0.540, 0.581, and 0.647%). The productive performance was measured for 30 weeks. The quality (34 and 50 wk old) and the amino acid profile of eggs (43 wk old) also were evaluated. A linear positive response was observed at higher Met+Cys levels for feed intake, number of eggs per housed bird, and digestible Met+Cys intake. Egg production, egg weight, egg mass, feed efficiency, and weight gain had their optimal values determined by the quadratic regression model at 0.638, 0.654, 0.647, 0.644, and 0.613% digestible Met+Cys, respectively. In the 34th wk, eggshell thickness decreased linearly at higher Met+Cys levels. In the 50th week, the optimal levels detected for eggshell thickness and percentage were 0.571 and 0.570% digestible Met+Cys, respectively. The percentages of proteins, branched-chain amino acids (leucine, isoleucine, and valine), histidine, and proline in eggs (albumen+yolk) showed a linear negative response in function of higher Met+Cys levels. Higher digestible Met+Cys levels (>0.630%) led to a good performance of layers, while lower Met+Cys levels improved the eggshell quality of layers in peak production. Optimal Met+Cys levels may change according to the price of the synthetic amino acid. [ABSTRACT FROM AUTHOR]

Specialized or secondary metabolites are small molecules of biological origin, often showing potent biological activities with applications in agriculture, engineering and medicine. Usually, the biosynthesis of these natural products is governed by sets of co-regulated and physically clustered genes known as biosynthetic gene clusters (BGCs). To share information about BGCs in a standardized and machine-readable way, the Minimum Information about a Biosynthetic Gene cluster (MIBiG) data standard and repository was initiated in 2015. Since its conception, MIBiG has been regularly updated to expand data coverage and remain up to date with innovations in natural product research. Here, we describe MIBiG version 4.0, an extensive update to the data repository and the underlying data standard. In a massive community annotation effort, 267 contributors performed 8304 edits, creating 557 new entries and modifying 590 existing entries, resulting in a new total of 3059 curated entries in MIBiG. Particular attention was paid to ensuring high data quality, with automated data validation using a newly developed custom submission portal prototype, paired with a novel peer-reviewing model. MIBiG 4.0 also takes steps towards a rolling release model and a broader involvement of the scientific community. MIBiG 4.0 is accessible online at https://mibig.secondarymetabolites.org/., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Purpose: This study aimed to identify the various clinical characteristics and ocular imaging features of ocular toxoplasmosis (OT) in HIV patients., Methods: Observational retrospective study of 52 eyes of 38 HIV patients with OT., Results: 68% of patients were male. 53.8% of eyes had keratic precipitates, anterior chamber cells were seen in 61.5%. 57.69% (n=30) eyes had de novo presentation of retinitis and 42.3% (n=22) of eyes had a retinochoroidal scar at presentation. Bilateral presentation was seen in 37% (n=14). Unifocal retinitis was noted in 38.4% (n=20) and 61.5% of eyes had multifocal (multizonal) retinitis (n=32). The most common multizonal involvement was noted in combined zone 1+2(n=14, 26.9%). The average horizontal and vertical measurements retinitis was 8.35 mm and 7.48 mm, respectively. The average area of retinitis was 66.2 mm 2 . Localised posterior vitreous detachment was seen in 83% of eyes and posterior hyaloid precipitates were noted in (n=7,39%) of eyes. Retinitis without retinal pigment epithelium elevation occurred in 88.8% of cases, compared with retinitis with retinal pigment epithelium elevation seen in 44.4% of eyes. 50% of eyes had thickened choroid beneath the retinitis lesion. Other unique signs of optical coherence tomography (OCT) noted were angular signs of Henle fibre layer hyper-reflectivity sign (n=1), halo lesions with central hypo (n=3) and choroidal excavation (n=1)., Conclusions: OT in HIV patients presents with large, bilateral, multifocal retinitis, generally devoid of haemorrhages and can rarely have central clearing. These lesions may present with or without a previous retinochoroidal scar. These lesions have horizontal spread and may not always have thickened choroid on OCT., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. No commercial re-use. See rights and permissions. Published by BMJ.)

