Peptidylarginine deiminase enzymes and citrullinated proteins in female reproductive physiology and associated diseases†.

Citrullination, the post-translational modification of arginine residues, is catalyzed by the four catalytically active peptidylarginine deiminase (PAD or PADI) isozymes and alters charge to affect target protein structure and function. PADs were initially characterized in rodent uteri and, since then, have been described in other female tissues including ovaries, breast, and the lactotrope and gonadotrope cells of the anterior pituitary gland. In these tissues and cells, estrogen robustly stimulates PAD expression resulting in changes in levels over the course of the female reproductive cycle. The best-characterized targets for PADs are arginine residues in histone tails, which, when citrullinated, alter chromatin structure and gene expression. Methodological advances have allowed for the identification of tissue-specific citrullinomes, which reveal that PADs citrullinate a wide range of enzymes and structural proteins to alter cell function. In contrast to their important physiological roles, PADs and citrullinated proteins are also involved in several female-specific diseases including autoimmune disorders and reproductive cancers. Herein, we review current knowledge regarding PAD expression and function and highlight the role of protein citrullination in both normal female reproductive tissues and associated diseases.

Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome.

Citrullination is a post-translational modification (PTM) in which positively charged peptidyl-arginine is converted into neutral peptidyl-citrulline by peptidylarginine deiminase (PAD or PADI) enzymes. The full protein citrullinome in many tissues is unknown. Herein, we used mass spectrometry and identified 107 citrullinated proteins in the lactation day 9 (L9) mouse mammary gland including histone H2A, α-tubulin, and ß-casein. Given the importance of prolactin to lactation, we next tested if it stimulates PAD-catalyzed citrullination using mouse mammary epithelial CID-9 cells. Stimulation of CID-9 cells with 5 µg/mL prolactin for 10 min induced a 2-fold increase in histone H2A citrullination and a 4.5-fold increase in α-tubulin citrullination. We next investigated if prolactin-induced citrullination regulates the expression of lactation genes ß-casein (Csn2) and butyrophilin (Btn1a1). Prolactin treatment for 12 h increased ß-casein and butyrophilin mRNA expression; however, this increase was significantly inhibited by the pan-PAD inhibitor, BB-Cl-amidine (BB-ClA). We also examined the effect of tubulin citrullination on the overall polymerization rate of microtubules. Our results show that citrullinated tubulin had a higher maximum overall polymerization rate. Our work suggests that protein citrullination is an important PTM that regulates gene expression and microtubule dynamics in mammary epithelial cells.

Local level epidemiological analysis of TB in people from a high incidence country of birth.

BACKGROUND: The setting for this analysis is the low tuberculosis (TB) incidence state of New South Wales (NSW), Australia. Local level analysis of TB epidemiology in people from high incidence countries-of-birth (HIC) in a low incidence setting has not been conducted in Australia and has not been widely reported. Local level analysis could inform measures such as active case finding and targeted earlier diagnosis. The aim of this study was to use a novel approach to identify local areas in an Australian state that have higher TB rates given the local areas' country of birth profiles. METHODS: TB notification data for the three year period 2006-2008 were analysed by grouping the population into those from a high-incidence country-of-birth and the remainder. RESULTS: During the study period there were 1401 notified TB cases in the state of NSW. Of these TB cases 76.5% were born in a high-incidence country. The annualised TB notification rate for the high-incidence country-of-birth group was 61.2/100,000 population and for the remainder of the population was 1.8/100,000. Of the 152 Local Government Areas (LGA) in NSW, nine had higher and four had lower TB notification rates in their high-incidence country-of-birth populations when compared with the high-incidence country-of-birth population for the rest of NSW. The nine areas had a higher proportion of the population with a country of birth where TB notification rates are >100/100,000. Those notified with TB in the nine areas also had a shorter length of stay in Australia than the rest of the state. The areas with higher TB notification rates were all in the capital city, Sydney. Among LGAs with higher TB notification rates, four had higher rates in both people with a high-incidence country of birth and people not born in a high-incidence country. The age distribution of the HIC population was similar across all areas, and the highest differential in TB rates across areas was in the 5-19 years age group. CONCLUSIONS: Analysing local area TB rates and possible explanatory variables can provide useful insights into the epidemiology of TB. TB notification rates that take country of birth in local areas into account could enable health services to strategically target TB control measures.

Population-wide active case finding and prevention for tuberculosis and leprosy elimination in Kiribati: the PEARL study protocol.

