Cell Protein Content Research Articles

LGI1 antibody-associated encephalitis without evidence of inflammation in CSF and brain MRI.

This study aimed to explore the frequency and distinct characteristics of adult patients with LGI1 antibody-associated encephalitis in the absence of inflammatory abnormalities in both routine CSF analysis and brain MRI. We conducted a retrospective study of adult patients with antibodies targeting LGI1 and then screened patients with no evidence of inflammation in brain MRI and normal results in routine CSF analysis, including white blood cell count, protein concentration, IgG, and oligoclonal bands. Among 80 patients with LGI1 antibody-associated encephalitis in our center, 31 (38.8%) fulfilled the screening criteria. For these patients, the onset age was 57.0 ± 14.7years, and 19 (61.3%) were female. Viral prodrome occurred in 5 patients (16.1%). Faciobrachial dystonic seizures (FBDS) were the most predominant symptom (38.7%), followed by seizure onset (22.6%) and memory deficits (19.4%). The sensitivity of antibody detection in serum was higher than CSF (96.8% vs. 48.4%, p < 0.001). Most patients (30/31, 96.8%) benefited from the first-line immunotherapy, and 23 patients (74.2%) achieved complete recovery, yet 3 patients (9.7%) had clinical relapses in 2-year follow-up after discharge. The patients had a higher prevalence of females (61.9% vs. 26.7%, p = 0.003) and were more frequently associated with FBDS during the disease course (38.7% vs. 10.2%, p = 0.004). However, there was no difference in treatment outcomes and recurrence ratio between the two groups (p = 0.144 and p = 0.515). Moreover, we divided all 80 patients into four groups according to antibody titer levels in serum and CSF at the time of diagnosis, respectively. WBC and protein concentrations in CSF showed no difference among the four groups. The absence of evidence of inflammation in routine CSF analysis and brain MRI did not rule out anti-LGI1 associated encephalitis. FBDS and the subacute onset of cognitive dysfunction should push forward with autoantibody testing for patients even without inflammatory abnormalities. The routine inflammatory indicators in CSF seemed to be unrelated to antibody titer levels.

Neuronal intermediate filament paraneoplastic autoimmunity complicating avelumab therapy of Merkel cell carcinoma

A 67-years-old woman developed subacute oculomotor nerve palsy and cerebellar gait instability while receiving avelumab as immunotherapy for Merkel cell carcinoma. Brain MRI revealed oculomotor nerve T2/FLAIR hyperintensity and contrast enhancement, CSF cell number and protein concentration were slightly increased. Antibodies against intracellular and surface antigens were excluded through commercial assays, but home-made immunohistochemistry on rat brain sections showed a “neurofilament-like” pattern. Antibodies against neuronal intermediate filament (NIF-IgG) were thus tested and resulted positive in both serum and CSF, confirming the diagnosis of NIF-IgG autoimmunity. Avelumab was discontinued and treatment with steroids and intravenous immunoglobulins led to partial improvement.

Influences of protein levels on the cerebrospinal fluid distribution of ceftazidime & ceftriaxone in the cerebrospinal fluid of patients with inflamed meningitis

IntroductionCeftazidime and ceftriaxone are used to treat various gram-negative pathogens, such as Streptococcus pneumoniae and Pseudomonas aeruginosa, and have shown excellent therapeutic efficacy against bacterial meningitis. However, there is insufficient information on the pharmacokinetic characteristics of their cerebrospinal distribution. Here, we investigated the association of clinical laboratory data in cerebrospinal fluid with ceftazidime and ceftriaxone concentration in the cerebrospinal fluid of patients with inflamed meningitis. MethodsCerebrospinal fluid samples were collected from eight adult patients with inflamed meningitis who intravenously received either ceftazidime or ceftriaxone (ceftazidime: a total of 25 samples from three patients, ceftriaxone: a total of 12 samples from five patients). Total cell number, protein concentration, and glucose concentration in the cerebrospinal fluid were retrospectively collected from electronic medical charts. All ceftazidime and ceftriaxone concentrations in the cerebrospinal fluid were determined using high-performance liquid chromatography. ResultsBoth ceftazidime and ceftriaxone concentrations in cerebrospinal fluid correlated with protein concentration in cerebrospinal fluid; however, no significant correlation was observed in total cell number and glucose concentration in cerebrospinal fluid. ConclusionsThis is the first report on the relationship between the cerebrospinal distribution of these antibiotics and clinical laboratory data in cerebrospinal fluid of adult patients with meningitis.

