Rapid genome-wide profiling of DNA methylation and genetic variation using guide positioning sequencing (GPS).

Whole-genome bisulfite sequencing (WGBS) has been extensively utilized for DNA methylation profiling over the past decade. However, it has shown limitations in terms of high costs and inefficiencies. The productivity and accuracy of DNA methylation detection rely critically on the optimization of methodologies and the continuous refinements of related sequencing platforms. Here, we describe a detailed protocol of guide positioning sequencing (GPS), a bisulfite-based, location-specific sequencing technology designed for comprehensive DNA methylation characterization across the genome. The fundamental principle of GPS lies in the substitution of dCTP with 5-methyl-dCTP (5 mC) at the 3'-end of DNA fragments by T4 DNA polymerase, which protects cytosines from bisulfite conversion to preserve the integrity of the base composition. This alteration allows the 3'-end to independently facilitate genetic variation profiling and guides the 5'-end, enriched with methylation information, to align more rapidly to the reference genome. Hence, GPS enables the concurrent detection of both genetic and epigenetic variations. Additionally, we provide an accessible description of the data processing, specifically involving certain software and scripts. Overall, the entire GPS procedure can be completed within a maximum of 15 days, starting with a low initial DNA input of 100-500 ng, followed by 4-5 days for library construction, 8-10 days for high-throughput sequencing (HTS) and data analysis, which can greatly facilitate the promotion and application of DNA methylation detection, especially for the rapid clinical diagnosis of diverse disease pathologies associated with concurrent genetic and epigenetic variations.

Membrane association and polar localization of the Legionella pneumophila T4SS DotO ATPase mediated by two nonredundant receptors.

The Legionella pneumophila Dot/Icm type IVB secretion system (T4BSS) is a large, multisubunit complex that exports a vast array of substrates into eukaryotic host cells. DotO, a distant homolog of the T4ASS ATPase VirB4, associates with the bacterial inner membrane despite lacking hydrophobic transmembrane domains. Employing a genetic approach, we found DotO's membrane association is mediated by three inner-membrane Dot/Icm components, IcmT, and a combined DotJ-DotI complex (referred to as DotJI). Although deletion of icmT or dotJI individually does not affect DotO's membrane association, the simultaneous inactivation of all three genes results in increased amounts of soluble DotO. Nevertheless, deleting each receptor separately profoundly affects positioning of DotO, disrupting its link with the Dot/Icm complex at the bacterial poles, rendering the receptors nonredundant. Furthermore, a collection of dotO point mutants that we isolated established that DotO's N-terminal domain interacts with the membrane receptors and is involved in dimerization, whereas DotO's C-terminal ATPase domain primarily contributes to the protein's formation of oligomers. Modeling data revealed the complex interaction between DotO and its receptors is responsible for formation of DotO's unique "hexamer of dimers" configuration, which is a defining characteristic of VirB4 family members.

Preclinical evaluation and pilot clinical study of [68Ga]Ga-NOTA-H006 for non-invasive PET imaging of 5T4 oncofetal antigen.