Objectives: Olfactory dysfunction is an overlooked adverse effect of radiation therapy. This study is designed to find the effect of radiation therapy on olfactory function in head and neck malignancy excluding tumors of nose and nasopharynx and correlate the olfactory changes with the radiotherapy dose., Method: This prospective observational study was done over a 2-year period in 34 participants with head and neck malignancies who underwent radiation therapy (RT). The participants olfaction was evaluated subjectively with Italian Nose Obstruction Symptom Evaluation (I-NOSE) scale and objectively by a modified I-Smell test which included an olfactory identification score and an olfactory threshold score at 5 time points. The beginning of RT ( T 0), at 2 weeks of RT( T 1), end of RT ( T 2), 1 month follow-up ( T 3), and 3-month follow-up ( T 4). The near maximum dose to the nasal cavity ( D 2% ) and mean dose to the nasal cavity ( D mean ) were calculated for all participants and correlated with olfactory function., Results: A total of 34 patients with head neck malignancy were recruited. The median I-NOSE score reached maximum at the end of radiation and decreased to baseline at 3 months follow-up ( P  < .001). The olfactory identification score, olfactory threshold score, and median combined olfactory score showed a significant decrease at the end of radiation therapy compared to Pre-radiation therapy values. There was a significant but incomplete recovery in the 3-month follow-up period ( P  < .001)., Conclusion: There was a significant deterioration in quality of life for olfaction, olfactory identification, and olfactory threshold at the completion of radiotherapy. At 3 months follow-up, though there was no complete recovery of olfaction, it did not have an adverse effect on the quality of life., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Controlling monoclonal antibody aggregation at the upstream stage itself can significantly reduce the burden on downstream processing and can improve the process yield. Hence, we have investigated the use of sugar osmolytes (glucose, mannose, sucrose and maltose) and formulation excipients (mannitol, polysorbate 20 and polysorbate 80) as medium additives to reduce protein aggregation during cell culture. Aggregate content in cell culture samples was estimated using a high-resolution size-exclusion chromatography technique, which efficiently resolved the antibody monomer and aggregates in the cell culture matrix i.e., without purification. Glucose, mannose, maltose and the polysorbates effectively reduced the mean aggregate content over the course of the culture. Sugar-based additives exhibited a higher degree of variation during aggregate quantitation as compared to polysorbate additives, rendering the latter a preferred additive. Therefore, this study demonstrated the potential of sugar osmolytes and formulation excipients as media additives during cell culture to reduce aggregate formation, without negatively impacting cell growth and antibody production, facilitated by the monitoring of aggregate content in cell culture samples without purification., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Conflict of interest I, as the corresponding author, certify on the behalf of the other authors that we have NO affiliations with or involvement in any organisation or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Cutaneous leishmaniasis exhibits a spectrum of clinical presentations dependent upon the parasites' persistence and host immunopathologic responses. Although cytolytic CD8 T cells cannot control the parasites, they significantly contribute to pathologic responses. In a murine model of cutaneous leishmaniasis, we previously found that NKG2D plays a role in the ability of cytolytic CD8 T cells to promote disease in leishmanial lesions. Here, we investigated whether NKG2D plays a role in human disease. We found that NKG2D and its ligands were expressed within lesions from L. braziliensis-infected patients and that IL-15 and IL-1β were factors driving NKG2D and NKG2D ligand expression, respectively. Blocking NKG2D reduced degranulation by CD8 T cells in a subset of patients. Additionally, our transcriptional analysis of patients' lesions found that patients who failed the first round of treatment exhibited higher expression of KLRK1, the gene coding for NKG2D, than those who responded to treatment. These findings suggest that NKG2D may be a promising therapeutic target for ameliorating disease severity in cutaneous leishmaniasis caused by L. braziliensis infection., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Sacramento et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Protein A chromatography is widely used for isolation of monoclonal antibodies (mAbs) from cell culture components. In this study, the effect of different process parameters of the Protein A purification namely, binding pH, elution pH, flow rate, neutralization pH and tween concentration, on the concentration and quality of the purified mAb were evaluated. Using design of experiments approach, the critical process parameters of protein A chromatography were identified and experimentally optimized. Their impact on quality attributes, such as size variants and charge variants, of the mAb was studied. Multivariate data analysis was subsequently performed using multiple linear regression and partial least squares regression methods. It was observed that the elution pH primarily governed the concentration of the purified mAb and the content of monomers and aggregates, while the tween concentration primarily influenced the main peak of the charge variants. This is the first study that evaluates the impact of tween concentration in buffers on the protein A chromatography purification step. These studies helped in identifying the design space and defining the target robust and optimal setpoints of the responses, which were subsequently verified experimentally. These setpoints not only passed the target criteria but also resulted in the highest recoveries during the investigation. Through this statistically-aided approach, an optimized and robust protein A chromatography process was rationally developed for purification of mAbs, while achieving the desired product quality. This study highlights the influence of multiple parameters of the protein A purification process on critical quality attributes of mAbs, such as the size and charge variants, which has been a very scarcely explored area., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Petrous apicitis and acute bacterial meningitis are uncommon in the present antibiotic era. The diagnosis of petrous apicitis is seldom considered unless there is cranial nerve palsy. A young child with aplasia cutis congenita presented with acute bacterial meningitis and an incidental opacified left mastoid in brain imaging. During the course, fever persisted, and high-resolution temporal bone imaging showed rapid progression to coalescent mastoiditis, petrous apicitis with erosions of tegmen tympani, and petrous apex. Other findings included bony dehiscences and thinning of left calvaria. Tympanomastoid exploration showed herniated brain and cerebrospinal fluid leak through tegmen tympani, which was closed with temporalis fascia graft. Herein, we report a rare presentation of petrous and tegmen erosion along with aplasia cutis congenita and discuss the challenges in diagnosis and management.

