Eight-year retrospective study of young adults in a diabetes transition clinic

The transition of people from paediatric to adult diabetes services is associated with worsening glycaemia and increased diabetes-related hospitalisation. This study compared the clinical characteristics of those with and without mental health conditions among attenders at a diabetes young adult clinic diabetes before and after changes in service delivery. Retrospective audit of 200 people with diabetes attending a Sydney public hospital over eight years corresponding to the period before (2012–2016) and after (2017–2018) restructuring of a clinic for young adults aged 16–25 years. Characteristics of those with and without mental health conditions (depression, anxiety, diabetes related distress, eating disorders), were compared. Among clinic attenders (type 1 diabetes n = 184, 83.2%), 40.5% (n = 89) had a mental health condition particularly, depression (n = 57, 64%), which was higher among Indigenous than non-Indigenous people (5.6% vs. 0.8%, p = 0.031) but similar between diabetes type. Over eight years, those with, compared with those without a mental health condition had higher haemoglobin A1c (HbA1c) at the last visit (9.4% (79 mmol/mol) vs. 8.7% (71 mmol/mol), p = 0.027), the proportion with diabetic ketoacidosis (DKA 60.7% vs. 42.7%, p = 0.009), smoking (38.4 vs. 13.6%, p = 0.009), retinopathy (9.0 vs. 2.3%, p = 0.025), multiple DKAs (28.4 vs. 16.0%, p = 0.031) were significantly higher. Having a mental health condition was associated with 2.02 (95% confidence intervals 1.1–3.7) fold increased risk of HbA1c ≥9.0% (75 mmol/mol). Changes to the clinic were not associated with improvements in mental health condition (39.0% vs. 32.4%, p = 0.096). In conclusion, we found that mental health conditions, particularly depression, are common in this population and are associated with diabetes complications. Diabetes type and clinic changes did not affect the reported mental health conditions. Additional strategies including having an in-house psychologist are required to reduce complication risks among those with mental health conditions

Proteinuria in early detection of human leptospirosis

Background: Leptospirosis is an infectious disease caused by spirochetes bacteria Leptospira spp. and is reported from all over the world. As the clinical signs and symptoms of Leptospirosis often are nonspecific and the disease is early mistaken for other major infectious febrile illness, laboratory test to confirm the clinical diagnosis thus is essential for optimal treatment and patient management.Methods: Serum and urine samples were collected from patients clinically suspected cases of Leptospirosis. Preparations of urine concentrate by precipitation and centrifugation.Results: It was interesting to note that immunoglobulins are present in the urine protein concentrate of patients with Leptospirosis on the day of admission in the hospital, with urine albumin reports either positive or negative. By ELISA test it was noted that antibodies present in urine and serum were of both IgM and IgG class against the Leptospiral antigens from three pathogenic serovars and one non-pathogenicserovars. In the immunospot test which was done and compared with standard ELISA test for serum antibodies using same antigen showed that antibodies present in urine protein concentrate, which was collected on the day of admission when patients come with suspecting symptoms of Leptospirosis.Conclusions: Proteinuria is the most frequent abnormality noted in all patients at some stage of illness. This is the first report on the presence of immunoglobulins in urine samples, which were found to be of IgM and IgG classes. These findings are of significant diagnostic potential as a simple immune-spot test can be done for detecting anti-leptospiral antibodies in urine samples of suspected cases. The present attempt was aimed at developing an immunospot test, a simple and rapid diagnostic test to detect Leptospirosis using urine samples of clinically suspected patients of the infection at the earliest. It was found to be in good correlation with standard ELISA method which is being used to detect serum antibodies in Leptospira infected patients using the same antigen

Effect of iron concentration on the expression and activity of catalase-peroxidases in mycobacteria

28-33Mycobacterial catalases are known to exist in different isoforms. We studied the influence of iron concentration on the expression and activity of the different isoforms in Mycobacterium bovis BCG, M. smegmatis, M. <span style="mso-bidi-font-family:Arial;mso-bidi-language: HI">fortuitum, M. kansasii and M. vaccae <span style="mso-bidi-language: HI">by growing them under iron-sufficient (4 µg Fe/mL) and iron-deficient (0.02 µg Fe/ml) conditions. Upon iron deprivation, significant differences were observed in the catalase/peroxidase activities in both quantitative spectrophotometric assays and in the activity staining in native gels. Notable feature was that the peroxidase activity showed a significant decrease upon iron deprivation in all the mycobacteria, except M. vaccae. Peroxidase activity in all the mycobacteria, irrespective of the iron status was susceptible to heat inactivation. However, the isoforms of catalase showed differences in their heat stability, indicating possible structural differences in these proteins. For example, M. bovis BCG expressed a heat labile catalase under iron-sufficient conditions, while a heat stable catalase band of similar mobility was expressed under iron-deprivation conditions. The study clearly indicates that iron plays an important role in the regulation of expression of the different isoforms of the catalase-peroxidases, </span

Expression and Characterization of an Iron-Regulated Hemin-Binding Protein, HbpA, from Leptospira interrogans Serovar Lai▿