PURPOSE: Trophoblast glycoprotein, the so-called 5T4, is an oncofetal antigen expressed in many different cancers. However, no 5T4-specific radioligand is employed in the clinic for non-invasive diagnosis. Thus, the aim of the current study was to develop a PET radiotracer for imaging 5T4 expression in preclinical and clinical stages. METHODS: A VHH library was constructed by camel immunization. The specificity of the VHHs toward 5T4 antigen was screened through phage display biopanning and periplasmic extract enzyme-linked immunosorbent assay. 1,4,7-Triazacyclononane-1,4,7-triacetate acid (NOTA) derivative was conjugated to the selected VHH. After radiolabeling, microPET/CT and ex vivo biodistribution were conducted using BxPC-3 and MDA-MB-468 tumor-bearing mice. Cold VHH was co-injected with the tracer to challenge its binding in vivo. For the pilot clinical study, PET/CT images were acquired at 1 h after injection of tracer in patients with pathologically confirmed primary and metastatic tumors. RESULTS: A library with a capacity of 1.2 × 1012 colony-forming units was constructed after successful camel immunization. Nb1-40 with a median effect concentration of 0.43 nM was selected. After humanization, the resulting H006 maintained a high affinity towards 5T4. [68Ga]Ga-NOTA-H006 with the molar activities of 6.48-54.2 GBq/µmol was prepared with high radiochemical purity (> 98%). Using [68Ga]Ga-NOTA-H006, microPET/CT revealed a clear visualization of 5T4 expression in BxPC-3 tumor-bearing mice. Ex vivo biodistribution showed that the highest tumor-to-blood ratio (∼ 3-fold) and tumor-to-muscle ratio (∼ 5-fold) were achieved at 60 min post-injection. Co-injection of the cold H006 at a dose of 1.5 mg/kg significantly reduced the tumor uptake (p < 0.0001). In the pilot clinical study, [68Ga]Ga-NOTA-H006 demonstrated its capacity to map 5T4-positive lesions in humans and yielded a mean effective dose of 3.4 × 10- 2 mSv/MBq. CONCLUSIONS: [68Ga]Ga-NOTA-H006, which can visualize 5T4 expression in vivo, has been successfully developed. This opens up opportunities for non-invasively studying 5T4 expression through nuclear medicine. Further clinical investigations are warranted to explore its clinical value in disease progression and companion diagnosis.

A dimeric holin/antiholin complex controls lysis by phage T4.

Lytic phages control the timepoint of host cell lysis by timing the holin-mediated release of cell wall-degrading endolysins. In phage T4, the antiholin RI inhibits the holin T, thereby preventing the early release of the T4 endolysin and lysis. The antiholin achieves lysis inhibition (LIN) in response to phage superinfections, thereby increasing the chance for lysis in an environment with a lower phage concentration. The holin T consists of a small N-terminal cytoplasmic domain, a transmembrane helix, and a periplasmic C-terminal domain. The antiholin is targeted to the periplasm by a cleavable signal peptide. Recently, the periplasmic soluble domains of the holin and the antiholin were found to form T2/RI2 tetramers in crystals. To investigate the functional relevance of this complex, we reconstituted LIN in a phage-free system, using only RI, T, and endolysin, and combined targeted mutagenesis with functional analyses. Inactivation of the RI signal peptide cleavage site did not abolish LIN, indicating that RI can function in a membrane-bound state, which argued against the tetramer. This led to analyses showing that only one of the two T/RI interfaces in the tetramer is physiologically relevant, which is also the only interaction site predicted by AlphaFold2. Some holin mutations at this interaction site prevented lysis, suggesting that the RI interaction likely acts by blocking the holin oligomerization required for hole formation. We conclude that LIN is mediated by a dimeric T/RI complex that, unlike the tetramer, can be easily formed when both partners are membrane-anchored.

Correlation of Solvent Interaction Analysis Signatures with Thermodynamic Properties and In Silico Calculations of the Structural Effects of Point Mutations in Two Proteins.

The partition behavior of single and double-point mutants of bacteriophage T4 lysozyme (T4 lysozyme) and staphylococcal nuclease A was examined in different aqueous two-phase systems (ATPSs) and studied by Solvent Interaction Analysis (SIA). Additionally, the solvent accessible surface area (SASA) of modeled mutants of both proteins was calculated. The in silico calculations and the in vitro analyses of the staphylococcal nuclease and T4 lysozyme mutants correlate, indicating that the partition analysis in ATPSs provides a valid descriptor (SIA signature) covering various protein features, such as structure, structural dynamics, and conformational stability.

[Mentally ill citizens as victims of violation of physical integrity and extermination under the NS regime based on the example of a German city]. / Psychisch kranke Bürgerinnen und Bürger als Opfer von Eingriffen in körperliche Unversehrtheit und Vernichtung in der NS-Zeit am Beispiel einer deutschen Stadt.