In this case-study, we demonstrate an approach for identifying correlations between nutrients/metabolites in the spent medium of CHO cell cultures and cell growth, mAb titre and critical quality attributes, using multivariate analyses, which can aid in selection of targets for medium and feed optimization. An extensive LC-MS-based method was used to analyse the spent medium composition. Partial least squares (PLS) model was used to identify correlations between nutrient composition and cell growth and mAb titre and orthogonal projections to latent structures (OPLS) model was used to determine the effect of the changing nutrient composition during the culture on critical quality attributes. The PLS model revealed that the initial concentrations of several amino acids as well as pyruvic acid and pyridoxine, governed the early cell growth, while the concentrations of TCA cycle intermediates and several vitamins highly influenced the stationary phase, in which mAb production was maximum. For the first time, with the help of the OPLS model, we were able to draw correlations between nutrients/metabolites during the culture and critical quality attributes, for example, optimizing the supply of certain amino acids and vitamins could reduce impurities while simultaneously increasing desirable glycoforms. The unique correlations obtained from such an exploratory analysis, utilizing conditions that are commonly adopted in early process development, present opportunities for optimizing the compositions of the growth media and the feed media for enhancing cell growth, mAb production and quality, thereby proving to be a useful preliminary step in bioprocess optimization., Supplementary Information: The online version contains supplementary material available at 10.1007/s10616-022-00561-z., Competing Interests: Competing interestsThe authors declare no competing interests.Conflict of interestThe authors have no relevant financial or non-financial interests to disclose., (© The Author(s), under exclusive licence to Springer Nature B.V. 2022, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Objectives: The aim of the study was to compare lecture-based teaching and simulation-based hybrid training for ENT induction and objectively assess the performance of trainees in a simulated environment., Methods: This is a prospective interventional study that included 60 interns in their rotatory internship with no prior exposure to ENT emergencies. The interns came in batches of 5‒6 for their 15-days ENT postings. On the first day, a pre-test questionnaire, lecture-based teaching on three scenarios and then allocation into one of the three simulation groups- Group A (Tracheostomy group), Group B (Nasogastric tube group), and Group C (Epistaxis group) was done. Hands-on simulation training was given only to the assigned group. At the end of 15-days, post-test questionnaire and an objective assessment of the three scenarios in a simulated environment was conducted. The same training was repeated for each batch of participants who attended the posting., Results: The participants had significant improvement in the post-test scores in all three scenarios (p <  0.05), and these improvements were marked in those who had received simulated training. On comparing simulation scores, the participants who received hands-on training on a particular scenario outperformed other (p <  0.05)., Conclusion: Simulation-based training improves cognition and overall confidence in managing ENT skills and emergencies. In simulation training, objective and standardized assessment is the key to achieve specific learning objectives to improve the psychomotor and cognitive skill., Level of Evidence: II., (Copyright © 2022 Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial. Published by Elsevier Editora Ltda. All rights reserved.)

