Risk factors and mortality rates of carbapenem-resistant Gram-negative bacterial infections in intensive care units.

Background: The prevalence of hospital-acquired infections caused by carbapenem-resistant gram-negative bacteria (CRGNB) is increasing worldwide. Several risk factors have been associated with such infections. The present study aimed to identify risk factors and determine the mortality rates associated with CRGNB infections in intensive care units. Methods: This retrospective case-control study was conducted at Erciyes University Hospital (Kayseri, Turkey) between January 2017 and December 2021. Demographic and laboratory data were obtained from the Infection Control Committee data and record system. Patients who had CRGNB infection 48-72 h after hospitalization were assigned to the case group, while those who were not infected with CRGNB during hospitalization formed the control group. Risk factors, comorbidity, demographic data, and mortality rates were compared between the two groups. Results: Approximately 1449 patients (8.97%) were monitored during the active follow-up period; of those, 1171 patients were included in this analysis. CRGNB infection developed in 14 patients (70.00%) who had CRGNB colonization at admission; in 162 (78.26%) were colonized during hospitalization, whereas 515 (54.56%) were not colonized. There was no significant difference in age, sex (male/female) or comorbidities. The total length of hospital stay was statistically significantly longer (P=0.001) in the case group (median: 24 [interquartile range: 3-378] days) than the control group (median: 16 [interquartile range: 3-135] days). The rates of colonization at admission (25.5%; vs. 10.6%, P=0.001) and mortality (64.4% vs. 45.8%, P=0.001) were also significantly higher in the cases than in the control group, respectively. In the univariate analysis, prolonged hospitalization, the time from intensive care unit admission to the development of infection, presence of CRGNB colonization at admission, transfer from other hospitals, previous antibiotic use, enteral nutrition, transfusion, hemodialysis, mechanical ventilation, tracheostomy, reintubation, central venous catheter, arterial catheterization, chest tube, total parenteral nutrition, nasogastric tube use, and bronchoscopy procedures were significantly associated with CRGNB infections (P <0.05). Multivariate analysis identified the total length of stay in the hospital (odds ratio [OR]=1.02; 95% confidence interval [CI]: 1.01 to 1.03; P=0.001), colonization (OR=2.19; 95% CI: 1.53 to 3.13; P=0.001), previous antibiotic use (OR=2.36; 95% CI: 1.53 to 3.62; P=0.001), intubation (OR=1.59; 95% CI: 1.14 to 2.20; P=0.006), tracheostomy (OR=1.42; 95% CI: 1.01 to 1.99; P=0.047), and central venous catheter use (OR=1.62; 95% CI: 1.20 to 2.19; P=0.002) as the most important risk factors for CRGNB infection. Conclusions: Colonization, previous use of antibiotics, and invasive interventions were recognized as the most important risk factors for infections. Future research should focus on measures for the control of these parameters.

The Outcome of Fungal Pneumonia with Hematological Cancer.

BACKGROUND: Fungal pneumonia is a common infectious complication of hematological cancer (HC) patients. In this retrospective study, the objective was set to identify the risk factors and outcome of fungal pneumonia in adult HC patients. MATERIALS AND METHODS: This retrospective study was conducted with adult (>16 years) HC patients from January 2017 and December 2018. RESULTS: During the study period, of 181 patients included 76 were diagnosed with fungal pneumonia. The most common HC was identified as acute myeloid leukaemia (40%). Of the participating patients, 52 (29%) were hematopoietic stem cell transplant (HSCT) recipients. The median age of patients with fungal pneumonia was significantly greater: 57 vs. 48 (odds ratio [OR]: 1.08) and they had longer hospitalization durations (OR: 1.14). Overall, 37 patients (20%) died, and 28-day mortality was significantly greater among patients with fungal pneumonia than without fungal pneumonia (33% vs. 11%). The most significant risk factors for mortality in fungal pneumonia were identified as need of intensive care unit (ICU) (OR: 191.2, P <0.001) and the need of vasopressor support (OR:81.6, P <0.012). ICU-mortality was (88%). CONCLUSION: Fungal pneumonia is a lethal complication in HC patients. Intensive care need is the most important predictive factor for mortality.

Risk prediction for candidemia in surgical intensive care unit patients.