The Metabolomic Pathways of the Senescence‐Associated Secretory Phenotype in C2C12 Myoblasts

Cellular senescence is considered a hallmark of aging that often occurs in tissues and cells in response to DNA damage. Most healthy cells become senescent after a fixed number of divisions or irreparable genomic damage, often because of external conditions such as circulating factors that are released by adjacent senescent cells. These secreted factors, known as the senescence‐associated secretory phenotype (SASP), are capable of in vivo reprogramming of otherwise healthy cells, ultimately inducing senescence. The involvement of cellular senescence in a variety of aging‐related diseases has been thoroughly examined, however the role of cellular senescence and the SASP in aging muscle remains understudied. Therefore, the objective of this study was to examine the role of metabolites as potential components of the SASP through an easily reproducible model of senescence in skeletal muscle myoblasts. C2C12 myoblasts were treated with an antitumour antibiotic, bleomycin, to cause DNA damage‐induced senescence, or with a vehicle control. Cells and associated media were collected 24 hours after treatment for metabolomic profiling using capillary‐electrophoresis – mass spectrometry (CE‐MS). Samples were compared to untreated myoblasts or fresh growth media and normalized to total cell count and protein content prior to analysis. Pathway analysis using the KEGG database revealed a substantial impact on amino acid metabolism within senescent cells, specifically phenylalanine and histidine metabolism. The media associated with senescent cells had a similar impact on amino acid metabolism, but also showed a significant impact on several polyunsaturated fatty acids, including arachidonic acid and docosahexaenoic acid. Notably, trimethylamine N‐oxide (TMAO), a metabolite previously shown to promote endothelial senescence/dysfunction and neuroinflammation, was detected at significantly higher levels in the media surrounding senescent myoblasts relative to vehicle‐treated controls (fold change = 2.878, p &lt; 0.05). These data suggest that DNA damage‐induced senescence significantly impacts the metabolome of skeletal muscle cells. Future work will aim to evaluate the targeted impact of candidate metabolites as potential senescence inducers.

Copper Bioremediation Ability of Ciliate Paramecium multimicronucleatum Isolated from Industrial Wastewater

The growing problems of environmental damage have been caused by the continuous outrush of heavy metals from industrial wastewater. To resolve this issue, bioremediation is playing a safe and eco-friendly role in the removal of these heavy metals from environmental wastewater bodies. It has provoked demand with regard to understanding the mechanisms of bioaccumulation and detoxification developed by the organisms living in the heavy metal-exposed industrial wastewater. The present investigation focuses on Paramecium multimicronucleatum, a ciliated protozoan isolated from industrial wastewater, with the objective of assessing its capabilities as an environmental bioremediator. Purified cell culture was maintained in bold basal salt medium and optimum growth conditions were determined. A maximum growth rate of 6.0–9.0 × 103 cells/mL at 25–30 °C and pH 7.0 was observed, and therefore revealed to be the optimal growth conditions for this species. It can tolerate 40–50 µg/mL of copper ion stress with little effect on growth rate as compared to control. It is able to uptake more than 80% of copper ions from the medium in 96 h. A significant twofold rise in glutathione content and non-protein thiols was recorded as an indication of a defensive mechanism in place to fight against the oxidative stress caused by the copper treatment. A notable increase of 50–70 µg/mL in total protein content of stressed cells in comparison to non-stressed was also observed as potential induction of some particular proteins for the purpose of resistance against copper stress.