BACKGROUND: Even 90 years after the National Socialist seizure of power and the beginning of the darkest chapter of German psychiatric history, examination of the medical historical past retains great relevance as an ongoing medical responsibility. The interventions in the lives of mentally ill people by the National Socialist regime are a firmly established part of medical historical research; however, little is known about how specific regions proceeded with affected citizens. OBJECTIVE: The aim of this study was therefore to identify transfer routes of the "euthanasia" transports and places of death of the victims from Bochum as well as their patient characteristics. MATERIALS AND METHODS: During the timeframe studied, inpatient care for mentally ill patients from the Westphalia region was provided in seven provincial institutions. There was no psychiatric clinic in Bochum at the time, so patients were distributed among these institutions. The investigation was based on the transfer lists of the Westphalian provincial institutions to the killing sites. RESULTS AND CONCLUSION: A total of 366 Bochum citizens affected by "euthanasia" transfers were identified. The transport lists were verified by admission and departure books, death lists, and patient files. The hereditary health files of the city archive were used to examine the relocated Bochum patients to determine whether they were victims of forced sterilization.

Copper sensing transcription factor ArsR2 regulates VjbR to sustain virulence in Brucella abortus.

Brucellosis, caused by the intracellular pathogen Brucella, is a major zoonotic infection that promotes reproductive disease in domestic animals and chronic debilitating conditions in humans. The ArsR family of transcriptional regulators plays key roles in diverse cellular processes, including metal ion homeostasis, responding to adverse conditions, and virulence. However, little is known about the function of ArsR family members in Brucella. Here, we identified ArsR2 as a nonclassical member of the family that lacks autoregulatory function, but which nevertheless plays a vital role in maintaining copper homeostasis in B. abortus. ArsR2 is a global regulator of 241 genes, including those involved in the VirB type IV secretion system (T4SS). Significantly, ArsR2 regulates T4SS production in B. abortus by targeting VjbR which encodes a LuxR-type family transcriptional regulator. Moreover, copper modulates transcriptional activity of ArsR2, but not of VjbR. Furthermore, deletion of arsR2 attenuated virulence in a mouse model. Collectively, these findings enhance understanding of the mechanism by which ArsR proteins regulate virulence gene expression in pathogenic Brucella species.

Treatment Preferences in Patients with Hypothyroidism: an Analysis of Eleven Randomized Controlled Trials.

INTRODUCTION: Levothyroxine (L-T4) monotherapy is the standard of care for the treatment of hypothyroidism. A minority of the L-T4-treated patients remain symptomatic and report better outcomes with combination therapy that contains liothyronine (L-T3) or with desiccated thyroid extract (DTE). GOAL: To assess patient preferences in the treatment of hypothyroidism. METHODS: A systematic review, meta-analysis, meta-regression, and network meta-analysis (NMA) of randomized controlled trials (RCTs) comparing treatments for adults with hypothyroidism (L-T4 vs. L-T4+L-T3 or DTE). Searches were conducted in PubMed, Embase, and Cochrane databases up to April 10, 2024. Data extraction and quality assessment were independently performed by four researchers. RESULTS: Eleven RCTs (eight cross-over studies) with a total of 1,135 patients were considered. Overall, 24% of patients preferred L-T4 versus 52 % who preferred L-T4+L-T3 or DTE; 24% had no preference. The meta-analysis confirmed the preference for combination therapy over L-T4 monotherapy (RR: 2.20, 95% CI: 1.38 to 3.52; p = 0.0009). Excluding four studies reduced the high heterogeneity (I2 = 81%) without affecting the results (RR: 1.97, 95% CI: 1.52 to 2.54; p < 0.00001; I2 = 24%). This preference profile remained when only crossover studies were considered (RR: 2.84, 95% CI: 1.50 to 5.39; p < 0.00001). Network meta-analysis confirmed the preference for DTE and L-T3+L-T4 versus L-T4 alone. CONCLUSION: Patients with hypothyroidism prefer combination therapy (L-T3+L-T4 or DTE) over L-T4 monotherapy. The strength of these findings justifies considering patient preferences in the setting of shared decision-making in the treatment of hypothyroidism.

The Banality of Data: Patient Records, Nursing, and Ideology.