INTRODUCTION: Population-wide interventions offer a pathway to tuberculosis (TB) and leprosy elimination, but 'real-world' implementation in a high-burden setting using a combined approach has not been demonstrated. This implementation study aims to demonstrate the feasibility and evaluate the effect of population-wide screening, treatment and prevention on TB and leprosy incidence rates, as well as TB transmission. METHODS AND ANALYSIS: A non-randomised 'screen-and-treat' intervention conducted in the Pacific atoll of South Tarawa, Kiribati. Households are enumerated and all residents ≥3 years, as well as children <3 years with recent household exposure to TB or leprosy, invited for screening. Participants are screened using tuberculin skin testing, signs and symptoms of TB or leprosy, digital chest X-ray with computer-aided detection and sputum testing (Xpert MTB/RIF Ultra). Those diagnosed with disease are referred to the National TB and Leprosy Programme for management. Participants with TB infection are offered TB preventive treatment and those without TB disease or infection, or leprosy, are offered leprosy prophylaxis. The primary study outcome is the difference in the annual TB case notification rate before and after the intervention; a similar outcome is included for leprosy. The effect on TB transmission will be measured by comparing the estimated annual risk of TB infection in primary school children before and after the intervention, as a co-primary outcome used for power calculations. Comparison of TB and leprosy case notification rates in South Tarawa (the intervention group) and the rest of Kiribati (the control group) before, during and after the intervention is a secondary outcome. ETHICS AND DISSEMINATION: Approval was obtained from the University of Sydney Human Research Ethics Committee (project no. 2021/127) and the Kiribati Ministry of Health and Medical Services (MHMS). Findings will be shared with the MHMS and local communities, published in peer-reviewed journals and presented at international conferences.

Correction: Effect of Carboxymethylation on the Rheological Properties of Hyaluronan.

[This corrects the article DOI: 10.1371/journal.pone.0162849.].

Effect of Carboxymethylation on the Rheological Properties of Hyaluronan.

Chemical modifications made to hyaluronan to enable covalent crosslinking to form a hydrogel or to attach other molecules may alter the physical properties as well, which have physiological importance. Here we created carboxymethyl hyaluronan (CMHA) with varied degree of modification and investigated the effect on the viscosity of CMHA solutions. Viscosity decreased initially as modification increased, with a minimum viscosity for about 30-40% modification. This was followed by an increase in viscosity around 45-50% modification. The pH of the solution had a variable effect on viscosity, depending on the degree of carboxymethyl modification and buffer. The presence of phosphates in the buffer led to decreased viscosity. We also compared large-scale production lots of CMHA to lab-scale and found that large-scale required extended reaction times to achieve the same degree of modification. Finally, thiolated CMHA was disulfide crosslinked to create hydrogels with increased viscosity and shear-thinning aspects compared to CMHA solutions.

EpiReview: tuberculosis in NSW, 2009-2011.

AIM: To describe the epidemiology of tuberculosis in NSW between 2009 and 2011 and compare with previous years. METHODS: Data from all cases of tuberculosis notified in NSW during this period were extracted from the Notifiable Conditions Information Management System. Descriptive analyses of notification data were undertaken. Incidence rates were calculated per 100000 population. RESULTS: Between 2009 and 2011, there were 1548 cases of tuberculosis notified in NSW, translating to an average annual notification rate of 7.2 per 100000 population for this period. A total of 89% (n=1371) of notified cases were overseas-born, and 1.6% (n=24) of cases were recorded as Aboriginal persons. The most common site of infection was the lung (60% of cases). Of notified cases, 68% were reported as having been tested for HIV, of which 3%(n=28) of cases had HIV/tuberculosis co-infection. There were 20 cases of multidrug-resistant tuberculosis, including one case of extensively drug-resistant tuberculosis. CONCLUSION: The notification rate of tuberculosis in NSW has remained relatively stable over the past two decades, though small incremental increases since 2003 are evident. Endemic transmission of tuberculosis within sub-groups of the NSW population, as well as the ongoing high endemnicity for tuberculosis in neighbouring countries, highlight the importance of tuberculosis control as a continued strategic priority for disease control in NSW.

Tuberculosis notifications in Australia, 2006.