Background: Renal transplant recipients (RTRs) are at increased risk of developing skin cancer; however, the role of immunosuppression is not yet fully understood. In this study, we evaluated the immunohistochemical changes in the skin of RTRs under three different immunosuppression regimens: mTOR inhibitors (mTORi), sirolimus or everolimus, mycophenolic acid (MPA) precursors such as mycophenolate sodium or mofetil, or azathioprine (AZA)., Methods: We evaluated biopsies of sun-exposed and sun-protected skin for immunohistochemical quantification of B lymphocytes (CD20 + ), T lymphocytes (CD3 + , CD4 + , and CD8 + ), and Langerhans cells (LCs) (CD1a + ) in 30 RTRs and 10 healthy controls. The RTRs were divided into three groups: mTORi (n = 10), MPA (n = 10), and AZA (n = 10)., Results: No differences were observed in the number of B lymphocytes. However, a significant decrease in the number of T lymphocytes and LCs was observed in both sun-protected and sun-exposed skin in the AZA and MPA groups, although to a lesser degree in the latter group. The skin of the mTORi group did not differ from that of the control group in terms of the number of B and T lymphocytes and LCs., Conclusions: Patients treated with mTORi exhibit preserved cellular elements related to cutaneous immune surveillance. The use of AZA induced a greater degree of skin immunosuppression than in the control group, as demonstrated by the decrease in T lymphocytes and LCs., (© 2022 European Academy of Dermatology and Venereology.)

There have been a variety of complications reported during and post- COVID infection. Recently, there has been an increase in sporadic cases of paranasal sinus mucormycosis in COVID- 19 patients. We report a case of COVID- 19 patient diagnosed to have orbital apex syndrome secondary to mucormycosis of nose and paranasal sinus requiring emergency endoscopic sinus surgery. Appropriate use of personal protective equipment and safety precautions taken by health care workers prevented the spread of the virus during surgery., Competing Interests: Conflicts of interestNone., (© Association of Otolaryngologists of India 2021.)

Glycosylation has been shown to define the safety and efficacy of biopharmaceuticals, thus classified as a critical quality attribute. However, controlling glycan heterogeneity has always been a major challenge owing to the multivariate factors that govern the glycosylation process. Conventional approaches for controlling glycosylation such as gene editing and metabolic control have succeeded in obtaining desired glycan profiles in accordance with the Quality by Design paradigm. Nonetheless, the development of smart algorithms and omics-enabled complete cell characterization has made it possible to predict glycan profiles beforehand, and manipulate process variables accordingly. This review thus discusses the various approaches available for control and prediction of glycosylation in biopharmaceuticals. Further, the futuristic goal of integrating such technologies is discussed in order to attain an automated and digitized continuous bioprocess for control of glycosylation. Given, control of a process as complex as glycosylation requires intense monitoring intervention, we examine the current technologies that enable automation. Finally, we discuss the challenges and the technological gap that currently limits incorporation of an automated process in routine bio-manufacturing, with a glimpse into the economic bearing., (© 2022 American Institute of Chemical Engineers.)

Benign paroxysmal positional vertigo (BPPV) is a common clinical disorder characterized by brief recurrent spells of vertigo often brought about by certain head position changes. General treatment for BPPV is by clinical examination by Dix-Hallpike maneuver, Video head impulse test (VHIT) is a novel test that enhances diagnostic opportunities and enables a clinician to precisely localize the site of vestibular disorders. This interested us to investigate its potential in diagnosis of BPPV.The aim of the present study is to assess the role of Video head impulse test (VHIT) in confirming the clinically diagnosed case of BPPV.All patients above the age of 18 years who were clinically diagnosed with BPPV underwent VHIT and results were correlated with clinical findings. Total 60 patients were studied in the period of 2016-2018.Among the 60 patients clinically diagnosed with BPPV, 41 were males and 19 were females. Majority of patients were in the age group of 51-60 years. Posterior canal is most commonly affected (97%) than anterior and lateral canals in BPPV. In unilateral posterior canal BPPV and bilateral posterior canal BPPV VOR (Vestibulo- ocular reflex) gains was reduced but were not statistically significant. Saccades were present only in 17 cases. There is no relationship between the presence of saccades, the canal involved and the side of the lesion.From the present study we conclude that the currently available equipment for VHIT is not useful in diagnosing BPPV. Also, strongly recommends advanced research on this to record minute changes in VOR gain., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© Association of Otolaryngologists of India 2021.)