Anti-LGI1 encephalitis recurring 3\xa0years after the first episode: a case report

BackgroundPatients with antibodies against leucine-rich glioma-inactivated 1 (LGI1) present with limbic encephalitis, which is clinically characterized by a subacute disturbance of memory and behavior, often experience seizures. Most patients have a monophasic course, often with hyponatremia.Case presentationHerein, we report a 67-year-old Japanese male presenting with involuntary neck movement, abnormal behavior and apraxia. He was disoriented to time and place and occasionally unable to follow directions. Laboratory tests revealed the patient had hyponatremia (131 mEq/L). Cerebrospinal fluid (CSF) analysis showed that the cell count (1/μL) and protein content (33 mg/dL) were in the normal ranges. Electroencephalography showed transient theta bursts in the right frontal lobe. Magnetic resonance imaging (MRI) of the brain demonstrated hyperintensities in the medial temporal lobe and basal forebrain on fluid-attenuated inversion recovery (FLAIR) without gadolinium enhancement. Anti-voltage-gated potassium channel (Anti-VGKC) complex antibodies were below the reference level for limbic encephalitis. Although the diagnosis was unknown, intravenous methylprednisolone therapy was effective. Three years later, the patient began to speak incoherently and became disoriented to time. FLAIR MRI of the brain revealed recurrence in the left medial temporal lobe. The patient’s serum sodium level was 131 mEq/L. After intravenous methylprednisolone therapy, he regained alertness. A CSF sample stored at the time of the first attack was assayed and the patient was found to be LGI1-positive and CASPR-2-negative, and the diagnosis of anti-LGI1 encephalitis was made.ConclusionsMonitoring serum sodium levels and the preserved samples from the first episode were useful for diagnosis.

Inactivation of Shigella flexneri by 405-nm Light-Emitting Diode Treatment and Possible Mechanism of Action.

Shigella flexneri, a common Gram-negative foodborne pathogen, is widely distributed in fresh-cut fruits and vegetables, unpasteurized milk, and food processing environments. The aims of this study were to evaluate the antibacterial effects of 405-nm light-emitting diode (LED) treatment on S. flexneri and to investigate the possible mechanism. The results showed that LED irradiation (360 min) reduced the number of S. flexneri in phosphate-buffered saline by 3.29 log colony-forming unit (CFU)/mL (initial bacterial count: 6.81 log CFU/mL). The cells in reconstituted infant formula, cells on fresh-cut carrot slices, and biofilm-associated cells on stainless steel surfaces were reduced by 1.83 log CFU/mL, 7.00 log CFU/cm2, and 4.35 log CFU/cm2 following LED treatment for 360, 120, and 120 min, respectively. LED treatment damaged both DNA and cell wall of S. flexneri and changed cell morphology and cell membrane permeability. In addition, LED treatment decreased total cell protein concentration of S. flexneri. These results indicated that 405-nm LED treatment effectively controlled S. flexneri contamination of foods and food contact surfaces and that the bacterial inactivation may be the result of damage to multiple cellular components. These findings highlight the potential of LED technology in controlling S. flexneri during food processing, storage, and preparation.

Antitumor Activity of Palladium(II) Complexes on DU-145 Cell Line in Vitro

Abstract In the area of non-platinum complexes, various complexes containing gold, copper, ruthenium, and palladium have shown a strong cytotoxic effect on different cancer cell lines. The aim of our study was to examine the cytotoxicity of the Pd(II) complexes (C1-C5) and the corresponding ligands (L1-L5) on the DU-145 prostate cancer cell line. Also, due to its clinical application, the cytotoxicity of cisplatin has been examined. Our findings showed that C1- C5 complexes and cisplatin show dose-dependent and strong cytotoxic effects against the DU-145 cell line in vitro. Furthermore, the results demonstrated that early apoptosis was induced by all five Pd(II) complexes. Also, the results showed that complexes C1, C3, and C5 induced G0/G1 phase arrest on DU- 145 cells. Pd(II) complex C2 induced S phase arrest, while C4 complex induced G2/M phase arrest on cancer cells. Additionally, all tested complexes significantly reduced the amount of antiapoptotic protein Bcl-2. Also, there was a significant increase in the concentration of proapoptotic Bax protein in DU-145 cells treated C1-C5 complexes. The results of our research demonstrated that Pd(II) complexes induced apoptosis via the mitochondrial pathway. Thus, it is crucial to further investigate the cytotoxicity of these Pd(II) complexes in vivo. Complex C2 might be a good candidate for a new generation of anticancer drugs.

LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p

BackgroundAccumulating evidence suggests that LINC00511 acts as an oncogenic long non-coding RNA (lncRNA) in various cancers, including lung adenocarcinoma (LUAD). Hence, we attempted to elucidate the potential role of LINC00511 in LUAD.MethodsLINC00511, miR-195-5p, and GCNT3 expression in LUAD was detected by qRT-PCR. Changes in the proliferation, migration, and invasion of LUAD cells after abnormal regulation of LINC00511, miR-195-5p, or GCNT3 were detected by CCK-8, BrdU, wound healing, and transwell assays. Bax and Bcl-2 protein expression was measured by western blotting. Additionally, we identified the targeting effects of LINC00511, miR-195-5p, and GCNT3 using luciferase and RNA immunoprecipitation (RIP) assays.ResultsLINC00511 and GCNT3 were found to be upregulated in LUAD, while miR-195-5p was downregulated. Silencing LINC00511 or GCNT3 decreased the proliferation, migration, invasion, and Bcl-2 protein content in LUAD cells and increased the expression of Bax. Interference with miR-195-5p promoted malignant proliferation of cancer cells. miR-195-5p expression was affected by LINC00511and targeted GCNT3.ConclusionSilencing LINC00511 promotes GCNT3 expression by inhibiting miR-195-5p and ultimately stimulates the malignant progression of LUAD.

A breathing mask attenuates acute airway responses to exercise in sub-zero environment in healthy subjects

PurposeCold air exposure is associated with increased respiratory morbidity and mortality. Repeated inhalation of cold and dry air is considered the cause of the high prevalence of asthma among winter endurance athletes. This study assessed whether a heat- and moisture-exchanging breathing device (HME) attenuates airway responses to high-intensity exercise in sub-zero temperatures among healthy subjects.MethodsUsing a randomized cross-over design, 23 healthy trained participants performed a 30-min warm-up followed by a 4-min maximal, self-paced running time trial in − 15 °C, with and without HME. Lung function was assessed pre- and immediately post-trials. Club cell protein (CC-16), 8-isoprostane, and cytokine concentrations were measured in plasma and urine pre- and 60 min post trials. Symptoms were assessed prior to, during, and immediately after each trial in the chamber.ResultsHME use attenuated the decrease in forced expiratory volume in 1 s (FEV1) post trials (∆FEV1: mean (SD) HME − 0.5 (1.9) % vs. no-HME − 2.7 (2.7) %, p = 0.002). HME also substantially attenuated the median relative increase in plasma-CC16 concentrations (with HME + 27% (interquartile range 9–38) vs no-HME + 121% (55–162), p < 0.001) and reduced airway and general symptom intensity, compared to the trial without HME. No significant changes between trials were detected in urine CC16, 8-isoprostane, or cytokine concentrations.ConclusionThe HME attenuated acute airway responses induced by moderate-to-maximal-intensity exercise in − 15 °C in healthy subjects. Further studies are needed to examine whether this HMEs could constitute primary prevention against asthma in winter endurance athletes.

Three-dimensional scaffolds for bioengineering of cartilage tissue

The cartilage tissue is neither supplied with blood nor innervated, so it cannot heal by itself. Thus, its reconstruction is highly challenging and requires external support. Cartilage diseases are becoming more common due to the aging population and obesity. Among young people, it is usually a post-traumatic complication. Slight cartilage damage leads to the spontaneous formation of fibrous tissue, not resistant to abrasion and stress, resulting in cartilage degradation and the progression of the disease. For these reasons, cartilage regeneration requires further research, including use of new type of biomaterials for scaffolds. This paper shows cartilage characteristics within its most frequent problems and treatment strategies, including a promising method that combines scaffolds and human cells. Structure and material requirements, manufacturing methods, and commercially available scaffolds were described. Also, the comparison of poly(L-lactide) (PLLA) and polyethersulfone (PES) 3D membranes obtained by a phase inversion method using nonwovens as a pore-forming additives were reported. The scaffolds' structure and the growth ability of human chondrocytes were compared. Scaffolds' structure, cells morphology, and protein presence in the membranes were examined with a scanning electron microscope. The metabolic activity of cells was tested with the MTT assay. The structure of the scaffolds and the growth capacity of human chondrocytes were compared. Obtained results showed higher cell activity and protein content for PES scaffolds than for PLLA. The PES membrane had better mechanical properties (e.g. ripping), greater chondrocytes proliferation, and thus a better secretion of proteins which build up the cartilage structure.