OBJECTIVE: Patients in surgical intensive care units are thought to be at the highest risk for developing candidemia, especially patients undergoing abdominal surgery. The present study aims to investigate risk factors for candidemia in patients with abdominal surgery. METHODS: A retrospective study was undertaken that involved patients admitted to the surgical ICU between January 2016 and January 2017. All postoperative adult patients (>18 years old) who underwent abdominal surgery were included in this study. RESULTS: During the one-year study period, 49 patients developed candidemia. Thirty-five of candida isolates were non-albicans strains. Of them, 25 (51%) isolates were Candida parapsilosis, eight (16.3%) isolates were C. glabrata, one (2%) isolate was C. tropicalis and one (2%) isolate was C. kefyr. The median age of all patients enrolled in this study was 60.5±15.6 years. In univariate analysis, the duration of the hospital stays, intensive care unit stay, type of surgery, respiratory failure, total parenteral nutrition, transfusion and use of central venous catheter were significantly higher in patients with candidemia. In multivariate analysis, duration of hospital and intensive care unit stay and use of central venous catheter was associated with an increased risk of candidemia. The mortality rate of case patients was 36.7%. CONCLUSION: Patients undergoing abdominal surgery are at increased risk of candidemia, especially the patients with prolonged intensive care unit/hospital stay and the patients with a central venous catheters. Antifungal prophylaxis may be considered for patients with increased risk.

Outcomes for haematological cancer patients admitted to an intensive care unit in a university hospital.

BACKGROUND: Haematological cancer (HC) patients are increasingly requiring intensive care (ICUs). The aim of this study was to investigate the outcome of HC patients in our ICU and evaluate 5 days-full support as a breakpoint for patients' re-assessment for support. METHODS: Retrospective study enrolling 112 consecutive HC adults, requiring ICU in January-December 2015. Patients' data were collected from medical records and Infection Control Committee surveillance reports. Logistic regression analysis was performed to identify independent risk factors for ICU mortality. RESULTS: Sixty-one were neutropenic, and 99 (88%) had infection at ICU admission. Acute myeloid leukaemia was diagnosed in 43%. Thirty-five (31%) were hematopoietic stem cell transplant recipients. Only 17 (15%) were in remission. Eighty-nine underwent mechanical ventilation on admission. Fifty-three patients acquired ICU-infection (35 bacteremia) being gram negative bacteria (Klebsiella pneumoniae and non-fermenters) the top pathogens. However, ICU-acquired infection had no impact on mortality. The overall ICU and 1-year survival rate was 27% (30 patients) and 7% (8 patients), respectively. Moreover, only 2/62 patients survived with APACHE II score ≥25. The median time for death was 4 days. APACHE II score ≥25 [OR:35.20], septic shock [OR:8.71] and respiratory failure on admission [OR:10.55] were independent risk factors for mortality in multivariate analysis. APACHE II score ≥25 was a strong indicator for poor outcome (ROC under curve 0.889). CONCLUSIONS: APACHE II score ≥25 and septic shock were criteria of ICU futility. Our findings support the full support of patients for 5 days and the need to implement a therapeutic limitations protocol.

An outbreak of bloodstream infection due to extensively resistant Acinetobacter baumannii among neonates.

BACKGROUND: Extensively resistant Acinetobacter baumannii has emerged and spread worldwide as a significant cause of health care-associated infections and outbreaks. It also causes life-threatening infections among neonates, including bacteremia. The aim of this study was to investigate an outbreak of A baumannii bacteremia (ABB) among neonates. MATERIALS AND METHODS: A retrospective, case-control study was conducted from July 2014 to July 2015 in a neonatal intensive care unit (NICU). Risk factors associated with ABB in univariate and multivariate analysis with logistic regression was performed. Molecular typing by pulsed field gel electrophoresis was used to confirm relatedness of bacteremic A baumannii strains. RESULTS: During the 5-year period (2011-2016), 68 patients in our NICU were diagnosed with BSI due to A baumannii. The case-control study included 41 case patients within the outbreak caused by a major epidemic clone and 108 control patients. Risk factors (by univariate analysis) associated with ABB were intubation, 14-day mortality, and use of peritoneal dialysis and an umbilical catheter. Multivariate analysis identified 14-day mortality (odds ratio, 5.75; 95% confidence interval, 2.58-12.79) and umbilical catheter use (odds ratio, 2.44; 95% confidence interval, 1.1-5.4) as independent risk factors for ABB. CONCLUSIONS: This outbreak of bacteremia due to resistant A baumannii affected 41 infants and was associated with 58% mortality. Control of the outbreak was achieved by implementing long-term sustained infection control measures within the unit.

