DFS Pattern Research Articles

Assessment of ameliorative effects of organic dietary interventions on neonicotinoid exposure rates in a Japanese population.

Neonicotinoid insecticides (NNIs) are a popular class of insecticides used in various pest management regimens worldwide. Biomonitoring studies continuously report high exposure rates of NNIs in various human populations across the globe. Yet, there is no validated countermeasure for combating the recent exponential rise in NNI exposure rates observed in human populations. The current study assessed the impacts of organic dietary interventions on NNI exposure rates in a Japanese population. A total of 103 volunteers were recruited into the study. Subjects were either served with Organic diets for 5 and 30days or conventional diets. A total of 919 repeated urine samples were collected from the participants and then subjected to LC-MS/MS analysis to determine urinary concentrations of 7 NNIs parent compounds and an NNI metabolite. Eight NNIs were detected; with a decreasing detection frequency (%Dfs) pattern; desmethyl-acetamiprid (dm-ACE) (64.96%)>dinotefuran (52.12%), imidacloprid (39.61%)>clothianidin (33.95%)>thiamethoxam (28.51%)>acetamiprid (12.62%)>nitenpyram (5.33%)>thiacloprid (2.83%). Dinotefuran, dm-ACE, and clothianidin recorded the highest concentrations in the subjects. The %Df of NNIs in the 5-days or 30-days organic diet group were lower than those of the conventional diet consumers. The organic diet group showed lower rates of multiple NNI exposures than those of the conventional diet consumers. The mean and median cumulative levels of NNIs (median IMIeq) were significantly lower in the organic diet group than the conventional diet group (p<0.0001). The estimated daily intakes (EDIs) of NNIs were higher in adults than children, but less than 1% of NNI cRfDs, except for clothianidin, which exhibited a %cRfD of 1.32 in children. Compared to the conventional diet group, the 5- and 30-day organic dietary intervention showed drastic reductions in NNI EDIs. Findings from the present study give credence to organic dietary interventions as potential ameliorative strategies for NNI exposure rates in human populations.

Low Prevalence of Anti-DFS70 Antibodies in Children With ANA-Associated Autoimmune Disease.

IntroductionAnti-DFS70 antibodies occur in healthy individuals with various medical conditions. Unlike other anti-nuclear autoantibodies (ANA), they are not associated with systemic autoimmune disease in adult patients. To date, only a few studies have addressed the prevalence and/or clinical relevance of anti-DFS70 autoantibodies in children with and without autoimmune disease.MethodsIncluded in this retrospective cross-sectional mono-centric study were 308 pediatric patients with suspected or known autoimmune conditions who had a positive ANA in indirect immune fluorescence (IIF) screening and who were screened for anti-DFS70 antibodies by extractable nuclear antigen antibodies (ENA) immunoblot. Patients were assigned to four different diagnostic categories according to their diagnosis in the corresponding medical record: (a) absence of autoimmune or rheumatic disease (noARD, n = 116); (b) suspected autoimmunity without definitive diagnosis (sAI, n = 48); (c) other rheumatic disease (ORD) (n = 115); and (d) ANA-associated autoimmune disease (AARD, n = 29).ResultsThe prevalence of anti-DFS70 antibodies in the overall cohort was 33.8%. Among children without ARD (46.6%, 54/116), prevalence was significantly higher than among children with ORD (23.7%, 27/115, p = 0.0003) or AARD (17.2%, 5/29, p = 0.0054). Among all of the anti-DFS70 positive patients with AARD, other autoantibodies were found in the ENA immunoblot. In contrast, among anti-DFS70 positive patients with ORD (11.5%, 4/27), sAI (33.3%, 6/18) and noARD (16.7%, 9/54), other autoantibodies infrequently were detected (p = 0.0005). Patients with uveitis rarely were positive for anti-DFS70 antibodies (7.7%, 1/13). No association was found between anti-DFS70 antibodies and a history of allergic conditions (p = 0.51). The concordance between a typical DFS pattern in IIF and the detection of anti-DFS70 antibodies by immunoblot was 59.3%.ConclusionAs with adults, the higher prevalence of anti-DFS70 among children without autoimmune disease confirms the mutual exclusion for this autoantibody in the pathogenesis of ARD. Among ANA-positive children, monospecific anti-DFS70 antibodies may help to discriminate between AARD and not-AARD-related conditions.

