Change in antimicrobial susceptibility of pathogens isolated from surgical site infections over the past decade in Japanese nation-wide surveillance study.

Inappropriate antimicrobial therapy for surgical site infections (SSIs) can lead to poor outcomes and an increased risk of antibiotic resistance. A nationwide survey was conducted in Japan from 2018 to 2019 to investigate the antimicrobial susceptibility of pathogens isolated from SSIs. The data were compared with those obtained in 2010 and 2014-2015 surveillance studies. Although the rate of detection of extended-spectrum ß-lactamase producing strains of Escherichia coli was increased from 9.5% in 2010 to 23% in 2014-2015, the incidence decreased to 8.7% in 2018-2019. Although high susceptibility rates were detected to piperacillin/tazobactam (TAZ), the geometric mean MICs were substantially higher than to meropenem (2.67 vs 0.08 µg/mL). By contrast, relatively low geometric mean MICs (0.397 µg/mL) were demonstrated for ceftolozane/TAZ. Although the MRSA incidence rate decreased from 72% in the first surveillance to 53% in the second, no further decrease was detected in 2018-2019. For the Bacteroides fragilis group species, low levels of susceptibility were observed for moxifloxacin (65.3%), cefoxitin (65.3%), and clindamycin (CLDM) (38.9%). In particular, low susceptibility against cefoxitin was demonstrated in non-fragilis Bacteroides, especially B. thetaiotaomicron. By contrast, low susceptibility rates against CLDM were demonstrated in both B. fragilis and non-fragilis Bacteroides species, and a steady decrease in susceptibility throughout was observed (59.3% in 2010, 46.9% in 2014-2015, and 38.9% in 2018-2019). In conclusion, Japanese surveillance data revealed no significant lowering of antibiotic susceptibility over the past decade in organisms commonly associated from SSIs, with the exception of the B. fragilis group.

[Successful Resection Using a Posterior Approach of a Tailgut Cyst in the Retrorectal Space].

We present the case of a Tailgut cyst occurring in the retrorectal space that was curatively resected using a posterior approach. A 40-year-old man presented to the Kochi Health Sciences Center with the chief complaint of perineal incongruity. Pelvic magnetic resonance imaging revealed a multilocular cystic lesion in the retrorectal space, with high signal intensity on T2-weighted imaging. After diagnosing a Tailgut cyst, we performed resection of the tumor using a posterior approach. The lesion was removed en bloc with the coccyx. Histopathologically, the lesion was diagnosed as a non-malignant Tailgut cyst, and the surgical margin was negative. The patient is currently doing well without recurrence at 20 months.

Presepsin in the prognosis of infectious diseases and diagnosis of infectious disseminated intravascular coagulation: a prospective, multicentre, observational study.

BACKGROUND: Few prospective studies have described the prognostic accuracy of presepsin for 28-day mortality during days 0 to 7, or its role in the diagnosis of disseminated intravascular coagulation (DIC) in patients with infection. OBJECTIVE: We aimed to evaluate the clinical usefulness of presepsin levels by comparing infection markers such as procalcitonin, interleukin-6 and C-reactive protein, as well as markers of DIC such as fibrin degradation products (FDPs) and D-dimer, from days 0 to 7. DESIGN: A prospective, multicentre, observational study. SETTING: Four medical institutions between June 2010 and June 2011. PATIENTS: A total of 191 patients who fulfilled at least one of the systemic inflammatory response syndrome (SIRS) criteria were enrolled in the study. MAIN OUTCOME MEASURES: The presepsin levels were evaluated for their diagnostic accuracy in discriminating between SIRS and sepsis, the prognostic accuracy for 28-day mortality from days 0 to 7 and the diagnostic accuracy for DIC in patients with infection by comparison with other infection markers. RESULTS: The diagnostic accuracy for discriminating between SIRS and sepsis from combining the presepsin and procalcitonin measurements [area under the curve (AUC), 0.91; likelihood ratio, 4.96] was higher than that of presepsin (AUC, 0.89; likelihood ratio, 4.75) or procalcitonin (AUC, 0.85; likelihood ratio, 3.18) alone. Not only the correlation coefficient between the presepsin level and the sequential organ failure assessment (SOFA) score but also the prognostic accuracy of presepsin for 28-day mortality increased with the elapsed time, and both were highest at day 7. The diagnostic accuracy for DIC generated by combining presepsin and FDP (AUC, 0.84; likelihood ratio, 3.57) was higher than that of FDP (AUC, 0.82; likelihood ratio, 2.64) or presepsin (AUC, 0.80; likelihood ratio, 2.94) alone. CONCLUSION: The prognosis and severity of infection may be assessed more accurately by measuring the presepsin levels until day 7. Presepsin is a useful diagnostic tool for DIC with infection.