Biochemical and Morphological Changes Triggered by Nitrogen Stress in the Oleaginous Microalga Chlorella vulgaris.

Oleaginous microalgae have been considered promising sources of biodiesel due to their high lipid content. Nitrogen limitation/starvation is one of the most prominent strategies to induce lipid accumulation in microalgae. Nonetheless, despite numerous studies, the mechanism underlying this approach is not well understood. The aim of this study was to investigate the effect of nitrogen limitation and starvation on biochemical and morphological changes in the microalga Chlorella vulgaris FACHB-1068, thereby obtaining the optimal nitrogen stress strategy for maximizing the lipid productivity of microalgal biomass. The results showed that nitrogen limitation (nitrate concentration < 21.66 mg/L) and starvation enhanced the lipid content but generally decreased the biomass productivity, pigment concentration, and protein content in algal cells. Comparatively, 3-day nitrogen starvation was found to be a more suitable strategy to produce lipid-rich biomass. It resulted in an increased biomass production and satisfactory lipid content of 266 mg/L and 31.33%, respectively. Besides, nitrogen starvation caused significant changes in cell morphology, with an increase in numbers and total size of lipid droplets and starch granules. Under nitrogen starvation, saturated fatty acids (C-16:0, C-20:0, and C-18:0) accounted for the majority of the total fatty acids (~80%), making C. vulgaris FACHB-1068 a potential feedstock for biodiesel production. Our work may contribute to a better understanding of the biochemical and morphological changes in microalgae under nitrogen stress. Besides, our work may provide valuable information on increasing the lipid productivity of oleaginous microalgae by regulating nitrogen supply.

The nonstructural protein NSs encoded by tomato zonate spot virus suppresses RNA silencing by interacting with NbSGS3.

Viral suppressors of RNA silencing (VSRs) are encoded by diverse viruses to counteract the RNA silencing‐mediated defence mounted by the virus‐infected host cells. In this study, we identified the NSs protein encoded by tomato zonate spot virus (TZSV) as a potent VSR, and used a potato virus X (PVX)‐based heterologous expression system to demonstrate TZSV NSs as a viral pathogenicity factor that intensified PVX symptoms in Nicotiana benthamiana. We then used a yeast two‐hybrid screen to identify the suppressor of gene silencing 3 protein of N. benthamiana (NbSGS3), a known component of the plant RNA silencing pathway, as an interaction partner of TZSV NSs. We verified this interaction in plant cells with bimolecular fluorescence complementation, subcellular colocalization, and co‐immunoprecipitation. We further revealed that the NSs–NbSGS3 interaction correlated with the VSR activity of TZSV NSs. TZSV NSs reduced the concentration of NbSGS3 protein in plant cells, probably through the ubiquitination and autophagy pathways. Interestingly, TZSV infection, but not NSs overexpression, significantly up‐regulated the NbSGS3 transcript levels. Our data indicate that TZSV NSs suppresses RNA silencing of the host plant and enhances TZSV pathogenicity through its interaction with NbSGS3. This study reveals a novel molecular mechanism of NSs‐mediated suppression of plant host antiviral defence.

Fate and inhibition of Bis (2-Ethylhexyl) phthalate in biophysical reactors for treating real landfill leachate