In an earlier study, based on the ferric enterobactin receptor FepA of Escherichia coli, we identified and modeled a TonB-dependent outer membrane receptor protein (LB191) from the genome of Leptospira interrogans serovar Lai. Based on in silico analysis, we hypothesized that this protein was an iron-dependent hemin-binding protein. In this study, we provide experimental evidence to prove that this protein, termed HbpA (hemin-binding protein A), is indeed an iron-regulated hemin-binding protein. We cloned and expressed the full-length 81-kDa recombinant rHbpA protein and a truncated 55-kDa protein from L. interrogans serovar Lai, both of which bind hemin-agarose. Assay of hemin-associated peroxidase activity and spectrofluorimetric analysis provided confirmatory evidence of hemin binding by HbpA. Immunofluorescence studies by confocal microscopy and the microscopic agglutination test demonstrated the surface localization and the iron-regulated expression of HbpA in L. interrogans. Southern blot analysis confirmed our earlier observation that the hbpA gene was present only in some of the pathogenic serovars and was absent in Leptospira biflexa. Hemin-agarose affinity studies showed another hemin-binding protein with a molecular mass of approximately 44 kDa, whose expression was independent of iron levels. This protein was seen in several serovars, including nonpathogenic L. biflexa. Sequence analysis and immunoreactivity with specific antibodies showed this protein to be LipL41

Role of sph2 Gene Regulation in Hemolytic and Sphingomyelinase Activities Produced by Leptospira interrogans.

Pathogenic members of the genus Leptospira are the causative agents of leptospirosis, a neglected disease of public and veterinary health concern. Leptospirosis is a systemic disease that in its severest forms leads to renal insufficiency, hepatic dysfunction, and pulmonary failure. Many strains of Leptospira produce hemolytic and sphingomyelinase activities, and a number of candidate leptospiral hemolysins have been identified based on sequence similarity to well-characterized bacterial hemolysins. Five of the putative hemolysins are sphingomyelinase paralogs. Although recombinant forms of the sphingomyelinase Sph2 and other hemolysins lyse erythrocytes, none have been demonstrated to contribute to the hemolytic activity secreted by leptospiral cells. In this study, we examined the regulation of sph2 and its relationship to hemolytic and sphingomyelinase activities produced by several L. interrogans strains cultivated under the osmotic conditions found in the mammalian host. The sph2 gene was poorly expressed when the Fiocruz L1-130 (serovar Copenhageni), 56601 (sv. Lai), and L495 (sv. Manilae) strains were cultivated in the standard culture medium EMJH. Raising EMJH osmolarity to physiological levels with sodium chloride enhanced Sph2 production in all three strains. In addition, the Pomona subtype kennewicki strain LC82-25 produced substantially greater amounts of Sph2 during standard EMJH growth than the other strains, and sph2 expression increased further by addition of salt. When 10% rat serum was present in EMJH along with the sodium chloride supplement, Sph2 production increased further in all strains. Osmotic regulation and differences in basal Sph2 production in the Manilae L495 and Pomona strains correlated with the levels of secreted hemolysin and sphingomyelinase activities. Finally, a transposon insertion in sph2 dramatically reduced hemolytic and sphingomyelinase activities during incubation of L. interrogans at physiologic osmolarity. Complementation of the mutation with the sph2 gene partially restored production of hemolytic and sphingomyelinase activities. These results indicate that the sph2 gene product contributes to the hemolytic and sphingomyelinase activities secreted by L. interrogans and most likely dominates those functions under the culture condition tested

Role of sph2 Gene Regulation in Hemolytic and Sphingomyelinase Activities Produced by Leptospira interrogans.

Pathogenic members of the genus Leptospira are the causative agents of leptospirosis, a neglected disease of public and veterinary health concern. Leptospirosis is a systemic disease that in its severest forms leads to renal insufficiency, hepatic dysfunction, and pulmonary failure. Many strains of Leptospira produce hemolytic and sphingomyelinase activities, and a number of candidate leptospiral hemolysins have been identified based on sequence similarity to well-characterized bacterial hemolysins. Five of the putative hemolysins are sphingomyelinase paralogs. Although recombinant forms of the sphingomyelinase Sph2 and other hemolysins lyse erythrocytes, none have been demonstrated to contribute to the hemolytic activity secreted by leptospiral cells. In this study, we examined the regulation of sph2 and its relationship to hemolytic and sphingomyelinase activities produced by several L. interrogans strains cultivated under the osmotic conditions found in the mammalian host. The sph2 gene was poorly expressed when the Fiocruz L1-130 (serovar Copenhageni), 56601 (sv. Lai), and L495 (sv. Manilae) strains were cultivated in the standard culture medium EMJH. Raising EMJH osmolarity to physiological levels with sodium chloride enhanced Sph2 production in all three strains. In addition, the Pomona subtype kennewicki strain LC82-25 produced substantially greater amounts of Sph2 during standard EMJH growth than the other strains, and sph2 expression increased further by addition of salt. When 10% rat serum was present in EMJH along with the sodium chloride supplement, Sph2 production increased further in all strains. Osmotic regulation and differences in basal Sph2 production in the Manilae L495 and Pomona strains correlated with the levels of secreted hemolysin and sphingomyelinase activities. Finally, a transposon insertion in sph2 dramatically reduced hemolytic and sphingomyelinase activities during incubation of L. interrogans at physiologic osmolarity. Complementation of the mutation with the sph2 gene partially restored production of hemolytic and sphingomyelinase activities. These results indicate that the sph2 gene product contributes to the hemolytic and sphingomyelinase activities secreted by L. interrogans and most likely dominates those functions under the culture condition tested