Presepsin as a powerful monitoring tool for the prognosis and treatment of sepsis: a multicenter prospective study.

Presepsin is a protein whose levels increase specifically in the blood of patients with sepsis. It is proposed as a diagnostic and prognostic marker for assessing the degree of sepsis severity. The present multicenter prospective study compared the clinical utility of presepsin with other conventional sepsis biomarkers including procalcitonin, interleukin-6, and C-reactive protein for evaluating the severity of sepsis during follow-up. Patients with sepsis (n = 103) admitted to the emergency room or intensive care unit were enrolled in this study and classified into 3 diagnostic groups: sepsis, severe sepsis, and septic shock. Blood samples were obtained from each patient on admission and after 1, 3, 5, and 7 days. The patients were further divided into the favorable and unfavorable prognosis groups on the basis of several indicators of sepsis severity (i.e., Sequential Organ Failure Assessment score, and Acute Physiology and Chronic Health Evaluation II score). The patients in the favorable prognosis group exhibited significant decreases in all biomarker levels on days 3 and 7 after admission. In the unfavorable prognosis group, only presepsin levels did not decrease significantly during follow-up. The period of antibiotics treatment in the unfavorable prognosis group was significantly longer than those in the favorable prognosis group (P < 0.05). The unfavorable prognosis group had significantly higher 28-day mortality than the favorable prognosis group (P < 0.05). Therefore, the results suggest that presepsin levels correlated with the severity of sepsis during follow-up in comparison with other conventional sepsis biomarkers.

[Examination of metallo-beta-lactamase-producing different types of Serratia marcescens detected in the same patient].

Metallo-beta-lactamase (MBL) producing Serratia marcescens isolate was recovered from a study patient in September, 2007 in whom MBL non-producing S. marcescens had been isolated 2 months previously. Two S. marcescens isolates recovered from the study patient showed the same pulsed-field gel electrophoresis (PFGE) pattern. Seven S. marcescens isolates were recovered from other patients in our hospital during August, 2007 and November, 2007. Five of the seven isolates produced MBL. All of the MBL-producing isolates showed the same PFGE pattern and harbored plasmids of the same size and bla(IMP) genes. The bla(IMP) genes were easily transferred to Escherichia coli DH5alpha by transformation of a plasmid purified from the MBL-producing isolate. Those transformation experiments suggested that bla(IMP) genes were encoded by the plasmid. From these observations, it was speculated that the MBL non-producing S. marcescens isolate recovered from the study patient had acquired the plasmid which encoded bla(IMP) genes and a monoclone of MBL-producing S. marcescens spread horizontally in our hospital.

A case of bladder and pelvic dead space inflammation successfully treated with endoscopic ultrasound drainage.

Key Clinical Message: Interventional endoscopic ultrasound (EUS) is effective not only for biopsy, but also for abscess drainage. We report the first use of EUS to drain inflammation of the bladder and pelvic dead space through the ileal conduit. EUS-guided drainage is effective in treating postoperative abscesses and should be employed more routinely. Abstract: The patient was a 77-year-old man with a vesicoureteral fistula. An ileal conduit was placed after abdominoperineal resection and partial bladder resection for local, postoperative recurrence of rectal cancer. During postoperative chemotherapy, the patient developed a high-grade fever and after a thorough examination, he was diagnosed with bladder and pelvic dead-space inflammation. All urine flowed through the ileal conduit, and it was assumed that secretions from the residual bladder and prostate gland had accumulated in the bladder and pelvic cavity, resulting in infection. A transcutaneous drain was inserted through the perineum and the infection was controlled, but it flared up again after the drain was removed. We concluded that long-term drainage was necessary and successfully controlled the infection by placing a plastic stent through the ileal conduit into the bladder and pelvic dead space under ultrasound endoscopy. This is the first report of ultrasound endoscopic drainage of an abscess through the ileal conduit.