ANA Pozitif Bireylerde ve ANA İlişkili Romatizmal Hastalıklarda Anti-DFS70 Antikorlarının ve DFS paterninin Değerlendirilmesi

Aim: In this study we aimed to find the frequency of anti-DFS70 antibodies and DFS pattern in ANA positive individuals and ANA associated rheumatic diseases (AARDs).Methods: In this study, 337 subjects who were evaluated in a rheumatology clinic with prediagnosis of rheumatic diseases with positive ANA test and had concurrent anti-extractable nuclear antigen (anti-ENA) antibodies results were retrospectively analyzed. Clinical diagnosis of patients and demographic characteristics were obtained from the patients' medical records.Results: A total of 337 subjects (305 women, 32 men) were included in this study. The mean age was 49.8 ± 14.2 years. Of the 337 participants, 111 (32.9%) had an IIF-DFS pattern and 226 (67.1%) had a non-DFS pattern. Anti-DFS70 antibodies were positive in 20.1% of individuals. Sixty eight individuals had AARDs. An IIF-DFS pattern was observed in 22.1% and a non-DFS pattern was observed in 77.9% of individuals with AARDs (p &amp;lt;0.05). Anti-DFS70 antibodies were positive in 13.2% of patients with AARDs. The frequency of AARDs was significantly lower in individuals with anti-DFS70 antibodies compared to individuals with other anti-ENAs antibodies (p &amp;lt;0.05).Conclusion: Anti-DFS70 antibodies may be present in patients with AARDs but AARDs are less prevalent in patients who had anti-DFS70 antibodies, compared with patients who had other anti-ENAs.

Dense fine speckled immunofluorescence pattern in a Chinese population: Prevalence and clinical association.

ObjectiveTo provide information on the prevalence and possible clinical association in a Chinese population for medical practice of the dense fine speckled pattern (DFS pattern).MethodsA retrospective study was conducted with patients who had the DFS pattern from June 2018 to December 2019 in West China Hospital.ResultsA total of 469 patients (1.27% of patients with positive anti‐nuclear antibody indirect immunofluorescence (ANA IIF) test results) revealed the DFS pattern, of which 92.96% had isolated DFS pattern and 23.67% had titers above/equal to 1:320. The average age of patients with the DFS pattern was 43.45 years, and females accounted for 76.97% of them. Ten different kinds of diseases made up the vast majority of the disease spectrum, in which inflammatory or infectious diseases (46.11%), mental diseases (21.45%), and systemic autoimmune rheumatic diseases (SARDs) (18.23%) ranked in the top three. The most common SARDs were rheumatoid arthritis (RA), undifferentiated connective tissue disease (UCTD), and systemic lupus erythematosus (SLE). Forty‐six patients (10.55%) had positive or suspicious extractable nuclear antigen (ENA) antibodies test results and a higher risk of suffering from SARDs. Forty‐seven patients would be missed if the DFS pattern with negative ENA antibodies test result was considered as exclusion criterion of SARDs.ConclusionsThe DFS pattern is basically isolated and with low titer. It is unwise to exclude the diagnosis of SARDs only depending on the appearance of the DFS pattern. Autoimmune diseases‐related antibodies, clinical information of patients, and long‐term follow‐up are of great importance to avoid missed or delayed diagnosis of SARDs.