This article explores nursing, patient records, and ideology within the context of the National Socialist "euthanasia" program (Aktion T4) in Germany and Austria from 1939 to 1941, which targeted individuals with mental and physical disabilities for systematic killing. Using Hannah Arendt's concept of the "banality of evil," it examines how ordinary individuals, including nurses, became agents of atrocity by adhering to bureaucratic orders. Jacques Ellul's Ethics of Technology framework is employed to analyze how National Socialist ideology manipulated technological processes to enhance efficiency in genocidal goals. Propaganda was crucial in garnering public support, blurring the lines between technology, ethics, and ideology. Archival research at documentation centers and national archives reveals methods for deciding who was killed, the role of family in medical records, and nurses' involvement in the T4 operation. Three narratives of T4 victims illustrate the personal impacts of these bureaucratic and ideological practices. The article reflects on contemporary nursing, emphasizing the importance of ethical standards and vigilance against data and misuse of technology in health care. This historical examination serves as a reminder of the potential consequences of depersonalization and blind adherence to institutional priorities, underscoring the need for critical engagement with the ethical dimensions of nursing practice.

Disrupting Social Violence Is Everybody's Business.

Our overarching theme for Creative Nursing Volume 30 is The Impact of Social Forces on Nursing and Health. The theme of this current journal issue was originally called Social Violence. In planning this issue, our Editorial Board had to confront head-on the many ways in which overt violence or the threat of violence pervades our lives, as nurses and as individuals. We broadened the title to Consequences and Disruptive Solutions for Social Violence, choosing to focus on what we can do to recognize the interlocking processes that enable, promote, reward, and sustain violent behavior, and to mitigate those effects when and where we can. The sobering content in this issue is the business of each of us. The realities of domestic violence and gun violence, the impact of othering and marginalization, the effects of substance use on family members and of students' experiences of trauma on the school nurses who care for them, and the lethal legacy of unthinking adherence to policies and procedures that is still relevant today-Awareness of this part of our lives and of our practice is where we must start.

Exploring the Effect of Thyroid Hormone on Serum Lipoprotein (a) Levels in Patients With Thyroid Hormone Dysfunction: A Systematic Review.

Genetic variations among people mainly determine the blood levels of lipoprotein (a) (Lp(a)), and it is relatively stable throughout one's lifetime. Nevertheless, there could still be other factors that control the Lp(a) level. Thyroid hormones are known to influence the serum lipid level by regulating the expression of key enzymes that are involved in lipid metabolism. Both hypo and hyperthyroidism are associated with changes in lipid levels. Even though thyroid hormone abnormalities have been shown to alter traditional lipid parameters like low-density lipoprotein (LDL-C), its influence on Lp(a) has not been established. This review aims to identify the relationship between Lp(a) and thyroid hormones by reviewing data from correlative studies and observing treatment-related Lp(a) level changes in thyroid disorders from interventional studies. We searched MEDLINE, Cochrane, and Google Scholar databases with predefined search criteria and search strategies for paper identification. Individual reviewers reviewed identified papers for selection. Finalized papers were reviewed for Lp(a) levels and their responses to treatment in patients with thyroid disorders to establish the relationship between Lp(a) and thyroid hormone. We concluded that the data were limited and sometimes contradicted one another to establish a clear relationship between Lp(a) and thyroid hormones. Even though correlative studies data showed strong indications that overt-hypothyroidism was associated with high Lp(a) levels, thyroid hormone replacement studies did not show any significant changes in Lp(a) levels compared to pre-treatment in patients with both overt-hypothyroidism and subclinical hypothyroidism. More clinical trials focusing on Lp(a) with longer periods of treatment and follow-up in thyroid patients are needed to establish the relationship between the two. The possibility of dose-related Lp(a) responses to thyroid hormone treatment should also be explored.

Male-specific bacteriophages and their potential on combating the spreading of T4SS-bearing antimicrobial resistance plasmids.

Antimicrobial resistance (AMR) has been recognized as an important health crisis in the twenty first century. Type IV secretion systems (T4SSs) play key roles in the dissemination of AMR plasmids. Novel strategies that combat AMR problem by targeting T4SS sprung up in recent years. Here, we focus on the strategy of male-specific phages that could target and kill bacteria carrying conjugative AMR plasmids encoding T4SSs. We reviewed the recent advances in male-specific phages, including anti-conjugation mechanisms, clinical isolation and identification methods, classification and characteristics, in vitro and in vivo anti-conjugation efficacy and improving strategies. Male-specific phages constitute exciting candidates for developing sustainable anti-resistance biocontrol applications.