Effective treatment of gallbladder neuroendocrine carcinoma with nivolumab.

An 89-year-old patient with gallbladder neuroendocrine neoplasms (GB-NENs) and lung cancer metastasis underwent nivolumab monotherapy, resulting in tumor shrinkage. Surgery and adjuvant nivolumab showed efficiency despite low expression of PD-L1.

Survival impact of surgical site infection in esophageal cancer surgery: A multicenter retrospective cohort study.

Aim: This study was performed to evaluate the oncological impact of surgical site infection (SSI) and pneumonia on long-term outcomes after esophagectomy. Methods: The Japan Society for Surgical Infection conducted a multicenter retrospective cohort study involving 407 patients with curative stage I/II/III esophageal cancer at 11 centers from April 2013 to March 2015. We investigated the association of SSI and postoperative pneumonia with oncological outcomes in terms of relapse-free survival (RFS) and overall survival (OS). Results: Ninety (22.1%), 65 (16.0%), and 22 (5.4%) patients had SSI, pneumonia, and both SSI and pneumonia, respectively. The univariate analysis demonstrated that SSI and pneumonia were associated with worse RFS and OS. In the multivariate analysis, however, only SSI had a significant negative impact on RFS (HR, 1.63; 95% confidence interval, 1.12-2.36; P = 0.010) and OS (HR, 2.06; 95% confidence interval, 1.41-3.01; P < 0.001). The presence of both SSI and pneumonia and the presence of severe SSI had profound negative oncological impacts. Diabetes mellitus and an American Society of Anesthesiologists score of III were independent predictive factors for both SSI and pneumonia. The subgroup analysis showed that three-field lymph node dissection and neoadjuvant therapy canceled out the negative oncological impact of SSI on RFS. Conclusion: Our study demonstrated that SSI, rather than pneumonia, after esophagectomy was associated with impaired oncological outcomes. Further progress in the development of strategies for SSI prevention may improve the quality of care and oncological outcomes in patients undergoing curative esophagectomy.

Usefulness of presepsin in the diagnosis of sepsis in a multicenter prospective study.

The clinical usefulness of presepsin for discriminating between bacterial and nonbacterial infections (including systemic inflammatory response syndrome) was studied and compared with procalcitonin (PCT) and interleukin-6 (IL-6) in a multicenter prospective study. Suspected sepsis patients (n = 207) were enrolled into the study. Presepsin levels in patients with systemic bacterial infection and localized bacterial infection were significantly higher than in those with nonbacterial infections. In addition, presepsin, PCT, and IL-6 levels in patients with bacterial infectious disease were significantly higher than in those with nonbacterial infectious disease (P < 0.0001, P < 0.0001, and P < 0.0001, respectively). The area under the receiver operating characteristic curve was 0.908 for presepsin, 0.905 for PCT, and 0.825 for IL-6 in patients with bacterial infectious disease and those with nonbacterial infectious disease. The cutoff value of presepsin for discrimination of bacterial and nonbacterial infectious diseases was determined to be 600 pg/ml, of which the clinical sensitivity and specificity were 87.8 % and 81.4 %, respectively. Presepsin levels did not differ significantly between patients with gram-positive and gram-negative bacterial infections. The sensitivity of blood culture was 35.4 %; that for presepsin was 91.9 %. Also there were no significant differences in presepsin levels between the blood culture-positive and -negative groups. Consequently, presepsin is useful for the diagnosis of sepsis, and it is superior to conventional markers and blood culture.

[Marked response to oral administration of UFT and leucovorin for liver metastases from colon cancer in an elderly patient].

A83 -year-old man was admitted to our hospital for the treatment of advanced ascending colon cancer with liver metastases. He had initially undergone an ileocecal resection for ascending colon cancer. Subsequently, we started oral administration of UFT/LV(UFT 400mg/day, LV 75 mg/day, and 4 weeks of therapy followed by a week suspension of treatment). After 3 courses, his tumors responded well to treatment, and CT showed marked regression of liver metastases. After 10 courses, liver metastases had almost disappeared. Two years passed without any adverse events since UFT/LV therapy was started. These findings suggest that UFT/LV therapy is very safe and effective for elderly patients with unresectable colorectal cancer.

Two cases of splenic neoplasms with differing imaging findings that required laparoscopic resection for a definitive diagnosis.