Anti-DFS70 antibodies detected by specific methods in patients with thrombosis or recurrent pregnancy loss: no evidence of an association

A dense fine speckled pattern (DFS) caused by antibodies to the DFS70 kDa nuclear protein is a relatively common finding while testing for anti-nuclear antibodies (ANA) by indirect immunofluorescence (IIF) on HEp-2 cells. However, despite many efforts and numerous studies, the clinical significance of anti-DFS70 antibodies is still unknown as they can be found in patients with various disorders and even in healthy subjects. In this study we aimed at verifying whether these antibodies are associated with thrombotic events or with unexplained recurrent pregnancy loss (RPL). We studied 443 patients with venous or arterial thrombosis or RPL and 244 controls by IIF on HEp-2 cells and by a DFS70-specific chemiluminescent immunoassay (CIA). The DFS pattern was observed in IIF in 31/443 (7.0%) patients and in 6/244 (2.5%) controls (p = 0.01) while anti-DFS70 specific antibodies were detected by CIA in 11 (2.5%) patients and in one (0.4%) control (p = 0.06). Positive samples, either by IIF or by CIA, were then assayed by a second DFS70-specific line-immunoassay (LIA) method: 83.3% of the CIA positive samples were confirmed DFS70 positive versus only 29.7% of the IIF positive samples. These findings show that IIF overestimates anti-DFS70 antibody frequency and that results obtained by specific CIA and LIA assays do not indicate that venous or arterial thrombosis or RPL are linked to a higher prevalence of anti-DFS70 antibodies.

Clinical value of anti-DFS70 antibodies in a cohort of patients undergoing routine antinuclear antibodies testing

BackgroundIt is has been proposed that the presence of anti-DFS70 antibodies could be used to eliminate a SARD diagnosis. However, anti-DFS70 antibodies were also observed at relatively high frequency in patients with SARD. The clinical significance of these antibodies is therefore not yet clear. In this study, we assessed the clinical significance of the presence of anti-DFS70 antibodies. MethodsA total of 3432 sera samples were obtained from patients who underwent routinely requested ANA screening in the clinical laboratory of a University Hospital, between June 2017 and June 2019. Antinuclear antibody testing was performed by indirect immunofluorescence (IIF) on Hep-2 cell substrates (Euroimmun, Germany), and anti-ENA were measured by LIA (Euroimmun, Germany). ResultsAmong 3432 serum samples tested for ANA, 57.3% were ANA positive by IIF. Participants had mean age of 46.4 and 74.8% of the participants were female. Only 11.4% of the study population had SARD. The frequency of DFS pattern by IIF was 8.1%. Analysis of the DFS pattern positive samples by LIA revealed the presence of anti-DFS70 autoantibodies in 67.5% of all DFS, AC-2 pattern ANAs. When using the results of the ANA IIF HEp-2 DFS pattern/Anti-DFS70 LIA, likelihood ratio (LR) for SARD was 0.33. When comparing the ANA IIF HEp-2 DFS pattern with anti-DFS70 LIA, the ANA IIF HEp-2 DFS pattern's LR was lower (0.63 vs 0.85). ConclusionsAlthough the DFS pattern cannot exclude the presence of SARD, the likelihood is lower than with other patterns. Therefore, anti-DFS70 antibodies represent an important biomarker that can aid in the interpretation of positive ANA patients and, therefore, should be included in test algorithms for ANA testing. The optimal test algorithm might be laboratory specific being dependent on referral patterns for ANA testing.

Complete peritonectomy versus selective peritonectomy in peritoneal carcinomatosis secondary to colorectal cancer: a prospective comparative analysis of outcomes