Biotherapeutics, such as mAbs and fusion proteins, are a major and rapidly growing class of pharmaceuticals. Majority of the biopharmaceuticals are glycoproteins, wherein about 1 to 30% of their molecular weight (MW) are contributed by the glycans. Determination of MW of heavily glycosylated proteins, such as Fc-fusion proteins, is seriously hampered by the physicochemical characteristics and heterogeneity of the attached carbohydrates. Glycosylation influences the expected size of the glycoprotein, which leads to disproportionate MW estimation, in size-dependent methods. Hence, in this study, we have demonstrated the advantages and limitations of four widely used MW estimation techniques for three proteins having varying levels of glycosylation. It was proven that glycosylation had least impact on MW determination by SEC-MALS and SV-AUC. However, MW estimation by LC-MS and SDS-PAGE was extensively hampered by the degree of glycosylation. It is, thus, essential to consider the structural characteristics of proteins while selecting a technique for determining their MW., (© 2022 Wiley-VCH GmbH.)

In the summer of 2020, 127 soybean [ Glycine max (L.) Merr] seedlings (V1-V3 stage) with reduced growth vigor were sampled as part of a bulk collection of seedling pathogens from Purdue's Agronomy Center for Research and Education in West Lafayette, Indiana. After rinsing off soil, one plant displayed prominent necrotic lesions on both cotyledons and the hypocotyl and rot of the roots. Root tissue segments measuring roughly 5 mm in length and adjacent to lesions were excised and surface sterilized in 0.6% NaOCl for 10 min, then in 70% ethanol for 2 min, rinsed thrice in sterile distilled H2O, and plated on dichloran-chloramphenicol-peptone agar (Andrews and Pitt 1986). Single-spore cultures were produced and grown on potato dextrose agar. The isolate (AC101) developed white aerial mycelium, rings of magenta coloration in the media, and pale orange sporodochia with age. Microscopic observation of two-week-old cultures grown on synthetic low-nutrient agar (NRRL Medium No. 4) in the dark at 28°C revealed 2-3 septate falcate macroconidia measuring 17.1 - 43.9 × 2.8 - 3.5 µm (avg. 29.4 × 3.1 µm, n=20); 0-1 septate straight to slightly curved microconidia measuring 3.9 - 8.6 × 1.9 - 2.5 µm (avg. 7.0 × 2.2 µm, n=20); and round chlamydospores borne singly or doubly with diameter measuring 6.1 - 14.2 µm (avg. 8.9 µm, n=20). These characteristics were consistent with descriptions of Fusarium commune K. Skovg., O'Donnell & Nirenberg (Skovgaard et al. 2003). DNA was extracted from aerial mycelium and the internal transcribed spacer (ITS) region using ITS1/ITS4 primers (White et al. 1990) (GenBank accession MW463361), the mitochondrial small subunit (mtSSU) rDNA using MS1/MS2 primers (White et al. 1990) (MW466537), and the translation elongation factor 1-alpha (TEF1α) gene using 983F/1567R primers (Rehner and Buckley 2005) (MW475296) were amplified and sequenced. Blast searches in GenBank showed that these sequences had 100% identity with corresponding sequences of F. commune (ITS: MN452698; mtSSU: AF362277; and TEF1α: KU171720). The matching mtSSU sequence was an accession from the original species description (Skovgaard et al. 2003). A pathogenicity test was conducted under greenhouse conditions (20-29°C, avg. 24°C) following the infested soil protocol of Ellis et al. (2013a). Ten seeds (cv. Williams) each were used in inoculated and mock-inoculated control treatments with one seed per foam cup. Root rot symptoms similar to, but more destructive than those observed in the field, were observed 14 days after planting on all inoculated plants but not on controls. Inoculated plants reached VE stage compared to controls which reached VC. Disease symptoms included severe necrotic lesions on the cotyledons, dark brown rot of the developing tap root, and brown hypocotyl lesions similar to field symptoms. F. commune was successfully reisolated from inoculated plants, but not from controls, as described above. F. commune has been reported to cause soybean root rot in China (Chang et al. 2018), Korea (Choi et al. 2020), as well as Iowa (Ellis et al. 2013b). To our knowledge this is the first report of F. commune infecting soybean seedlings in the state of Indiana. The expanded distribution of this soybean pathogen warrants heightened attention for its control.