Low T3 vs low T3T4 euthyroid sick syndrome in septic shock patients: A prospective observational cohort study.

BACKGROUND: Both phases of euthyroid sick syndrome (ESS) are associated with worse prognosis in septic shock patients. Although there are still no indications for supplementation therapy, there is no evidence that both phases (initial and prolonged) are adaptive or that only prolonged is maladaptive and requires supplementation. AIM: To analyze clinical, hemodynamic and laboratory differences in two groups of septic shock patients with ESS. METHODS: A total of 47 septic shock patients with ESS were divided according to values of their thyroid hormones into low T3 and low T3T4 groups. The analysis included demographic data, mortality scores, intensive care unit stay, mechanical ventilation length and 28-day survival and laboratory with hemodynamics. RESULTS: The Simplified Acute Physiology Score II score (P = 0.029), dobutamine (P = 0.003) and epinephrine requirement (P = 0.000) and the incidence of renal failure and multiple organ failure (MOF) (P = 0.000) were significantly higher for the low T3T4. Hypoalbuminemia (P = 0.047), neutrophilia (P = 0.038), lymphopenia (P = 0.013) and lactatemia (P = 0.013) were more pronounced on T2 for the low T3T4 group compared to the low T3 group. Diastolic blood pressure at T0 (P = 0.017) and T1 (P = 0.007), as well as mean arterial pressure at T0 (P = 0.037) and T2 (P = 0.033) was higher for the low T3 group. CONCLUSION: The low T3T4 population is associated with higher frequency of renal insufficiency and MOF, with worse laboratory and hemodynamic parameters. These findings suggest potentially maladaptive changes in the chronic phase of septic shock.

The Association Between Surgical Site Infection and Prognosis of T4 Colorectal Cancer.

OBJECTIVES: Patients with T4 colorectal cancer have poor prognosis, wherein no prognostic factors have been established. Surgical site infection (SSI) has been reported to be one of the risk factors for colorectal cancer recurrence. In this study, we evaluated the relationship between SSI occurrence and prognosis of T4 colorectal cancer and the prognostic impact of the site of SSI occurrence. METHODS: We examined 100 patients with T4 colorectal cancer who underwent radical surgery between April 2002 and December 2017, in a retrospective case-control study, excluding stage IV cases, and classified them into two groups: without SSI (non-SSI) and with SSI (SSI). The five-year relapse-free survival (RFS) and overall survival (OS) were calculated and compared between the two groups. The relationship between prognosis and the SSI site was also assessed according to the SSI site in the incisional/deep and organ/space SSI groups.  Results: The without SSI and with SSI groups included 73 and 27 patients, respectively. The five-year RFS was 55.1% and 22.2% in the without SSI and with SSI groups, respectively (hazard ratio (HR), 2.224; 95% confidence interval (CI), 1.269-3.898; P=0.005). The five-year OS was 67.0% and 38.4% in the without SSI and with SSI groups, respectively (HR, 2.366; 95% CI, 1.223-4.575; P=0.010). The patients in the with SSI group had a significantly poorer prognosis compared with the without SSI group. By SSI site, the prognosis was significantly worse in patients with SSI in the incisional/deep SSI group. CONCLUSIONS: In T4 colorectal cancer, SSI occurrence was a high-risk factor for recurrence and may be a prognostic factor. This result suggested that patients with SSI occurrence may require close postoperative follow-up and appropriate adjuvant chemotherapy.

Minimally Invasive Surgery: Is It a Risk Factor for Postoperative Peritoneal Metastasis in pT4 Colon Cancer?