Introduction: Cytoreductive surgery (CRS) with the intention to achieve complete cytoreduction is by far the most important factor deciding the outcome of peritoneal surface malignancy (PSM). There are a lot of grey areas with respect to the surgical technique in the management of PSM. In spite of doing selective disease-directed peritonectomy, fluoroscopic imaging and microscopy of remaining peritoneum have shown the presence of disease that is not visible to the naked eye. The aim of this study was to assess the morbidity and mortality, recurrence pattern, and oncological outcomes of the extent of parietal peritonectomy with CRS and hyperthermic intraperitoneal chemotherapy (HIPEC) for colorectal carcinoma. Methods: Patients diagnosed with peritoneal surface malignancy from various diseases underwent complete parietal Peritonectomy (CPP) or selective parietal peritonectomy (SPP) with cytoreduction to achieve complete cytoreduction (CC score 0) followed by HIPEC done with the Performer HT (RanD) system. All data prospectively entered in the HIPEC registry were analyzed with main focuses on morbidity, mortality and oncological outcomes (DFS, OS, and recurrence pattern). Results: Of the 163 cases with peritoneal carcinomatosis secondary to colorectal cancer that were analyzed, 128 were females and 35 males. ECOG was 0 in 145 and 1 in 11 patients. Prior surgical scores of 0, 1, 2, and 3 occurred in 101, 18, 38, and 6 patients respectively. Cases were 20 upfront, 94 interval, and 49 recurrent. Of the patients, 70 and 93 patients CPP and SPP respectively. CPP group had higher PCI (18.5 vs 8), longer duration of surgery (11 vs 9), more blood loss (1050 vs 600 ml) and increased hospital stay (14 vs 11) when compared with the SPP group. The number of diaphragmatic resections, bowel resections, anastomosis, and stoma were the same among both groups but the CPP group had more splenectomies and multivisceral resections. The CPP group required more prolonged ventilation. Overall morbidity was increased in the CPP group (61.4% vs 32.3%), with pulmonary and intra-abdominal collections being important. With a median follow up of 45 months, the CPP group had a DFS and OS of 26 and 47 months, respectively, whereas the SPP group had 21 and 44 months, respectively. Most of the recurrences in the CPP group were in lymph nodes (55%), liver (18%), and extra-abdominal (27%), whereas in SPP the most common sites of recurrence were peritoneal (45%), nodal (30%) and extra-abdominal (25%). Conclusion: CPP can be done with acceptable morbidity. Complete parietal peritonectomy lead to a change in the pattern of recurrence moving from peritoneal to systemic recurrence and has shown better outcomes. One has to be watchful about fluid collections post surgery. Although the CPP group had better DFS, there was no significant difference in OS.

Improved accuracy in DFS pattern interpretation using a novel HEp-2 ELITE system.

Accurate interpretation of DFS70 (dense fine speckled 70) and mixed antinuclear antibodies (ANAs) patterns can be challenging using conventional HEp-2 immunofluorescence (IIF) method. We evaluated a novel HEp-2 IIF substrate (HEp-2 ELITE/DFS70-KO) composed of a mixture of engineered HEp-2 devoid of the DFS70 autoantigen and conventional HEp-2 cells. The study assessed the utility of the new substrate in ANA screening and its advantages. One thousand and five consecutive routine samples sent for ANA screening were tested on both standard HEp-2 and the HEp-2 ELITE DFS70 KO substrates (ImmuGlo ANA HEp-2 and HEp-2 ELITE/DFS70-KO, Trinity Biotech, Buffalo, NY). Anti-DFS70 antibody specificity was additionally determined by immunoblot (IB). Clinical and serological data were included in the analysis of the overall impact of the novel HEp-2 substrate on DFS pattern interpretation. Of the 22 cases suspected as positive for DFS pattern alone or in combination with homogeneous or speckled patterns on conventional HEp-2 cells, 17 were interpreted with a higher accuracy using the new HEp-2 ELITE method as positive for DFS70 (monospecific DFS70 (10), mixed DFS70 (7)), speckled (3), and DFS (2) patterns. The new substrate was not only useful in deciphering unclear mixed ANA patterns but also highly sensitive in detecting DFS70 pattern in comparison to the DFS70 positivity obtained using IB.

A better definition of the anti-DFS70 antibody screening by IIF methods

BackgroundAnti-DFS70 antibodies have been recently included in a new testing algorithm for patients with suspicion of connective tissue diseases (CTDs). This algorithm enables to assess the probability of having a CTD in patients with a positive antinuclear antibodies (ANA) result. The aim of the study was to analyze the the inter-method agreement between three different HEp-2 cell substrates for anti-DFS70 detection, focusing on two novel IIF methods that assess the presence of monospecific anti-DFS70 antibodies. MethodsImmunological and clinical records of 29 patients who were double positive for anti-DFS70 autoantibodies using chemiluminescence assay (CIA) and Immunoblot (IB) were studied. The IIF on HEp-2 cells were determined using slides from Inova Diagnostics, Euroimmun and Immco. The capability to detect isolated anti-DFS70 antibodies was compared using immunoadsorption on NOVA Lite HEp-2 Select (Inova Diagnostics) and the HEp-2 ELITE/DFS70 knockout test (Immco). ResultsThe three substrates had very good sensitivity for detecting patients with anti-DFS staining pattern (93.1%, 79.3% and 72.4% for Euroimmun, Immco and Inova respectively). Most of the patients had full inhibition of DFS pattern (65.5%) by immunoabsorption test. Also, the 55.2% of the subjects were positive for monospecific DFS pattern using HEp-2 ELITE/DFS70 knockout test. However, the correlation between the full inhibition by immunoadsorption and the monospecific DFS pattern in knockout cells was very low (kappa: 0.22). ConclusionThe evaluation of monospecific anti-DFS70 antibodies is clinically fundamental and challenging using traditional HEp-2 IIF. Results obtained in this study support the hypothesis that the lack of standardization across IIF kits along with the subjectivity of user interpretation among other factors contribute to the overall reduction in the agreement.

