Ocular manifestations of HIV infection at Chris Hani Baragwanath Academic Hospital, Johannesburg, South Africa.

BACKGROUND: The pattern of HIV-associated eye disease has changed with ongoing advancements in highly active antiretroviral therapy (HAART). HIV-infected individuals now live longer, enabling us to observe the long-term effects of HIV and HAART on the eye. There are few recent studies on HIV-related ocular disease in sub-Saharan Africa. OBJECTIVES: To describe the ocular manifestations of HIV in patients attending the Nthabiseng HIV clinic at Chris Hani Baragwanath Academic Hospital, Johannesburg, South Africa. METHODS: A cross-sectional study was conducted in 2021 and 2022 using convenience sampling of patients at the HIV clinic. The participants' clinical history was taken, their files were reviewed, and they underwent ocular examination. Correlation between eyes was managed by taking disease in one eye as the presence of disease in the participant. Descriptive statistics were used to summarise participant characteristics. Univariate and multivariate logistic regression models were used to assess the odds ratio (OR) of developing HIV-associated ocular diseases, and a p-value of <0.05 was used to define statistical significance. RESULTS: There were 182 participants (139 females and 43 males), with a mean (standard deviation) age of 48.9 (10.6) years. The most common anterior segment diagnoses were conjunctival microangiopathy (34.6%), pinguecula (31.3%) and cataracts (30.2%), while the most common posterior segment finding was peripheral retinal scarring with features in keeping of previous cytomegalovirus retinitis (24.2%). Notably, only 1.1% of patients had HIV retinopathy. A CD4 count <200 cells/µL showed an increased OR for cataracts (OR 4.24; p=0.003) and any anterior segment diagnoses (OR 10.05; p=0.029), while a CD4 count ≥200 cells/µL showed an increased risk of conjunctival microangiopathy (OR 2.14; p=0.017). CONCLUSION: With the advent of HAART, ocular manifestations of HIV are changing and the incidence of severe ocular opportunistic infections and HIV retinopathy has decreased precipitously. Although this study has shown that patients with a CD4 count <200 cells/µL are at increased risk of developing anterior ocular manifestations of HIV, including cataracts, these diseases are relatively innocuous or easily treatable. Routine ocular screening of HIV patients seems to be substantially less important now than it was in the pre-HAART era.

Epithelial Regeneration After Doxorubicin Arises Primarily From Early Progeny of Active Intestinal Stem Cells.

BACKGROUND & AIMS: aISCs (aISCs) are sensitive to acute insults including chemotherapy and irradiation. Regeneration after aISC depletion has primarily been explored in irradiation (IR). However, the cellular origin of epithelial regeneration after doxorubicin (DXR), a common chemotherapeutic, is poorly understood. METHODS: We monitored DXR's effect on aISCs by enumerating Lgr5-eGFP+ and Olfm4+ crypts, cleaved caspase-3 (CASP3+) immunofluorescence, and time-lapse organoid imaging. Lineage tracing from previously identified regenerative cell populations (Bmi1+, Hopx+, Dll1+, and Defa6+) was performed with DXR damage. Lineage tracing from aISCs was compared with lineage tracing from early progeny cells (transit-amplifying cells arising from aISCs 1 day predamage) in the context of DXR and IR. We compared stem cell and DNA damage response (DDR) transcripts in isolated aISCs and early progeny cells 6 and 24 hours after DXR. RESULTS: Epithelial regeneration after DXR primarily arose from early progeny cells generated by aISCs. Early progeny cells upregulated stem cell gene expression and lacked apoptosis induction (6 hours DXR: 2.5% of CASP3+ cells, p<0.0001). aISCs downregulated stem cell gene expression and underwent rapid apoptosis (6 hours DXR: 63.4% of CASP3+ cells). There was minimal regenerative contribution from Bmi1+, Hopx+, Dll1+, and Defa6+-expressing populations. In homeostasis, 48.4% of early progeny cells were BrdU+, and expressed low levels of DDR transcripts. CONCLUSIONS: We show that DXR effectively depleted aISCs in the small intestine and subsequent epithelial regeneration depended on nonquiescent early progeny cells of aISCs. The chemoresistant phenotype of the early progeny cells may rely on a dampened DDR in contrast to aISCs' robust DDR, which facilitates expeditious apoptosis.