The National Notifiable Disease Surveillance System received 1,201 tuberculosis (TB) notifications in 2006, of which 1,142 were new cases and 59 were relapses. The incidence of TB in Australia was 5.8 cases per 100,000 population in 2006 up from 5.3 per 100,000 in 2005, but still below 6 per 100,000 as it has been since 1985. Eighty-five per cent of TB notifications in 2006 were in people born outside Australia. The incidence in people born overseas and Indigenous Australians were 20.7 and 6.6 cases per 100,000 population, respectively. By contrast, the incidence of TB in the non-Indigenous Australian-born population was 0.9 cases per 100,000 population. Household or other close contact was reported as the most common risk factor for TB infection. The number of cases of TB reported in health care workers increased in 2006; these were mostly in health care workers born in TB-endemic countries and there were no reports of TB transmission in Australian health care settings. Outcome data of the 2005 TB cohort indicates that treatment success was attained in more than 95% of cases. Progress towards TB elimination in Australia will rely on continued TB awareness, maintenance of high standards of TB diagnostic and control practices, and promoting regional and global TB control activities.

Tuberculosis notifications in Australia, 2005.

The National Notifiable Disease Surveillance System received 1,072 tuberculosis (TB) notifications in 2005, of which 1,022 were new cases and 50 were relapses. The incidence of TB in Australia was 5.3 cases per 100,000 population in 2005 and has remained at a stable rate since 1985. The high-incidence groups remain people born overseas and Indigenous Australians at 20.6 and 5.9 cases per 100,000 population, respectively. By contrast, the incidence of TB in the non-Indigenous Australian-born population was 0.8 cases per 100,000 population. Rates in the Australian-born, both Indigenous and non-Indigenous have been declining since 1991, while rates in the overseas-born have been increasing. TB control in Australia relies on pre-migration screening and provision of free and effective treatment.

Tuberculosis notifications in Australia, 2004.

The National Notifiable Disease Surveillance System received 1,076 tuberculosis (TB) notifications in 2004, of which 1,043 were new cases and 33 were relapses. The incidence of TB in Australia has remained at a stable rate since 1985 and was 5.4 cases per 100,000 population in 2004. The high-incidence groups remain people born overseas and Indigenous Australians at 21.7 and 8.1 cases per 100,000 population, respectively. By contrast, the incidence of TB in the non-Indigenous Australian-born population was 1.0 cases per 100,000 population. Comparison of the 2004 TB notification data against the performance indicators set by National Tuberculosis Advisory Committee highlights that enhanced TB control measures should be considered among these high-risk groups.

Tuberculosis notifications in Australia, 2002.

In 2002, there were 1,028 cases of tuberculosis (TB) reported to the National Notifiable Disease Surveillance System, of which 997 were new cases, 30 were relapses and 1 unknown. The incidence rate of TB in Australia in 2002 was 5.2 cases per 100,000 population. The highest incidence of TB was reported in people born overseas (20.2 cases per 100,000 population), followed by Indigenous Australians (8.5 cases per 100,000 population). By contrast, the incidence rate of TB in the nonIndigenous Australian-born population was 1.1 cases per 100,000 population. This pattern of TB incidence rates amongst the sub-populations of Australia has been observed for over 10 years. The rates were evaluated against the performance indicators set by the National Tuberculosis Advisory Committee to ensure that Australia's record of TB control is maintained and improved.

Tuberculosis notifications in Australia, 2003.

The National Notifiable Disease Surveillance System (NNDSS) received 982 tuberculosis (TB) notifications in 2003, of which 947 were new cases, 33 were relapses and two were cases with unknown history. The incidence of TB in Australia has remained at a stable rate since 1985 and was 4.9 cases per 100,000 population in 2003. The high-incidence groups remain people born overseas and Indigenous Australians at 19.9 and 8.7 cases per 100,000 population, respectively. By contrast the incidence in non-Indigenous Australians was 0.9 per 100,000. Comparison of the 2003 TB notification data against the performance indicators set by National Tuberculosis Advisory Committee highlights that enhanced TB control measures should be considered among these high-risk groups.

Tuberculosis notifications in Australia, 2001.

In 2001, there were 997 cases of tuberculosis (TB) reported to the National Notifiable Diseases Surveillance System, of which, 967 were new cases of TB and 30 cases were relapses. The incidence rate of TB in Australia in 2001 was 5.1 cases per 100,000 population. The highest incidence of TB was reported in people born overseas (19.3 cases per 100,000 population), followed by Indigenous Australians (9.8 cases per 100,000 population). In contrast, the incidence rate of TB in the non-Indigenous Australian-born population was 1.0 cases per 100,000 population. This pattern of TB incidence rates amongst the sub-populations of Australia has been observed for over 10 years. Eighty-six per cent of TB cases completed treatment in 2001. Treatment was unsuccessful in 7 cases and only 22 cases defaulted. The National Tuberculosis Advisory Committee has published a National Strategic Plan with performance indicators to ensure that this enviable record of TB control is maintained and improved.