Recognition and Relevance of Anti-DFS70 Autoantibodies in Routine Antinuclear Autoantibodies Testing at a Community Hospital.

Antinuclear autoantibodies (ANA) displaying a dense fine speckled pattern (DFS, ICAP AC-2) on HEp-2 cells are frequently observed in clinical laboratory referrals, often associated with anti-DFS70 specificity. Anti-DFS70 positive patients rarely develop systemic autoimmune rheumatic disease (SARD), especially in the absence of clinical evidence or additional anti-extractable nuclear antigen (ENA) antibodies, prompting suggestions that an isolated DFS70-specific ENA may be an exclusionary finding for SARD. In this study, the frequency and diagnostic significance of anti-DFS70 autoantibodies was investigated in a community hospital cohort of patients undergoing routine ANA testing. ANA screening was performed by HEp-20-10-based indirect immunofluorescence, followed by ENA profiling using a multiparametric line immunoassay (LIA). Of 6,511 patient samples tested for ANA in 2016, the DFS pattern was identified in 1,758 (27.0%), 720 (41.0%) of which were anti-DFS70 positive by LIA. Of these, 526 (73.1%) revealed isolated anti-DFS70 reactivity, while 194 (26.9%) showed additional ENA specificities. Among 1,038 anti-DFS70 negative or borderline samples, 778 (75.0%) were ENA profile negative, while the remaining 260 (25.0%) showed a varied presence of other ENA specificities. Chart reviews of patients with an isolated anti-DFS70 ANA affirmed that ANA-related SARD is rare in the absence of clinical evidence or other ENA specificities, there being no case thus far identified. Rheumatoid arthritis patients occasionally had an isolated anti-DFS70 ANA and were positive for rheumatoid factor and anti-cyclic citrullinated peptide antibodies. In conclusion, the recognition of a DFS ANA pattern using a mitotic-rich HEp-2 substrate, followed by confirmation of anti-DFS70 specificity should be a routine ANA testing service. Use of an expanded ENA profile and clinical correlation is necessary to affirm the “isolation” of anti-DFS70 as the cause of an ANA. Recognition of isolated anti-DFS70 ANA enables reassurance of patients that SARD is unlikely, thus avoiding referral for more extensive testing. The presence of significant elevations of other ENAs may reflect SARD and warrants close clinical correlation and follow-up.

Anti-DFS70 autoantibodies in undifferentiated connective tissue diseases subjects: what's on the horizon?

The main objective was to determine the prevalence of anti-dense fine speckled (DFS70) antibodies in a stable population of undifferentiated connective tissue disease (UCTD) to better define their potential role. Immunological and clinical records of 91 long-standing UCTD patients were studied. DFS pattern was determined using the IIF ANA test on HEp-2 cells and anti-DFS70 antibodies were tested by chemiluminescence assay and by DFS70 line immunoassay. Twelve (13.2%) of 91 serum samples were positive for anti-DFS70 antibodies by chemiluminescence assay and line immunoassay. There was no statistical significance between the prevalence of anti-ENA and anti-DNA autoantibodies in patients with and without anti-DFS70 antibodies. No differences were found in the clinical characteristics of both groups. The presence of the anti-DFS70 antibodies was related to the younger age class. The high prevalence of anti-DFS70 antibodies in the UCTD patients suggested the potential role of these autoantibodies as a marker in the evolution of UCTD to CTD.