In this study, the performance of combined Non-Thermal Plasma ( as a pre-treatment) and granular sludge sequencing batch reactor ( as a post-treatment) was evaluated for decontamination of real landfill leachate containing Bis (2-Ethylhexyl) phthalate. Special emphasis was placed on the biodegradability of leachate due to its high chemical stability and generation of intermediate species. A plasma reactor with a useful volume of 0.5 L, quartz material was utilized by this study for the pre-treatment process. The biological treatment efficiency was investigated using three granular sludge sequencing batch reactors, each containing 1.2 L with different apparatuses. The Hamadan landfill leachate sample was diluted with synthetic wastewater at 20%, 50%, and 100% v/v ratios. The applied influent Bis (2-Ethylhexyl) phthalate concentrations were 2, 4, 6, 8, 10, 15, and 20 mg/l. Organic loading rate in terms of COD and Bis (2-Ethylhexyl) phthalate concentrations, as well as reactor hydraulic retention time, were worked out to assess the processes performance (COD, nitrogen, phosphorus, and Bis (2-Ethylhexyl) phthalate removal efficiencies). Total Organic Carbon and 5-day Biochemical Oxygen Demand were also measured at the influent and effluent of the NTP reactor over a range of retention time (5, 10, 15, 20, 30, 45, and 60 min) to determine the degree of biodegradability enhancement. Cell protein concentration was also measured using a nanotherape device to assess the effect of DEHP inhibitor on bacterial growth. The main results arrived at from the overall assessment of the processes are that the BOD5/COD ratio of Non-Thermal Plasma reactor effluent at all leachate dilution, i.e., 20%, 50%, and 100% (v/v ratios) considerably increased, reaching 1.09 as compared to 0.33 in the raw influent. The findings indicated that the longer hydraulic retention time affected the granular sludge sequencing batch reactor efficiency. Further, cell protein concentration decreased by increasing the leachate contribution of inlet COD and increased Bis (2-Ethylhexyl) phthalate. It is concluded that Non-Thermal Plasma, as a highly competitive waste treatment technology coupled with aerobic granular sludge, could be considered for the biodegradation of landfill leachate containing chemically stable and recalcitrant organics not treatable by conventional techniques.

Laboratory diagnosis of canine uroperitoneum based on cellular and biochemical characteristics of serum and abdominal fluid.

Literature on the laboratory diagnosis of uroperitoneum is scarce, and it is mostly based on the biochemical findings of cavitary fluid and serum. Cell count and protein concentrations measurements are rarely used and available studies on this subject are based on a relatively small cohort of individuals. We aimed to use a large sample pool of dogs to establish cutoff points for biochemical analytes in cavitary fluids and serum for the diagnosis of uroperitoneum. We also sought to evaluate the general classification of these cavitary fluids. In a retrospective and prospective study, 180 canine abdominal effusion cases were evaluated, 30 of which were uroperitoneum (uroperitoneum group, UG) and 150 with other etiologies (non-uroperitoneum group, NUG). The results showed that 83.3% of UG and 12.7% of NUG abdominal fluid cases were not classified as transudates or exudates. The use of specific cutoffs for fluid creatinine concentrations (≥2.1 mg/dL) and fluid:serum creatinine ratios (Cf: Cs ≥ 1.25) in these unclassified effusions resulted in an accuracy of 99.0% for the laboratory diagnosis of uroperitoneum. The adoption of a new set of criteria and cutoffs based on the combination of parameters such as TP, TNCC, fluid creatinine and Cf: Cs improves the diagnosis of uroperitoneum in dogs.

Abstract P4-06-06: Role of lymphatic SEC62 expression in prediction of treatment response in patients undergoing neoadjuvant chemotherapy for primary breast cancer