Efficacy of loading dose of colistin in Acinetobacter baumannii ventilator-associated pneumonia.

Colistin loading dose (LD) has been postulated as an advance in therapy. The clinical, microbiological effectiveness and nephrotoxicity of adding an LD to systemic colistin in ventilator-associated pneumonia (VAP) caused by multidrug-resistant (MDR) Acinetobacter baumannii remain unknown. In this quasi experimental study, the efficacy, outcomes and nephrotoxicity in 30 adults who received intravenous colistin with LD for MDR A. baumannii ventilator associated pneumonia were compared with 22 in absence of LD. Adding LD, the clinical cure rate at 14 days of therapy increased from 47.6% to 56.7% (p>0.397). No significant differences in bacteriological clearance (80 vs 81%), ICU mortality (50% vs 54.2%) or ICU length of stay (median: 32 vs 36 days) were identified. Mortality increased (76.2% vs 35.5%, p=0.004) in patients with nephrotoxicity, with age (median 67.0 vs. 50.0 years, p=0.002) being the only risk factor for nephrotoxicity. The nephrotoxicity rate increased from 27.3% in absence of LD to 35.3% with LD and SOFA <8, and 69.2% (p= 0.065) with LD and SOFA >7. Overall, nephrotoxicity was more severe in the LD group according to RIFLE criteria (p=0.015). Adding LD to systemic colistin for MDR A. baumannii VAP had no significant effect on clinical cure rates, bacteriologic clearance or pre-defined outcomes. However, the nephrotoxicity rate increased with LD, with special risk in adults with high organ failure development or advanced age. Further evidence regarding the risks and benefits of LD is required. The development of newer agents and strategies is urgently needed.

Is surveillance for colonization of carbapenem-resistant gram-negative bacteria important in adult bone marrow transplantation units?

BACKGROUND: The aim of this study was to investigate the rate of carbapenem-resistant gram-negative bacilli (CRGNB) colonization and to analyze the risk factors associated with CRGNB colonization. METHODS: This prospective study was conducted in adult patients hospitalized in hematopoietic stem cell transplantation (HSCT) units over a period of 8 months. Rectal swab samples were obtained from each participant every Monday, and patients CRGNB positive on admission were excluded. RESULTS: Of 185 participants, the median age was 47 years, and 59.5% were men. CRGNB colonization was detected in 21 (11.4%) patients. The most commonly isolated CRGNB were Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Multivariate analysis revealed that busulfan use (11.9 times), fludarabine use (6.4 times), transfer from another hospital (7.8 times), transfer between units (9.3 times), and central venous catheterization (5.1 times) were risk factors for CRGNB colonization. During the study period, febrile neutropenia (FN) developed in 9 (56.2%) of the 21 colonized patients, and 1 patient died. CONCLUSIONS: Screening of patients for CRGNB colonization may have a role in preventing the spread of CRGNB. However, the empirical antimicrobial treatment for FN in patients with CRGNB colonization did not change, and their mortality rates were similar.

Epidemiology and cost implications of candidemia, a 6-year analysis from a developing country.

Surveillance of candidemia is essential to monitor trends in species distribution and change in the incidence and antifungal resistance. In this study, we aimed to investigate prevalence, resistance rates, antifungal utilization and costs. A 6-year retrospective analysis of the data belonging to patients with candidemia hospitalized between 2010 and 2016 was performed. The annual usage of fluconazole and caspofungin and the usage of these antifungals in different units were described in defined daily doses (DDD) per 1000 patient days. In total, 351 patients of candidemia were included. Median age of the patients was 45 (0-88) and 55.1% of them were male. Overall, 48.1% of the candidemia episodes (169/351) were due to C. albicans, followed by C. parapsilosis (25.1%), C. glabrata (11.7%). Length of hospital stay was longer with a median of 20 days among patients with non-albicans candidemia. Presence of a central venous catheter was found to be an associated risk for candidemia caused by non-albicans strains. Annual incidence of candidemia increased from 0.10 to 0.30 cases/1000 patient days. Antifungal use was increased over years correlated with the cost paid for it. The policy against candidemia should be specified by each institution with respect to candidemia prevalence, resistance rates, antifungal use and costs.