ANA Testing: What should we know about the methods, indication and interpretation?

AbstractThough ANA is a common test requested in several settings, one may not be aware of the potential traps for interpretation. Nowadays, there is a trend for autoantibodies diagnostics to move from traditional time honored manual methods to high throughput automated platforms. Nevertheless, the clinical significance and assay performance characteristics may be different from those “historical” methods. Though indirect immunofluorescence is the gold standard method for ANA tests, different laboratories vary in the slides (from different cell lines and commercial source, e.g., Hep 2, Hep 2000, etc.), screening dilutions, terminology, reporting format and expertise. Hence, discrepancy in results among different laboratories is not uncommon and could be confusing. Knowing the assay characteristic and limitations helps proper results interpretation and facilitate patient’s management. Indeed, the titer and pattern by indirect immunofluorescence do provide valuable information in screening patients. In particular, DFS pattern with the associated anti-DFS70 antibodies has been shown to have a role to risk stratify cases referred for suspected autoimmune rheumatic disease.

Disease-free survival of women 70 years or older with stage I-II BC treated with BCS and RT.

e268 Background: Breast RT is the standard of care for women undergoing BCS. Recent trends suggest omitting RT for women &gt; 70 years of age. Our retrospective study evaluated the DFS and failure pattern of women &gt; 70 treated with BCS and RT compared with their younger cohorts. Methods: Between 2009-2011, 101 women with Stage I-II BC were treated with BCS and RT at a physician-owned, community-based cancer center. Clinical, pathologic and treatment characteristics were extracted from patients’ medical records. Results: Of the 101 patients, 71 comprised the &lt; 70 cohort and 30 comprised the &gt; 70 cohort. Patients in the older cohort were more likely to have Stage I tumors (80% vs 64%), but the cohorts were otherwise balanced in terms of T-size, grade, ER/PR and Her2 status. RT was typically delivered in a first course of 50.4 Gy to the whole breast followed by a 10 Gy boost. Median DFS for the older cohort was 36 months (range, 1-66) and 41 months for the younger cohort (range, 1-74). There was only one event in the older cohort, a patient who developed DM at 40 months. In the younger cohort one patient developed LR at 54 months, and 4 developed DM at 18, 33, 51 and 54 months. Conclusions: In our community-based study, women &gt; 70 who received BCS and RT experienced excellent DFS and local control. The three-year median DFS observed in the older women in our study suggests that a long-term view should be taken when treating older patients and that biological age rather than chronological age is a better determinant of when cancer therapies should be withheld.

Frequency of dense fine speckled pattern in immunofluorescence screening test.

The presence of antinuclear antibodies (ANA), directed against intracellular antigens, is a distinctive feature of systemic autoimmune rheumatic diseases (SARDs). The standard test for antinuclear antibody screening is the indirect immunofluorescence (IIF). Anti-dense fine speckled 70 (anti-DFS70) antibodies were initially identified as an ANA IIF pattern from a patient with interstitial cystitis, but they were later associated with various other conditions. The objective of the study was to determine the frequency of anti-DFS70 antibodies in a cohort of patients undergoing routine ANA testing. From January 2011 to January 2012, a total of 5800 serum samples were screened for ANA by IIF (Euroimmune AG, Lübeck, Germany). DFS pattern was searched. ANA were present in 1302 (22.4%) of all patients. There were 16 (1.2%) anti-DFS70 antibody-positive patients. The number of females and males who have anti-DFS70 antibody was eleven and five, respectively. All of the samples presented a titer of ≥1/320. There was one patient with SARD from the rheumatology department. Another 15 patients were from gastroenterology, endocrinology, and general internal medicine. Although a distinctive clinical association has not been reported, anti-DFS70 have been proposed as a significant biomarker for the exclusion of SARD. The present study is a preliminary study. There is a need for a reliable assay to ensure reactivity to DFS70 and screening large populations.

Serum Autoantibodies: From Identification to Clinical Relevance

Serum Autoantibodies: From Identification to Clinical Relevance