INTRODUCTION AND IMPORTANCE: Splenic tumors are rare and are sometimes found incidentally. In such cases, laboratory tests and imaging studies should be performed based on the diagnostic algorithm to determine whether the tumor is benign or malignant. However, we clinicians sometimes encounter challenging cases. Herein we experienced two challenging cases of splenic tumor which we could not correctly diagnosis preoperatively. CASE PRESENTATION: Case 1: A female in her 80s presented to our surgical department to undergo follow-up examinations for Stage IIIa ascending colon cancer. A follow-up CT scan showed marked enlargement of the splenic tumor which suggested metastatic cancer. We performed laparoscopic splenectomy. Case 2: A healthy female in her 50s presented to our internal medicine department to undergo a workup after multiple splenic tumors. A follow-up CT scan showed that the tumors had grown slightly. We could not completely rule out a malignant tumor. She rejected further follow-up study and chose splenectomy. CLINICAL DISCUSSION: We experienced two cases of splenic hemangioma with different clinical presentations and imaging findings. Although some studies have reported that biopsying a splenic tumor is a safe and effective way of distinguishing among splenic tumors, in our country splenic biopsies are seldom performed due to fears of causing intraabdominal bleeding or tumor dissemination. Clinicians should consider whether it would be better to perform follow up with a biopsy or splenectomy as a definitive treatment on a case-by-case basis. CONCLUSION: Laparoscopic splenectomy can be used for definitive management in cases involving malignancy or an uncertain etiology.

Ascending colon stenosis caused by repeated diverticulitis that clinically mimicked advanced colon cancer: A case report.

INTRODUCTION: We experienced a rare case of right-sided large bowl obstruction (LBO) of the colon caused by chronic diverticulitis, which was challenging to diagnose. PRESENTATION OF CASE: A young male was admitted to our department with a fever, diarrhea, and right-sided lateral abdominal pain for several days. CT showed a thickened ascending colon wall with stenosis and adjacent retroperitoneal inflammation without marked diverticula. The next day, he developed severe abdominal pain, and perforation was suspected. We chose the "interval definitive surgery"; at that time, intestinal decompression and laparoscopic drainage. Colonoscopy showed an edematous membrane, but no cancerous lesions or diverticula. Hemi-colectomy was performed after 10 days' nutritional therapy. No postoperative complication occurred. The histopathology showed that the pathogenesis was chronic diverticulitis. DISCUSSION: There have been few reported cases of right-sided LBO caused by diverticulitis, but it is important to be aware that benign disease, such as chronic diverticulitis, can cause LBO. Initial conservative therapy and nutritional therapy produced a correct diagnosis and good outcomes. CONCLUSION: Performing "interval surgery" allowed us to make an accurate diagnosis and may help to prevent surgical complications in rare cases of right-sided LBO due to diverticulitis.

Video. Advantages of the laparoscopic approach for intersphincteric resection.