Purpose: To describe disease characteristics and outcomes of Vogt-Koyanagi-Harada (VKH) syndrome in paediatric patients., Study Design: Retrospective chart analysis., Methods: a Retrospective: Analysis of all patients ≤16 years with VKH syndrome was done. Clinical presentations, complications, recurrences and outcomes in cases of paediatric VKH were reviewed., Results: 72 eyes of 36 patients with a mean age at presentation of 13.7 ± 2.34 years were assessed. Mean duration of symptoms and follow up were 9.88 ± 17.3 weeks and 55 months respectively. Clinical signs at presentation included anterior chamber cells >2+(34/72eyes, 47.2%), granulomatous keratic precipitates (6 eyes, 8.3%), posterior synechiae (35 eyes,48.6%), disc edema (46 eyes, 63.8%), neurosensory retinal detachments (44 eyes, 61.1%) and 'sunset-glow' fundus (9 eyes, 12.5%). Best corrected visual acuity (BCVA) at the time of presentation was 1.3logMAR or a Snellens equivalent of 20/400 which improved to 0.51logMAR (Snellens equivalent of 20/63) at last follow up. Remission was achieved in 61.1% cases. More than half of our patients developed one or more complications., Conclusion: VKH in paediatric patients poses a challenge due to a delayed presentation and paediatric VKH patients have a worse visual acuity at the time of presentation as compared to adult age groups. Rates of remission may be low along with high risk of complications and hence there is a need for prolonged immunosuppression.

Background: Oral Squamous Cell Carcinoma (OSCC) is the most widely reported cancer worldwide. Liquid biopsy, a method that relies on identification of tumor-associated cells and/or cell free nucleic acids from body fluids is becoming increasingly popular in cancer diagnostics. The aim of the study was to evaluate the feasibility of Oral Rub and Rinse (ORR) technique in determining the genetic changes in common biomarkers of oral cancer such as TP53 using DNA obtained from saliva of oral cancer patients., Methods: A total of 15 oral cancer patients were recruited in the study and pre-surgical saliva samples were collected using the ORR technique. Tissue samples included in the study were obtained during the surgical excision of the cancerous oral lesion. Genomic DNA was isolated from the salivary cell plug and the tissues and the TP53 gene was amplified by PCR. The PCR products of all the exons of TP53 (Exons 2 to 11) were electrophoresed on agarose gel, purified and sequenced by Sanger method. The obtained sequences were compared with the reference sequence of TP53 gene., Statistical Analysis Used: Descriptive statistics were used and reported as frequency and percentage., Results: Capillary sequencing of TP53 gene from tissue DNA revealed the presence of codon 72 c.215C>G (p.Pro72Arg) polymorphism in 10 patients (67%) and a heterozygous mutation at codon 172 c.514 G>T (p.Val172Phe) in 2 patients (13%). Among the 10 samples that showed codon 72 polymorphism, matched salivary DNA was available for 6 samples and 4 out of these showed same genetic change at codon 72. Similarly, of the 3 samples that showed codon 172 mutation, matched salivary DNA was available for 1 sample and the mutation status was identical., Conclusion: The results suggest a potential for clinical applications of ORR technique as an alternative to invasive tissue biopsy for detection of genetic changes in candidate biomarkers in oral cancer.