Incidence and economic burden of prosthetic joint infections in a university hospital: A report from a middle-income country.

In this study, we aimed to evaluate the incidence and economic burden of prosthetic joint infections (PJIs) in a university hospital in a middle-income country. Surveillance data between April 2011 and April 2013 in the Orthopedic Surgery Department was evaluated. Patients (>16 years old) who had primary arthroplasty in Erciyes University were included in the study, and patients with preoperative infection were excluded. Patients were followed up during their stay in the hospital and during readmission to the hospital for PJI by a trained Infection Control Nurse. During the study period, 670 patients were followed up. There were 420 patients (62.7%) with total hip arthroplasty (THA), 241 (36.0%) with total knee arthroplasty (TKA) and 9 (1.3%) with shoulder arthroplasty (SA). The median age was 64, and 70.6% were female. The incidence of PJI was 1.2% (5/420) in THA, 4.6% (11/241) in TKA and 0% (0/9) in SA. PJI was significantly more prevalent in TKA (p=0.029). All of the PJIs showed early infection, and the median time for the development of PJI was 23.5 days (range 7-120 days). The median total length of the hospital stay was seven times higher in PJI patients than patients without PJI (49 vs. 7 days, p=0.001, retrospectively). All hospital costs were 2- to 24-fold higher in patients with PJI than in those without PJI (p=0.001). In conclusion, the incidence and economic burden of PJI was high. Implementing a national surveillance system and infection control protocols in hospitals is essential for the prevention of PJI and a cost-effective solution for the healthcare system in low-middle-income countries.

Predictive factors for percutaneous and mucocutaneous exposure among healthcare workers in a developing country.

The aim of this study is to determine the risk factors for percutaneous and mucocutaneous exposures in healthcare workers (HCW) in one of the largest centers of a middle income country, Turkey. This study has a retrospective design. HCWs who presented between August 2011 and June 2013, with Occupational Exposures (OEs) (cases) and those without (controls) were included. Demographic information was collected from infection control committee documents. A questionnaire was used to ask the HCWs about their awareness of preventive measures. HCWs who work with intensive work loads such as those found in emergency departments or intensive care units have a higher risk of OEs. Having heavy workloads and hours increases the risk of percutaneous and mucocutaneous exposures. For that reason the most common occupation groups are nurses and cleaning staff who are at risk of OEs. Increasing work experience has reduced the frequency of OEs.

Saprochaete capitata as an emerging fungus among patients with haematological malignencies.

Saprochaete capitata is a very rare pathogen that causes invasive disease particularly in patients with haematological malignancies. We recognised a clustering of S. capitata fungaemia in recent years. So, we report our 6-year surveillance study of fungaemia among patients with haematological malignancies and haematopoietic stem cell transplant. We performed a retrospective and observational study. Hospitalised patients aged >18 years with haematological malignancies were included in the study. A total of 51 fungaemia episodes of 47 patients were analysed. The characteristics of fungaemia in patients with S. capitata compared to patients with candidemia. Median duration of neutropenia was 21.5 days in patients with S. capitata fungaemia, whereas this duration was significantly shorter in patients with candidemia (8 days). Interval between first and last positive culture was significantly longer in patients with S. capitata fungaemia (P < 0.05). Previous use of caspofungin was significantly more common in patients with S. capitata fungaemia. Thirty-day mortality was found 40% for patients with candidemia, whereas it was 39% for patients with S. capitata. In conclusion, despite its limitations this study showed that a novel and more resistant yeast-like pathogen become prevalent due to use of caspofungin in patients with long-lasting neutropenia which was the most noteworthy finding of this 6-year surveillance study.

Increasing frequency of Pseudomonas aeruginosa infections during tigecycline use.