BACKGROUND: Intersphincteric resection (IRS) is a surgical technique used to preserve sphincter function, mainly cases of low rectal cancer located less than 5 cm from the anal verge [1, 2]. There have been reports of laparoscopic ISR [3, 4], but discussion of the specific techniques used in this laparoscopic surgical procedure have not been sufficient. For better outcomes of this sophisticated procedure, extreme care must taken to prevent perforation of the rectal wall and to preserve the external sphincter muscle. The most difficult steps for ISR are the circular dissection and separation of the internal sphincter muscle from the external sphincter and puborectalis using the perineal approach. The authors' techniques and the advantages of laparoscopic ISR are shown by a video presentation of three rectal tumor cases. Also, the perioperative outcomes for the patients who underwent laparoscopic ISR with this technique are described. METHODS: From January 2006 to September 2009, laparoscopic ISR with total mesorectal excision was performed for 15 patients (10 men and 5 women). The median age of the patients was 60.5 years. The T categories of the tumor node metastasis (TNM) classification for the rectal cancers were Tis for two patients, T1 for one patient, T2 for four patients, and T3 for eight patients. The median distance from the anal verge to the tumor in this series was 3.7 cm. The Tis cases had large laterally spreading tumors that could not be removed by endoscopic submucosal dissection. The T1 case presented in the video had a small tumor just above the dentate line that had developed in the presence of chronic ulcerative colitis. Because this case required total proctocolectomy and ileal pouch anal anastomosis, local resection was not used (Table 1). Table 1 Patients' clinical characteristics (2006.1-2009.8) No. of patients: 15 Gender (M/F):10/5 Age: years (range): 60.5 (31-75) pT*: Tis (n=2), T1 (n=1), T2 (n=4), T3 (n=8) Distance from anal verge: cm (range): 3.7 (2-5) * Pathological T categories of the tumor node metastagis (TNM) classification CASES: The 68-year-old man in case 1 had a large, laterally spreading rectal tumor. The 61-year-old man in case 2 had rectal cancer, with a tumor located 4 cm from the anal verge. Laparoscopic surgery was performed after neoadjuvant chemoradiotherapy. The 71-year-old woman in case 3 had T1 rectal cancer, with a tumor located just above the dentate line. After dissection of the intersphincteric space, the prolapsing technique was used. METHODS: In the male patients, the rectum with the mesorectum was first dissected to the anal hiatus, initially on the posterior side along the avascular plane. Second, Denonvilliers' fascia was dissected, and the seminal vesicle was exposed. The third step was dissection of the lateral tissues followed by incision of Denonvilliers' fascia with the rectal wall exposure and care taken not to injure the neurovascular bundle (Fig. 1). Along this dissection plane, the puborectalis could be reached and intersphincteric space entered from the lateral side of the rectal wall (Fig. 2). The final step was dissection of the hiatal ligament at the posterior side of the rectum. Nearly circular dissection of the intersphincteric space could be completed. The difficulties associated with the perineal approach were reduced by this abdominal approach, and the tumor could be exteriorized easily. Fig. 1 After incission of the Denonvilliers' fascia at the lateral side of the seminal vesicle puborectalis muscle can be reached at the lateral side of the rectum. Fig. 2 Adhesion line between the puborectalis muscle and rectal wall is enposed. Intersphinecteric space can be entered along this dissection plane at the lateral side of the rectum. RESULTS: The mean duration of surgery was 386 min, and the mean blood loss was 108 ml. The mean postoperative hospital stay was 18 days. The diverting ileostomy was closed at a mean of 7.3 postoperative months. No remarkable perioperative complication was encountered (Table 2). Table 2 Perioperative outcomes (n=15) Duration of surgery: min (range) 386 (319-510) Blood loss: ml (range) 108 (0-180) Postoperative hospital stay: days (range) 18 (11-31) COMPLICATIONS: n (range) Anastomotic leakage 1 Stricture of the anastomosis 1 Pelvic abscess 1 Postoperative period until the stoma closure (months) 7.3 (3-16) CONCLUSION: Laparoscopic ISR enabled reduction of the difficulties associated with the perineal approach. An advantage of laparoscopic ISR is the ability clearly to visualize anatomic structures in the deep pelvic cavity.

A patient with esophageal hemangioma treated by endoscopic mucosal resection: a case report and review of the literature.

In a 58-year-old male, upper digestive endoscopy revealed a protruding lesion in the esophagus on a medical examination. The patient was referred to the Department of Surgery in our hospital to undergo surgery. On the initial consultation, upper digestive endoscopy showed a smooth, soft, black purple, type II protruding lesion measuring approximately 25 mm at 35 cm apart from the incisor. For diagnostic treatment and patient's request, endoscopic mucosal resection (EMR) was performed. The resected specimen measured 25 mm x 25 mm. The histological findings suggested cavernous hemangioma. To treat esophageal hemangioma, esohagectomy, tumor enucleation, or sclerotherapy has been performed. However, recently, thorough preoperative examination, such as endoscopic ultrasonography (EUS), has facilitated endoscopic resection, such as EMR.

Diagnostic accuracy of procalcitonin and presepsin for infectious disease in patients with acute kidney injury.