BACKGROUND: Performing laparoscopic surgery for T4 colon cancer remains controversial because of concerns about whether its oncologic outcomes are comparable to those of open surgery, and postoperative peritoneal metastasis (PM) has been reported to occur more frequently in laparoscopic colectomy for T4 colon cancer. We investigated whether minimally invasive surgery (MIS) demonstrated a higher PM rate than open surgery and analyzed the risk factors for PM in pT4 colon cancer. METHODS: This study included 392 patients with pT4 colon cancer who underwent curative surgery at a referral hospital between January 2000 and December 2018. Patients with previous neoadjuvant therapy, synchronous malignancy, metastasis, or those who underwent hyperthermic intraperitoneal chemotherapy were excluded. RESULTS: The MIS group had fewer high-risk clinical features, such as tumors too large for endoscope admission or complications like perforation and fistula. The group also exhibited shorter operative time, intraoperative blood loss, multivisceral resection, hospital stay, fewer postoperative complications, smaller tumor size, lower pT4b ratio, and higher pN+ rates. Multivariate analysis revealed that high-risk clinical features, MIS, pT4b, pN+, tumor size < 5 cm, high histological grade, lymphovascular invasion, and postoperative complications were significant risk factors for PM. During the median 59-month follow-up, the 5-year cumulative incidence of PM was elevated in the MIS group (17.5% vs. 8.2%; P = 0.057). No significant differences were observed in the 5-year overall and disease-free survival rates. CONCLUSIONS: Minimally invasive surgery increases the risk of postoperative PM in patients with pT4 colon cancer. Surgeons may require thorough tumor staging and radical resection to prevent PM.

Anaplasma phagocytophilum effector EgeA facilitates infection by hijacking TANGO1 and SCFD1 from ER-Golgi exit sites to pathogen-occupied inclusions.

The obligatory intracellular bacterium Anaplasma phagocytophilum causes human granulocytic anaplasmosis, an emerging zoonosis. Anaplasma has limited biosynthetic and metabolic capacities, yet it effectively replicates inside of inclusions/vacuoles of eukaryotic host cells. Here, we describe a unique Type IV secretion system (T4SS) effector, ER-Golgi exit site protein of Anaplasma (EgeA). In cells infected by Anaplasma, secreted native EgeA, EgeA-GFP, and the C-terminal half of EgeA (EgeA-C)-GFP localized to Anaplasma-containing inclusions. In uninfected cells, EgeA-C-GFP localized to cis-Golgi, whereas the N-terminal half of EgeA-GFP localized to the ER. Pull-down assays identified EgeA-GFP binding to a transmembrane protein in the ER, Transport and Golgi organization protein 1 (TANGO1). By yeast two-hybrid analysis, EgeA-C directly bound Sec1 family domain-containing protein 1 (SCFD1), a host protein of the cis-Golgi network that binds TANGO1 at ER-Golgi exit sites (ERES). Both TANGO1 and SCFD1 localized to the Anaplasma inclusion surface. Furthermore, knockdown of Anaplasma EgeA or either host TANGO1 or SCFD1 significantly reduced Anaplasma infection. TANGO1 and SCFD1 prevent ER congestion and stress by facilitating transport of bulky or unfolded proteins at ERES. A bulky cargo collagen and the ER-resident chaperon BiP were transported into Anaplasma inclusions, and several ER stress marker genes were not up-regulated in Anaplasma-infected cells. Furthermore, EgeA transfection reduced collagen overexpression-induced BiP upregulation. These results suggest that by binding to the two ERES proteins, EgeA redirects the cargo-adapted ERES to pathogen-occupied inclusions and reduces ERES congestion, which facilitates Anaplasma nutrient acquisition and reduces ER stress for Anaplasma survival and proliferation.

Is the acquired hypothyroidism a risk factor for developing psychiatric disorders?

Hypothyroidism is a prevalent thyroid condition in which the thyroid gland fails to secrete an adequate amount of thyroid hormone into the bloodstream. This condition may develop due to genetic or acquired factors. The most frequent cause of acquired hypothyroidism is chronic autoimmune thyroiditis, also known as Hashimoto's disease. Acquired hypothyroidism is diagnosed when patients present with overt hypothyroidism (also known as clinical hypothyroidism), as they exhibit increased TSH and decreased T3 and T4 serum levels. This article examines the prevalence of psychiatric disorders among patients diagnosed with acquired hypothyroidism with or without Levothyroxine treatment. We discuss the available evidence indicating that acquired hypothyroidism may be a risk factor for psychiatric disorders, and the effectiveness of thyroid treatment in relieving psychiatric symptoms. Additionally, we provide critical details on thyroid hormone cutoff values reported in the literature, their potential clinical importance, and their correlation with psychiatric symptoms. Finally, we examined the various mechanisms by which acquired hypothyroidism can lead to depression. The high rate of comorbidity between hypothyroidism and psychiatric disorders deserves special attention, indicating the importance of consistent monitoring and timely identification of psychiatric symptoms to prevent disease exacerbation and facilitate therapeutic management. On the other hand, several mechanisms underlie the strong association between depression and acquired hypothyroidism. Deeper research into these mechanisms will allow knowledge of the pathophysiology of depression in patients with acquired hypothyroidism and will provide clues to design more precise therapeutic strategies for these patients.