INTRODUCTION: The widespread use of tigecycline raises the question of increasing infection rates of Pseudomonas aeruginosa (PA) in ICUs which are not affected by this antibiotic. OBJECTIVE: The aim of this study was to determine if treatment with tigecycline is a risk factor for PA infection in ICU patients. METHODOLOGY: A retrospective and observational study was conducted at Erciyes University Hospital, Turkey, between 2008 and 2010. The Erciyes University Hospital is a 1300-bed tertiary care facility. The patients included in this study were hospitalized in four adult ICUs. Patients with PA infections (case group) were compared with patients with nosocomial infection other than PA (control group). RESULTS: A total of 1,167 patients with any nosocomial infections were included in the study. Two hundred and seventy eight (23.8%) of the patients had PA infection during their ICU stay. Fifty nine patients (21.2%) in the case group received tigecycline before developing PA infections, which were found to be significantly more frequent than in the controls (p < 0.01). Multivariate analysis showed that risk factors for PA infection were previous tigecycline use (4 times), external ventricular shunt (4.2 times), thoracic drainage catheter (2.5 times) and tracheostomy (1.6 times). CONCLUSION: Our results contribute to the need for new studies to determine the safety of tigecycline use, especially for the treatment of critically ill patients. Since tigecycline seems to be an alternative for the treatment of multidrug resistant (MDR) microorganisms, rational use of this antibiotic in ICU patients is essential.

Economic evaluation of appropriate duration of antibiotic prophylaxis for prevention of neurosurgical infections in a middle-income country.

BACKGROUND: Preoperative antibiotic prophylaxis is one of the preventive measures for surgical site infections (SSIs). Very little data about the cost effectiveness of the appropriate duration of antibiotic prophylaxis in low- and middle-income countries are available. We aim to assess the cost effectiveness of the use of antibiotic prophylaxis for <24 hours to prevent neurosurgical infections in a middle-income country, Turkey. METHODS: A 1-year prospective study was performed between June 2012 and June 2013. During this study period patients were followed-up on for the development of SSI by means of hospital and postdischarge surveillance. Patients included in the study group received appropriate duration of antibiotic prophylaxis (<24 hours), and the duration of prophylaxis was longer in the control group. The antibiotic costs per patient, including prophylaxis and treatment, were calculated. RESULTS: A total of 822 operations consisting of craniotomy (n = 558), spinal fusion (n = 220), and ventricular shunt (n = 44) were included in the study. The study group included 488 (59.4%) patients who underwent operations with appropriate duration (<24 hours) of antibiotic prophylaxis. Prophylactic antibiotic cost per patient was significantly lower in the study group ($3.35 and $20.41, respectively). The SSI rates did not differ between the 2 groups: 3.5% (17/488) in the study group and 3.6 (12/822) in the control group (P > .05). CONCLUSION: This cost-analysis study demonstrates that prolonged antibiotic prophylaxis correlates with increased burden of cost, but it is not preventive for SSI.

Evaluation of the effectiveness of an infection control program in adult intensive care units: a report from a middle-income country.

BACKGROUND: The rates of hand hygiene improvement and health care-associated infections (HAIs) were evaluated after the introduction in 2004 of an infection surveillance and prevention program at a university teaching hospital in a low- to middle-income country. METHODS: Data on hand hygiene compliance, HAI rate, multiresistant organisms, and antibiotic consumption in 4 adult intensive care units (ICUs; medical, general surgery, anesthesiology and reanimation, and neurosurgery) were collected retrospectively for each year from 2004 to 2012. Negative binomial regression modeling with a log link was used to adjust for overdispersion in observations, and the first year of observations served as the baseline for comparing changes in incidence rate ratio (IRR) over the subsequent years. RESULTS: Total hand hygiene compliance improved from 30.5% in 2004 to 43.5% by 2010 (IRR, 1.3; P <.0001) and reached 63.8% by 2012 (IRR, 1.9; P < .0001). The HAI rate was 42.6/1,000 patient-days at baseline and increased significantly thereafter until 2012, when it decreased by 20% to 33.6/1,000 patient-days (IRR, 0.8; P = .001). The rate of central line-associated bloodstream infection was 7.85 (95% confidence interval [CI], 5.89-10.26)/1,000 catheter-days in 2004 and increased to 12.4 (95% CI, 9.98-14.39)/1,000 catheter-days in 2012 (IRR, 1.5; P = .024). The rate of ventilator-associated pneumonia remained stable from the 2004 baseline rate of 31.66/1,000 ventilator-days to the 2012 rate of 24.04/ 1,000 ventilator-days (IRR, 0.88; P = .574). The rate of catheter-associated urinary tract infection remained relatively stable between 2004 and 2012 (from 7.92/1,000 catheter-days to 4.97/1,000 catheter-days; P = .101). The rate of methicillin-resistant Staphylococcus aureus infection was 6.24/1,000 patient-days at baseline and decreased significantly to 0.73/1,000 patient-days by 2007 (IRR, 0.13; P <.001) and continued to remain below 2/1,000 patient-days for the next 5 years. The rate of Pseudomonas aeruginosa infection decreased significantly from 8.66/1,000 patient-days in 2004 to 6.09/1,000 patient-days in 2010 (IRR, 0.72; P = .026) and to 5.44/1,000 patient-days by 2012 (IRR, 0.63; P = .002). The rate of Acinetobacter baumannii infection was 14.3/1,000 patient-days at baseline, decreased significantly by 2005 (IRR, 0.73; P = .012), fluctuated between 2006 and 2010, and then decreased significantly to 10.44/1,000 patient-days in 2011 (IRR, 0.74; P = .007) and then to 7.6/1,000 patient-days in 2012 (IRR, 0.53; P < .001). Antibiotic consumption did not decrease noticeably over the 9-year study period. CONCLUSIONS: Hand hygiene improved in all of the ICUs evaluated. Measuring changes in HAI rates in a single health care setting can be statistically challenging, and a bias in the detection rates is not uncommon in the early years of a new infection prevention program. Here, for the first time, implementation of an infection surveillance and prevention program was associated with a reduction in HAI rate.