Purpose: To document the clinical features, systemic association, and treatment outcome of patients with a combined retinal vein and artery occlusion (CRVAO) and review of literature., Methods: A retrospective chart review of patients diagnosed with CRVAO at a tertiary eye care center. Patient's demographic details and associated ocular and systemic factors were recorded. Treatment included laser photocoagulation, anti-vascular endothelial growth factor (VEGF) intravitreal injection or transscleral cyclophotocoagulation (TSCPC), alone or in combination. At last, follow- up treatment response was measured in visual acuity status, regression of neovascularization, and control of intraocular pressure (IOP). All cases reported in the current decade were analyzed and compared with this study., Results: Seventeen eyes with CRVAO accounted for 0.3% of total vascular occlusion (total 5151 patients were seen in this period). The mean age was 48.12 ± 17.5 years (range: 12-87 years) and there were 9 females. Nine eyes had CRVO + CRAO; 6 eyes had BRVO + BRAO, and one patient each had CRVO + BRAO and CRAO + BRVO. Fluorescein angiography (FA) showed delayed 'arm to retina' time (>20 seconds) in all 10 eyes and delayed arteriovenous transit time in 9 out of 10 eyes. Optical coherence tomography (OCT) showed hypereflective inner retinal layers (16 eyes) and neurosensory detachment (7 eyes). The most common systemic associations were hypertension and dyslipidemia (n = 7 people; 41.18%) each. Four patients (23.5%) had a plaque in carotid arteries with normal 2D echocardiography. Ten (59%) eyes were treated with intravitreal bevacizumab + laser; four (23.5%) eyes were treated with laser only, and three (17.6%) eyes were treated with laser + anti-VEGF + TSCPC. At last follow up, vision improved in 9 (52.9%) eyes; stable in 3 (17.7%) eyes, and reduced to perception of light in 5 (29.4%) eyes., Conclusion: Combined CRVAO is a rare emergency leading to acute vision loss. Early diagnosis and treatment for ocular complications and systemic evaluation for cardiovascular risk factors are needed., Competing Interests: None

Psoriasis is a chronic, inflammatory disease affecting the skin and joints. The pathogenesis of this disease is associated with genetic, environmental and immunological factors, especially unbalanced T cell activation and improper keratinocyte differentiation. Psoriatic lesion infiltrate is composed of monocytes and T cells, and most studies have focused on the participation of T cells in the pathogenesis of this disease. Here we investigated the contribution of mononuclear phagocytes in the immunopathology observed in psoriatic patients. Significant increases in the levels of TNF, IL-1β, CXCL9, as well as the soluble forms of CD14 and CD163, were observed within the lesions of psoriatic patients compared to skin biopsies obtained from healthy individuals. Moreover, we found an association between the levels of CCL2, a monocyte attractant chemokine, and disease severity. In conclusion, our findings suggest a potential role for mononuclear phagocytes in the pathogenesis of psoriasis., (Copyright © 2020 Costa, Paixão, Viana, Rocha, Saldanha, da Mota, Machado, Pagliari, de Oliveira, Arruda, Carvalho and Carvalho.)

Background: Skin lesions from patients infected with Leishmania braziliensis has been associated with inflammation induced by cytotoxic CD8+ T cells. In addition, CD8+ T cell-mediated cytotoxicity has not been linked to parasite killing. Meanwhile, the cytotoxic role played by natural killer (NK) cells in cutaneous leishmaniasis (CL) remains poorly understood., Methods: In this study, we observed higher frequencies of NK cells in the peripheral blood of CL patients compared with healthy subjects, and that NK cells expressed more interferon-γ, tumor necrosis factor (TNF), granzyme B, and perforin than CD8+ T cells., Results: We also found that most of the cytotoxic activity in CL lesions was triggered by NK cells, and that the high levels of granzyme B produced in CL lesions was associated with larger lesion size. Furthermore, an in vitro blockade of granzyme B was observed to decrease TNF production., Concclusions: Our data, taken together, suggest an important role by NK cells in inducing inflammation in CL, thereby contributing to disease immunopathology., (© The Author(s) 2019. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Iris metastases secondary to primary malignancy are rare and usually have poor overall survival. We report a case of a man aged 60 years who presented with chief complaints of progressive pain and redness of right eye of 5 days duration. Slit-lamp examination revealed a greyish white irregular nodular mass, arising from anterior iris surface reaching up to mid-periphery. With a clinical suspicion of iris metastasis, systemic investigations were requested. CT scan of chest reported a well-defined lobulated lesion in the lower lobe of lung with presence of multiple satellite nodules in both lungs suggestive of primary malignant neoplasm of lung. Tissue diagnosis was obtained through an iris lesion biopsy, which was opted for relative ease of approach. Histopathology and immunohistochemistry confirmed the diagnosis of iris metastasis secondary to non-keratinising squamous cell carcinoma of lung. Local intravitreal injection of antivascular endothelial growth factor was administered for tumour regression along with systemic chemotherapy., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2019. No commercial re-use. See rights and permissions. Published by BMJ.)