Procalcitonin (PCT) and presepsin (PSEP) are sepsis markers, but their diagnostic accuracy may be compromised in acute kidney injury (AKI). We evaluated their diagnostic accuracy in patients with/without AKI. This retrospective study comprised 91 patients with at least one criterion of systematic inflammatory response syndrome. AKI markers plasma neutrophil gelatinase-associated lipocalin (NGAL), plasma cystatin C (CysC), and estimated glomerular filtration rate (eGFR) were measured upon hospital admission and on days 1, 3, 5, and 7. Patients were divided into non-AKI and AKI groups. APACHE II severity scores were determined. PCT and PSEP levels were increased significantly in non-AKI and AKI patients with infection. NGAL, CysC, and eGFR in patients with infection were associated with PCT, PSEP, and APACHE II score, and levels of PCT and PSEP were correlated significantly with disease severity. PCT and PSEP are useful markers of bacterial infections in AKI but different thresholds should be applied.

Laparoscopic permanent sigmoid stoma creation through the extraperitoneal route.

About 50% of patients who have a permanent stoma experience some degree of parastomal hernia formation. To prevent this complication, the extraperitoneal route is considered to be more effective than the transperitoneal route in the case of open colorectal surgery. This technique also has superiority in avoiding postoperative intestinal obstruction. Although laparoscopic surgery for rectal cancer has not been proved to be as safe as open surgery by a randomized-controlled trial, some studies have shown the equality of long-term results with laparoscopic low anterior resection and laparoscopic abdominoperineal resection. It is anticipated that cases of laparoscopic abdominoperineal resection will increase in the near future. However, a laparoscopic technique for creation of a permanent stoma has hardly been discussed. Most operative procedures for laparoscopic stoma creation have been performed with transperitoneal route, which may cause parastomal hernia and/or intestinal obstruction. This report describes a laparoscopic technique for permanent sigmoid stoma creation through the extraperitoneal approach.

Resected xanthogranulomatous pancreatitis.

Xanthogranulomatous changes in the pancreas are extremely rare. A 66-year-old man presented with a 2-year history of epigastralgia. Computed tomography scan revealed a 4-cm low-density area around the body of the pancreas. Magnetic resonance imaging demonstrated that the mass appeared hyperintense on a T2-weighted image and isointense on a T1-weighted image. Based on a diagnosis of invasive ductal carcinoma of the pancreas, distal pancreatectomy and splenectomy were performed. Sections examined from the mass showed an aggregation of many foamy histiocytes, lymphocytes, and plasma cells. The surrounding pancreatic tissue showed fibrosis and chronic inflammation. These findings suggested a xanthogranulomatous inflammation, and resulted in a diagnosis of xanthogranulomatous pancreatitis.

New laparoscopic double-stapling technique.

BACKGROUND: Laparoscopic surgery for colon cancer has been shown by several randomized, controlled trials to be an acceptable alternative to open surgery; however, laparoscopic rectal surgery has not been evaluated in a randomized trial. One of the most serious problems associated with laparoscopic rectal surgery are bowel clamping, irrigation, and transection of the rectum, and laparoscopic rectal surgery has not been as reliable as open rectal surgery. MATERIALS AND METHODS: We present our new technique, the laparoscopic double-stapling technique, which eliminates these problems. This technique uses curved Doyen forceps introduced through the wound just above pubis symphysis for clamping the rectal wall at the anal side of the tumor. An endolinear stapler (length 60 mm) is inserted through the same wound, applied at the rectal wall parallel and caudal to the Doyen forceps, and transects the rectum under pneumoperitoneum. We used this technique for eight cases of rectal surgery. RESULTS AND DISCUSSION: The laparoscopic double-stapling technique provided secure bowel clamping and rectal irrigation. The number of cartridges used in laparoscopic double-stapling technique cases was not more than 2, with an average of 1.6 per patient. None of the laparoscopic double-stapling technique cases experienced major complications. CONCLUSION: We consider that many cases of rectal cancer that are suitable for laparoscopic low anterior resection can undergo laparoscopic surgery by using this technique, which will improve the quality of rectal surgery.

Molecular loops in the galactic center: evidence for magnetic flotation.

The central few hundred parsecs of the Milky Way host a massive black hole and exhibit very violent gas motion and high temperatures in molecular gas. The origin of these properties has been a mystery for the past four decades. Wide-field imaging of the (12)CO (rotational quantum number J = 1 to 0) 2.6-millimeter spectrum has revealed huge loops of dense molecular gas with strong velocity dispersions in the galactic center. We present a magnetic flotation model to explain that the formation of the loops is due to magnetic buoyancy caused by the Parker instability. The model has the potential to offer a coherent explanation for the origin of the violent motion and extensive heating of the molecular gas in the galactic center.