A nightmare for haematology clinics: extensively drug-resistant (XDR) Acinetobacter baumannnii.

The purpose of our study was to share experience on demographic characteristics and clinical outcome of the patients infected with extensively drug-resistant Acinetobacter baumannii (XDRAB) in haematology clinics, focusing on the period with a sudden increase in the number of XDRAB cases. A regular patient-based infection control programme was set up in haematology clinics and haematopoietic stem cell transplant centre starting from 2008. An infection control nurse visited all patients daily. A form including demographic data and laboratory results were recorded for all patients. The source of infections was identified according to the criteria proposed by the Centers for Disease Control and Prevention. While haematology ward-acquired XRDAB was rare before 2012, between January 2012 and July 2013, 29 A. baumannii infection episodes were detected in 28 patients. All but one isolate were MDR and 72.4% (21 out of 29) were XDR. Blood cultures revealed A. baumannii in 26 out of 29 episodes. While the haematological malignancy was relapsing or not under remission in 15 patients, four patients were under remission, and 10 patients were newly diagnosed. The mortality rate was 81.2%. All patients with a poor outcome died in the first week after the index blood culture was performed. In 16 out of 29 episodes, the patients died before the culture results became available. Colistin was initiated for the treatment in 11 out of 29 episodes. Three patients received colistin combined with sulbactam or sulbactam containing beta-lactams; the remaining eight patients who received colistin monotherapy were already under carbapenems. In conclusion, XDRAB infections can easily become nightmares for haematology clinics without any reliable treatment option.

Prevaccination screening of health-care workers for immunity to measles, rubella, mumps, and varicella in a developing country: What do we save?

A structured questionnaire was administered to health-care workers (HCWs). The HCWs were also screened for measles, rubella, mumps, and varicella (MMRV) using serological methods. One thousand two hundred and fifty-five HCWs were tested. Of the HCWs examined, 94% were immune to measles, 97% to rubella, 90% to mumps and 98% to varicella. The positive predictive values of histories of measles, mumps, rubella and varicella were 96%, 93%, 100% and 98%, respectively. The negative predictive values of histories of measles, mumps, rubella and varicella were 13%, 17%, 5% and 2%, respectively. The cost of vaccination without screening was significantly more expensive (cost difference: €24,385) for varicella, although vaccination without screening was cheap (cost difference: €5693) for MMR. Although the use of cheaper vaccines supports the implementation of vaccination programs without screening, the cost of vaccination should not be calculated based only on the direct costs. The indirect costs associated with lost work time due to vaccination and its side effects and the direct costs of potential side effects should be considered. However, if prescreening is not conducted, some HCWs (2-7%) would be unprotected against these contagious illnesses because of the unreliability of their MMRV history. In conclusion, the screening of HCWs before vaccination continues to be advisable.