Botulinum neurotoxin injections in essential infantile esotropia-a comparative study with surgery in large-angle deviations.

PURPOSE: To compare botulinum neurotoxin (BNT) injections to surgery as first-line therapy in large-angle essential infantile esotropia (IE). PATIENTS AND METHODS: Children between the ages 6 months and 6 years with IE of ≥40 prism dioptres (PD) were randomised to either a maximum of three BNT injections or surgical intervention of bimedial rectus muscle recession for angles ≤60 PD and augmented with BNT injection in angles >60 PD. Time taken for each procedure was documented. Orthophoria or misalignment of ≤10 PD was regarded as a complete response (CR). Follow-up visits were done at 3, 6, 12 and 24 weeks. RESULTS: Mean (SD) age and baseline angle of esotropia were 26.9 (14.5) months and 61.9 PD (12.8), respectively, for the overall cohort. The proportion of children who achieved CR was significantly higher in the surgery arm compared to the BNT injection arm (OR = 4.01, 95% CI 1.74-9.22) but the time taken was six times longer (p < 0.0001). In the BNT arm, 55.2% of children aged ≤24 months and 16% of children >24 months achieved CR. In children with esotropia ≤60 PD, CR was achieved in 50% while those with esotropia >60 PD CR was achieved in 25%. CONCLUSION: Surgery remains the gold standard for treatment of esotropia but BNT injection is a safe and effective alternative in children ≤24 m and with smaller angles of esotropia ≤60 PD in resource-limited centres.

Inhibition of vascular endothelial growth factor A expression in mouse granulosa cells by lentivector-mediated RNAi.

Vascular endothelial growth factor (VEGF) has been found responsible for the induction of proliferation and differentiation in granulosa cells. We constructed four short hairpin RNA (shRNA) expression plasmids targeting the mouse VEGFA gene, and examined their effect on VEGF expression in mouse granulosa cells (MGC) in vitro. Four different shRNA oligonucleotides targeting the coding sequence of mouse VEGFA mRNA and one negative control (shNC) were designed and cloned into a pGPU6/GFP/Neo siRNA expression vector, and transiently transfected into MGC. At 48 h post-transfection, total RNA was extracted from the cells and subjected to qRT-PCR analysis. The most effective interference vector, shVEGF1487 was chosen for lentiviral construction. The recombinant plasmid was then transfected into 293FT cells via Lipofectamine(TM) 2000-mediated gene transfer, for the production of lentivirus, and then concentrated via ultracentrifugation. This lentiviral vector was then used for the transduction of MGC. VEGFA gene expression, apoptosis genes and VEGFA receptor genes were detected by qRT-PCR, the VEGFA protein level in culture media by ELISA assay and protein levels in MGC by Western blot analysis. The four VEGFA expression plasmids were successfully constructed and the most effective interference vector, shVEGF1487, was chosen for lentiviral production and MGC transduction. There was significant knockdown of the VEGFA gene, receptor genes and apoptosis genes for all the shVEGF constructs, compared with the shNC and Mock controls. The lentiviral vector also gave significant knockdown of the VEGFA gene. Protein levels were lower for most of the shVEGFs based on Western blot analysis with exception of VEGF1359; in this case, it was higher than shNC but lower than for the Mock group. Lentivector-transduced MGC also gave lower levels of protein. We conclude that shVEGF expression plasmids and lentivector carrying RNAi are promising tools for the inhibition of VEGF, the corresponding receptor genes, and apoptosis gene expression in MGC.

Doxycycline-regulated growth hormone gene expression system for swine.