Abstract Background Chromosome 3q26 amplifications have been shown to represent a frequent alteration in various cancer entities including breast cancer. SEC62 - a 3q26 encoded gene - was identified as a potential onco- and tumor-driver gene for the pathogenesis of breast cancer. Sec62 seems to induce an increased stress tolerance, enhanced cell migration and proliferation in SEC62 overexpressing cells. Patients with SEC62 overexpressing breast cancer have been shown to have a higher rate of lymph node metastasis and poorer overall prognosis. Hence, we aspired to further evaluate the function of lymphatic SEC62 expression and its predictive role on the response to neoadjuvant chemotherapy. Objectives We aimed to assess the role of lymph node SEC62 expression as a predictive biomarker for response to neoadjuvant chemotherapy, besides we sought to evaluate potential differences in SEC62 expression levels in metastatic lymph node core biopsies and final pathology specimen according to breast cancer subtype. Material &amp; Methods All nodal positive patients treated for primary breast cancer with neoadjuvant chemotherapy at the Department of Gynaecology and Obstetricts, Saarland University Hospital, Homburg, Germany between 01/2009 and 12/2019 were enrolled in this retrospective study. We analyzed SEC62 expression in tissue samples by immunohistochemistry (IHC). Sec62 immunoreactivity was evaluated using the immunoreactive score (IRS) according to Remmele and Stegner (0-12). For the assessment of the Sec62 protein content of the tumor cells, we rated Sec62 “low” for a score of 0-8, and “high” for 9-12 as described in previous publications. IHC was performed on initial lymph node core biopsy (CB) tissue (pre-treatment) or sentinel node biopsy specimen and on definite pathology specimen (PS) obtained during axillary dissection after completion of neoadjuvant chemotherapy.SEC62 expressions in both samples were compared and correlated with response to neoadjuvant chemotherapy, evaluated pathologically using the semi-quantitative response to neoadjuvant chemotherapy scoring system by Sinn et al. ranging from 0 (no effect) to 4 (no tumor detectable). Results 103 patients were assessed for eligibility. 15 cases were excluded, due to incomplete clinical information or insufficient slide quality, leaving 88 patients for final analysis. Median patient age was 55 (32-78) years and median response to neoadjuvant chemotherapy score of 2 (0-4). 11 (12.5%) patients had luminal A, 31 (35.2%) luminal B, 21 (23.9%) Her2/neu positive and 25 (28.4%) triple negative breast cancer. Median SEC62 expression in lymph node core biopsies was significantly higher compared to expression in final specimen (p&amp;lt;0.01). While no differences in basic expression of SEC62 in CB and the response to neoadjuvant chemotherapy were detected, patients with an IRS score ≤ 8 in PS and patients with a difference of IRS score ≥6 between CB and PS showed a significantly higher median response to neoadjuvant chemotherapy score compared to other patients (p&amp;lt; 0.01). Analyzing median Sec62 IHC expression in final pathology specimen, we observed significantly lower Sec62 scores for Her2/neu positive and triple negative patients compared to other subtypes (p&amp;lt;0.01). Median SEC62 expression showed the highest decrease between CB and PS in Her2/neu positive and triple negative patients. Regarding response to neoadjuvant chemotherapy, the effects described above could be observed in all breast cancer subtypes, but were strongest in triple negative patients. Conclusion We identified lymph node SEC62 expression as a promising biomarker for prediction of response to neoadjuvant chemotherapy in breast cancer patients. This effect seems to be strongest in patients with triple negative breast cancer. Citation Format: Julia Zimmermann, Anna Jung, Merle Doerk, Mariz Kasoha, Barbara Linxweiler, Maximilian Linxweiler, Mathias Wagner, Rainer Bohle, Marc Radosa, Erich Solomayer, Julia C Radosa. Role of lymphatic SEC62 expression in prediction of treatment response in patients undergoing neoadjuvant chemotherapy for primary breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-06-06.

Salt-alkalization may potentially promote Microcystis aeruginosa blooms and the production of microcystin-LR

The development of saline-alkali lands has contributed to the increasing discharge of alkaline salt-laden wastewater, which poses a threat to aquatic organisms. However, the comprehensive effect of alkaline salt on Microcystis aeruginosa, a harmful cyanobacterium, remains unclear. In this study, the growth, physiology, cell ultrastructure and production of microcystin-LR (MC-LR) in Microcystis aeruginosa exposed to four levels of alkaline salt stress were evaluated. The growth of Microcystis aeruginosa was stimulated at an electrical conductivity (EC) of 2.5 mS/cm compared to the control, as supported by the increased cell density, photosynthetic pigment and protein contents. Microcystis aeruginosa could tolerate a certain level of alkaline salt (i.e., EC of 5 mS/cm) via increasing photosynthetic pigment contents to protect cells from alkaline salt stress, but the antioxidant defence system and cell ultrastructure were not affected. When EC increased to 7.5 mS/cm, alkaline salt caused oxidative stress and toxicity in Microcystis aeruginosa, as evidenced by analysis of the integrated biomarker response (IBR). Furthermore, the photosynthetic pigment and protein contents decreased, and cell apoptosis associated with ultrastructural changes was observed. Therefore, we propose that EC of 7.5 mS/cm is a threshold for growth of Microcystis aeruginosa. Additionally, the intracellular MC-LR content was stimulated by alkaline salt, and the highest value was observed at EC of 2.5 mS/cm. The extracellular MC-LR content increased with the increasing alkaline salt concentration. When EC was 7.5 mS/cm, the extracellular MC-LR content was significantly higher than in the control and was associated with the upregulated mcyH gene. This study recommends that more attention should be paid to the risk of Microcystis aeruginosa bloom and microcystin-LR pollution in lakes located in salinization regions.