Surgical Technique for Compartmental Resection of T4b Gingivobuccal Cancer.

Management of advanced gingivo-buccal complex cancers involving the masticatory space (T4b) is often managed by compartment resection. The oncological safety of the procedure is now clearly established. Based on the origin and epicenter of the tumor there are two classes of compartmental resection. Those tumors arising from the tuberosity of the maxilla and/or upper gingival sulcus region; the resection involves the tumor, posterior maxilla, and the ipsilateral infratemporal fossa. These tumors can be resected by mandibulotomy approach, preserving the mandible. This constitutes class-1 infratemporal fossa resection. The class-2 infratemporal fossa resection is applied for those tumors arising from the retromolar trigone and/or lower gingivo-buccal sulcus region. In this class, the mandible and often the overlying cheek skin needs to be sacrificed, in addition to the contents of the infratemporal fossa and the posterior maxilla. Both the classes of resections are carried out in an orderly fashion following well-defined steps. These sequential steps maximize the exposure of inaccessible structures, enables protection of critical structures as well as minimizes blood loss. This manuscript describes the surgical steps for the two classes of compartmental resection of the infratemporal fossa for advanced gingivo-buccal complex cancers involving the masticatory space.

Roles of the components of the cag-pathogenicity island encoded type IV secretion system in Helicobacter pylori.

The Helicobacter pylori (H. pylori) cytotoxin-associated gene pathogenicity island (cagPAI) encodes 31 genes that assemble the cag type IV secretion system (T4SS) apparatus, which includes structures such as the outer membrane core complex, periplasmic ring, inner membrane complex and bacterial hairs. These proteins interact with each other to inject CagA into the host gastric epithelium. There are also individual unique functions that help H. pylori interfere with host cellular pathways, modulate the immune response and colonize the host for a long time. However, the functions of some of the proteins remain unclear. This review summarizes what is known about the structure and function of these auxiliary components and discusses their role in H. pylori pathogenesis.

[Box: see text].

Expression of cytokine and Apoptosis-Associated genes in mice bone Marrow-Derived Macrophages stimulated with Brucella recombinant type IV secretion effectors.

BACKGROUND: Brucellosis is an economically important infectious caused by most commonly by Brucella. Detection of infected animals at the early stage is important for controlling the disease. The diagnostic antigens, usually protein antigens, have attracted much interest. However, the accurate mechanism of immune response is still unknown. The secretory effectors (BPE005, BPE275, and BPE123) of the type IV secretion system (T4SS) were involved in the intracellular circulation process of Brucella and the immune responses of the host. METHODS: Genes encoding three B. abortus effector proteins (BPE005, BPE275, and BPE123) of T4SS were cloned and the recombinant proteins were expressed and purified. The purified recombinant proteins were named rBPE005, rBPE275 and rBPE123. Then, the expressions of Th1- and Th2-related cytokine genes were analyzed in mice bone marrow-derived macrophages (BMDMs) after stimulation with rBPE005, rBPE275, and rBPE123. Furthermore, four apoptosis-associated genes (Caspase-3, Caspase-8, Bax, and Bcl-2) were also detected to explore the damage of the proteins to the cells. RESULTS: Expressions of all Th1- and Th2-related cytokine genes were induced with three proteins, and different cytokine expression patterns induced by each protein depend on the stimulation time and dose of protein. However, expressions of apoptosis-related genes did not change. CONCLUSION: These results showed that the secreted antigens of Brucella induced an immune reaction via the production of Th1- and Th2-type cytokines in BMDMs without exerting any damage on the cells.