Although growth hormone gene transgenic animals are much larger than normal animals, they manifest differences that have adverse effects on survival due to overexpression of growth hormone. We developed a stable pig embryonic fibroblast cell line expressing pig growth hormone (pGH) using the Tet-On system, with which we can conditionally manipulate expression of pGH in vivo. Inducible expression of pGH was achieved by combining reserve Tet-controlled transcriptional activator and tetracycline-responsive element in a single plasmid. The mRNA expression of pGH was significantly increased compared to the non-induced group by about 10-fold. The controlled secretion of pGH induced by doxycycline was further tested in stably transfected cells. We conclude that inducible GH expression can be achieved in pig embryonic fibroblasts.

Uso da ecocardiografia contrastada para avaliação de tumores e trombos / Contrast echocardiography for the evaluation of tumors and thrombi

A ecocardiografia contrastada baseia-se na injeção endovenosa de microbolhas que são marcadores de fluxo sangüíneo e aumentam o sinal ultra-sonográfico. O uso de agentes de contraste melhora a opacificação das cavidades cardíacas e o delineamento dos bordos endocárdicos, além de permitir a avaliação da perfusão. Recentemente, a ecocardiografia contrastada tem sido empregada na avaliação de massas cardíacas. Neste artigo são relatados um caso de mixoma atrial esquerdo (tumor benigno), um caso de metástase de adenocarcinoma de pulmão (tumor maligno) e um caso de trombo avaliados pela ecocardiografia contrastada, demonstrando que esse método tem o potencial para diagnosticar diferentes tipos de massas cardíacas.

Contrast echocardiography is based on intravenous injection of microbubbles that act as blood flow tracers and increase ultra-sound signal. Contrast agents have shown to improve cardiac cavities opacification and endocardial border delineation in addition to helping perfusion evaluation. Contrast echocardiography has recently been used to evaluate cardiac masses. In this report we will describe three cases evaluated by contrast echocardiography: a left atrial myxoma (benign tumor), a lung adenocarcinoma metastasis (malign tumor), and one thrombus. Contrast echocardiography showed to be valuable in the diagnosis of the different types of cardiac masses.

Contrast echocardiography for the evaluation of tumors and thrombi.

Contrast echocardiography is based on intravenous injection of microbubbles that act as blood flow tracers and increase ultra-sound signal. Contrast agents have shown to improve cardiac cavities opacification and endocardial border delineation in addition to helping perfusion evaluation. Contrast echocardiography has recently been used to evaluate cardiac masses. In this report we will describe three cases evaluated by contrast echocardiography: a left atrial myxoma (benign tumor), a lung adenocarcinoma metastasis (malign tumor), and one thrombus. Contrast echocardiography showed to be valuable in the diagnosis of the different types of cardiac masses.

Vasoespasmo coronariano induzido pela ecocardiografia sob estresse pela dobutamina-atropina / Coronary spasm induced by dobutamine-atropine stress echocardiography

Relatamos caso de mulher de 45 anos de idade, com antecedentes de hipertensão arterial sistêmica e tabagismo, submetida a ecocardiografia sob estresse pela dobutamina-atropina para investigação de doença arterial coronariana. No pico do estresse, a paciente apresentou dor precordial súbita e de forte intensidade. O eletrocardiograma de doze derivações revelou elevação do segmento ST nas derivações DII, DIII, aVF, V5 e V6 e depressão do segmento ST nas derivações DI, aVL, V2 e V3. Pela monitoração das imagens ecocardiográficas foi observado aparecimento de discinesia do septo inferior e acinesia da parede inferior do ventrículo esquerdo. O exame foi interrompido imediatamente, a paciente foi medicada e evoluiu com melhora da dor precordial e das alterações de motilidade segmentar. A angiografia coronariana revelou lesões coronarianas irregulares com menos de 50 por cento de obstrução do diâmetro luminal. Trata-se de um caso de vasoespasmo coronariano induzido por estimulação alfa-adrenérgica durante a ecocardiografia sob estresse pela dobutamina-atropina.