Design of an integrated continuous downstream process for acid-sensitive monoclonal antibodies based on a calcium-dependent Protein A ligand

Monoclonal antibodies (mAb) are used as therapeutics and for diagnostics of a variety of diseases, and novel antibodies are continuously being developed to find treatments for new diseases. Therefore, the manufacturing process must accommodate a range of mAb characteristics. Acid-sensitive mAbs can severely compromise product purity and yield in the purification process due to the potential formation of aggregates. To address this problem, we have developed an integrated downstream process for the purification of pH-sensitive mAbs at mild conditions. A calcium-dependent Protein A-based ligand, called ZCa, was used in the capture step in a 3-column periodic counter-current chromatography operation. The binding of ZCa to antibodies is regulated by calcium, meaning that acidic conditions are not needed to break the interaction and elute the antibodies. Further, the virus inactivation was achieved by a solvent/detergent method, where the pH could remain unchanged. The polishing steps included a cation and an anion exchange chromatography step, and screening of the capture and polishing steps was performed to allow for a seamless integration of the process steps. The process was implemented at laboratory scale for 9 days obtaining a high yield, and a consistently pure drug substance, including high reduction values of the host cell protein and DNA concentrations, as well as aggregate levels below the detection limit, which is attributed to the mild conditions used in the process.

Clinical Efficacy of Dexamethasone in the Treatment of Patients with Tuberculous Meningitis: A Meta-Analysis.

Objective This study aimed to systematically evaluate the clinical efficacy of dexamethasone in patients with tuberculous meningitis (TBM) through meta-analysis. Method PubMed, Web of Science, Embase, China National Knowledge Infrastructure (CNKI), and the Wanfang Databases were searched, and all relevant Chinese and English literature from 2000 to 2021 were retrieved from each database. We collected randomized controlled trials of conventional antituberculosis drugs combined with dexamethasone treatment (treatment group) and conventional antituberculosis drug treatment or combined with placebo treatment (control group) in TBM patients. Meta-analysis was performed with Stata16.0 software. Results A total of 1645 articles were retrieved, and 11 articles were finally included in the study. Meta-analysis results showed that the treatment group had a significantly higher response rate and lower incidence of adverse reactions compared with the control group. Additionally, compared with the control group, the postoperative cerebrospinal fluid cell count, protein content, and glucose in the treatment group were significantly lower, while the chloride level increased. Conclusion Conventional antituberculosis drugs combined with dexamethasone therapy can improve cerebrospinal fluid cell count, protein content, glucose, and chloride levels in patients with TBM. This treatment can improve the treatment effective rate and reduce the incidence of adverse reactions, which is considered an effective treatment for TBM. Our results provide strong evidence for enhancing existing treatment regimens and developing novel combination therapy to improve TBM treatment efficacy.

Inhibitory mechanisms of cinnamic acid on the growth of Geotrichum citri-aurantii

Geotrichum citri-aurantii is the leading causes for sour rot diseases of postharvest citrus fruit, which results in a big loss of commercial value of citrus fruit. Cinnamic acid (CA) is widely used for the preservation of agricultural products due to its antiseptic and bactericidal effect. The effects of CA on the growth of G. citri-aurantii and its potential inhibitory mechanisms were investigated in this study. The results showed a complete inhibition in the growth of G. citri-aurantii caused by 400 mg L−1 CA in vitro, however, in vivo, the disease incidence in the citrus fruit decreased to 38.31% after the application of 4000 mg L−1. Scanning electron microscopy results showed that CA caused a shrinking and distortion of G. citri-aurantii hyphae, and transmission electron microscopy analysis revealed that the intracellular components were greatly leaked and degraded. When treated with 800 mg L−1 CA, 37.72% of G. citri-aurantii spores showed a destruction of plasma membrane integrity. The leakage of electrolyte and the intercellular constituents exosmosis results showed that CA led to a significant increase of the cell membrane permeability. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the protein test kit assay showed that the number of the protein bands reduced after 3 h of CA treatment compared with the control, suggesting that small-molecular and macromolecular substances had been leaked out of the cells. Soluble protein contents of the fungal cells treated with CA significantly dropped. All these results indicated that CA was effective in protecting citrus fruit from postharvest sour rot by damaging the cell membrane of G. citri-aurantii.