This is the report on a 45-year-old female, with a history of systemic arterial hypertension and cigarette smoking, submitted to dobutamine-atropine stress echocardiography for the investigation of coronary artery disease. At stress peak, the patient reported sudden, highly intense precordial pain. The 12-lead electrocardiogram showed ST segment elevation in DII, DIII, aVF, V5 and V6, and depression in DI, aVL, V2 and V3. Echocardiographic imaging monitoring showed dyskinesia of inferior septum and akinesia of inferior wall. The test was interrupted immediately. The patient was medicated and improved her precordial pain condition as well as wall motion abnormalities. Coronary angiography showed irregular coronary lesions with <50 percent luminal diameter obstruction. It is a case of coronary spasm induced by alpha-adrenergic stimulation during dobutamine-atropine stress echocardiography.

Coronary spasm induced by dobutamine-atropine stress echocardiography.

This is the report on a 45-year-old female, with a history of systemic arterial hypertension and cigarette smoking, submitted to dobutamine-atropine stress echocardiography for the investigation of coronary artery disease. At stress peak, the patient reported sudden, highly intense precordial pain. The 12-lead electrocardiogram showed ST segment elevation in DII, DIII, aVF, V5 and V6, and depression in DI, aVL, V2 and V3. Echocardiographic imaging monitoring showed dyskinesia of inferior septum and akinesia of inferior wall. The test was interrupted immediately. The patient was medicated and improved her precordial pain condition as well as wall motion abnormalities. Coronary angiography showed irregular coronary lesions with <50% luminal diameter obstruction. It is a case of coronary spasm induced by alpha-adrenergic stimulation during dobutamine-atropine stress echocardiography.

Expression of protease-activated receptor-2 by osteoblasts.

Osteoblasts express protease-activated receptor-1 (PAR-1), which is activated by thrombin or by synthetic peptides corresponding to the new "tethered ligand" N-terminus of PAR-1 created by receptor cleavage. Both thrombin and human PAR-1-activating peptide stimulate an elevation of [Ca2+]i in the human SaOS-2 osteoblast-like cell line, but the peptide stimulates receptor-mediated Ca+ entry, whereas thrombin does not. Stimulation of proliferation in rat primary osteoblast-like cells is greater in response to rat PAR-1-activating peptide than to thrombin. Because the PAR-1-activating peptides are now known to activate PAR-2, the current study was undertaken to investigate whether osteoblasts express this receptor and, if so, whether this could account for the observed discrepancies between responses of osteoblasts to thrombin and to PAR-1-activating peptides. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemical studies demonstrated expression of PAR-2 by primary cultures of rat calvarial osteoblast-like cells. In immunohistochemical studies of embryonic mouse bones, osteoblasts showed positive staining for the presence of PAR-2. Activators of PAR-2 include trypsin, mast cell tryptase, gingipain-R, and synthetic peptides corresponding to the PAR-2 tethered ligand sequence. Treatment of primary rat osteoblast-like cells with rat PAR-2-activating peptide (SLIGRL), or SaOS-2 cells with human PAR-2-activating peptide (SLIGKV), caused a dose-dependent increase in [Ca2+]i. Trypsin or gingipain-R also induced an increase in intracellular calcium concentration, and caused reciprocal cross desensitization. Activators of PAR-2 caused a sharp peak in [Ca2+]i followed by a sustained plateau; [Ca2+]i returned to baseline levels upon treatment with ethylene-glycol tetraacetic acid (EGTA). Treatment of rat osteoblast-like cells in vitro with SLIGRL did not affect thymidine incorporation or endogenous alkaline phosphatase activity. The results presented here demonstrate that osteoblasts express PAR-2, and that such expression is able to account for the observed discrepancies between thrombin and PAR-1-activating peptides in their ability to evoke calcium entry, but not proliferative responses.